development of novel therapeutic antibodies via computational … · 2018-07-24 · b cell epitope...
TRANSCRIPT
Development of Novel Therapeutic Antibodies via Computational Design
by Yue Liu PhDAb Studio Inc
wwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
A novel biotech platform company with promising therapeutic antibody programs
We have invented the following three novel platforms via computational design
1) ldquoImbalancedrdquo bispecific antibody platform2) ldquoSerialrdquo internalization antibody platform3) ldquoCatalyticrdquo antibody platform
We deeply believe in the ldquoquality by designrdquo concept and wish our work can help addressingunmet clinical needs
Unique feature of Ab Studio Inc
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Introduction of Ab Studiorsquos novel therapeutic antibody discovery platforms
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Myeloma cellsB cell
PEGElectro Fusion
Hybridoma
Screening
Platform I- Hybridoma
Secreting Ag specific Ab
Platform II- Phage display
Wash Elute
Screening
Fv modelingAlign VHVL interface structure to the real
VHVL interface in pdb database
Basic technologies
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Hybridoma + Computational Design
1) Apply computer aided modeling to identify epitopes exposed on 3D surface of antigen
2) Based on target biology 3D information and bioinformatics (such as cross-species homology) design and apply appropriate strategy for immunization
3) Based on target biology and target protein structure design andapply high throughput binding assay functional assay and epitopebin assay to screen the best functional antibody
4) Align the sequences of several ldquobest functional antibodiesrdquo to get ldquobest developable antibodiesrdquo by screening candidates withhigh ldquohuman scorerdquo + less immunogenicity epitopes + less PTMs
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Unique features for phage display technology
1) In house developed 10e9 human naiumlve ScFV phage library witha restriction enzyme site between VH and VL Therefore VH andVL can be replaced separately (suitable for common light chain screening)
2) Leading phage screening not only based on antigen binding ability pre and post heating but also based on ScFV seqsrsquo otherdevelop-ability features After building up a Fv model structure we study
bull Aggregation surface on Fv modelbull PTMs on Fv modelbull Immunogenecity epitopes inside ScFV sequencebull ldquoNaturalityrdquo of VHVL interface by aligning the query ScFVrsquos VHVL
interface to that of real 3D antibody structures in the pdb database
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Antibody Humanization andOptimization
We balance
Humanization percentage
CDR Immunogenicity
Antigen binding affinity
Antibody expression level
Aggregation potential
Antibody stability
Heterogenicity
Workflow of Antibody Humanization and Optimization
Modeling of Mabhomo model vs hetero model
BioinformaticsPTM analysisAggregation surface analysisPatch analysisT cell epitope searchB cell epitope searchMHC II epitope search
Human germlineacceptor selection
CDR graftingBack mutation design
BioinformaticsPTM analysisAggregation surface analysisPatch analysisT cell epitope searchB cell epitope searchMHC II epitope searchHumanization percentage
Lead Selection on Bench1st round Binding based on ELISAor FACS2nd round Bioanalytics includingBiacore DLSDSCCEIEF
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Molecular Modeling and 3D Surface Analysis
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Immunogenecity Epitope Scan
Novel platform 1
Imbalanced Bispecific Antibody Technology Platform
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
The
Zoo
of B
ispe
cific An
tibo
dy Fo
rmats
Ab Studio Inc
your partner for therapeutic antibody development
http
sww
wn
cbin
lmn
ihgo
vpm
carticlesPM
C5
29
75
37
Ulrich Brinkmann
wwwantibodystudiocom
Only a few platforms meet the following criterias
1) Free to operate2) No extra immunogenicity and
developability concerns3) No PKPD concerns
ldquoKnob into holerdquo + Common LC formatis an one meets the above criterias
Ab Studio Inc
your partner for therapeutic antibody development
Imbalanced Bispecific Antibody Technology Platform
What is an imbalanced bispecific antibody
- A ldquoKnob-into-hole plus common LCrdquo version allowing two arms to have very differentbinding affinity to each onersquos own antigen (natural IgG structure and half life)
Why ldquoimbalancedrdquo
- Imbalanced binding affinity is required for some bispecific therapeutic antibodiesrsquoMechanism of Action
wwwantibodystudiocom
Bispecific involves a CD3 binder
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CD3 binding Cancer antigen binding CD3
Cancer antigen
Bispecific
wwwantibodystudiocom
Bispecific against a CSA (cancer specific antigen) and a CAA (cancer associated antigen)
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CSA binding CAA binding
Bispecific
CSA Cancer specific antigen
CAA Cancer associated antigen Normal cell
Normal cell
Normal cell
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Bispecific against one target for CNS and another target on BBB
Ab Studio Inc
your partner for therapeutic antibody development
Brain target
cell
ldquoKnob-into-holerdquo
Common light chain
CNS target binding BBB binding
Bispecific
BBB
Brain target
cell
wwwantibodystudiocom
How ldquoimbalanced bispecific antibodiesrdquo are made at Ab Studio
(lsquoArsquo is the corresponding high affinity arm and lsquoBrsquo is the corresponding low affinity arm)
VL Design
1) If VL-A and VL-B light chain homology gt80 design a VL-C based on VL-Avia computer aided design Maintain VH-AVL-Crsquos affinity as close as that ofVH-AVL-A allow VH-BVL-C to have reduced affinity compared to VH-BVL-B
2) If VL-A and VL-B light chain homology lt80 construct phage ScFV libraryVHs of the ScFV library are human germline VH libraryVLs of the ScFV library are VL-A and its mutants derived from error prone PCR at less than 20 of mutation frequency
VH Design
Use computer aided design to differentiate the biochemical and biophysical featuresof VH-A and VH-B in order to isolate lsquoABrsquo better from lsquoAArsquo and lsquoBBrsquo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Case Study CD20CD3 Bispecific
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chainComputer aided design
CD3 CD20 CD3
Cancer antigen
Bispecific
Biophysical and biochemical features were differentiated for reducing manufacture cost
wwwantibodystudiocom
Difference to other CD20CD3 bispecific ADCCCDC effector function is maintained
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Current CD20+ Cancer Immuno-Therapy
Mechanism of action ADCC CDC ApoptosisRepresentatives Rituxan and its biosimilar biobetterLimitation Other than antigen escape different genetic makeup of patientsrsquo effector cells could cause different response to these therapies
Mechanism of action CAR-T TCR-T T cell engagementRepresentatives CD20CD3 bispecific (ndashADCCCDC)Limitation Other than antigen escape cancer cells with T cell immune suppressors such as PDL1 overexpressed may have worse response to T cell activation based therapies
Novel ldquoimbalancedrdquo CD20CD3 based cancer therapy
By making a CD20CD3 bispecific antibody with CD3binding affinity significantly reduced we hope toldquosafelyrdquo maintain the effector function and reduce cytokine storm Therefore the novel ldquoimbalancedrdquo CD20CD3 bispecific would have all mechanisms as described above
Binding Ability of Designed Abs Compared to That of Parental Abs
After computational design CD20 homodimer (hole-hole) IgG contains designed VH sequence and common VL
sequence showed similar binding capacity for CD20 compared to that of parental CD20 IgG while CD3
homodimer (knob-knob) IgG contains designed VH sequence and common VL sequence showed significant
reduced binding capacity for CD3 compared to that of parental CD3 IgG
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has Good Efficacy and Safety in vitro
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Day 0 IV injection with ldquoAbsPBS + Raji-Luc cell + human PBMCrdquo first imaging 15min post injection G1 PBS G2 BsAb G3 Rituximab
1 died3 died
Day 0 Day 2 Day 13 Day 22 Day 28
CD20CD3 ldquoImbalancedrdquo BsAb has better Tumor Killing Potency than Rituxan in vivo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
A novel biotech platform company with promising therapeutic antibody programs
We have invented the following three novel platforms via computational design
1) ldquoImbalancedrdquo bispecific antibody platform2) ldquoSerialrdquo internalization antibody platform3) ldquoCatalyticrdquo antibody platform
We deeply believe in the ldquoquality by designrdquo concept and wish our work can help addressingunmet clinical needs
Unique feature of Ab Studio Inc
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Introduction of Ab Studiorsquos novel therapeutic antibody discovery platforms
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Myeloma cellsB cell
PEGElectro Fusion
Hybridoma
Screening
Platform I- Hybridoma
Secreting Ag specific Ab
Platform II- Phage display
Wash Elute
Screening
Fv modelingAlign VHVL interface structure to the real
VHVL interface in pdb database
Basic technologies
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Hybridoma + Computational Design
1) Apply computer aided modeling to identify epitopes exposed on 3D surface of antigen
2) Based on target biology 3D information and bioinformatics (such as cross-species homology) design and apply appropriate strategy for immunization
3) Based on target biology and target protein structure design andapply high throughput binding assay functional assay and epitopebin assay to screen the best functional antibody
4) Align the sequences of several ldquobest functional antibodiesrdquo to get ldquobest developable antibodiesrdquo by screening candidates withhigh ldquohuman scorerdquo + less immunogenicity epitopes + less PTMs
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Unique features for phage display technology
1) In house developed 10e9 human naiumlve ScFV phage library witha restriction enzyme site between VH and VL Therefore VH andVL can be replaced separately (suitable for common light chain screening)
2) Leading phage screening not only based on antigen binding ability pre and post heating but also based on ScFV seqsrsquo otherdevelop-ability features After building up a Fv model structure we study
bull Aggregation surface on Fv modelbull PTMs on Fv modelbull Immunogenecity epitopes inside ScFV sequencebull ldquoNaturalityrdquo of VHVL interface by aligning the query ScFVrsquos VHVL
interface to that of real 3D antibody structures in the pdb database
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Antibody Humanization andOptimization
We balance
Humanization percentage
CDR Immunogenicity
Antigen binding affinity
Antibody expression level
Aggregation potential
Antibody stability
Heterogenicity
Workflow of Antibody Humanization and Optimization
Modeling of Mabhomo model vs hetero model
BioinformaticsPTM analysisAggregation surface analysisPatch analysisT cell epitope searchB cell epitope searchMHC II epitope search
Human germlineacceptor selection
CDR graftingBack mutation design
BioinformaticsPTM analysisAggregation surface analysisPatch analysisT cell epitope searchB cell epitope searchMHC II epitope searchHumanization percentage
Lead Selection on Bench1st round Binding based on ELISAor FACS2nd round Bioanalytics includingBiacore DLSDSCCEIEF
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Molecular Modeling and 3D Surface Analysis
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Immunogenecity Epitope Scan
Novel platform 1
Imbalanced Bispecific Antibody Technology Platform
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
The
Zoo
of B
ispe
cific An
tibo
dy Fo
rmats
Ab Studio Inc
your partner for therapeutic antibody development
http
sww
wn
cbin
lmn
ihgo
vpm
carticlesPM
C5
29
75
37
Ulrich Brinkmann
wwwantibodystudiocom
Only a few platforms meet the following criterias
1) Free to operate2) No extra immunogenicity and
developability concerns3) No PKPD concerns
ldquoKnob into holerdquo + Common LC formatis an one meets the above criterias
Ab Studio Inc
your partner for therapeutic antibody development
Imbalanced Bispecific Antibody Technology Platform
What is an imbalanced bispecific antibody
- A ldquoKnob-into-hole plus common LCrdquo version allowing two arms to have very differentbinding affinity to each onersquos own antigen (natural IgG structure and half life)
Why ldquoimbalancedrdquo
- Imbalanced binding affinity is required for some bispecific therapeutic antibodiesrsquoMechanism of Action
wwwantibodystudiocom
Bispecific involves a CD3 binder
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CD3 binding Cancer antigen binding CD3
Cancer antigen
Bispecific
wwwantibodystudiocom
Bispecific against a CSA (cancer specific antigen) and a CAA (cancer associated antigen)
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CSA binding CAA binding
Bispecific
CSA Cancer specific antigen
CAA Cancer associated antigen Normal cell
Normal cell
Normal cell
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Bispecific against one target for CNS and another target on BBB
Ab Studio Inc
your partner for therapeutic antibody development
Brain target
cell
ldquoKnob-into-holerdquo
Common light chain
CNS target binding BBB binding
Bispecific
BBB
Brain target
cell
wwwantibodystudiocom
How ldquoimbalanced bispecific antibodiesrdquo are made at Ab Studio
(lsquoArsquo is the corresponding high affinity arm and lsquoBrsquo is the corresponding low affinity arm)
VL Design
1) If VL-A and VL-B light chain homology gt80 design a VL-C based on VL-Avia computer aided design Maintain VH-AVL-Crsquos affinity as close as that ofVH-AVL-A allow VH-BVL-C to have reduced affinity compared to VH-BVL-B
2) If VL-A and VL-B light chain homology lt80 construct phage ScFV libraryVHs of the ScFV library are human germline VH libraryVLs of the ScFV library are VL-A and its mutants derived from error prone PCR at less than 20 of mutation frequency
VH Design
Use computer aided design to differentiate the biochemical and biophysical featuresof VH-A and VH-B in order to isolate lsquoABrsquo better from lsquoAArsquo and lsquoBBrsquo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Case Study CD20CD3 Bispecific
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chainComputer aided design
CD3 CD20 CD3
Cancer antigen
Bispecific
Biophysical and biochemical features were differentiated for reducing manufacture cost
wwwantibodystudiocom
Difference to other CD20CD3 bispecific ADCCCDC effector function is maintained
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Current CD20+ Cancer Immuno-Therapy
Mechanism of action ADCC CDC ApoptosisRepresentatives Rituxan and its biosimilar biobetterLimitation Other than antigen escape different genetic makeup of patientsrsquo effector cells could cause different response to these therapies
Mechanism of action CAR-T TCR-T T cell engagementRepresentatives CD20CD3 bispecific (ndashADCCCDC)Limitation Other than antigen escape cancer cells with T cell immune suppressors such as PDL1 overexpressed may have worse response to T cell activation based therapies
Novel ldquoimbalancedrdquo CD20CD3 based cancer therapy
By making a CD20CD3 bispecific antibody with CD3binding affinity significantly reduced we hope toldquosafelyrdquo maintain the effector function and reduce cytokine storm Therefore the novel ldquoimbalancedrdquo CD20CD3 bispecific would have all mechanisms as described above
Binding Ability of Designed Abs Compared to That of Parental Abs
After computational design CD20 homodimer (hole-hole) IgG contains designed VH sequence and common VL
sequence showed similar binding capacity for CD20 compared to that of parental CD20 IgG while CD3
homodimer (knob-knob) IgG contains designed VH sequence and common VL sequence showed significant
reduced binding capacity for CD3 compared to that of parental CD3 IgG
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has Good Efficacy and Safety in vitro
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Day 0 IV injection with ldquoAbsPBS + Raji-Luc cell + human PBMCrdquo first imaging 15min post injection G1 PBS G2 BsAb G3 Rituximab
1 died3 died
Day 0 Day 2 Day 13 Day 22 Day 28
CD20CD3 ldquoImbalancedrdquo BsAb has better Tumor Killing Potency than Rituxan in vivo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Introduction of Ab Studiorsquos novel therapeutic antibody discovery platforms
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Myeloma cellsB cell
PEGElectro Fusion
Hybridoma
Screening
Platform I- Hybridoma
Secreting Ag specific Ab
Platform II- Phage display
Wash Elute
Screening
Fv modelingAlign VHVL interface structure to the real
VHVL interface in pdb database
Basic technologies
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Hybridoma + Computational Design
1) Apply computer aided modeling to identify epitopes exposed on 3D surface of antigen
2) Based on target biology 3D information and bioinformatics (such as cross-species homology) design and apply appropriate strategy for immunization
3) Based on target biology and target protein structure design andapply high throughput binding assay functional assay and epitopebin assay to screen the best functional antibody
4) Align the sequences of several ldquobest functional antibodiesrdquo to get ldquobest developable antibodiesrdquo by screening candidates withhigh ldquohuman scorerdquo + less immunogenicity epitopes + less PTMs
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Unique features for phage display technology
1) In house developed 10e9 human naiumlve ScFV phage library witha restriction enzyme site between VH and VL Therefore VH andVL can be replaced separately (suitable for common light chain screening)
2) Leading phage screening not only based on antigen binding ability pre and post heating but also based on ScFV seqsrsquo otherdevelop-ability features After building up a Fv model structure we study
bull Aggregation surface on Fv modelbull PTMs on Fv modelbull Immunogenecity epitopes inside ScFV sequencebull ldquoNaturalityrdquo of VHVL interface by aligning the query ScFVrsquos VHVL
interface to that of real 3D antibody structures in the pdb database
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Antibody Humanization andOptimization
We balance
Humanization percentage
CDR Immunogenicity
Antigen binding affinity
Antibody expression level
Aggregation potential
Antibody stability
Heterogenicity
Workflow of Antibody Humanization and Optimization
Modeling of Mabhomo model vs hetero model
BioinformaticsPTM analysisAggregation surface analysisPatch analysisT cell epitope searchB cell epitope searchMHC II epitope search
Human germlineacceptor selection
CDR graftingBack mutation design
BioinformaticsPTM analysisAggregation surface analysisPatch analysisT cell epitope searchB cell epitope searchMHC II epitope searchHumanization percentage
Lead Selection on Bench1st round Binding based on ELISAor FACS2nd round Bioanalytics includingBiacore DLSDSCCEIEF
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Molecular Modeling and 3D Surface Analysis
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Immunogenecity Epitope Scan
Novel platform 1
Imbalanced Bispecific Antibody Technology Platform
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
The
Zoo
of B
ispe
cific An
tibo
dy Fo
rmats
Ab Studio Inc
your partner for therapeutic antibody development
http
sww
wn
cbin
lmn
ihgo
vpm
carticlesPM
C5
29
75
37
Ulrich Brinkmann
wwwantibodystudiocom
Only a few platforms meet the following criterias
1) Free to operate2) No extra immunogenicity and
developability concerns3) No PKPD concerns
ldquoKnob into holerdquo + Common LC formatis an one meets the above criterias
Ab Studio Inc
your partner for therapeutic antibody development
Imbalanced Bispecific Antibody Technology Platform
What is an imbalanced bispecific antibody
- A ldquoKnob-into-hole plus common LCrdquo version allowing two arms to have very differentbinding affinity to each onersquos own antigen (natural IgG structure and half life)
Why ldquoimbalancedrdquo
- Imbalanced binding affinity is required for some bispecific therapeutic antibodiesrsquoMechanism of Action
wwwantibodystudiocom
Bispecific involves a CD3 binder
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CD3 binding Cancer antigen binding CD3
Cancer antigen
Bispecific
wwwantibodystudiocom
Bispecific against a CSA (cancer specific antigen) and a CAA (cancer associated antigen)
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CSA binding CAA binding
Bispecific
CSA Cancer specific antigen
CAA Cancer associated antigen Normal cell
Normal cell
Normal cell
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Bispecific against one target for CNS and another target on BBB
Ab Studio Inc
your partner for therapeutic antibody development
Brain target
cell
ldquoKnob-into-holerdquo
Common light chain
CNS target binding BBB binding
Bispecific
BBB
Brain target
cell
wwwantibodystudiocom
How ldquoimbalanced bispecific antibodiesrdquo are made at Ab Studio
(lsquoArsquo is the corresponding high affinity arm and lsquoBrsquo is the corresponding low affinity arm)
VL Design
1) If VL-A and VL-B light chain homology gt80 design a VL-C based on VL-Avia computer aided design Maintain VH-AVL-Crsquos affinity as close as that ofVH-AVL-A allow VH-BVL-C to have reduced affinity compared to VH-BVL-B
2) If VL-A and VL-B light chain homology lt80 construct phage ScFV libraryVHs of the ScFV library are human germline VH libraryVLs of the ScFV library are VL-A and its mutants derived from error prone PCR at less than 20 of mutation frequency
VH Design
Use computer aided design to differentiate the biochemical and biophysical featuresof VH-A and VH-B in order to isolate lsquoABrsquo better from lsquoAArsquo and lsquoBBrsquo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Case Study CD20CD3 Bispecific
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chainComputer aided design
CD3 CD20 CD3
Cancer antigen
Bispecific
Biophysical and biochemical features were differentiated for reducing manufacture cost
wwwantibodystudiocom
Difference to other CD20CD3 bispecific ADCCCDC effector function is maintained
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Current CD20+ Cancer Immuno-Therapy
Mechanism of action ADCC CDC ApoptosisRepresentatives Rituxan and its biosimilar biobetterLimitation Other than antigen escape different genetic makeup of patientsrsquo effector cells could cause different response to these therapies
Mechanism of action CAR-T TCR-T T cell engagementRepresentatives CD20CD3 bispecific (ndashADCCCDC)Limitation Other than antigen escape cancer cells with T cell immune suppressors such as PDL1 overexpressed may have worse response to T cell activation based therapies
Novel ldquoimbalancedrdquo CD20CD3 based cancer therapy
By making a CD20CD3 bispecific antibody with CD3binding affinity significantly reduced we hope toldquosafelyrdquo maintain the effector function and reduce cytokine storm Therefore the novel ldquoimbalancedrdquo CD20CD3 bispecific would have all mechanisms as described above
Binding Ability of Designed Abs Compared to That of Parental Abs
After computational design CD20 homodimer (hole-hole) IgG contains designed VH sequence and common VL
sequence showed similar binding capacity for CD20 compared to that of parental CD20 IgG while CD3
homodimer (knob-knob) IgG contains designed VH sequence and common VL sequence showed significant
reduced binding capacity for CD3 compared to that of parental CD3 IgG
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has Good Efficacy and Safety in vitro
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Day 0 IV injection with ldquoAbsPBS + Raji-Luc cell + human PBMCrdquo first imaging 15min post injection G1 PBS G2 BsAb G3 Rituximab
1 died3 died
Day 0 Day 2 Day 13 Day 22 Day 28
CD20CD3 ldquoImbalancedrdquo BsAb has better Tumor Killing Potency than Rituxan in vivo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Myeloma cellsB cell
PEGElectro Fusion
Hybridoma
Screening
Platform I- Hybridoma
Secreting Ag specific Ab
Platform II- Phage display
Wash Elute
Screening
Fv modelingAlign VHVL interface structure to the real
VHVL interface in pdb database
Basic technologies
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Hybridoma + Computational Design
1) Apply computer aided modeling to identify epitopes exposed on 3D surface of antigen
2) Based on target biology 3D information and bioinformatics (such as cross-species homology) design and apply appropriate strategy for immunization
3) Based on target biology and target protein structure design andapply high throughput binding assay functional assay and epitopebin assay to screen the best functional antibody
4) Align the sequences of several ldquobest functional antibodiesrdquo to get ldquobest developable antibodiesrdquo by screening candidates withhigh ldquohuman scorerdquo + less immunogenicity epitopes + less PTMs
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Unique features for phage display technology
1) In house developed 10e9 human naiumlve ScFV phage library witha restriction enzyme site between VH and VL Therefore VH andVL can be replaced separately (suitable for common light chain screening)
2) Leading phage screening not only based on antigen binding ability pre and post heating but also based on ScFV seqsrsquo otherdevelop-ability features After building up a Fv model structure we study
bull Aggregation surface on Fv modelbull PTMs on Fv modelbull Immunogenecity epitopes inside ScFV sequencebull ldquoNaturalityrdquo of VHVL interface by aligning the query ScFVrsquos VHVL
interface to that of real 3D antibody structures in the pdb database
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Antibody Humanization andOptimization
We balance
Humanization percentage
CDR Immunogenicity
Antigen binding affinity
Antibody expression level
Aggregation potential
Antibody stability
Heterogenicity
Workflow of Antibody Humanization and Optimization
Modeling of Mabhomo model vs hetero model
BioinformaticsPTM analysisAggregation surface analysisPatch analysisT cell epitope searchB cell epitope searchMHC II epitope search
Human germlineacceptor selection
CDR graftingBack mutation design
BioinformaticsPTM analysisAggregation surface analysisPatch analysisT cell epitope searchB cell epitope searchMHC II epitope searchHumanization percentage
Lead Selection on Bench1st round Binding based on ELISAor FACS2nd round Bioanalytics includingBiacore DLSDSCCEIEF
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Molecular Modeling and 3D Surface Analysis
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Immunogenecity Epitope Scan
Novel platform 1
Imbalanced Bispecific Antibody Technology Platform
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
The
Zoo
of B
ispe
cific An
tibo
dy Fo
rmats
Ab Studio Inc
your partner for therapeutic antibody development
http
sww
wn
cbin
lmn
ihgo
vpm
carticlesPM
C5
29
75
37
Ulrich Brinkmann
wwwantibodystudiocom
Only a few platforms meet the following criterias
1) Free to operate2) No extra immunogenicity and
developability concerns3) No PKPD concerns
ldquoKnob into holerdquo + Common LC formatis an one meets the above criterias
Ab Studio Inc
your partner for therapeutic antibody development
Imbalanced Bispecific Antibody Technology Platform
What is an imbalanced bispecific antibody
- A ldquoKnob-into-hole plus common LCrdquo version allowing two arms to have very differentbinding affinity to each onersquos own antigen (natural IgG structure and half life)
Why ldquoimbalancedrdquo
- Imbalanced binding affinity is required for some bispecific therapeutic antibodiesrsquoMechanism of Action
wwwantibodystudiocom
Bispecific involves a CD3 binder
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CD3 binding Cancer antigen binding CD3
Cancer antigen
Bispecific
wwwantibodystudiocom
Bispecific against a CSA (cancer specific antigen) and a CAA (cancer associated antigen)
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CSA binding CAA binding
Bispecific
CSA Cancer specific antigen
CAA Cancer associated antigen Normal cell
Normal cell
Normal cell
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Bispecific against one target for CNS and another target on BBB
Ab Studio Inc
your partner for therapeutic antibody development
Brain target
cell
ldquoKnob-into-holerdquo
Common light chain
CNS target binding BBB binding
Bispecific
BBB
Brain target
cell
wwwantibodystudiocom
How ldquoimbalanced bispecific antibodiesrdquo are made at Ab Studio
(lsquoArsquo is the corresponding high affinity arm and lsquoBrsquo is the corresponding low affinity arm)
VL Design
1) If VL-A and VL-B light chain homology gt80 design a VL-C based on VL-Avia computer aided design Maintain VH-AVL-Crsquos affinity as close as that ofVH-AVL-A allow VH-BVL-C to have reduced affinity compared to VH-BVL-B
2) If VL-A and VL-B light chain homology lt80 construct phage ScFV libraryVHs of the ScFV library are human germline VH libraryVLs of the ScFV library are VL-A and its mutants derived from error prone PCR at less than 20 of mutation frequency
VH Design
Use computer aided design to differentiate the biochemical and biophysical featuresof VH-A and VH-B in order to isolate lsquoABrsquo better from lsquoAArsquo and lsquoBBrsquo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Case Study CD20CD3 Bispecific
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chainComputer aided design
CD3 CD20 CD3
Cancer antigen
Bispecific
Biophysical and biochemical features were differentiated for reducing manufacture cost
wwwantibodystudiocom
Difference to other CD20CD3 bispecific ADCCCDC effector function is maintained
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Current CD20+ Cancer Immuno-Therapy
Mechanism of action ADCC CDC ApoptosisRepresentatives Rituxan and its biosimilar biobetterLimitation Other than antigen escape different genetic makeup of patientsrsquo effector cells could cause different response to these therapies
Mechanism of action CAR-T TCR-T T cell engagementRepresentatives CD20CD3 bispecific (ndashADCCCDC)Limitation Other than antigen escape cancer cells with T cell immune suppressors such as PDL1 overexpressed may have worse response to T cell activation based therapies
Novel ldquoimbalancedrdquo CD20CD3 based cancer therapy
By making a CD20CD3 bispecific antibody with CD3binding affinity significantly reduced we hope toldquosafelyrdquo maintain the effector function and reduce cytokine storm Therefore the novel ldquoimbalancedrdquo CD20CD3 bispecific would have all mechanisms as described above
Binding Ability of Designed Abs Compared to That of Parental Abs
After computational design CD20 homodimer (hole-hole) IgG contains designed VH sequence and common VL
sequence showed similar binding capacity for CD20 compared to that of parental CD20 IgG while CD3
homodimer (knob-knob) IgG contains designed VH sequence and common VL sequence showed significant
reduced binding capacity for CD3 compared to that of parental CD3 IgG
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has Good Efficacy and Safety in vitro
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Day 0 IV injection with ldquoAbsPBS + Raji-Luc cell + human PBMCrdquo first imaging 15min post injection G1 PBS G2 BsAb G3 Rituximab
1 died3 died
Day 0 Day 2 Day 13 Day 22 Day 28
CD20CD3 ldquoImbalancedrdquo BsAb has better Tumor Killing Potency than Rituxan in vivo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Hybridoma + Computational Design
1) Apply computer aided modeling to identify epitopes exposed on 3D surface of antigen
2) Based on target biology 3D information and bioinformatics (such as cross-species homology) design and apply appropriate strategy for immunization
3) Based on target biology and target protein structure design andapply high throughput binding assay functional assay and epitopebin assay to screen the best functional antibody
4) Align the sequences of several ldquobest functional antibodiesrdquo to get ldquobest developable antibodiesrdquo by screening candidates withhigh ldquohuman scorerdquo + less immunogenicity epitopes + less PTMs
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Unique features for phage display technology
1) In house developed 10e9 human naiumlve ScFV phage library witha restriction enzyme site between VH and VL Therefore VH andVL can be replaced separately (suitable for common light chain screening)
2) Leading phage screening not only based on antigen binding ability pre and post heating but also based on ScFV seqsrsquo otherdevelop-ability features After building up a Fv model structure we study
bull Aggregation surface on Fv modelbull PTMs on Fv modelbull Immunogenecity epitopes inside ScFV sequencebull ldquoNaturalityrdquo of VHVL interface by aligning the query ScFVrsquos VHVL
interface to that of real 3D antibody structures in the pdb database
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Antibody Humanization andOptimization
We balance
Humanization percentage
CDR Immunogenicity
Antigen binding affinity
Antibody expression level
Aggregation potential
Antibody stability
Heterogenicity
Workflow of Antibody Humanization and Optimization
Modeling of Mabhomo model vs hetero model
BioinformaticsPTM analysisAggregation surface analysisPatch analysisT cell epitope searchB cell epitope searchMHC II epitope search
Human germlineacceptor selection
CDR graftingBack mutation design
BioinformaticsPTM analysisAggregation surface analysisPatch analysisT cell epitope searchB cell epitope searchMHC II epitope searchHumanization percentage
Lead Selection on Bench1st round Binding based on ELISAor FACS2nd round Bioanalytics includingBiacore DLSDSCCEIEF
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Molecular Modeling and 3D Surface Analysis
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Immunogenecity Epitope Scan
Novel platform 1
Imbalanced Bispecific Antibody Technology Platform
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
The
Zoo
of B
ispe
cific An
tibo
dy Fo
rmats
Ab Studio Inc
your partner for therapeutic antibody development
http
sww
wn
cbin
lmn
ihgo
vpm
carticlesPM
C5
29
75
37
Ulrich Brinkmann
wwwantibodystudiocom
Only a few platforms meet the following criterias
1) Free to operate2) No extra immunogenicity and
developability concerns3) No PKPD concerns
ldquoKnob into holerdquo + Common LC formatis an one meets the above criterias
Ab Studio Inc
your partner for therapeutic antibody development
Imbalanced Bispecific Antibody Technology Platform
What is an imbalanced bispecific antibody
- A ldquoKnob-into-hole plus common LCrdquo version allowing two arms to have very differentbinding affinity to each onersquos own antigen (natural IgG structure and half life)
Why ldquoimbalancedrdquo
- Imbalanced binding affinity is required for some bispecific therapeutic antibodiesrsquoMechanism of Action
wwwantibodystudiocom
Bispecific involves a CD3 binder
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CD3 binding Cancer antigen binding CD3
Cancer antigen
Bispecific
wwwantibodystudiocom
Bispecific against a CSA (cancer specific antigen) and a CAA (cancer associated antigen)
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CSA binding CAA binding
Bispecific
CSA Cancer specific antigen
CAA Cancer associated antigen Normal cell
Normal cell
Normal cell
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Bispecific against one target for CNS and another target on BBB
Ab Studio Inc
your partner for therapeutic antibody development
Brain target
cell
ldquoKnob-into-holerdquo
Common light chain
CNS target binding BBB binding
Bispecific
BBB
Brain target
cell
wwwantibodystudiocom
How ldquoimbalanced bispecific antibodiesrdquo are made at Ab Studio
(lsquoArsquo is the corresponding high affinity arm and lsquoBrsquo is the corresponding low affinity arm)
VL Design
1) If VL-A and VL-B light chain homology gt80 design a VL-C based on VL-Avia computer aided design Maintain VH-AVL-Crsquos affinity as close as that ofVH-AVL-A allow VH-BVL-C to have reduced affinity compared to VH-BVL-B
2) If VL-A and VL-B light chain homology lt80 construct phage ScFV libraryVHs of the ScFV library are human germline VH libraryVLs of the ScFV library are VL-A and its mutants derived from error prone PCR at less than 20 of mutation frequency
VH Design
Use computer aided design to differentiate the biochemical and biophysical featuresof VH-A and VH-B in order to isolate lsquoABrsquo better from lsquoAArsquo and lsquoBBrsquo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Case Study CD20CD3 Bispecific
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chainComputer aided design
CD3 CD20 CD3
Cancer antigen
Bispecific
Biophysical and biochemical features were differentiated for reducing manufacture cost
wwwantibodystudiocom
Difference to other CD20CD3 bispecific ADCCCDC effector function is maintained
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Current CD20+ Cancer Immuno-Therapy
Mechanism of action ADCC CDC ApoptosisRepresentatives Rituxan and its biosimilar biobetterLimitation Other than antigen escape different genetic makeup of patientsrsquo effector cells could cause different response to these therapies
Mechanism of action CAR-T TCR-T T cell engagementRepresentatives CD20CD3 bispecific (ndashADCCCDC)Limitation Other than antigen escape cancer cells with T cell immune suppressors such as PDL1 overexpressed may have worse response to T cell activation based therapies
Novel ldquoimbalancedrdquo CD20CD3 based cancer therapy
By making a CD20CD3 bispecific antibody with CD3binding affinity significantly reduced we hope toldquosafelyrdquo maintain the effector function and reduce cytokine storm Therefore the novel ldquoimbalancedrdquo CD20CD3 bispecific would have all mechanisms as described above
Binding Ability of Designed Abs Compared to That of Parental Abs
After computational design CD20 homodimer (hole-hole) IgG contains designed VH sequence and common VL
sequence showed similar binding capacity for CD20 compared to that of parental CD20 IgG while CD3
homodimer (knob-knob) IgG contains designed VH sequence and common VL sequence showed significant
reduced binding capacity for CD3 compared to that of parental CD3 IgG
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has Good Efficacy and Safety in vitro
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Day 0 IV injection with ldquoAbsPBS + Raji-Luc cell + human PBMCrdquo first imaging 15min post injection G1 PBS G2 BsAb G3 Rituximab
1 died3 died
Day 0 Day 2 Day 13 Day 22 Day 28
CD20CD3 ldquoImbalancedrdquo BsAb has better Tumor Killing Potency than Rituxan in vivo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Unique features for phage display technology
1) In house developed 10e9 human naiumlve ScFV phage library witha restriction enzyme site between VH and VL Therefore VH andVL can be replaced separately (suitable for common light chain screening)
2) Leading phage screening not only based on antigen binding ability pre and post heating but also based on ScFV seqsrsquo otherdevelop-ability features After building up a Fv model structure we study
bull Aggregation surface on Fv modelbull PTMs on Fv modelbull Immunogenecity epitopes inside ScFV sequencebull ldquoNaturalityrdquo of VHVL interface by aligning the query ScFVrsquos VHVL
interface to that of real 3D antibody structures in the pdb database
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Antibody Humanization andOptimization
We balance
Humanization percentage
CDR Immunogenicity
Antigen binding affinity
Antibody expression level
Aggregation potential
Antibody stability
Heterogenicity
Workflow of Antibody Humanization and Optimization
Modeling of Mabhomo model vs hetero model
BioinformaticsPTM analysisAggregation surface analysisPatch analysisT cell epitope searchB cell epitope searchMHC II epitope search
Human germlineacceptor selection
CDR graftingBack mutation design
BioinformaticsPTM analysisAggregation surface analysisPatch analysisT cell epitope searchB cell epitope searchMHC II epitope searchHumanization percentage
Lead Selection on Bench1st round Binding based on ELISAor FACS2nd round Bioanalytics includingBiacore DLSDSCCEIEF
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Molecular Modeling and 3D Surface Analysis
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Immunogenecity Epitope Scan
Novel platform 1
Imbalanced Bispecific Antibody Technology Platform
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
The
Zoo
of B
ispe
cific An
tibo
dy Fo
rmats
Ab Studio Inc
your partner for therapeutic antibody development
http
sww
wn
cbin
lmn
ihgo
vpm
carticlesPM
C5
29
75
37
Ulrich Brinkmann
wwwantibodystudiocom
Only a few platforms meet the following criterias
1) Free to operate2) No extra immunogenicity and
developability concerns3) No PKPD concerns
ldquoKnob into holerdquo + Common LC formatis an one meets the above criterias
Ab Studio Inc
your partner for therapeutic antibody development
Imbalanced Bispecific Antibody Technology Platform
What is an imbalanced bispecific antibody
- A ldquoKnob-into-hole plus common LCrdquo version allowing two arms to have very differentbinding affinity to each onersquos own antigen (natural IgG structure and half life)
Why ldquoimbalancedrdquo
- Imbalanced binding affinity is required for some bispecific therapeutic antibodiesrsquoMechanism of Action
wwwantibodystudiocom
Bispecific involves a CD3 binder
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CD3 binding Cancer antigen binding CD3
Cancer antigen
Bispecific
wwwantibodystudiocom
Bispecific against a CSA (cancer specific antigen) and a CAA (cancer associated antigen)
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CSA binding CAA binding
Bispecific
CSA Cancer specific antigen
CAA Cancer associated antigen Normal cell
Normal cell
Normal cell
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Bispecific against one target for CNS and another target on BBB
Ab Studio Inc
your partner for therapeutic antibody development
Brain target
cell
ldquoKnob-into-holerdquo
Common light chain
CNS target binding BBB binding
Bispecific
BBB
Brain target
cell
wwwantibodystudiocom
How ldquoimbalanced bispecific antibodiesrdquo are made at Ab Studio
(lsquoArsquo is the corresponding high affinity arm and lsquoBrsquo is the corresponding low affinity arm)
VL Design
1) If VL-A and VL-B light chain homology gt80 design a VL-C based on VL-Avia computer aided design Maintain VH-AVL-Crsquos affinity as close as that ofVH-AVL-A allow VH-BVL-C to have reduced affinity compared to VH-BVL-B
2) If VL-A and VL-B light chain homology lt80 construct phage ScFV libraryVHs of the ScFV library are human germline VH libraryVLs of the ScFV library are VL-A and its mutants derived from error prone PCR at less than 20 of mutation frequency
VH Design
Use computer aided design to differentiate the biochemical and biophysical featuresof VH-A and VH-B in order to isolate lsquoABrsquo better from lsquoAArsquo and lsquoBBrsquo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Case Study CD20CD3 Bispecific
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chainComputer aided design
CD3 CD20 CD3
Cancer antigen
Bispecific
Biophysical and biochemical features were differentiated for reducing manufacture cost
wwwantibodystudiocom
Difference to other CD20CD3 bispecific ADCCCDC effector function is maintained
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Current CD20+ Cancer Immuno-Therapy
Mechanism of action ADCC CDC ApoptosisRepresentatives Rituxan and its biosimilar biobetterLimitation Other than antigen escape different genetic makeup of patientsrsquo effector cells could cause different response to these therapies
Mechanism of action CAR-T TCR-T T cell engagementRepresentatives CD20CD3 bispecific (ndashADCCCDC)Limitation Other than antigen escape cancer cells with T cell immune suppressors such as PDL1 overexpressed may have worse response to T cell activation based therapies
Novel ldquoimbalancedrdquo CD20CD3 based cancer therapy
By making a CD20CD3 bispecific antibody with CD3binding affinity significantly reduced we hope toldquosafelyrdquo maintain the effector function and reduce cytokine storm Therefore the novel ldquoimbalancedrdquo CD20CD3 bispecific would have all mechanisms as described above
Binding Ability of Designed Abs Compared to That of Parental Abs
After computational design CD20 homodimer (hole-hole) IgG contains designed VH sequence and common VL
sequence showed similar binding capacity for CD20 compared to that of parental CD20 IgG while CD3
homodimer (knob-knob) IgG contains designed VH sequence and common VL sequence showed significant
reduced binding capacity for CD3 compared to that of parental CD3 IgG
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has Good Efficacy and Safety in vitro
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Day 0 IV injection with ldquoAbsPBS + Raji-Luc cell + human PBMCrdquo first imaging 15min post injection G1 PBS G2 BsAb G3 Rituximab
1 died3 died
Day 0 Day 2 Day 13 Day 22 Day 28
CD20CD3 ldquoImbalancedrdquo BsAb has better Tumor Killing Potency than Rituxan in vivo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Antibody Humanization andOptimization
We balance
Humanization percentage
CDR Immunogenicity
Antigen binding affinity
Antibody expression level
Aggregation potential
Antibody stability
Heterogenicity
Workflow of Antibody Humanization and Optimization
Modeling of Mabhomo model vs hetero model
BioinformaticsPTM analysisAggregation surface analysisPatch analysisT cell epitope searchB cell epitope searchMHC II epitope search
Human germlineacceptor selection
CDR graftingBack mutation design
BioinformaticsPTM analysisAggregation surface analysisPatch analysisT cell epitope searchB cell epitope searchMHC II epitope searchHumanization percentage
Lead Selection on Bench1st round Binding based on ELISAor FACS2nd round Bioanalytics includingBiacore DLSDSCCEIEF
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Molecular Modeling and 3D Surface Analysis
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Immunogenecity Epitope Scan
Novel platform 1
Imbalanced Bispecific Antibody Technology Platform
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
The
Zoo
of B
ispe
cific An
tibo
dy Fo
rmats
Ab Studio Inc
your partner for therapeutic antibody development
http
sww
wn
cbin
lmn
ihgo
vpm
carticlesPM
C5
29
75
37
Ulrich Brinkmann
wwwantibodystudiocom
Only a few platforms meet the following criterias
1) Free to operate2) No extra immunogenicity and
developability concerns3) No PKPD concerns
ldquoKnob into holerdquo + Common LC formatis an one meets the above criterias
Ab Studio Inc
your partner for therapeutic antibody development
Imbalanced Bispecific Antibody Technology Platform
What is an imbalanced bispecific antibody
- A ldquoKnob-into-hole plus common LCrdquo version allowing two arms to have very differentbinding affinity to each onersquos own antigen (natural IgG structure and half life)
Why ldquoimbalancedrdquo
- Imbalanced binding affinity is required for some bispecific therapeutic antibodiesrsquoMechanism of Action
wwwantibodystudiocom
Bispecific involves a CD3 binder
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CD3 binding Cancer antigen binding CD3
Cancer antigen
Bispecific
wwwantibodystudiocom
Bispecific against a CSA (cancer specific antigen) and a CAA (cancer associated antigen)
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CSA binding CAA binding
Bispecific
CSA Cancer specific antigen
CAA Cancer associated antigen Normal cell
Normal cell
Normal cell
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Bispecific against one target for CNS and another target on BBB
Ab Studio Inc
your partner for therapeutic antibody development
Brain target
cell
ldquoKnob-into-holerdquo
Common light chain
CNS target binding BBB binding
Bispecific
BBB
Brain target
cell
wwwantibodystudiocom
How ldquoimbalanced bispecific antibodiesrdquo are made at Ab Studio
(lsquoArsquo is the corresponding high affinity arm and lsquoBrsquo is the corresponding low affinity arm)
VL Design
1) If VL-A and VL-B light chain homology gt80 design a VL-C based on VL-Avia computer aided design Maintain VH-AVL-Crsquos affinity as close as that ofVH-AVL-A allow VH-BVL-C to have reduced affinity compared to VH-BVL-B
2) If VL-A and VL-B light chain homology lt80 construct phage ScFV libraryVHs of the ScFV library are human germline VH libraryVLs of the ScFV library are VL-A and its mutants derived from error prone PCR at less than 20 of mutation frequency
VH Design
Use computer aided design to differentiate the biochemical and biophysical featuresof VH-A and VH-B in order to isolate lsquoABrsquo better from lsquoAArsquo and lsquoBBrsquo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Case Study CD20CD3 Bispecific
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chainComputer aided design
CD3 CD20 CD3
Cancer antigen
Bispecific
Biophysical and biochemical features were differentiated for reducing manufacture cost
wwwantibodystudiocom
Difference to other CD20CD3 bispecific ADCCCDC effector function is maintained
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Current CD20+ Cancer Immuno-Therapy
Mechanism of action ADCC CDC ApoptosisRepresentatives Rituxan and its biosimilar biobetterLimitation Other than antigen escape different genetic makeup of patientsrsquo effector cells could cause different response to these therapies
Mechanism of action CAR-T TCR-T T cell engagementRepresentatives CD20CD3 bispecific (ndashADCCCDC)Limitation Other than antigen escape cancer cells with T cell immune suppressors such as PDL1 overexpressed may have worse response to T cell activation based therapies
Novel ldquoimbalancedrdquo CD20CD3 based cancer therapy
By making a CD20CD3 bispecific antibody with CD3binding affinity significantly reduced we hope toldquosafelyrdquo maintain the effector function and reduce cytokine storm Therefore the novel ldquoimbalancedrdquo CD20CD3 bispecific would have all mechanisms as described above
Binding Ability of Designed Abs Compared to That of Parental Abs
After computational design CD20 homodimer (hole-hole) IgG contains designed VH sequence and common VL
sequence showed similar binding capacity for CD20 compared to that of parental CD20 IgG while CD3
homodimer (knob-knob) IgG contains designed VH sequence and common VL sequence showed significant
reduced binding capacity for CD3 compared to that of parental CD3 IgG
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has Good Efficacy and Safety in vitro
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Day 0 IV injection with ldquoAbsPBS + Raji-Luc cell + human PBMCrdquo first imaging 15min post injection G1 PBS G2 BsAb G3 Rituximab
1 died3 died
Day 0 Day 2 Day 13 Day 22 Day 28
CD20CD3 ldquoImbalancedrdquo BsAb has better Tumor Killing Potency than Rituxan in vivo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Workflow of Antibody Humanization and Optimization
Modeling of Mabhomo model vs hetero model
BioinformaticsPTM analysisAggregation surface analysisPatch analysisT cell epitope searchB cell epitope searchMHC II epitope search
Human germlineacceptor selection
CDR graftingBack mutation design
BioinformaticsPTM analysisAggregation surface analysisPatch analysisT cell epitope searchB cell epitope searchMHC II epitope searchHumanization percentage
Lead Selection on Bench1st round Binding based on ELISAor FACS2nd round Bioanalytics includingBiacore DLSDSCCEIEF
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Molecular Modeling and 3D Surface Analysis
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Immunogenecity Epitope Scan
Novel platform 1
Imbalanced Bispecific Antibody Technology Platform
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
The
Zoo
of B
ispe
cific An
tibo
dy Fo
rmats
Ab Studio Inc
your partner for therapeutic antibody development
http
sww
wn
cbin
lmn
ihgo
vpm
carticlesPM
C5
29
75
37
Ulrich Brinkmann
wwwantibodystudiocom
Only a few platforms meet the following criterias
1) Free to operate2) No extra immunogenicity and
developability concerns3) No PKPD concerns
ldquoKnob into holerdquo + Common LC formatis an one meets the above criterias
Ab Studio Inc
your partner for therapeutic antibody development
Imbalanced Bispecific Antibody Technology Platform
What is an imbalanced bispecific antibody
- A ldquoKnob-into-hole plus common LCrdquo version allowing two arms to have very differentbinding affinity to each onersquos own antigen (natural IgG structure and half life)
Why ldquoimbalancedrdquo
- Imbalanced binding affinity is required for some bispecific therapeutic antibodiesrsquoMechanism of Action
wwwantibodystudiocom
Bispecific involves a CD3 binder
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CD3 binding Cancer antigen binding CD3
Cancer antigen
Bispecific
wwwantibodystudiocom
Bispecific against a CSA (cancer specific antigen) and a CAA (cancer associated antigen)
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CSA binding CAA binding
Bispecific
CSA Cancer specific antigen
CAA Cancer associated antigen Normal cell
Normal cell
Normal cell
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Bispecific against one target for CNS and another target on BBB
Ab Studio Inc
your partner for therapeutic antibody development
Brain target
cell
ldquoKnob-into-holerdquo
Common light chain
CNS target binding BBB binding
Bispecific
BBB
Brain target
cell
wwwantibodystudiocom
How ldquoimbalanced bispecific antibodiesrdquo are made at Ab Studio
(lsquoArsquo is the corresponding high affinity arm and lsquoBrsquo is the corresponding low affinity arm)
VL Design
1) If VL-A and VL-B light chain homology gt80 design a VL-C based on VL-Avia computer aided design Maintain VH-AVL-Crsquos affinity as close as that ofVH-AVL-A allow VH-BVL-C to have reduced affinity compared to VH-BVL-B
2) If VL-A and VL-B light chain homology lt80 construct phage ScFV libraryVHs of the ScFV library are human germline VH libraryVLs of the ScFV library are VL-A and its mutants derived from error prone PCR at less than 20 of mutation frequency
VH Design
Use computer aided design to differentiate the biochemical and biophysical featuresof VH-A and VH-B in order to isolate lsquoABrsquo better from lsquoAArsquo and lsquoBBrsquo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Case Study CD20CD3 Bispecific
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chainComputer aided design
CD3 CD20 CD3
Cancer antigen
Bispecific
Biophysical and biochemical features were differentiated for reducing manufacture cost
wwwantibodystudiocom
Difference to other CD20CD3 bispecific ADCCCDC effector function is maintained
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Current CD20+ Cancer Immuno-Therapy
Mechanism of action ADCC CDC ApoptosisRepresentatives Rituxan and its biosimilar biobetterLimitation Other than antigen escape different genetic makeup of patientsrsquo effector cells could cause different response to these therapies
Mechanism of action CAR-T TCR-T T cell engagementRepresentatives CD20CD3 bispecific (ndashADCCCDC)Limitation Other than antigen escape cancer cells with T cell immune suppressors such as PDL1 overexpressed may have worse response to T cell activation based therapies
Novel ldquoimbalancedrdquo CD20CD3 based cancer therapy
By making a CD20CD3 bispecific antibody with CD3binding affinity significantly reduced we hope toldquosafelyrdquo maintain the effector function and reduce cytokine storm Therefore the novel ldquoimbalancedrdquo CD20CD3 bispecific would have all mechanisms as described above
Binding Ability of Designed Abs Compared to That of Parental Abs
After computational design CD20 homodimer (hole-hole) IgG contains designed VH sequence and common VL
sequence showed similar binding capacity for CD20 compared to that of parental CD20 IgG while CD3
homodimer (knob-knob) IgG contains designed VH sequence and common VL sequence showed significant
reduced binding capacity for CD3 compared to that of parental CD3 IgG
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has Good Efficacy and Safety in vitro
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Day 0 IV injection with ldquoAbsPBS + Raji-Luc cell + human PBMCrdquo first imaging 15min post injection G1 PBS G2 BsAb G3 Rituximab
1 died3 died
Day 0 Day 2 Day 13 Day 22 Day 28
CD20CD3 ldquoImbalancedrdquo BsAb has better Tumor Killing Potency than Rituxan in vivo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Molecular Modeling and 3D Surface Analysis
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Immunogenecity Epitope Scan
Novel platform 1
Imbalanced Bispecific Antibody Technology Platform
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
The
Zoo
of B
ispe
cific An
tibo
dy Fo
rmats
Ab Studio Inc
your partner for therapeutic antibody development
http
sww
wn
cbin
lmn
ihgo
vpm
carticlesPM
C5
29
75
37
Ulrich Brinkmann
wwwantibodystudiocom
Only a few platforms meet the following criterias
1) Free to operate2) No extra immunogenicity and
developability concerns3) No PKPD concerns
ldquoKnob into holerdquo + Common LC formatis an one meets the above criterias
Ab Studio Inc
your partner for therapeutic antibody development
Imbalanced Bispecific Antibody Technology Platform
What is an imbalanced bispecific antibody
- A ldquoKnob-into-hole plus common LCrdquo version allowing two arms to have very differentbinding affinity to each onersquos own antigen (natural IgG structure and half life)
Why ldquoimbalancedrdquo
- Imbalanced binding affinity is required for some bispecific therapeutic antibodiesrsquoMechanism of Action
wwwantibodystudiocom
Bispecific involves a CD3 binder
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CD3 binding Cancer antigen binding CD3
Cancer antigen
Bispecific
wwwantibodystudiocom
Bispecific against a CSA (cancer specific antigen) and a CAA (cancer associated antigen)
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CSA binding CAA binding
Bispecific
CSA Cancer specific antigen
CAA Cancer associated antigen Normal cell
Normal cell
Normal cell
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Bispecific against one target for CNS and another target on BBB
Ab Studio Inc
your partner for therapeutic antibody development
Brain target
cell
ldquoKnob-into-holerdquo
Common light chain
CNS target binding BBB binding
Bispecific
BBB
Brain target
cell
wwwantibodystudiocom
How ldquoimbalanced bispecific antibodiesrdquo are made at Ab Studio
(lsquoArsquo is the corresponding high affinity arm and lsquoBrsquo is the corresponding low affinity arm)
VL Design
1) If VL-A and VL-B light chain homology gt80 design a VL-C based on VL-Avia computer aided design Maintain VH-AVL-Crsquos affinity as close as that ofVH-AVL-A allow VH-BVL-C to have reduced affinity compared to VH-BVL-B
2) If VL-A and VL-B light chain homology lt80 construct phage ScFV libraryVHs of the ScFV library are human germline VH libraryVLs of the ScFV library are VL-A and its mutants derived from error prone PCR at less than 20 of mutation frequency
VH Design
Use computer aided design to differentiate the biochemical and biophysical featuresof VH-A and VH-B in order to isolate lsquoABrsquo better from lsquoAArsquo and lsquoBBrsquo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Case Study CD20CD3 Bispecific
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chainComputer aided design
CD3 CD20 CD3
Cancer antigen
Bispecific
Biophysical and biochemical features were differentiated for reducing manufacture cost
wwwantibodystudiocom
Difference to other CD20CD3 bispecific ADCCCDC effector function is maintained
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Current CD20+ Cancer Immuno-Therapy
Mechanism of action ADCC CDC ApoptosisRepresentatives Rituxan and its biosimilar biobetterLimitation Other than antigen escape different genetic makeup of patientsrsquo effector cells could cause different response to these therapies
Mechanism of action CAR-T TCR-T T cell engagementRepresentatives CD20CD3 bispecific (ndashADCCCDC)Limitation Other than antigen escape cancer cells with T cell immune suppressors such as PDL1 overexpressed may have worse response to T cell activation based therapies
Novel ldquoimbalancedrdquo CD20CD3 based cancer therapy
By making a CD20CD3 bispecific antibody with CD3binding affinity significantly reduced we hope toldquosafelyrdquo maintain the effector function and reduce cytokine storm Therefore the novel ldquoimbalancedrdquo CD20CD3 bispecific would have all mechanisms as described above
Binding Ability of Designed Abs Compared to That of Parental Abs
After computational design CD20 homodimer (hole-hole) IgG contains designed VH sequence and common VL
sequence showed similar binding capacity for CD20 compared to that of parental CD20 IgG while CD3
homodimer (knob-knob) IgG contains designed VH sequence and common VL sequence showed significant
reduced binding capacity for CD3 compared to that of parental CD3 IgG
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has Good Efficacy and Safety in vitro
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Day 0 IV injection with ldquoAbsPBS + Raji-Luc cell + human PBMCrdquo first imaging 15min post injection G1 PBS G2 BsAb G3 Rituximab
1 died3 died
Day 0 Day 2 Day 13 Day 22 Day 28
CD20CD3 ldquoImbalancedrdquo BsAb has better Tumor Killing Potency than Rituxan in vivo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Immunogenecity Epitope Scan
Novel platform 1
Imbalanced Bispecific Antibody Technology Platform
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
The
Zoo
of B
ispe
cific An
tibo
dy Fo
rmats
Ab Studio Inc
your partner for therapeutic antibody development
http
sww
wn
cbin
lmn
ihgo
vpm
carticlesPM
C5
29
75
37
Ulrich Brinkmann
wwwantibodystudiocom
Only a few platforms meet the following criterias
1) Free to operate2) No extra immunogenicity and
developability concerns3) No PKPD concerns
ldquoKnob into holerdquo + Common LC formatis an one meets the above criterias
Ab Studio Inc
your partner for therapeutic antibody development
Imbalanced Bispecific Antibody Technology Platform
What is an imbalanced bispecific antibody
- A ldquoKnob-into-hole plus common LCrdquo version allowing two arms to have very differentbinding affinity to each onersquos own antigen (natural IgG structure and half life)
Why ldquoimbalancedrdquo
- Imbalanced binding affinity is required for some bispecific therapeutic antibodiesrsquoMechanism of Action
wwwantibodystudiocom
Bispecific involves a CD3 binder
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CD3 binding Cancer antigen binding CD3
Cancer antigen
Bispecific
wwwantibodystudiocom
Bispecific against a CSA (cancer specific antigen) and a CAA (cancer associated antigen)
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CSA binding CAA binding
Bispecific
CSA Cancer specific antigen
CAA Cancer associated antigen Normal cell
Normal cell
Normal cell
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Bispecific against one target for CNS and another target on BBB
Ab Studio Inc
your partner for therapeutic antibody development
Brain target
cell
ldquoKnob-into-holerdquo
Common light chain
CNS target binding BBB binding
Bispecific
BBB
Brain target
cell
wwwantibodystudiocom
How ldquoimbalanced bispecific antibodiesrdquo are made at Ab Studio
(lsquoArsquo is the corresponding high affinity arm and lsquoBrsquo is the corresponding low affinity arm)
VL Design
1) If VL-A and VL-B light chain homology gt80 design a VL-C based on VL-Avia computer aided design Maintain VH-AVL-Crsquos affinity as close as that ofVH-AVL-A allow VH-BVL-C to have reduced affinity compared to VH-BVL-B
2) If VL-A and VL-B light chain homology lt80 construct phage ScFV libraryVHs of the ScFV library are human germline VH libraryVLs of the ScFV library are VL-A and its mutants derived from error prone PCR at less than 20 of mutation frequency
VH Design
Use computer aided design to differentiate the biochemical and biophysical featuresof VH-A and VH-B in order to isolate lsquoABrsquo better from lsquoAArsquo and lsquoBBrsquo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Case Study CD20CD3 Bispecific
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chainComputer aided design
CD3 CD20 CD3
Cancer antigen
Bispecific
Biophysical and biochemical features were differentiated for reducing manufacture cost
wwwantibodystudiocom
Difference to other CD20CD3 bispecific ADCCCDC effector function is maintained
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Current CD20+ Cancer Immuno-Therapy
Mechanism of action ADCC CDC ApoptosisRepresentatives Rituxan and its biosimilar biobetterLimitation Other than antigen escape different genetic makeup of patientsrsquo effector cells could cause different response to these therapies
Mechanism of action CAR-T TCR-T T cell engagementRepresentatives CD20CD3 bispecific (ndashADCCCDC)Limitation Other than antigen escape cancer cells with T cell immune suppressors such as PDL1 overexpressed may have worse response to T cell activation based therapies
Novel ldquoimbalancedrdquo CD20CD3 based cancer therapy
By making a CD20CD3 bispecific antibody with CD3binding affinity significantly reduced we hope toldquosafelyrdquo maintain the effector function and reduce cytokine storm Therefore the novel ldquoimbalancedrdquo CD20CD3 bispecific would have all mechanisms as described above
Binding Ability of Designed Abs Compared to That of Parental Abs
After computational design CD20 homodimer (hole-hole) IgG contains designed VH sequence and common VL
sequence showed similar binding capacity for CD20 compared to that of parental CD20 IgG while CD3
homodimer (knob-knob) IgG contains designed VH sequence and common VL sequence showed significant
reduced binding capacity for CD3 compared to that of parental CD3 IgG
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has Good Efficacy and Safety in vitro
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Day 0 IV injection with ldquoAbsPBS + Raji-Luc cell + human PBMCrdquo first imaging 15min post injection G1 PBS G2 BsAb G3 Rituximab
1 died3 died
Day 0 Day 2 Day 13 Day 22 Day 28
CD20CD3 ldquoImbalancedrdquo BsAb has better Tumor Killing Potency than Rituxan in vivo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Novel platform 1
Imbalanced Bispecific Antibody Technology Platform
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
The
Zoo
of B
ispe
cific An
tibo
dy Fo
rmats
Ab Studio Inc
your partner for therapeutic antibody development
http
sww
wn
cbin
lmn
ihgo
vpm
carticlesPM
C5
29
75
37
Ulrich Brinkmann
wwwantibodystudiocom
Only a few platforms meet the following criterias
1) Free to operate2) No extra immunogenicity and
developability concerns3) No PKPD concerns
ldquoKnob into holerdquo + Common LC formatis an one meets the above criterias
Ab Studio Inc
your partner for therapeutic antibody development
Imbalanced Bispecific Antibody Technology Platform
What is an imbalanced bispecific antibody
- A ldquoKnob-into-hole plus common LCrdquo version allowing two arms to have very differentbinding affinity to each onersquos own antigen (natural IgG structure and half life)
Why ldquoimbalancedrdquo
- Imbalanced binding affinity is required for some bispecific therapeutic antibodiesrsquoMechanism of Action
wwwantibodystudiocom
Bispecific involves a CD3 binder
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CD3 binding Cancer antigen binding CD3
Cancer antigen
Bispecific
wwwantibodystudiocom
Bispecific against a CSA (cancer specific antigen) and a CAA (cancer associated antigen)
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CSA binding CAA binding
Bispecific
CSA Cancer specific antigen
CAA Cancer associated antigen Normal cell
Normal cell
Normal cell
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Bispecific against one target for CNS and another target on BBB
Ab Studio Inc
your partner for therapeutic antibody development
Brain target
cell
ldquoKnob-into-holerdquo
Common light chain
CNS target binding BBB binding
Bispecific
BBB
Brain target
cell
wwwantibodystudiocom
How ldquoimbalanced bispecific antibodiesrdquo are made at Ab Studio
(lsquoArsquo is the corresponding high affinity arm and lsquoBrsquo is the corresponding low affinity arm)
VL Design
1) If VL-A and VL-B light chain homology gt80 design a VL-C based on VL-Avia computer aided design Maintain VH-AVL-Crsquos affinity as close as that ofVH-AVL-A allow VH-BVL-C to have reduced affinity compared to VH-BVL-B
2) If VL-A and VL-B light chain homology lt80 construct phage ScFV libraryVHs of the ScFV library are human germline VH libraryVLs of the ScFV library are VL-A and its mutants derived from error prone PCR at less than 20 of mutation frequency
VH Design
Use computer aided design to differentiate the biochemical and biophysical featuresof VH-A and VH-B in order to isolate lsquoABrsquo better from lsquoAArsquo and lsquoBBrsquo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Case Study CD20CD3 Bispecific
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chainComputer aided design
CD3 CD20 CD3
Cancer antigen
Bispecific
Biophysical and biochemical features were differentiated for reducing manufacture cost
wwwantibodystudiocom
Difference to other CD20CD3 bispecific ADCCCDC effector function is maintained
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Current CD20+ Cancer Immuno-Therapy
Mechanism of action ADCC CDC ApoptosisRepresentatives Rituxan and its biosimilar biobetterLimitation Other than antigen escape different genetic makeup of patientsrsquo effector cells could cause different response to these therapies
Mechanism of action CAR-T TCR-T T cell engagementRepresentatives CD20CD3 bispecific (ndashADCCCDC)Limitation Other than antigen escape cancer cells with T cell immune suppressors such as PDL1 overexpressed may have worse response to T cell activation based therapies
Novel ldquoimbalancedrdquo CD20CD3 based cancer therapy
By making a CD20CD3 bispecific antibody with CD3binding affinity significantly reduced we hope toldquosafelyrdquo maintain the effector function and reduce cytokine storm Therefore the novel ldquoimbalancedrdquo CD20CD3 bispecific would have all mechanisms as described above
Binding Ability of Designed Abs Compared to That of Parental Abs
After computational design CD20 homodimer (hole-hole) IgG contains designed VH sequence and common VL
sequence showed similar binding capacity for CD20 compared to that of parental CD20 IgG while CD3
homodimer (knob-knob) IgG contains designed VH sequence and common VL sequence showed significant
reduced binding capacity for CD3 compared to that of parental CD3 IgG
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has Good Efficacy and Safety in vitro
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Day 0 IV injection with ldquoAbsPBS + Raji-Luc cell + human PBMCrdquo first imaging 15min post injection G1 PBS G2 BsAb G3 Rituximab
1 died3 died
Day 0 Day 2 Day 13 Day 22 Day 28
CD20CD3 ldquoImbalancedrdquo BsAb has better Tumor Killing Potency than Rituxan in vivo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
The
Zoo
of B
ispe
cific An
tibo
dy Fo
rmats
Ab Studio Inc
your partner for therapeutic antibody development
http
sww
wn
cbin
lmn
ihgo
vpm
carticlesPM
C5
29
75
37
Ulrich Brinkmann
wwwantibodystudiocom
Only a few platforms meet the following criterias
1) Free to operate2) No extra immunogenicity and
developability concerns3) No PKPD concerns
ldquoKnob into holerdquo + Common LC formatis an one meets the above criterias
Ab Studio Inc
your partner for therapeutic antibody development
Imbalanced Bispecific Antibody Technology Platform
What is an imbalanced bispecific antibody
- A ldquoKnob-into-hole plus common LCrdquo version allowing two arms to have very differentbinding affinity to each onersquos own antigen (natural IgG structure and half life)
Why ldquoimbalancedrdquo
- Imbalanced binding affinity is required for some bispecific therapeutic antibodiesrsquoMechanism of Action
wwwantibodystudiocom
Bispecific involves a CD3 binder
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CD3 binding Cancer antigen binding CD3
Cancer antigen
Bispecific
wwwantibodystudiocom
Bispecific against a CSA (cancer specific antigen) and a CAA (cancer associated antigen)
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CSA binding CAA binding
Bispecific
CSA Cancer specific antigen
CAA Cancer associated antigen Normal cell
Normal cell
Normal cell
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Bispecific against one target for CNS and another target on BBB
Ab Studio Inc
your partner for therapeutic antibody development
Brain target
cell
ldquoKnob-into-holerdquo
Common light chain
CNS target binding BBB binding
Bispecific
BBB
Brain target
cell
wwwantibodystudiocom
How ldquoimbalanced bispecific antibodiesrdquo are made at Ab Studio
(lsquoArsquo is the corresponding high affinity arm and lsquoBrsquo is the corresponding low affinity arm)
VL Design
1) If VL-A and VL-B light chain homology gt80 design a VL-C based on VL-Avia computer aided design Maintain VH-AVL-Crsquos affinity as close as that ofVH-AVL-A allow VH-BVL-C to have reduced affinity compared to VH-BVL-B
2) If VL-A and VL-B light chain homology lt80 construct phage ScFV libraryVHs of the ScFV library are human germline VH libraryVLs of the ScFV library are VL-A and its mutants derived from error prone PCR at less than 20 of mutation frequency
VH Design
Use computer aided design to differentiate the biochemical and biophysical featuresof VH-A and VH-B in order to isolate lsquoABrsquo better from lsquoAArsquo and lsquoBBrsquo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Case Study CD20CD3 Bispecific
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chainComputer aided design
CD3 CD20 CD3
Cancer antigen
Bispecific
Biophysical and biochemical features were differentiated for reducing manufacture cost
wwwantibodystudiocom
Difference to other CD20CD3 bispecific ADCCCDC effector function is maintained
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Current CD20+ Cancer Immuno-Therapy
Mechanism of action ADCC CDC ApoptosisRepresentatives Rituxan and its biosimilar biobetterLimitation Other than antigen escape different genetic makeup of patientsrsquo effector cells could cause different response to these therapies
Mechanism of action CAR-T TCR-T T cell engagementRepresentatives CD20CD3 bispecific (ndashADCCCDC)Limitation Other than antigen escape cancer cells with T cell immune suppressors such as PDL1 overexpressed may have worse response to T cell activation based therapies
Novel ldquoimbalancedrdquo CD20CD3 based cancer therapy
By making a CD20CD3 bispecific antibody with CD3binding affinity significantly reduced we hope toldquosafelyrdquo maintain the effector function and reduce cytokine storm Therefore the novel ldquoimbalancedrdquo CD20CD3 bispecific would have all mechanisms as described above
Binding Ability of Designed Abs Compared to That of Parental Abs
After computational design CD20 homodimer (hole-hole) IgG contains designed VH sequence and common VL
sequence showed similar binding capacity for CD20 compared to that of parental CD20 IgG while CD3
homodimer (knob-knob) IgG contains designed VH sequence and common VL sequence showed significant
reduced binding capacity for CD3 compared to that of parental CD3 IgG
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has Good Efficacy and Safety in vitro
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Day 0 IV injection with ldquoAbsPBS + Raji-Luc cell + human PBMCrdquo first imaging 15min post injection G1 PBS G2 BsAb G3 Rituximab
1 died3 died
Day 0 Day 2 Day 13 Day 22 Day 28
CD20CD3 ldquoImbalancedrdquo BsAb has better Tumor Killing Potency than Rituxan in vivo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Ab Studio Inc
your partner for therapeutic antibody development
Imbalanced Bispecific Antibody Technology Platform
What is an imbalanced bispecific antibody
- A ldquoKnob-into-hole plus common LCrdquo version allowing two arms to have very differentbinding affinity to each onersquos own antigen (natural IgG structure and half life)
Why ldquoimbalancedrdquo
- Imbalanced binding affinity is required for some bispecific therapeutic antibodiesrsquoMechanism of Action
wwwantibodystudiocom
Bispecific involves a CD3 binder
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CD3 binding Cancer antigen binding CD3
Cancer antigen
Bispecific
wwwantibodystudiocom
Bispecific against a CSA (cancer specific antigen) and a CAA (cancer associated antigen)
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CSA binding CAA binding
Bispecific
CSA Cancer specific antigen
CAA Cancer associated antigen Normal cell
Normal cell
Normal cell
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Bispecific against one target for CNS and another target on BBB
Ab Studio Inc
your partner for therapeutic antibody development
Brain target
cell
ldquoKnob-into-holerdquo
Common light chain
CNS target binding BBB binding
Bispecific
BBB
Brain target
cell
wwwantibodystudiocom
How ldquoimbalanced bispecific antibodiesrdquo are made at Ab Studio
(lsquoArsquo is the corresponding high affinity arm and lsquoBrsquo is the corresponding low affinity arm)
VL Design
1) If VL-A and VL-B light chain homology gt80 design a VL-C based on VL-Avia computer aided design Maintain VH-AVL-Crsquos affinity as close as that ofVH-AVL-A allow VH-BVL-C to have reduced affinity compared to VH-BVL-B
2) If VL-A and VL-B light chain homology lt80 construct phage ScFV libraryVHs of the ScFV library are human germline VH libraryVLs of the ScFV library are VL-A and its mutants derived from error prone PCR at less than 20 of mutation frequency
VH Design
Use computer aided design to differentiate the biochemical and biophysical featuresof VH-A and VH-B in order to isolate lsquoABrsquo better from lsquoAArsquo and lsquoBBrsquo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Case Study CD20CD3 Bispecific
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chainComputer aided design
CD3 CD20 CD3
Cancer antigen
Bispecific
Biophysical and biochemical features were differentiated for reducing manufacture cost
wwwantibodystudiocom
Difference to other CD20CD3 bispecific ADCCCDC effector function is maintained
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Current CD20+ Cancer Immuno-Therapy
Mechanism of action ADCC CDC ApoptosisRepresentatives Rituxan and its biosimilar biobetterLimitation Other than antigen escape different genetic makeup of patientsrsquo effector cells could cause different response to these therapies
Mechanism of action CAR-T TCR-T T cell engagementRepresentatives CD20CD3 bispecific (ndashADCCCDC)Limitation Other than antigen escape cancer cells with T cell immune suppressors such as PDL1 overexpressed may have worse response to T cell activation based therapies
Novel ldquoimbalancedrdquo CD20CD3 based cancer therapy
By making a CD20CD3 bispecific antibody with CD3binding affinity significantly reduced we hope toldquosafelyrdquo maintain the effector function and reduce cytokine storm Therefore the novel ldquoimbalancedrdquo CD20CD3 bispecific would have all mechanisms as described above
Binding Ability of Designed Abs Compared to That of Parental Abs
After computational design CD20 homodimer (hole-hole) IgG contains designed VH sequence and common VL
sequence showed similar binding capacity for CD20 compared to that of parental CD20 IgG while CD3
homodimer (knob-knob) IgG contains designed VH sequence and common VL sequence showed significant
reduced binding capacity for CD3 compared to that of parental CD3 IgG
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has Good Efficacy and Safety in vitro
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Day 0 IV injection with ldquoAbsPBS + Raji-Luc cell + human PBMCrdquo first imaging 15min post injection G1 PBS G2 BsAb G3 Rituximab
1 died3 died
Day 0 Day 2 Day 13 Day 22 Day 28
CD20CD3 ldquoImbalancedrdquo BsAb has better Tumor Killing Potency than Rituxan in vivo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Bispecific involves a CD3 binder
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CD3 binding Cancer antigen binding CD3
Cancer antigen
Bispecific
wwwantibodystudiocom
Bispecific against a CSA (cancer specific antigen) and a CAA (cancer associated antigen)
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CSA binding CAA binding
Bispecific
CSA Cancer specific antigen
CAA Cancer associated antigen Normal cell
Normal cell
Normal cell
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Bispecific against one target for CNS and another target on BBB
Ab Studio Inc
your partner for therapeutic antibody development
Brain target
cell
ldquoKnob-into-holerdquo
Common light chain
CNS target binding BBB binding
Bispecific
BBB
Brain target
cell
wwwantibodystudiocom
How ldquoimbalanced bispecific antibodiesrdquo are made at Ab Studio
(lsquoArsquo is the corresponding high affinity arm and lsquoBrsquo is the corresponding low affinity arm)
VL Design
1) If VL-A and VL-B light chain homology gt80 design a VL-C based on VL-Avia computer aided design Maintain VH-AVL-Crsquos affinity as close as that ofVH-AVL-A allow VH-BVL-C to have reduced affinity compared to VH-BVL-B
2) If VL-A and VL-B light chain homology lt80 construct phage ScFV libraryVHs of the ScFV library are human germline VH libraryVLs of the ScFV library are VL-A and its mutants derived from error prone PCR at less than 20 of mutation frequency
VH Design
Use computer aided design to differentiate the biochemical and biophysical featuresof VH-A and VH-B in order to isolate lsquoABrsquo better from lsquoAArsquo and lsquoBBrsquo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Case Study CD20CD3 Bispecific
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chainComputer aided design
CD3 CD20 CD3
Cancer antigen
Bispecific
Biophysical and biochemical features were differentiated for reducing manufacture cost
wwwantibodystudiocom
Difference to other CD20CD3 bispecific ADCCCDC effector function is maintained
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Current CD20+ Cancer Immuno-Therapy
Mechanism of action ADCC CDC ApoptosisRepresentatives Rituxan and its biosimilar biobetterLimitation Other than antigen escape different genetic makeup of patientsrsquo effector cells could cause different response to these therapies
Mechanism of action CAR-T TCR-T T cell engagementRepresentatives CD20CD3 bispecific (ndashADCCCDC)Limitation Other than antigen escape cancer cells with T cell immune suppressors such as PDL1 overexpressed may have worse response to T cell activation based therapies
Novel ldquoimbalancedrdquo CD20CD3 based cancer therapy
By making a CD20CD3 bispecific antibody with CD3binding affinity significantly reduced we hope toldquosafelyrdquo maintain the effector function and reduce cytokine storm Therefore the novel ldquoimbalancedrdquo CD20CD3 bispecific would have all mechanisms as described above
Binding Ability of Designed Abs Compared to That of Parental Abs
After computational design CD20 homodimer (hole-hole) IgG contains designed VH sequence and common VL
sequence showed similar binding capacity for CD20 compared to that of parental CD20 IgG while CD3
homodimer (knob-knob) IgG contains designed VH sequence and common VL sequence showed significant
reduced binding capacity for CD3 compared to that of parental CD3 IgG
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has Good Efficacy and Safety in vitro
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Day 0 IV injection with ldquoAbsPBS + Raji-Luc cell + human PBMCrdquo first imaging 15min post injection G1 PBS G2 BsAb G3 Rituximab
1 died3 died
Day 0 Day 2 Day 13 Day 22 Day 28
CD20CD3 ldquoImbalancedrdquo BsAb has better Tumor Killing Potency than Rituxan in vivo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Bispecific against a CSA (cancer specific antigen) and a CAA (cancer associated antigen)
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
CSA binding CAA binding
Bispecific
CSA Cancer specific antigen
CAA Cancer associated antigen Normal cell
Normal cell
Normal cell
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Bispecific against one target for CNS and another target on BBB
Ab Studio Inc
your partner for therapeutic antibody development
Brain target
cell
ldquoKnob-into-holerdquo
Common light chain
CNS target binding BBB binding
Bispecific
BBB
Brain target
cell
wwwantibodystudiocom
How ldquoimbalanced bispecific antibodiesrdquo are made at Ab Studio
(lsquoArsquo is the corresponding high affinity arm and lsquoBrsquo is the corresponding low affinity arm)
VL Design
1) If VL-A and VL-B light chain homology gt80 design a VL-C based on VL-Avia computer aided design Maintain VH-AVL-Crsquos affinity as close as that ofVH-AVL-A allow VH-BVL-C to have reduced affinity compared to VH-BVL-B
2) If VL-A and VL-B light chain homology lt80 construct phage ScFV libraryVHs of the ScFV library are human germline VH libraryVLs of the ScFV library are VL-A and its mutants derived from error prone PCR at less than 20 of mutation frequency
VH Design
Use computer aided design to differentiate the biochemical and biophysical featuresof VH-A and VH-B in order to isolate lsquoABrsquo better from lsquoAArsquo and lsquoBBrsquo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Case Study CD20CD3 Bispecific
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chainComputer aided design
CD3 CD20 CD3
Cancer antigen
Bispecific
Biophysical and biochemical features were differentiated for reducing manufacture cost
wwwantibodystudiocom
Difference to other CD20CD3 bispecific ADCCCDC effector function is maintained
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Current CD20+ Cancer Immuno-Therapy
Mechanism of action ADCC CDC ApoptosisRepresentatives Rituxan and its biosimilar biobetterLimitation Other than antigen escape different genetic makeup of patientsrsquo effector cells could cause different response to these therapies
Mechanism of action CAR-T TCR-T T cell engagementRepresentatives CD20CD3 bispecific (ndashADCCCDC)Limitation Other than antigen escape cancer cells with T cell immune suppressors such as PDL1 overexpressed may have worse response to T cell activation based therapies
Novel ldquoimbalancedrdquo CD20CD3 based cancer therapy
By making a CD20CD3 bispecific antibody with CD3binding affinity significantly reduced we hope toldquosafelyrdquo maintain the effector function and reduce cytokine storm Therefore the novel ldquoimbalancedrdquo CD20CD3 bispecific would have all mechanisms as described above
Binding Ability of Designed Abs Compared to That of Parental Abs
After computational design CD20 homodimer (hole-hole) IgG contains designed VH sequence and common VL
sequence showed similar binding capacity for CD20 compared to that of parental CD20 IgG while CD3
homodimer (knob-knob) IgG contains designed VH sequence and common VL sequence showed significant
reduced binding capacity for CD3 compared to that of parental CD3 IgG
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has Good Efficacy and Safety in vitro
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Day 0 IV injection with ldquoAbsPBS + Raji-Luc cell + human PBMCrdquo first imaging 15min post injection G1 PBS G2 BsAb G3 Rituximab
1 died3 died
Day 0 Day 2 Day 13 Day 22 Day 28
CD20CD3 ldquoImbalancedrdquo BsAb has better Tumor Killing Potency than Rituxan in vivo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Bispecific against one target for CNS and another target on BBB
Ab Studio Inc
your partner for therapeutic antibody development
Brain target
cell
ldquoKnob-into-holerdquo
Common light chain
CNS target binding BBB binding
Bispecific
BBB
Brain target
cell
wwwantibodystudiocom
How ldquoimbalanced bispecific antibodiesrdquo are made at Ab Studio
(lsquoArsquo is the corresponding high affinity arm and lsquoBrsquo is the corresponding low affinity arm)
VL Design
1) If VL-A and VL-B light chain homology gt80 design a VL-C based on VL-Avia computer aided design Maintain VH-AVL-Crsquos affinity as close as that ofVH-AVL-A allow VH-BVL-C to have reduced affinity compared to VH-BVL-B
2) If VL-A and VL-B light chain homology lt80 construct phage ScFV libraryVHs of the ScFV library are human germline VH libraryVLs of the ScFV library are VL-A and its mutants derived from error prone PCR at less than 20 of mutation frequency
VH Design
Use computer aided design to differentiate the biochemical and biophysical featuresof VH-A and VH-B in order to isolate lsquoABrsquo better from lsquoAArsquo and lsquoBBrsquo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Case Study CD20CD3 Bispecific
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chainComputer aided design
CD3 CD20 CD3
Cancer antigen
Bispecific
Biophysical and biochemical features were differentiated for reducing manufacture cost
wwwantibodystudiocom
Difference to other CD20CD3 bispecific ADCCCDC effector function is maintained
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Current CD20+ Cancer Immuno-Therapy
Mechanism of action ADCC CDC ApoptosisRepresentatives Rituxan and its biosimilar biobetterLimitation Other than antigen escape different genetic makeup of patientsrsquo effector cells could cause different response to these therapies
Mechanism of action CAR-T TCR-T T cell engagementRepresentatives CD20CD3 bispecific (ndashADCCCDC)Limitation Other than antigen escape cancer cells with T cell immune suppressors such as PDL1 overexpressed may have worse response to T cell activation based therapies
Novel ldquoimbalancedrdquo CD20CD3 based cancer therapy
By making a CD20CD3 bispecific antibody with CD3binding affinity significantly reduced we hope toldquosafelyrdquo maintain the effector function and reduce cytokine storm Therefore the novel ldquoimbalancedrdquo CD20CD3 bispecific would have all mechanisms as described above
Binding Ability of Designed Abs Compared to That of Parental Abs
After computational design CD20 homodimer (hole-hole) IgG contains designed VH sequence and common VL
sequence showed similar binding capacity for CD20 compared to that of parental CD20 IgG while CD3
homodimer (knob-knob) IgG contains designed VH sequence and common VL sequence showed significant
reduced binding capacity for CD3 compared to that of parental CD3 IgG
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has Good Efficacy and Safety in vitro
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Day 0 IV injection with ldquoAbsPBS + Raji-Luc cell + human PBMCrdquo first imaging 15min post injection G1 PBS G2 BsAb G3 Rituximab
1 died3 died
Day 0 Day 2 Day 13 Day 22 Day 28
CD20CD3 ldquoImbalancedrdquo BsAb has better Tumor Killing Potency than Rituxan in vivo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
How ldquoimbalanced bispecific antibodiesrdquo are made at Ab Studio
(lsquoArsquo is the corresponding high affinity arm and lsquoBrsquo is the corresponding low affinity arm)
VL Design
1) If VL-A and VL-B light chain homology gt80 design a VL-C based on VL-Avia computer aided design Maintain VH-AVL-Crsquos affinity as close as that ofVH-AVL-A allow VH-BVL-C to have reduced affinity compared to VH-BVL-B
2) If VL-A and VL-B light chain homology lt80 construct phage ScFV libraryVHs of the ScFV library are human germline VH libraryVLs of the ScFV library are VL-A and its mutants derived from error prone PCR at less than 20 of mutation frequency
VH Design
Use computer aided design to differentiate the biochemical and biophysical featuresof VH-A and VH-B in order to isolate lsquoABrsquo better from lsquoAArsquo and lsquoBBrsquo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Case Study CD20CD3 Bispecific
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chainComputer aided design
CD3 CD20 CD3
Cancer antigen
Bispecific
Biophysical and biochemical features were differentiated for reducing manufacture cost
wwwantibodystudiocom
Difference to other CD20CD3 bispecific ADCCCDC effector function is maintained
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Current CD20+ Cancer Immuno-Therapy
Mechanism of action ADCC CDC ApoptosisRepresentatives Rituxan and its biosimilar biobetterLimitation Other than antigen escape different genetic makeup of patientsrsquo effector cells could cause different response to these therapies
Mechanism of action CAR-T TCR-T T cell engagementRepresentatives CD20CD3 bispecific (ndashADCCCDC)Limitation Other than antigen escape cancer cells with T cell immune suppressors such as PDL1 overexpressed may have worse response to T cell activation based therapies
Novel ldquoimbalancedrdquo CD20CD3 based cancer therapy
By making a CD20CD3 bispecific antibody with CD3binding affinity significantly reduced we hope toldquosafelyrdquo maintain the effector function and reduce cytokine storm Therefore the novel ldquoimbalancedrdquo CD20CD3 bispecific would have all mechanisms as described above
Binding Ability of Designed Abs Compared to That of Parental Abs
After computational design CD20 homodimer (hole-hole) IgG contains designed VH sequence and common VL
sequence showed similar binding capacity for CD20 compared to that of parental CD20 IgG while CD3
homodimer (knob-knob) IgG contains designed VH sequence and common VL sequence showed significant
reduced binding capacity for CD3 compared to that of parental CD3 IgG
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has Good Efficacy and Safety in vitro
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Day 0 IV injection with ldquoAbsPBS + Raji-Luc cell + human PBMCrdquo first imaging 15min post injection G1 PBS G2 BsAb G3 Rituximab
1 died3 died
Day 0 Day 2 Day 13 Day 22 Day 28
CD20CD3 ldquoImbalancedrdquo BsAb has better Tumor Killing Potency than Rituxan in vivo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Case Study CD20CD3 Bispecific
Ab Studio Inc
your partner for therapeutic antibody development
T cell
Cancer cell
ldquoKnob-into-holerdquo
Common light chainComputer aided design
CD3 CD20 CD3
Cancer antigen
Bispecific
Biophysical and biochemical features were differentiated for reducing manufacture cost
wwwantibodystudiocom
Difference to other CD20CD3 bispecific ADCCCDC effector function is maintained
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Current CD20+ Cancer Immuno-Therapy
Mechanism of action ADCC CDC ApoptosisRepresentatives Rituxan and its biosimilar biobetterLimitation Other than antigen escape different genetic makeup of patientsrsquo effector cells could cause different response to these therapies
Mechanism of action CAR-T TCR-T T cell engagementRepresentatives CD20CD3 bispecific (ndashADCCCDC)Limitation Other than antigen escape cancer cells with T cell immune suppressors such as PDL1 overexpressed may have worse response to T cell activation based therapies
Novel ldquoimbalancedrdquo CD20CD3 based cancer therapy
By making a CD20CD3 bispecific antibody with CD3binding affinity significantly reduced we hope toldquosafelyrdquo maintain the effector function and reduce cytokine storm Therefore the novel ldquoimbalancedrdquo CD20CD3 bispecific would have all mechanisms as described above
Binding Ability of Designed Abs Compared to That of Parental Abs
After computational design CD20 homodimer (hole-hole) IgG contains designed VH sequence and common VL
sequence showed similar binding capacity for CD20 compared to that of parental CD20 IgG while CD3
homodimer (knob-knob) IgG contains designed VH sequence and common VL sequence showed significant
reduced binding capacity for CD3 compared to that of parental CD3 IgG
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has Good Efficacy and Safety in vitro
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Day 0 IV injection with ldquoAbsPBS + Raji-Luc cell + human PBMCrdquo first imaging 15min post injection G1 PBS G2 BsAb G3 Rituximab
1 died3 died
Day 0 Day 2 Day 13 Day 22 Day 28
CD20CD3 ldquoImbalancedrdquo BsAb has better Tumor Killing Potency than Rituxan in vivo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Current CD20+ Cancer Immuno-Therapy
Mechanism of action ADCC CDC ApoptosisRepresentatives Rituxan and its biosimilar biobetterLimitation Other than antigen escape different genetic makeup of patientsrsquo effector cells could cause different response to these therapies
Mechanism of action CAR-T TCR-T T cell engagementRepresentatives CD20CD3 bispecific (ndashADCCCDC)Limitation Other than antigen escape cancer cells with T cell immune suppressors such as PDL1 overexpressed may have worse response to T cell activation based therapies
Novel ldquoimbalancedrdquo CD20CD3 based cancer therapy
By making a CD20CD3 bispecific antibody with CD3binding affinity significantly reduced we hope toldquosafelyrdquo maintain the effector function and reduce cytokine storm Therefore the novel ldquoimbalancedrdquo CD20CD3 bispecific would have all mechanisms as described above
Binding Ability of Designed Abs Compared to That of Parental Abs
After computational design CD20 homodimer (hole-hole) IgG contains designed VH sequence and common VL
sequence showed similar binding capacity for CD20 compared to that of parental CD20 IgG while CD3
homodimer (knob-knob) IgG contains designed VH sequence and common VL sequence showed significant
reduced binding capacity for CD3 compared to that of parental CD3 IgG
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has Good Efficacy and Safety in vitro
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Day 0 IV injection with ldquoAbsPBS + Raji-Luc cell + human PBMCrdquo first imaging 15min post injection G1 PBS G2 BsAb G3 Rituximab
1 died3 died
Day 0 Day 2 Day 13 Day 22 Day 28
CD20CD3 ldquoImbalancedrdquo BsAb has better Tumor Killing Potency than Rituxan in vivo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Binding Ability of Designed Abs Compared to That of Parental Abs
After computational design CD20 homodimer (hole-hole) IgG contains designed VH sequence and common VL
sequence showed similar binding capacity for CD20 compared to that of parental CD20 IgG while CD3
homodimer (knob-knob) IgG contains designed VH sequence and common VL sequence showed significant
reduced binding capacity for CD3 compared to that of parental CD3 IgG
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has Good Efficacy and Safety in vitro
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Day 0 IV injection with ldquoAbsPBS + Raji-Luc cell + human PBMCrdquo first imaging 15min post injection G1 PBS G2 BsAb G3 Rituximab
1 died3 died
Day 0 Day 2 Day 13 Day 22 Day 28
CD20CD3 ldquoImbalancedrdquo BsAb has better Tumor Killing Potency than Rituxan in vivo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
CD20CD3 ldquoImbalancedrdquo BsAb has Good Efficacy and Safety in vitro
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Day 0 IV injection with ldquoAbsPBS + Raji-Luc cell + human PBMCrdquo first imaging 15min post injection G1 PBS G2 BsAb G3 Rituximab
1 died3 died
Day 0 Day 2 Day 13 Day 22 Day 28
CD20CD3 ldquoImbalancedrdquo BsAb has better Tumor Killing Potency than Rituxan in vivo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Day 0 IV injection with ldquoAbsPBS + Raji-Luc cell + human PBMCrdquo first imaging 15min post injection G1 PBS G2 BsAb G3 Rituximab
1 died3 died
Day 0 Day 2 Day 13 Day 22 Day 28
CD20CD3 ldquoImbalancedrdquo BsAb has better Tumor Killing Potency than Rituxan in vivo
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
CD20CD3 ldquoImbalancedrdquo BsAb has good Developability
Re-CE-SDS
Non-Re-CE-SDS
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Summary
1) Computational designed Imbalanced CD20CD3 BsMab has good efficacy safety and developability in vitro and in animal model
2) This imbalanced CD20CD3 BsMab has potential to treat Rituxan resistant difficult to treat and relapsed CD20+ cancer (unmet clinical needs fast track for filing IND)
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Development and Screening of Therapeutic Antibodieswith Internalization Potential
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
bull Computer aided design for serial reduced internalizationbull In house developed high-throughput screening assay
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Case Study PDL1CDx Bispecific Antibody with Internalization Potential
Cancer cell
ldquoKnob-into-holerdquo
Common light chain
PDL1 binding CDx bindingBispecific
PDL1
CDx
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
0
5
10
15
20
25
30
35
40
45
Cell only Cell+PEP-ZAP
Cell+CDx
Cell+PD-L1
Cell+Bispecific
Cell+CDx
+PEP-ZAP
Cell+PD-L1
+PEP-ZAP
Cell+Bispecific+PEP-ZAP
PEP-ZAP Mediated Cell Death of Target Cancer Cell (18 amp 40 hrs incubation)
18 hours 40 hours
PDL1CDx Bispecific Antibody Induces more Cell Death than PDL1 and CDx Homodimer in the Presence of Pep-ZAP
How PEP-ZAP works
Disease Cell
Antigen
Primary antibody produced by hybridoma
Peptide fused to a translation inhibitor
Pep-ZAP is a Fc binding peptide conjugated to a translation inhibitor protein cell death is thereadout of antibody internalization
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Biological Significance
This imbalanced bispecific may induce target cancer cell death at three different levels
1) Blocking PD1PDL1 interaction
2) Induce PDL1 internalization
3) When conjugated to a drug kill target cancer cells an Antibody drug conjugate
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much
Ab Studio Inc
your partner for therapeutic antibody developmentwwwantibodystudiocom
Thank you very much