MSD -Meso Scale Discovery

Applications include:

Signal transduction path way evaluation

ROCK

Rho

FAK CrK

Pax PYK2

GTP Gα-O

GEFs

CDC42 Rac

MEKs

PAK MKKs

ASK1

JNK

P38 IP3R

PKC

BIMP1

BCL10

NFκB IκBs

P

IKKs

IκBsDegradation

Src

SOSGRB2SHC

RasGEFs

Raf1

MEKs

ERKs

PLC-β

DAG

NFκB

IKKs

PκB

IP3

Ca+2Ca+2

Ca+2

Calm

P

Caln

NFAT

NFAT

P

PI3K

Gβ Gγ

LARG

BRaf

MEKs

ERKs

cAMP

EPACPKA

PDZRhoGEF

Gα-1 GTP

Rap1

p115RhoGEF

RT

KS

CREB

Gα12/13

SRF

GTP

CREB

SRF ELK-1

c-Jun

c-Fos

STAT

NFκBNFAT

Gene Expression

Gα Gβ

© 2013 CDT all rights reserved

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EFTECHNICAL BRIEF:

MSD -Meso Scale DiscoveryD I S C O V E R Y T E C H N O L O G I E S

The Meso Scale Discovery (MSD) platform provides a faster alternative to traditional ELISAs for performing sand-wich immunoassays. The MSD assay platform utilizes Ruthenium (II) tris-bipyridine-(4-methylsulfone) [Ru(bpy)3] that, once conjugated to the analyte, serves as the tracer in competitive assays. The Ru(bpy)3-based tag undergoes a rapid redox reaction that emits light in the presence of an applied voltage. Assay kits for several eicosanoids have been adapted to the MSD platform. The Confluence Discovery Technologies (CDT) team has access to dozens of kits avail-able across therapeutic areas. An optimized MSD electro-chemiluminescent assay can quantitate host cell proteins with a simple protocol few wash steps and low cost. The MSD assay offers several technical advantages as an assay platform:

•Capture: 10-100 fold increase compared to ELISA

•Dynamic Range: 3-4 log vs. 1-1.5 log range for ELISA

•Sensitivity: Detection of analytes at sub-picogram levels

•Reduced Sample Volume: 5-25 µl volume vs. 50-100 µl for ELISA

•Speed: The entire assay can be performed in 3 hours

•Multiplex Capability: Simultaneous measurements for up to 10 analytes

Applications

The CDT team has a wide range of experience in develop-ing and validating MSD assays to suit many different requirements. Our highly trained and knowledgeable scien-tists will assist with experimental design to develop and vali-date assays to meet client-specified criteria. We also perform data analysis and interpretation of MSD data to enable our clients to obtain the best insights from large, complex data outputs.

ROCK

Rho

FAK CrK

Pax PYK2

GTP Gα-O

GEFs

CDC42 Rac

MEKs

PAK MKKs

ASK1

JNK

P38 IP3R

PKC

BIMP1

BCL10

NFκB IκBs

P

IKKs

IκBsDegradation

Src

SOSGRB2SHC

RasGEFs

Raf1

MEKs

ERKs

PLC-β

DAG

NFκB

IKKs

PκB

IP3

Ca+2Ca+2

Ca+2

Calm

P

Caln

NFAT

NFAT

P

PI3K

Gβ Gγ

LARG

BRaf

MEKs

ERKs

cAMP

EPACPKA

PDZRhoGEF

Gα-1 GTP

Rap1

p115RhoGEF

RT

KS

CREB

Gα12/13

SRF

GTP

CREB

SRF ELK-1

c-Jun

c-Fos

STAT

NFκBNFAT

Gene Expression

Gα Gβ

© 2013 CDT all rights reserved

CDTv1.8.13

Confluence Discovery Technologies, Inc.4320 Forest Park Avenue

Saint Louis, MO 63108Phone: 314-932-4032 x305

Fax: 314-932-4038www.confluencediscovery.com

CDTinfo@confluencestl.com

D I S C O V E R Y T E C H N O L O G I E S

CAPABILITIESD I S C O V E R Y T E C H N O L O G I E S

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Leaders in biochemistry and mechanistic enzymology, our senior team has access to a suite of capabilities and technologies enabling: • Drug:Target interaction • Enzyme:Inhibitor mechanisms • Direct binding kinetics • Compound mechanism of action

Con�uence o�ers a spectrum of cell and molecular biology capabilities: • Identi�cation of biologically important signal transduction pathways • Quantitation of e�cacy and selectivity • Understanding safety issues linked to pathways • De-convolution to isolate pathway nodes • Evaluation of multiple targets within the pathway • Identi�cation of key pathway point(s) for therapeutic intervention • Target validation, cloning, expression, and puri�cation • Custom assay development • High-throughput screening

Our team delivers when “OFF the SHELF” is not an option: • Assay development, optimization, and validation • Multiplex technology platforms and instrumentation • Complex data analysis • Specialty screens for preclinical technical application • Trouble shooting and problem solving

Our team provides clients with the expertise to: • Identify target, mechanism, and disease biomarkers • Select the appropriate biomarkers for a drug discovery program • Execute biomarker assay design, development, validation, and interpretation for pre-clinical and clinical studies • Seamless assay transfer from pre-clinical to clinical studies

Our medicinal and computational chemists are experts in designing and delivering novel chemical entities through: • Schrödinger® enabled computational drug design • High throughput hit prioritization • Chemical series prioritization • Structure activity relationship generation • Custom synthesis