establishing a testing strategy for a qbd development product mary cromwell director, protein...

Establishing a Testing Strategy

for a QbD Development Product

Mary Cromwell

Director, Protein Analytical Chemistry

Genentech

CMC Strategy Forum

July 20, 2010

Bethesda, MD

A-MAb Case Study: Elements of the Control Strategy

Raw Material Control

Procedural Controls

Process Parameter Controls

In-Process Testing

Specifications (Lot Release and Stability)

Characterizationand Comparability Testing

Process Monitoring

Process Control

Testing

Continual Process Verification

Today’s Discussion

Key Tools and Information Used to Develop a Testing Strategy for a QbD Product

CQA Identification

CQA Acceptance Criteria

Testing Strategy

Process Impact Score

Stability Impact Score

Testing StrategyRobustness Assessment

QA Impact Score

QA Uncertainty Score

Decision TreeRRF

TableQA Specific

Tim

e, In

form

atio

n

CQA Identification


Testing Strategy




QA Impact Score


Decision TreeRRF

TableQA Specific

CQA Identification


Testing Strategy




QA Impact Score


Decision TreeRRF

TableQA Specific

Decision TreeRRF

Decision TreeRRF

TableQA Specific

TableQA Specific

Tim

e, In

form

atio

n

Quality Attribute Impact Score

1 Based on a relevant potency assay and dependent on assay variability2 Based on serum exposure (AUC) or FcRn binding. PD considered if information available3 Based on effects observed in clinical studies

Impact &

Rating

Biologica

l Activity1PK2 Immunogenicity3

Safety3

(Potential or Observed)

Very High

(20)

>100%

change

>40% change

on PK

ATA (Anti-Therapeutic

Antibodies) detected that may

life threatening

Irreversible or life-

threatening AEs and/or life

threatening loss of efficacy

High

(16)

40-100%

change

20-40%

change with

impact on PD

ATA detected that may be

associated with non-life

threatening loss of efficacy

Reversible AEs and/or loss

of efficacy that is not life

threatening

Moderate

(12)

20-40%

change

20-40%

change with

no impact on

PD

ATA detected with effect that can

be managed by clinical treatment

(i.e. dose titration, medication,

etc.)

AE that can be managed by

clinical treatment (i.e. dose

titration, medication, etc.)

Low

(4)

<20%

change

<20% change

with no

impact on PD

ATA detected with effect on PK

or PD, but no effect on safety or

efficacy

Safety or efficacy effect

with minimal clinical

significance

None (2)No

change

No impact on

PK or PD

ATA not detected or ATA

detected with no effect on PK,

PD, safety, or efficacy

No effect on safety of

efficacy

P. Motchnik

CQA Acceptance Criteria (CQA-AC) and Target Ranges (CQA-TR)

• CQA-AC: – Numerical limits that must be met for the product to be

considered acceptable

– Based on non-clinical and clinical experience, platform knowledge and literature (with appropriate justification of applicability of data used)

– Based on patient impact only - process capability is not considered

– May change as product gains more clinical experience, not as a function of additional manufacturing experience

• CQA-TR:– Constrained range of the CQA-AC to ensure that the process

will always deliver product within the CQA-AC

Attribute Testing Strategy Risk Ranking and Filtering Tool

Quality Attribute

Impact Score

Process or Stability Impact Score

X == ATS (1)

(1) Attribute Testing Strategy

Performed for:• DS manufacturing process• DP manufacturing process• DS storage• DP storage

2, 4, 12, 16, 20 1, 2, 4, 10 2-200

Defines Testing Strategy:• No Testing• “Comparability and Monitoring”• Control System (Release, Stability, and/or In-Process)

Process Impact Scoring Decision Tree: Used for DS and DP Manufacturing Processes

PC/PV Outcome

Start: For each Quality Attribute

Process ImpactScore = 2



Difference:Actual Result to

CQA-TR

Highly RobustDefault

representativeprocess

model exists

No

Yes

QA Impact Score

of 2 or 4

Abundance0-1%*

*specific for product-related impurities; for HMWS, Abundance threshold is <0.1%



Yes

No


No

Yes

N. McKnight

Stability Impact Scoring Decision Tree

Start: for each Quality Attribute

Can moleculeform attribute?


Rate of changerelative to CQA-AC




No

Yes

Slow Fast

Moderate

R. Wong

(< 11%*) (>33%*)

(11-33%*)*of allowable range;Assessed to expiry at recommendedstorage temperature and for allowableexcursions

Attribute Testing Score: Defines Testing Strategy

Attribute Testing Strategy (ATS) Scoring Matrix

CQA Impact Score

Process or Stability Impact Score

2

(Very Low) 4

(Low) 12

(Moderate) 16

(High) 20

(Very High)

1 2a 4a 12 16 20

2 4a 8a 24 32 40

4 8 16 48 64 80

10 20a 40a 120 160 200

ATS Score:

< 21 No testing required.

21-50 Comparability and Process Monitoring testing required.

> 50 Control system testing required.

“Comparability and Monitoring” (CaM)

• Attribute class to be tested as part of:

• Comparability exercises– Performed to support site transfer, version changes, scale changes– Provides streamlined testing

>Testing includes appropriate (DS or DP) tests designated “CaM” in the Testing Strategy as

well as Control System testing (IP, Lot Release, Stability)

>Choice of tests based on risk associated with change; only CaM attributes known to be

impacted by particular step that is changing will be tested

• Process Monitoring– Continuous Process Verification

>Subset of CaM attributes

>Frequency of monitoring may be attribute dependent

>Control System testing

>Key Performance Indicators

Application of Attribute Testing Strategy Tool

MAb 1: Background

• Target Product Profile• Immunology indication• Drug will be administered IV monthly for six months• Doses up to 5X of proposed marketed dose given in clinical trials• Very low immunogenicity rate• Safety profile well established

• Product characterization• Effector function required for potency

– CDC (terminal galactose distribution)– ADCC (afucosylation)

• CDR deamidation, fragmentation, aggregation impact potency– Deamidation increases ~ 6% on DP storage (allowable range = 9%)– Fragmentation increases ~1% on DP storage (allowable range = 5.8%)– Aggregation does not change on storage of DS or DP

ATS for Acidic VariantsParameter Rationale

CQA Impact Score 12 Based on potency impact in CDC and ADCC assays

CQA Acceptance Criteria 45% Based on level required to maintain potency 80%.

CQA Target Range for DS Process

34% Based on level required to ensure that DP meets acceptance criterion at end of shelf life

DS Process 10 Worst-case conditions across all operations result in a batch that exceeds the CQA-TR; Restriction Relationship and Adaptive Design Space employed

DP Process 4 Changes during ambient handling and excursions

DS Stability 2 No changes observed throughout development

Process Impact Scores

DP Stability 10 Significant changes observed over shelf life and during excursions

Testing Strategy (CQA Impact score X Process Stability Impact score)

DS Process 120 Control system testing

DP Process 48 “Comparability and Monitoring” testing

DS Stability 24 “Comparability and Monitoring” testing

Attribute Testing Strategy Scores

DP Stability 120 Control system testing

ATS for CHOP (Application to Process-Related Impurities)

Parameter Rationale

CQA Impact Score 20 Based on potential for Immunogenicity

CQA Acceptance Criteria ≤ 25 ppm Based on dosing at 5x proposed to-be-marketed dose with no difference in observed ATA incidence


≤ 23 ppm Truncated standard 5% CQA-AC reduction.

DS Process 2 Highly robust

DP Process 1 Will not involve cells

DS Stability 1 Will not change on stability Process Impact Scores

DP Stability 1 WIll not change on stability

Testing Strategy (CQA impact score X process/stability impact score)

DS Process 40 “Comparability and Monitoring” testing

DP Process 20

DS Stability 20


DP Stability 20

No testing

ATA anti-therapeutic antibodies; CDC complement-dependent cytotoxicity; CQA Critical Quality Attribute; CQA-TR Critical Quality Attribute Target Range; DS Drug Substance; DP Drug Product; NA not applicable.

ATS for Fragmentation (application of Abundance Filter)

Parameter Rationale

CQA Impact Score 20 Based on potency impact in CDC and ADCC assays; altered PK

CQA Acceptance Criteria 5.8% Fab Ensures acceptable potency and PK


NA Abundance of 1% indicates that establishing a target range is not required.

DS Process 1 Abundance <1%

DP Process 1 Abundance < 1%

DS Stability 2 No changes observed throughout development Process Impact Scores

DP Stability 4 Change of ~1% observed during recommended storage/allowable excursions


DS Process 20

DP Process 20

No testing

DS Stability 40 “Comparability and Monitoring” testing


DP Stability 80 Control System testing required

ADCC antibody-dependent cellular cytotoxicity; CDC complement-dependent cytotoxicity; CQA Critical Quality Attribute; DS Drug Substance; DP Drug Product; NA not applicable; PK pharmacokinetics; NA = Not applicable.

ATS for Afucosylation (Stability)

Parameter Rationale

CQA Impact Score 20 Based on potency impact in ADCC assay

CQA Acceptance Criteria 1.6-12.1% Based on clinical experience and levels to maintain ADCC potency


1.7-11.5% Standard 5% CQA-AC reduction applied to both limits

DS Process 10 Significant process variability observed

DP Process 1 Will not be impacted by DP Processing

DS Stability 1 Will not change on stability Process Impact Scores

DP Stability 1 Will not change on stability


DS Process 200 Control system testing

DP Process 20

DS Stability 20


DP Stability 20

No testing

ADCC antibody-dependent cellular cytotoxicity; CDC complement-dependent cytotoxicity; CQA Critical Quality Attribute; DS Drug Substance; DP Drug Product; NA not applicable.

Key Tools and Information Used to Develop a Testing Strategy for a QbD Product

CQA Identification


Testing Strategy




QA Impact Score


Decision TreeRRF

TableQA Specific

Tim

e, In

form

atio

n

CQA Identification


Testing Strategy




QA Impact Score


Decision TreeRRF

TableQA Specific

CQA Identification


Testing Strategy




QA Impact Score


Decision TreeRRF

TableQA Specific

Decision TreeRRF

Decision TreeRRF

TableQA Specific

TableQA Specific

Tim

e, In

form

atio

n

Robustness Assessment of Testing Strategy

• Testing Strategy Tool used to develop the proposed Control System• Is proposed testing strategy of sufficiently low risk?• Do proposed methods provide adequate control?

• Robustness Assessment Tool considers• ATS score (reflecting CQA Impact and Process/Stability control)• Sensitivity of method used for analysis• Testing Strategy (Control System, “CaM”, no testing)

• Expected to be iterative process: Unacceptable score indicates that• Testing strategy may need to change• A more sensitive method may be required for testing attribute• Manufacturing process may need to provide greater process control• Shelf-life/allowable excursions may need to be shortened

Robustness Assessment of Testing Strategy

Attribute Testing Score

X TestingStrategy Score

= RobustnessScore

2-200 2, 4, 6, 10 4 - 2000

LR Lot release; IP In-Process.

No testing 10

The attribute is monitored directly, collectively, or via a surrogate assay ONLY during “Comparability and Monitoring” testing.

6

The impact of an attribute is measured in a surrogate assay, is measured collectively, or is measured directly with a relatively insensitive assay (IP or LR/Stability).

4

Attribute is measured directly with a suitably sensitive assay (IP or LR/Stability).

2

DescriptionScore

Testing Strategy Scoring for Robustness Assessment RRF

LR Lot release; IP In-Process.

No testing 10

The attribute is monitored directly, collectively, or via a surrogate assay ONLY during “Comparability and Monitoring” testing.

6

The impact of an attribute is measured in a surrogate assay, is measured collectively, or is measured directly with a relatively insensitive assay (IP or LR/Stability).

4

Attribute is measured directly with a suitably sensitive assay (IP or LR/Stability).

2

DescriptionScore

Testing Strategy Scoring for Robustness Assessment RRF

Score </= 400 indicates Robust Control Strategy

Robustness Assessment for Afucosylation: Glycan Assay

• Control of afucosylation by CE-glycan assay is ROBUST

Attribute Process ATS

Score Testing Strategy

Test Method

Testing Strategy

Score Robustness

Score Assess-

ment

DS Process

200 Control System

2 400

DP Process

20 No testing 10 200

DS Stability


Afucosyl-ation

DP Stability

20 No testing

CE Glycan Assay

10 200

Robust

ATS Attribute Testing Strategy; CE capillary electrophoresis; DP Drug Product; DS Drug Substance.

Robustness Assessment for Afucosylation: ADCC Potency Assay

• Control of afucosylation by ADCC potency assay isNOT ROBUST

• high impact CQA• low process control


Score Testing Strategy

Test Method

Testing Strategy

Score Robustness

Score Assess-

ment

DS Process

200 Control System

4 800 Not

Robust

DP Process


DS Stability


Afucosyl-ation

DP Stability

20 No testing

ADCC Potency

10 200

Robust

ATS Attribute Testing Strategy; CE capillary electrophoresis; DP Drug Product; DS Drug Substance.

Robustness Assessment: Size Variants


Score Testing Strategy Test Method

Testing Strategy

Score Robustness

Score Assess-

ment

DS Process

48 288

DP Process

24 144

DS Stability

24 144

Aggregates

DP Stability

24

Comparability and Monitoring

Size-Exclusion Chromatography

6

144

Robust

DS Process


DP Process


DS Stability

24 Comparability and Monitoring

6 144

Fragments

DP Stability

48 Control System

Size-Exclusion Chromatography

2 96

Robust

ATS Attribute Testing Strategy; DP Drug Product; DS Drug Substance.

Proposed Control System for MAb 1: Drug Product

In-Process Testing

Attribute Test Name

Sterility Product Sterility Test (USP method)

Bioburden Bioburden Testing of In-Process Samples

Endotoxin Standard Procedure for limulus Amebocyte Lysate (LAL) Assay

pH pH

Osmolalitya Osmolality

Polysorbate 20 Polysorbate Content

Protein Concentration UV Spec Scan Using Gravimetric Sample Preparation

CFU colony forming unit; UV ultraviolet visible.

a direct measure of osmolality, indirect measure of excipient concentrations

Proposed Control System for MAb 1: Drug Product

Attribute Test Name

Identity CZE

Appearance COC

Acidic Variantsa IEC

Size Variantsa SEC

Subvisible Particulatesa Particulate Analysis

Endotoxin (LAL) Endotoxin (LAL)

Sterility Sterility

Sterilitya Dye Leak Test

Biological Activitya CDC Potency

Fill Volume Volume in Container

Lot Release and Stability

CDC complement-dependent cytotoxicity; COC clarity, opalescence, and coloration; CZE capillary zone electrophoresis; IEC ion-exchange chromatography; LAL limulus amebocyte lysate; SEC size-exclusion chromatography. a Tests that will be performed on stability.

Comparability and Monitoring Testing: Drug Product

Attribute Robustness Score Detection by Control System

Assay for Drug Product?

Acidc Variants 288 CDC Potency Assay

Oxidation 384 CDC Potency Assay

Proposed Control System: MAb 1 Drug Substance

In Process TestingAttribute Test Name

Mycoplasma detection, culture method Mycoplasma

Mycoplasma detection, indicator cell/DNAF procedure

General Viral Screening Assay Virus

Rodent Parvovirus PCR

Bioburden Bioburden

Endotoxin Endotoxin (LAL)

Afucosylation (%G0F/% total G0)

CE-Glycan

pH pH

Osmolalitya Osmolality

Polysorbate 20 Polysorbate Content

Protein Concentration

UV Spec Scan

CE = capillary electrophoresis.a Direct measure of osmolality, indirect measure of excipient concentrations

Proposed Control System: Drug Substance

Lot Release and Stability

Attribute Test Name

Identity Peptide Map

Acidic variants Ion-Exchange Chromatography


Sterility Sterility

Biological Activitya CDC Potency

a Test will be performed on stability.

Comparability and/or Monitoring Testing: Drug Substance

Attribute Robustness Score Detection by Control System Assay for Drug Substance?

Fragments (stability)a 240 (240) CDC Potency Assay, IEC

Acidic Variants (stability)a (240) CDC Potency Assay, IEC

CHOP 240 No

Reduced MAb 384 CDC Potency Assay

Leached Protein A 192 No

Glycan Distribution 288 CE-Glycan Assay

Non-glycosylated Heavy Chain 144 CDC Potency Assay

DNA 144 No

Aggregates 144 CDC Potency

Protein Conformation 240 CDC Potency, IEC

Approach Applied to a Second MAb

• MAb 2 Background• Binds to and blocks receptor on cell; effector function not required for MOA

• Oncology indication• Molecule is exceptionally stable; forced degradation studies cannot generate deamidation, oxidation, or glycation of CDR sites

–Limited number of CQAs requiring control–Therefore, limited number of CPPs to constrain Design Space

MAb 2 Proposed Control System

Attribute Test Name

Identity CZE

Appearance COC

Size Variantsa SEC

Subvisible Particulatesa Particulate Analysis


Sterility Sterility

Sterilitya Dye Leak Test

Biological Activitya Potency

Fill Volume Volume in Container

Attribute Test Name

Identity Peptide Map


Sterility Sterility

Biological Activitya Potency

Drug Substance CoA/Stability

Drug Product CoA/Stability

Summary

• Risk-based tools developed to define Testing Strategy and Control System• Utilizes knowledge of

– CQA Impact on potency, immunogenicity, safety, and PK/PD– Process Impact – Stability Impact– Specificity and Sensitivity of analytical method

• Assigns attributes to three categories of testing– Control System (Lot release, stability, in-process)– “Comparability and Monitoring”– No testing

• Robustness assessment for control of CQAs– Iterative process– Evaluates risk that control strategy is insufficient

Summary, continued…

• Use of filters in assessment of testing strategy• Abundance filter as part of Process Impact, not CQA ID• Default process impact score for low impact quality attributes

– Balances wide CQA Acceptance Criteria with potentially little process impact knowledge

Advantages of the QbD Approach to Developing a Control Strategy

• Clear, logical approach – applicable to all types of biologics

• Control Strategy development relies on significant process and product knowledge

• Clear rationale for selection of attributes to test and the type of testing strategy applied can lead to streamlined Control System testing

• Understanding impact of each process step to CQA levels leads to targeted testing for comparability

– Only test those attributes impacted by changed steps

Acknowledgements

Jerry DongLynn Gennaro Yung-Hsiang KaoParamjit KaurDaniel KelatiBrian KelleyLynne KrummenReed HarrisKathy HsiaRaquel IversonKim Latimer

Nadja Alt (Roche)Bernd Hilger (Roche)

Joseph MarhoulNathan McKnightPaul MotchnikDave ReifsnyderSofia RibeiroNatalie Saldou-HoltzCristina SanchezDieter SchmalzingRon TaticekPin-Yee WongRita Wong

MAb 1: Attributes that Require No Testing for Drug Substance

Attribute Robustness Score Detection by Control System Assay for Drug Substance?

Fragments (stability)a 240 (240) CDC Potency Assay, IEC

Acidic Variants (stability)a (240) CDC Potency Assay, IEC

CHOP 240 No

Reduced MAb 384 CDC Potency Assay

Leached Protein A 192 No

Glycan Distribution 288 CE-Glycan Assay

Non-glycosylated Heavy Chain 144 CDC Potency Assay

DNA 144 No

Aggregates 144 CDC Potency

Protein Conformation 240 CDC Potency, IEC

MAb 1: DS Testing Strategy for All Attributes

Control System Testing

Attribute Assay In-Process Lot Release Stability

Process Monitoring and Comparability

No Testing

Product-Related Variants

Afucosylated glycans

CE-glycan X

Glycan distribution CE-glycan X

Fragments SEC X

Aggregates SEC X

Acidic variants IEC X X

Non-glycosylated heavy chain

CE-SDS X

Oxidation (Trp and Met)

CDC Potency X X (stability)

Reduced MAb CE-SDS (NR) X

Protein Conformation

X

Cys22-Cys96 free thiol

X

N-terminal extension IEC X

C-terminal variants CZE X

Non-functional Region Deamidation

IEC X

Non-functional Region glycation

IEC X

MAb 1: DS Testing Strategy for All Attributes, cont’d

Process Related Impurities

Mycoplasma X

Rodent Parvovirus X

CHOP X

Leached Protein A X

DNA X

Bioburden Bioburden X X X

Endotoxin LAL X X X

Cell Culture and Purification Raw Materials

NMR, ICP X

Leachables GC-MS X

Control System Testing

Attribute Assay In-Process Lot

Release Stability

Process Monitoring and Comparability

No Testing

Statutory Requirements/Stability-Impacting Attributes

Identity Peptide Map X

Protein Concentration

UV Spec Scan X

Osmolality Osmolality X

pH pH X

Acetate Concentration

Osmolality X

Trehalose Concentration

Osmolality X

Polysorbate Concentration

Polysorbate 20 Content

X

establishing a testing strategy for a qbd development product mary cromwell director, protein...

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