use of prior knowledge for biotech products: applications...

This document provides an outline of a presentation and is incomplete without the accompanying oral commentary and discussion. Conclusions and/ or potential strategies contained herein are NOT necessarily endorsed by Pfizer management. Any implied strategy herein would be subject to management, regulatory and legal review and approval before implementation.

Use of Prior Knowledge for Biotech Products: Applications and Examples

Ranga GodavartiBioProcess R&D, Pfizer

EBE-CMC Strategy Forum, CopenhagenMay 4, 2015

Use of prior knowledge is beneficial to Industry, Regulators and Patients

• Enable products to clinic faster, providing patients faster access to therapies

• Improves process development and technical transfer success– Enables right sized experimentation plans – Areas of risk can be understood early and focused on– Enables appropriate validation approaches

• Prior knowledge in biotech products: Applications & case studies– Modular viral clearance for early clinical products– Late stage process characterization studies– Other potential applications

Pfizer Confidential │ 2

Prior knowledge applications: Use of modular viral clearance to support Phase 1 and 2 CTAs


• Monoclonal antibody (mAb) manufacturing processes are very similar and consistent with significant viral clearance data accumulated over the years

• Additional bracketing VC studies demonstrated robust clearance for each unit operation over broad set of factors

- mAb isotype (IgG1 and IgG2): 14 different antibodies- Broad range of mAb concentration- Multiple buffer conditions (pH, salt, buffer type)- Wide range in load impurity content

• Bracketing design included several parameters set to ‘worse case’- Examples: Temperature, Detergent conc., pH, Impurity levels, Load challenge

• Module = Robust bracketed viral clearance unit operation- Independent of the other unit operations

Modular VC proposal: Apply modular viral clearance data from bracketing studies to other programs when specific criteria are met

Pfizer’s Standard Mab Process Includes Four Orthogonal, Robust Viral Clearance Modules

• Four orthogonal modules evaluated for viral clearance• One or more modules may be used in combination with product specific data as needed for overall VC package

Protein A

Triton X-100 Detergent Treatment

Virus Retentive Filter

Capture Column

Low pH

TMAE HiCap AEX

Retrovirus and parvovirus removal (binds impurities)

Retrovirus inactivation (by low pH)

Retrovirus inactivation (by Octoxynol-9)

Retrovirus and parvovirus removal (sized based retention)

• Potential worst case condition: low viability & high cell density

Triton X-100 (Octoxynol-9) treatment provides robust virus inactivation under all conditions

mAb 1 2 3 4mAb Type IgG1 IgG1 IgG2 IgG2

Viability (%) 40% 99% 60% 90%Cell density (cells/mL) 2.0e7 1.6e7 2.0e7 1.8e7

Triton X-100 (% v/v) 0.18 0.18 0.18 0.18

Temperature (°C) 15 15 15 15

Time (min) X-MuLV LRV

X-MuLVLRV

X-MuLV LRV

X-MuLV LRV

0 >3.36 >3.53 >2.91 >4.4630 >3.36 >3.53 >2.91 >4.4690 >5.5 >5.68 >5.06 >6.61

120 >5.5 >5.68 >5.06 >6.61

3-FactorsTested

Worst Case

Modular Claim - Lowest LRV observed: > 5.06 LRV

Viral Inactivation by Low pH results in robust clearance under all conditions

Mab Type Salt level BufferProduct

Concentration [g/L]

Time 60 min

Run 1 Run 2

IgG1 Low Glycine 20 7.38 6.99IgG1 Low Glycine 20 6.1 6.34IgG2 Low Glycine 20 >7.33 7.14IgG2 Low Acetate 20 6.54 >6.95IgG1 High Glycine 20 5.82 6.34IgG1 High Glycine 20 5.69 5.68IgG2 High Glycine 20 5.94 6.24IgG2 High Acetate 20 7.55 >7.71IgG1 Low Glycine 5 6.56 N/AIgG1 Low Glycine 5 6.9 7.09IgG2 Low Glycine 5 6.67 6.51IgG2 Low Acetate 5 7.41 6.78

Modular Claim - Lowest LRV observed: 5.68 LRV

Viral filtration results in complete retrovirus removal for all conditions

MabIsotype

Salt Level pH

Product Concentration

[g/L]

Pool X-MuLV

Run 1 Run 2

IgG1 10 5.5 15 >6.96 >6.92 IgG1 10 8.1 15 >6.96 >6.96 IgG1 100 5.5 15 >6.76 >6.75 IgG1 100 8.1 10 >7.07 >7.07 IgG1 50 6.8 15 >7.37 >7.37 IgG2 10 5.5 10 >6.51 >6.52 IgG2 10 8.1 15 >5.95 >5.96 IgG2 100 5.5 15 >6.24 >6.23 IgG2 100 8.1 15 >6.10 >6.10 IgG2 50 6.8 10 >6.68 >6.69

Modular Claim - Lowest LRV observed: > 5.95 LRV

Platform AEX Viral Clearance: Robust Removal of Retrovirus and Parvovirus for over 20 mAbs

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• Virus removal tested extensively over the course of numerous projects using the AEX chromatography step operated in weak partitioning mode

Retrovirus RemovalParvovirus

Removal

Range Evaluated Historically:Chloride: 10 - 120 mM Conductivity: 1 - 12 mS/cmpH: 7.0 - 8.4

No LRV less than 4.0 log10

Virus clearance by AEX Chromatography results in robust clearance under all conditions

99

mAbType

Load pH

Load Chloride

(mM)Kp

MuLV LRV

Run 1 Run 2

IgG1 7.1 75 0.3 (FT) > 6.61 > 6.62IgG1 8.4 10 5.0 > 6.16 > 6.16IgG1 7.3 10 5.0 > 6.57 > 6.57IgG1 8.4 75 5.0 > 6.38 > 6.38IgG2 7.1 75 < 0.01 (FT) > 6.29 > 6.29IgG2 8.7 10 2.8 > 4.57 > 4.57IgG2 6.9 75 0.04 (FT) > 6.53 > 6.53IgG2 8.5 38 5.0 5.10 5.54

Modular Claim - Lowest LRV observed: 5.10 LRV

AEX Load Impurity Impurity Range

HCP 170 - 29,000 ppm

DNA 100 - 1380 pg/mg

HMMS 2 - 14%

Pfizer’s Modular Viral Clearance Package Summary

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Process Step Mechanism Log10 Reduction of Retrovirus

Module 1: Triton X-100 Detergent Treatment Inactivation ≥ 5.06

Module 2: Low pH Treatment Inactivation 5.68

Module 3: Virus Retaining Filtration Removal ≥ 5.69

Module 4: AEX Chromatography Removal 5.10

• CTA would typically leverage 3 of these modules• Triton, Low pH and VRF would provide ≥16.43 LRV of retrovirus clearance• Safety margin based on process specific RVLP, yield, titer and dosage

• CTA may leverage 2 modules and include process specific data for 1 unit operation• If certain process steps are outside of the bracketed control space

Example of Application of MVC Approach to ‘mAb X’

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Unit Operation Operating Condition and/or Control Range

Does the ‘mAb X’ process meet criteria?

Module Retrovirus Clearance (LRV)

Triton X-100 (Module 1)

Cell Density 0 - 9e7 cells/mL Yes

≥ 5.06Cell Viability 40 - 100% Yes

Time 120 minutes Yes

Temperature 15 - 25ºC Yes

Triton X-100 Concentration 4 - 6 mL/L Yes

Low pH (Module 2)

pH ≤ 3.60 Yes

5.68

Salt or Ion Levels 10 - 150 mM Yes

Buffer: Glycine or Acetate Yes

Time ≥ 60 minutes Yes

Temperature 15 - 25ºC Yes

Protein Concentration 5 - 20 g/L Yes

Planova 20N Virus Retaining Filtration (Module 3)

Pressure 10 - 14 psi Yes

≥ 5.69Volumetric Loading ≤ 200 L/m2 Yes

pH 5.5 - 8.1 Yes

Salt or Ions: 10 - 100 mM Yes

Protein Concentration ≤ 15 g/L Yes

PF-06664178 Cumulative Retrovirus Clearance (log10 removal value) ≥ 16.43

Application of Prior Knowledge in process characterization can result in efficient, effective study designs

• Risk assessment leverages prior knowledge for mAbs in clinical pipeline and/or commercial products utilizing similar platform process

• Process characterization studies include multi-variate DOEs focusing on only parameters with higher risk to impact CQAs– Prior knowledge from similar processes leveraged to

• Eliminate some low risk factors that have no impact to CQAs• Predict outcomes of higher risk factors that potentially have impact to

CQAs– Streamlined and efficient experimental plans


Application of Prior Knowledge in process characterization – MabSelect example

Pre-scored C&E Matrices are confirmed for every project: Sample MabSelect Protein A output

Application of Prior Knowledge in process characterization experimental design

Prior KnowledgemAb 1 mAb2 mAb3 mAb4 mAb5 mAb 6

Unit Op 1 Factor 1Factor 2Factor 3Factor 4Factor 5

Factor 1Factor 2Factor 3Factor 4Factor 5




Factor 1Factor 2Factor 3


• No impact to CQAs for all mAbs tested• Document prior knowledge and considered during risk assessment for mAb 6• mAb 6 design focused on most impactful factors

Summary: Prior knowledge potentially reduces need for product specific data in multiple areas of development

• Prior knowledge from technically sound data can enable faster development and access of medicines to patients with less reliance on product specific data– May includes literature and/or company-specific data

• Other potential applications– Process validation– Setting product specifications– Assessing criticality of quality attributes

• Example: abundance of literature on C-terminal lysines– Analytical Method development– Sterile filtration process development for early phase mAbs– Lyophilization process development


use of prior knowledge for biotech products: applications...

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