Analysis and Structure Characterization of Monoclonal Antibodies

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On 6 January 2003, 129 attendees participated in the second Well-Characterized Biotechnology Product (WCBP) Chemistry and Manufacturing Controls (CMC) strategy forum, titled “Analysis and Structure Characterization of Monoclonal Antibodies (MAbs),” held in San Francisco to discuss lot release and characterization test issues specific to MAbs (1).

The objective of the meeting was twofold: to identify a “core” set of assays most useful for lot-release testing of MAbs and to define a mechanism for selecting appropriate potency tests. Two separate workshops were held as a part of the strategy forum discussing these topics in detail. The purpose of this article is to describe the discussion that occurred and to define the “core” set of assays that are most frequently used for MAb characterization and lot-release testing.

Two Sessions
The morning session featured talks by three experts using a case study format to provide an industry perspective and approach to the test methods used for characterization and release testing of MAbs. The goal of the roundtable discussions was to identity those “critical quality attributes” required to demonstrate product consistency for lot release — and the corresponding test methods used to determine those criteria. So the first discussion focused on examining the physicochemical quality attributes of MAbs and test methods required for their characterization. The second discussion examined the question of which physicochemical test methods should be used to assess the quality attributes of MAbs for lot release.

The morning panel members were Weseley Wang (Amgen), Mark Plucinsky (Centocor), Wassim Nashabeh (Genentech), Rohin Mhatre (Biogen), Keith Webber (FDA-CDER), and Brooks Sunday (Schering-Plough).

Panelists in the afternoon session were Mark Schenerman (MedImmune, Inc.), Steve Kozlowski (FDA-CDER), Kathryn Stein (Macrogenics), and Hélène Gazzano-Santoro (Genentech).

The goal of this session was to discuss the issues regarding development and selection of an appropriate potency assay(s) supporting commercialization. The group was asked to try to find a potential algorithm for making decisions about what type of potency testing should be done and when during the course of product development such decisions should be made. The categories of assays discussed included animal-based potency tests, cell culture–based bioassays, enzymatic assays, and binding assays (ELISAs).

Analytical Methods Discussed

CGE: capillary gel electrophoresis
CIEF: capillary isoelectric focusing
CZE: capillary zone electrophoresis
ELISA: enzyme-linked immunosorbent assay
ESI-MS: electrospray-ionization mass spectrometry
FACS: fluorescence-activated cell sorting
HIC: hydrophobic-interaction chromatography
IEC: ion-exchange chromatography
IEF: isoelectric focusing
SDS-PAGE: sodium-dodecyl sulfate polyacrylamide gel electrophoresis
SEC: size-exclusion chromatography

Reference

1 Schenerman MA, et al. Analysis and Structure Characterization of Monoclonal Antibodies. BioProcess Int. 2(2) 2004: 42–52; www.bioprocessintl.com/download- PDF/?article=17215&post=cmc-strategy-forum-report- 20120046&spon=false&partner=.

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