Institute for International Research

Course Description

In this course, you will learn the fundamentals of protein structural analysis using modern analytical technologies. We will review the post-translational modifications commonly observed on recombinant proteins produced from manufacturing cell lines and discuss the potential impact of the structural heterogeneities on biological activity. In addition, biophysical methods used for characterization of aggregates and application of target binding and cell-based functional assays will be discussed. We will review application examples on characterization of monoclonal antibodies and evaluate the use of key techniques in comparison and in combination. We will discuss the impact of cell line selection and process parameters on product quality profile as well as considerations of effects of sample preparation, storage and handling of process intermediates. The objective is to provide participants with key technical information along with perspectives to enable them to apply the technologies to their own projects and evolve their own analytical strategy to support the various stages of product development.

Who Should Attend?

This course is designed for scientists, engineers and managers in the life science industry whose job functions involve analytical methods development, upstream and downstream in-process monitoring, formulation, quality control and research and development of recombinant therapeutic proteins.

Complete Course Agenda

Introduction to protein structure and post-translational modifications

Protein chemistry techniques: Highlight key technical basics, instrumentation and review application examples

• Capillary electrophoresis: CE-SDS, CE-IEF, CZE with UV and LIF detection, CE-MS
• Enzymatic methods and HPLC
• Peptide mapping
• Amino acid analysis
• Protein sequencing: N-terminal and C-terminal chemistries
• Mass spectrometry
• Biophysical characterization and other methods: spectral methods, light scattering, calorimetry, analytical ultracentrifugation, and field flow fractionation

Additional analytical topics:

• Glycosylation analysis
• Activity assays: Immunoassays, binding assays, cell-based assays
• Assays for impurities
• Sample preparation, automation work flow

Characterization of recombinant protein products:

• Quality control of biotechnology products: ICH guidance documents and CMC strategy considerations
• Monoclonal antibody analysis
  • Antibody domain structure and subclasses
  • Common sources of product heterogeneity
  • Influence of product heterogeneity on activity and pharmacokinetics
  • Product variant analysis: assay application examples
• Additional recombinant protein analysis

Bioprocess impact on product quality

• Cell line selection: choice of host cell, screening for production cell clone
• Upstream process variables and control
• Downstream process impact: aggregation and impurities
• Formulation and storage

Selecting characterization methods to support various stages of product development

Course Instructor

Christine P. Chan, Ph.D., Senior Manager, Technology Development, Genzyme Corporation
Christine Chan is a protein biochemist with over 20 years of experience in the biopharmaceutical industry. She specializes in the analysis of recombinant products produced from mammalian cells for vaccines and biologics development. She has extensive hands-on experience with classical protein chemistry methods including Edman sequencing, amino acid analysis and protein purification, as well as the newer techniques such as capillary electrophoresis and mass spectrometry. She is experienced in highthroughput screening using plate-based assays as well as cell-based assays. Before joining Genzyme in December 2008, Dr. Chan was group leader for an analytical group in the Department of Bioprocess Research & Development at Merck & Co., responsible for quantitation and characterization assays supporting cell line and process development for therapeutic protein product development. Dr. Chan obtained her Ph.D. from the University of California-Davis and did postdoctoral work at the Howard Hughes Medical Institute at the University of Washington on growth factor signal transduction and protein phosphorylation.