Detection of Protein Heterogeneity by HPLC
Yasutoshi Kawai, PhD
Friday, March 30, 2012
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Chapters
Detection of Protein Heterogeneity by HPLC
Presentation Outline
Introduction
Protein Heterogeneity Results From...
Heterogeneity in Therapeutic Antibodies
Size Exclusion Chromatography (SEC)
SEC Chromatograms of Immunoglobulins
Schematic Diagram of IgG
Separation of Pepsin Digested IgG by SEC
Separation of Papain Digested IgG by SEC
Separation of IgG Monomer and Dimer by SEC
Separation of IgG Monomer, Dimer, and Fragments by Novel SEC Columns
Separation of IgG Aggregates by SEC
Separation of PEGylated IgGs by SEC
Reversed Phase Chromatography RPC
Separation of Papain Digested IgG Fragments by RPC
Separation of Heavy and Light Chains of IgG by RPC
Separation of PEGylated Lysozymes by RPC
Hydrophobic Interaction Chromatography (HIC)
Separation of Deamidation Variants of mAbs by HIC
Separation of Methionine Oxidized Variants of mAbs by HIC
Ion Exchange Chromatography IEC
IEC (WCX) Chromatograms if Commercially Available Therapeutic IgGs
Separation of Lysine Variants of IgG by IEC (WCX)
Separation of Deamidation Variants of Ribo-nuclease A by IEC (WCX)
Conclusions
Thank You!
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