Quantification of monoclonal antibodies in complex mixtures by protein G high-performance liquid affinity chromatography.
High-performance liquid affinity chromatography (HPLAC) utilizing Protein G as a ligand has been evaluated for rapid quantification of monoclonal antibodies (MAbs) in various solutions. The results obtained by HPLAC agreed to within 10% of a standard enzyme-linked immunospecific assay (ELISA). A standard curve was prepared by injection of known amounts of a purified murine IgG1 with the elution peak area analyzed by computer integration software. Accuracy of quantification was independent of the injection volume, solution compositions, or mouse IgG subclass. A method is described for using Protein G HPLAC to determine murine IgG levels in various complex mixtures within 15 min, compared to the ELISA which required 5 h.
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Department of Recovery Process R & D, Genentech, Inc., South San Francisco, California 94080.
Vetterlein
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D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay; D019204:GTP-Binding Proteins; D007074:Immunoglobulin G / Q000276:immunology; D008024:Ligands; D051379:Mice; D012680:Sensitivity and Specificity; D012984:Software
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D008024:Ligands; D019204:GTP-Binding Proteins