Two-dimensional fluorescence difference gel electrophoresis for comparison of affinity and non-affinity based downstream processing of recombinant monoclonal antibody |
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Authors: | Julita K Grzeskowiak Anne Tscheliessnig Poh Choo Toh Janet Chusainow Yih Yean Lee Niki Wong Alois Jungbauer |
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Institution: | 1. Department of Biotechnology, University of Natural Resources and Applied Life Sciences, Vienna, Muthgasse 18, A-1190 Vienna, Austria;2. Bioprocessing Technology Institute, A*STAR (Agency for Science, Technology and Research), 20 Bioplois Way, No 06-01 Centros, Singapore 138668, Singapore |
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Abstract: | Although Staphylococcus Protein A (SpA) affinity chromatography is the state of the art capture step for antibody purification, non-affinity methods are more economical. We used two-dimensional fluorescence difference gel electrophoresis (2-D DIGE) to evaluate the purification of a recombinant IgG1 antibody from cultured cells, with two different processes: (1) SpA capture followed by cation-exchange chromatography (CEX); and (2) CEX capture, followed by anion exchanger, then hydrophobic interaction chromatography. Efficiencies were similar in sodium dodecylsulphate polyacrylamide gel electrophoresis and size-exclusion chromatography; however, 2-D DIGE revealed higher efficiency with SpA than with CEX capture. Thus, 2-D DIGE is a valuable tool for downstream process development. |
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Keywords: | 2-D DIGE IgG Recombinant antibody Staphylococcus Protein A chromatography Affinity chromatography Ion exchange chromatography |
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