期刊
PROTEIN EXPRESSION AND PURIFICATION
卷 66, 期 1, 页码 58-65出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.pep.2009.01.007
关键词
2-D DIGE; IgG; Recombinant antibody; Staphylococcus Protein A chromatography; Affinity chromatography
类别
资金
- Biomedical Research Council of A*STAR (Agency for Science, Technology and Research), Singapore
Two-dimensional fluorescence difference gel electrophoresis (2-D DIGE) is an established method for assessing protein expression strategies, understanding pathogenesis mechanisms, characterizing biomarkers, and controlling therapeutic processes. We applied 2-D DIGE to facilitate the development of a purification process for a recombinant IgG(1) antibody against Rhesus D antigen expressed by Chinese hamster ovary cells. The variability of two expression clones as well as the influence of cell viability on the host-cell protein pattern was assessed quantitatively. Up to 800 different spots were identified. 2-D DIGE showed that differences in cell viability had more influence on the protein expression pattern than did the expression clone itself After purification of the IgG from different culture supernatants, the protein patterns on 2-D DIGE were identical, indicating the validity of purification scheme. (C) 2009 Elsevier Inc. All rights reserved.
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