2-D DIGE to expedite downstream process development for human  monoclonal antibody purification.
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 IgG1 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.
Department of Biotechnology, University of Natural Resources and Applied Life Sciences Vienna, Muthgasse 18, Vienna 1190, Austria.
Julita K
JK
Grzeskowiak
Anne
A
Tscheliessnig
Poh Choo
PC
Toh
Janet
J
Chusainow
Yih Yean
YY
Lee
Niki
N
Wong
Alois
A
Jungbauer
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D016466:CHO Cells; D002470:Cell Survival; D002845:Chromatography / Q000379:methods; D006224:Cricetinae; D003412:Cricetulus; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005453:Fluorescence; D006801:Humans; D015203:Reproducibility of Results
D000911:Antibodies, Monoclonal