Efficient method for production of high yields of Fab fragments in Drosophila S2 cells

被引：17
|
作者：
Backovic, Marija [1 ,2 ]
Johansson, Daniel X. [3 ]
Klupp, Barbara G. [4 ]
Mettenleiter, Thomas C. [4 ]
Persson, Mats A. A. [3 ]
Rey, Felix A. [1 ,2 ]
机构：
[1] Inst Pasteur, Dept Virol, Unite Virol Struct, F-75724 Paris 15, France
[2] CNRS, Unite Rech Assoc, F-75724 Paris, France
[3] Karolinska Inst, Karolinska Univ Hosp, Ctr Mol Med, Dept Med, S-17176 Stockholm, Sweden
[4] Friedrich Loeffler Inst, Inst Mol Biol, Greifswald, Germany
来源：
PROTEIN ENGINEERING DESIGN & SELECTION | 2010年 / 23卷 / 04期
基金：
瑞典研究理事会;
关键词：
Drosophila S2 cells; expression; Fab; insect cells; monoclonal antibodies; RABBIT GAMMA-GLOBULIN; PSEUDORABIES VIRUS; ANTIBODY FRAGMENTS; INSECT CELLS; RECOMBINANT ANTIBODIES; REGULATED EXPRESSION; N-GLYCOSYLATION; SCHNEIDER CELLS; CRYSTALLIZATION; PROTEIN;
D O I：
10.1093/protein/gzp088
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
Fab molecules are used as therapeutic agents, and are invaluable tools in structural biology. We report here a method for production of recombinant Fab in Drosophila S2 cells for use in structural biology. Stably transfected S2 cell lines expressing the Fab were created within weeks. The recombinant Fab was secreted, and after affinity and size exclusion chromatography, 16 mg of pure protein were obtained from a liter of cell culture. The Fab was functional and formed a complex with its cognate antigen as demonstrated by co-precipitation and size exclusion chromatography. Biochemical characterization indicated that the Fab from S2 cells is less extensively glycosylated than the Fab obtained by digestion of antibody produced in hybridoma cells, a feature that may be advantageous for the purposes of crystallogenesis. Taken together, obtaining recombinant Fab from the S2 cells has been a faster and considerably more cost-effective method compared with the enzymatic digestion of the monoclonal antibody.
引用
收藏
页码：169 / 174
页数：6
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