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

Protein Engineering, Design & Selection : PEDS
Marija BackovicFélix A Rey

Abstract

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.

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Citations

Apr 28, 2011·Journal of Natural Products·Thaís B LimaOctávio L Franco
Dec 15, 2010·Proceedings of the National Academy of Sciences of the United States of America·Marija BackovicFelix A Rey
Dec 14, 2011·Protein Engineering, Design & Selection : PEDS·Allissia A GilmartinThomas Krey
Apr 5, 2013·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Alexander W TarrThomas Krey
Jun 17, 2014·Regulatory Toxicology and Pharmacology : RTP·Satin G SawantKurt A Black
Jun 22, 2012·Nature Materials·Joseph N Grima, Roberto Caruana-Gauci
Jun 25, 2016·Nature·Giovanna Barba-SpaethFélix A Rey
Sep 28, 2020·Antiviral Research·Ricardo Sanchez-VelazquezMargus Varjak
Aug 29, 2021·Vaccines·Claudia GandolfoMaria Grazia Cusi

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