title: Efficient method for production of high yields of Fab fragments in Drosophila S2 cells.
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.
authors.affiliation: Institut Pasteur, Unité de Virologie Structurale, Département de Virologie and CNRS Unité de Recherche Associée 3015, Paris, France. marija@pasteur.fr
authors.forename: Marija
authors.initials: M
authors.lastname: Backovic
authors.forename: Daniel X
authors.initials: DX
authors.lastname: Johansson
authors.forename: Barbara G
authors.initials: BG
authors.lastname: Klupp
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Mettenleiter
authors.forename: Mats A A
authors.initials: MA
authors.lastname: Persson
authors.forename: Felix A
authors.initials: FA
authors.lastname: Rey
mesh_terms: D000818:Animals; D002460:Cell Line; D004330:Drosophila / Q000166:cytology / Q000235:genetics* / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection
chemical_list: D007140:Immunoglobulin Fab Fragments; D011993:Recombinant Fusion Proteins