Growth at sub-optimal temperatures allows the production of functional, antigen-binding Fab fragments in Escherichia coli |
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Authors: | S Cabilly |
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Institution: | Department of Biology, Technion-Israel Institute of Technology, Haifa. |
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Abstract: | Expression in Escherichia coli of recombinant genes coding for the kappa-chain and the Fd fragment of an antibody directed against carcinoembryonic antigen gives rise to Fab dimers. These Fab fragments possess antibody activity, as demonstrated by enzyme-linked immunosorbent assay as well as by ligand competition assay. Effective production of soluble Fab in Escherichia coli was achieved by a decrease in the growth temperature. Following a one step purification by anion exchange chromatography, the bacterially-produced Fab retains its activity at 4 degrees C for at least two months. The relatively simple methodology described in this study should be useful for the design and production of antibodies in bacteria. |
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