Growth at sub-optimal temperatures allows the production of functional, antigen-binding Fab fragments in Escherichia coli.
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.
Cabilly
S
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Department of Biology, Technion-Israel Institute of Technology, Haifa.
D000936:Antigen-Antibody Complex; D002272:Carcinoembryonic Antigen / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development; D005803:Genes, Immunoglobulin; D005822:Genetic Vectors; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D015183:Restriction Mapping; D013696:Temperature
D000936:Antigen-Antibody Complex; D002272:Carcinoembryonic Antigen; D007140:Immunoglobulin Fab Fragments; D011994:Recombinant Proteins