ABSTRACT
Antibodies are a target of very diverse biotechnological interest. They are used widely in the laboratory, and hardly any branch of the biology or biochemistry can do without them; be it for qualitative assays (e.g., Western blots, microdiffusion tests, or immunohistology), quantitative assays [radioimmunoassays or enzyme-linked immunosorbent assays]. The cosecretion of both chains making up the antibody fragment can also be guaranteed by the covalent linking of both chains of the Fv fragment to form a single-chain Fv fragment. Several research groups have established protocols, and Fab fragments have been refolded at 10-40% yield, and scFv fragments have usually been refolded at 10-20% yield. By using a phosphorylcholine affinity column, originally developed for the whole antibody, the recombinant Fab fragment, the Fv fragment, the scFv fragments, and various bivalent miniantibodies, all could be purified directly from E. coli.
