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  • Title: A genetic approach to the generation of antibodies with enhanced catalytic activities.
    Author: Lesley SA, Patten PA, Schultz PG.
    Journal: Proc Natl Acad Sci U S A; 1993 Feb 15; 90(4):1160-5. PubMed ID: 8094556.
    Abstract:
    A hydrolytic catalytic antibody, generated against a nitrophenyl phosphonate transition state analogue, has been cloned and expressed in Escherichia coli for use as a model system to demonstrate the feasibility of using genetic selections to enhance catalytic activity. Conditions were found that permit the secretion of active recombinant antibody into the periplasm of a strain of E. coli deficient in the biotin biosynthetic genes (delta bio-gal). A number of substrates were synthesized that, upon hydrolysis by the antibody, yield free biotin, which is required for cell growth. The substrates and selections can be used to identify mutants of the antibody with altered activities. This approach should be generalizable to a wide number of hydrolytic reactions including the selective cleavage of peptide, polysaccharide, phosphodiester, and ester bonds.
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