Construction and characterization of a single-chain catalytic antibody.

Gibbs, R A; Posner, B A; Filpula, D R; Dodd, S W; Finkelman, M A; Lee, T K; Wroble, M; Whitlow, M et al. · Proc Natl Acad Sci U S A · 1991

basic_science · Level V

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Abstract

The antigen-binding (Fab) fragment of the catalytic monoclonal antibody NPN43C9 has recently been cloned by using bacteriophage lambda. By inserting the variable regions of this Fab coding sequence into a (NH2)-VL-linker-VH-(COOH) construct (where VL and VH represent the heavy and light chain variable regions), we have assembled a recombinant gene encoding a catalytic single-chain antigen-binding protein. This protein has been expressed in Escherichia coli and exhibits the same catalytic parameters as the parent monoclonal antibody NPN43C9. Single-chain forms of catalytic antibodies may prove valuable for structural and site-directed mutagenesis studies as well as for large-scale applications of catalytic antibodies.

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