Construction and characterization of a single-chain catalytic antibody. |
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| Authors: | R A Gibbs B A Posner D R Filpula S W Dodd M A Finkelman T K Lee M Wroble M Whitlow S J Benkovic |
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| Affiliation: | Department of Chemistry, Pennsylvania State University, University Park 16802. |
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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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