Construction and characterization of a single-chain catalytic antibody

Richard A. Gibbs, Bruce A. Posner, David R. Filpula, Steven W. Dodd, Malcolm A J Finkelman, Timothy K. Lee, Marie Wroble, Marc Whitlow, Stephen J. Benkovic

Research output: Contribution to journalArticle

52 Scopus citations

Abstract

The antigen-binding (Fab) fragment of the catalytic monoclonal antibody NPN43C9 has recently been cloned by using bacteriophage λ. 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.

Original languageEnglish (US)
Pages (from-to)4001-4004
Number of pages4
JournalProceedings of the National Academy of Sciences of the United States of America
Volume88
Issue number9
Publication statusPublished - 1991

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Keywords

  • Amide hydrolysis
  • Antibody cloning
  • Escherichia coli expression
  • Single-chain antigen-binding protein
  • Single-chain Fv

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

Gibbs, R. A., Posner, B. A., Filpula, D. R., Dodd, S. W., Finkelman, M. A. J., Lee, T. K., ... Benkovic, S. J. (1991). Construction and characterization of a single-chain catalytic antibody. Proceedings of the National Academy of Sciences of the United States of America, 88(9), 4001-4004.