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
DOIs
StatePublished - Jan 1 1991

Keywords

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

ASJC Scopus subject areas

  • General

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    Gibbs, R. A., Posner, B. A., Filpula, D. R., Dodd, S. W., Finkelman, M. A. J., Lee, T. K., Wroble, M., Whitlow, M., & 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. https://doi.org/10.1073/pnas.88.9.4001