Engineering human monoclonal antibody fragments: a recombinant enzyme-linked Fab

New Microbiol. 1995 Apr;18(2):127-33.

Abstract

A new plasmid vector, pCRP, allowing the expression of human recombinant monoclonal antibody Fab fragments fused with a bacterial acid phosphatase has been constructed. pCRP can accept heavy- and light-chain cDNAs cloned from combinatorial antibody libraries displayed on filamentous phages with the pCombIII system and is able to direct expression to soluble Fabs in which the carboxy-terminus of the heavy chain is fused to the amino-terminus of the mature PhoN nonspecific acid phosphatase of Providencia stuartii. Using the pCRP vector, we expressed two different human recombinant Fabs cloned from combinatorial libraries (one anti-tetenus toxoid and the other anti-HIV-1 gp120) fused with the acid phosphatase. In both cases chimeric antibodies were obtained which retained the antigen-binding ability and the enzymatic activity. Similar Fab-enzyme fusions can be successfully used, even unpurified, in enzyme immunoassays.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acid Phosphatase / genetics
  • Acid Phosphatase / metabolism
  • Antibodies, Monoclonal / biosynthesis*
  • Antibodies, Monoclonal / genetics
  • Base Sequence
  • Gene Library
  • Genes, Bacterial / genetics
  • Genetic Vectors / genetics
  • HIV Envelope Protein gp120 / immunology
  • HIV-1
  • Humans
  • Immunoglobulin Fab Fragments / biosynthesis*
  • Immunoglobulin Fab Fragments / genetics
  • Molecular Sequence Data
  • Providencia / enzymology
  • Recombinant Fusion Proteins / biosynthesis*
  • Tetanus Toxoid / immunology

Substances

  • Antibodies, Monoclonal
  • HIV Envelope Protein gp120
  • Immunoglobulin Fab Fragments
  • Recombinant Fusion Proteins
  • Tetanus Toxoid
  • Acid Phosphatase