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

R. Burioni, P. Plaisant, M. L. Riccio, G. M. Rossolini, R. Santangelo, A. Vannini, G. Satta

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)127-133
Number of pages7
JournalNew Microbiologica
Volume18
Issue number2
Publication statusPublished - Apr 1995

ASJC Scopus subject areas

  • Microbiology
  • Microbiology (medical)

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