Bacteriophage cloning and Escherichia coli expression of a human IgM Fab

Hum Antibodies Hybridomas. 1992 Apr;3(2):81-5.

Abstract

We have combined the molecular biology methods of the polymerase chain reaction and recombinant DNA cloning in bacteriophage lambda to express a human IgM Fab in Escherichia coli using genes derived from an Epstein-Barr virus transformed cell line. This method comprises three cDNA amplifications and a single cloning step, culminating in the stable overexpression of mammalian heterodimeric recombinant protein in a prokaryotic host.

MeSH terms

  • Bacteriophage lambda / genetics
  • Base Sequence
  • Cloning, Molecular
  • DNA / genetics
  • Escherichia coli / genetics
  • Gene Expression
  • Genes, Immunoglobulin*
  • Humans
  • Hybridomas / immunology
  • Immunoglobulin Fab Fragments / genetics*
  • Immunoglobulin M / genetics*
  • Molecular Sequence Data
  • Polymerase Chain Reaction / methods

Substances

  • Immunoglobulin Fab Fragments
  • Immunoglobulin M
  • DNA