Production of specific antibodies against protein A fusion proteins

EMBO J. 1986 Sep;5(9):2393-8. doi: 10.1002/j.1460-2075.1986.tb04509.x.

Abstract

The gene for Staphylococcal protein A was fused to the coding sequence of bacterial beta-galactosidase, alkaline phosphatase and human insulin-like growth factor I (IGF-I). The fusion proteins, expressed in bacteria, were purified by affinity chromatography on IgG-Sepharose and antibodies were raised in rabbits. All three fusion proteins elicited specific antibodies against both the inserted protein sequences and the protein A moiety. In the case of IGF-I, the protein A moiety in the fusion protein may act as an adjuvant since native IGF-I alone is a poor immunogen. The results suggest that the protein A fusion system can be used for efficient antibody production against peptides or proteins expressed from cloned or synthetic genes. To facilitate such gene fusions a set of optimized vectors have been constructed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkaline Phosphatase / analysis
  • Amino Acid Sequence
  • Antibodies*
  • Antigen-Antibody Complex
  • Base Sequence
  • Escherichia coli / genetics
  • Genes
  • Humans
  • Insulin-Like Growth Factor I / analysis
  • Recombinant Fusion Proteins / analysis*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology
  • Recombinant Proteins / analysis*
  • Staphylococcal Protein A / analysis*
  • Staphylococcal Protein A / immunology
  • beta-Galactosidase / analysis

Substances

  • Antibodies
  • Antigen-Antibody Complex
  • Recombinant Fusion Proteins
  • Recombinant Proteins
  • Staphylococcal Protein A
  • Insulin-Like Growth Factor I
  • Alkaline Phosphatase
  • beta-Galactosidase