Analysis of epitopes of mouse monoclonal antibodies against human alpha-fetoprotein

Tumour Biol. 2001 Jul-Aug;22(4):254-61. doi: 10.1159/000050624.

Abstract

Thirty-six monoclonal antibodies (MAbs) against human alpha-fetoprotein (AFP) were analyzed for the location of their epitopes by reacting them with a set of yeast recombinant AFP proteins using ELISA. Recombinant AFP proteins containing either one, two or all three domains, i.e. domain I, domain III, domain I-II, domain II-III and domain I-II-III, were produced and secreted into the culture medium of yeast cells harboring the expression plasmids. Epitopes of 13 MAbs were localized on domain I and 17 others were on domain III. However, the exact location of the epitopes of the remaining 6 MAbs could not be defined. The epitope of an antibody, namely AFY6, which was located in domain I, was successfully mapped on an octapeptide, C175KAENAVE182, using synthesized overlapping octapeptides.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology*
  • Enzyme-Linked Immunosorbent Assay
  • Epitopes / immunology*
  • Escherichia coli
  • Humans
  • Mice
  • Protein Conformation
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / immunology
  • alpha-Fetoproteins / chemistry
  • alpha-Fetoproteins / immunology*

Substances

  • Antibodies, Monoclonal
  • Epitopes
  • Recombinant Proteins
  • alpha-Fetoproteins