Site-directed mutagenesis of HLA molecules reveals the functional epitope of a human HLA-A1/A36-specific monoclonal antibody

HLA. 2023 Feb;101(2):138-142. doi: 10.1111/tan.14895. Epub 2022 Nov 25.

Abstract

Eplet 44KM is currently listed in the HLA Epitope Registry but does not adhere to the eplet definition of an amino acid configuration within a 3.5 Å radius. Eplet 44KM has been previously redefined to the antibody-verified reactivity pattern 44K/150V/158V, based on reactivity analysis of monoclonal antibody VDK1D12. Since the three residues are always simultaneously present on common HLA alleles, methods to define which residue is crucial for antibody-induction and binding are limited. In this proof-of-concept study, we performed site-directed mutagenesis to narrow down the antibody-verified reactivity pattern 44K/150V/158V to a single amino acid and defined 44K as the eplet or functional epitope of mAb VDK1D12.

Keywords: antibody verification; antigenicity; eplet; immunogenicity; monoclonal antibody; reactivity pattern; site-directed mutagenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Amino Acids
  • Antibodies, Monoclonal* / chemistry
  • Antibody Specificity
  • Epitopes
  • HLA-A Antigens
  • HLA-A1 Antigen*
  • Histocompatibility Testing
  • Humans
  • Mutagenesis, Site-Directed

Substances

  • Antibodies, Monoclonal
  • Epitopes
  • HLA-A1 Antigen
  • HLA-A Antigens
  • Amino Acids