Hitchhiker antigens: inconsistent ChIP results, questionable immunohistology data, and poor antibody performance may have a common factor

Biochem Cell Biol. 2013 Dec;91(6):378-94. doi: 10.1139/bcb-2013-0059. Epub 2013 Oct 23.

Abstract

Questionable data and poor antibody performance may have a common factor: antigens "hitchhiking" on the very antibodies designed to target them. Here I focus on histone hitchhikers and their antibodies, given the impact of chromatin immunoprecipitation on our understanding of DNA regulation. Caused by a lack of stringency during antibody purification, hitchhikers will impede important advances in chromatin research and therapeutics derived from that research, if similar circumstances in the study of lupus decades ago are any guide. Evidence of this phenomenon is reviewed, purification modifications for antibody manufacturing are suggested, and a histone hitchhiker detection procedure is provided.

Publication types

  • Review

MeSH terms

  • Animals
  • Antibody Affinity
  • Antibody Specificity
  • Antigen-Antibody Complex / chemistry
  • Antigen-Antibody Complex / immunology
  • Artifacts*
  • Blotting, Western / standards*
  • Chromatin / chemistry
  • Chromatin Immunoprecipitation / standards*
  • DNA / genetics
  • DNA / metabolism
  • DNA Replication
  • Histones / analysis*
  • Histones / genetics
  • Histones / immunology
  • Humans
  • Protein Binding

Substances

  • Antigen-Antibody Complex
  • Chromatin
  • Histones
  • DNA