Electrochromatography: a method for automatic immunoaffinity chromatography on porous membranes

Immunol Lett. 1994 May;40(2):133-8. doi: 10.1016/0165-2478(94)90184-8.

Abstract

Electrochromatography (ECHR) exploits a very high electro-osmotic counterflow developed in porous membranes at discontinuous electrophoresis. This counterflow exceeds considerably the anodic migration of any negatively charged protein and is used as a 'conveyer belt' for sequential transfer of immunoreagents to the specific adsorbents (antigens or antibodies) fixed on the nitrocellulose membrane. This approach was applied for simultaneous detection of two antigens (alpha-fetoprotein and carcino-embryonic antigen) in one sample, for determination of subfractions of alpha-fetoprotein, different in their epitope specificity, and for detection of L chains with certain idiotype on the background of heterogeneous L fraction. ECHR was used also for the partition of different antibodies to DNA adducts, demonstrating the possibility of applying this method to the study of DNA-binding proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal
  • Automation
  • Carcinoembryonic Antigen / analysis*
  • Carcinoembryonic Antigen / immunology
  • Chromatography, Affinity / methods*
  • DNA / immunology
  • DNA Damage
  • DNA-Binding Proteins / analysis
  • Electrochemistry
  • Epitopes / analysis
  • Epitopes / immunology
  • Humans
  • Membranes, Artificial*
  • alpha-Fetoproteins / analysis*
  • alpha-Fetoproteins / immunology

Substances

  • Antibodies, Monoclonal
  • Carcinoembryonic Antigen
  • DNA-Binding Proteins
  • Epitopes
  • Membranes, Artificial
  • alpha-Fetoproteins
  • DNA