Electrochromatography: a method for automatic immunoaffinity chromatography on porous membranes.
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.
Abelev
G I
GI
Cancer Research Center, Russian Academy of Medical Sciences, Moscow.
Karamova
E R
ER
Lazarevich
N L
NL
Kiseleva
V I
VI
Poverenny
A M
AM
D000911:Antibodies, Monoclonal; D001331:Automation; D002272:Carcinoembryonic Antigen / Q000032:analysis* / Q000276:immunology; D002846:Chromatography, Affinity / Q000379:methods*; D004247:DNA / Q000276:immunology; D004249:DNA Damage; D004268:DNA-Binding Proteins / Q000032:analysis; D004563:Electrochemistry; D000939:Epitopes / Q000032:analysis / Q000276:immunology; D006801:Humans; D008567:Membranes, Artificial; D000509:alpha-Fetoproteins / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D002272:Carcinoembryonic Antigen; D004268:DNA-Binding Proteins; D000939:Epitopes; D008567:Membranes, Artificial; D000509:alpha-Fetoproteins; D004247:DNA