Immunoadsorption of IgG onto second antibody covalently attached to Amino-Dylark beads for radioimmunoassays.
We present a solid-phase immobilization method for radioligand assays, using an immunoadsorption coating procedure of anti-triiodothyronine rabbit IgG (anti-T3 IgG) onto second antibody (sheep anti-rabbit IgG) covalently bound to Amino-Dylark beads. The second antibody was in excess, compared with the first antibody, thus eliminating reproducibility problems between immunoadsorptions. Beads coated with second antibody can be used to immobilize a variety of antigen-specific first antibodies. The amount of anti-T3 antibody required for solid-phase T3 radioimmunoassay (RIA) was only 10% more, per assay tube, than that utilized in liquid-phase T3 RIA, in which polyethylene glycol solution was the separation reagent; characteristics of assay performance were comparable. The immobilization procedure requires high-titer antisera or antigen-specific IgG and seems advantageous because of the decrease in antibody requirements without significant modification of antibody functionality.
Kakabakos
S E
SE
National Center for Scientific Research Demokritos, Athens, Greece.
Evangelatos
G P
GP
Ithakissios
D S
DS
D000888:Antibodies, Anti-Idiotypic; D005029:Ethylenediamines; D005663:Furans; D006801:Humans; D007074:Immunoglobulin G; D007163:Immunosorbent Techniques; D008299:Maleic Anhydrides; D008863:Microspheres; D011092:Polyethylene Glycols; D011137:Polystyrenes; D011863:Radioimmunoassay / Q000706:statistics & numerical data; D014284:Triiodothyronine / Q000097:blood / Q000276:immunology*
D000888:Antibodies, Anti-Idiotypic; D005029:Ethylenediamines; D005663:Furans; D007074:Immunoglobulin G; D008299:Maleic Anhydrides; D011137:Polystyrenes; D014284:Triiodothyronine; D011092:Polyethylene Glycols; C031234:ethylenediamine; C025704:Styromal