Properties of immunoadsorbents prepared by antigen coupling to glutaraldehydeactivated polyacrylamide gel, BrCN-activated Sepharose and by copolymerization of antigens by glutaraldehyde
Komissarenko, S.V.; Avrameas, S.
Ukrainskii Biokhimicheskii Zhurnal 50(4): 500-511
1978
ISSN/ISBN: 0201-8470 PMID: 98883 Document Number: 134825
Three types of immunoadsorbents were synthetized by antigen coupling to glutaraldehyde-activated polyacrylamide gel, BrCN-activated sepharose 4B and protein insolubilization using glutaraldehyde as a cross-linking agent. Different concentrations of rabbit gamma globulin, bovine serum albumin, soybean trypsin inhibitor and bovine ribonuclease were used as antigens and some properties of such immunoadsorbents were studied. Using 125J labelled antigens it was shown that glutaraldehyde-activated polyacrylamide gel (Bio-Gel P-300) couples 81-94% of the antigen added in a concentration of 0.5-4.0 mg.ml-1 gel and BrCN-activated sepharose 4B bounds 62-88% of the labelled antigen in a concentration of 5.0-20.0 mg.ml-1 gel. These antigen derivatives as well as those obtained by glutaraldehyde protein insolubilization permitted 54-88% of antibodies added with the immune sera to be isolated. There was a significant antigen leakage from sepharose immunoadsorbents after several antisera treatments or after half a year storage but despite of this antigen desorption all types of the immunoadsorbents studied preserved their antibody isolation capacity. Immune sera immunoglobulins nonspecific binding by immunoadsorbents was less on sepharose than Bio-Gel antigen derivatives.