[The use of immunoaffinity chromatography for isolation of carcinoembryonal antigen].
Shcherban' Ai, Lukashevich Lv, Marchuk Si, Nezhinets Ea
Abstract
Shcherban' Ai, Lukashevich Lv, Marchuk Si, Nezhinets Ea
Abstract
A procedure for cancer-embryonal antigen isolation involved extraction of the antigen by chloric acid, affinity chromatography on CNBr-Sepharose 4B containing immobilized antibodies and gel filtration on Sephadex G-200. Contaminating proteins, adsorbed on the immunosorbent were eluted with 0.05 M phosphate buffer, pH 7.4 containing 0.3% sodium cholate and 1 M NaCl, while cancer-embryonal antigen--with 0.1 M glycyl-HCl buffer, pH 2.2. After purification of the preparation on Sephadex G-200 homogenous fraction of the antigen, as shown by means of polyacrylamide gel electrophoresis in presence of SDS and by immunoblotting, was obtained. Yield of the antigen was about 40%.
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A procedure for cancer-embryonal antigen isolation involved extraction of the antigen by chloric acid, affinity chromatography on CNBr-Sepharose 4B containing immobilized antibodies and gel filtration on Sephadex G-200. Contaminating proteins, adsorbed on the immunosorbent were eluted with 0.05 M phosphate buffer, pH 7.4 containing 0.3% sodium cholate and 1 M NaCl, while cancer-embryonal antigen--with 0.1 M glycyl-HCl buffer, pH 2.2. After purification of the preparation on Sephadex G-200 homogenous fraction of the antigen, as shown by means of polyacrylamide gel electrophoresis in presence of SDS and by immunoblotting, was obtained. Yield of the antigen was about 40%.
Key concepts: Chromatography, Chemistry, Sephadex, Elution, Size-exclusion chromatography, Antigen, Affinity chromatography, Sepharose