使用生物素标记的癌胚抗原和抗生物素蛋白作为沉淀剂在溶液相中测定单克隆抗体的表位特异性和亲和力。
Determination of epitope specificities and affinities of monoclonal antibodies in solution phase using biotin-labeled carcinoembryonic antigen and avidin as precipitating agent.
作者信息
Krüger U, Wickert L, Wagener C
机构信息
Institute of Clinical Chemistry and Pathobiochemistry, University of Technology, Aachen, F.R.G.
出版信息
J Immunol Methods. 1989 Feb 8;117(1):25-32. doi: 10.1016/0022-1759(89)90114-2.
A generally applicable method for the selective precipitation of antigens from solution phase is presented. The method is based on the rapid and quantitative binding of avidin to biotin-labeled antigen in the presence of biotinylated carrier protein. Using carcinoembryonic antigen (CEA) as a model antigen biotin-labeled CEA was precipitated to greater than 97% at amounts of up to 5000 ng/tube. The method was applied to the determination of epitope specificities and affinities of monoclonal anti-CEA antibodies in solution phase. Non-specific precipitation did not exceed 2% for radiolabeled Fab fragments and 6% for radiolabeled IgG. Because antigen-antibody binding takes place in solution, artifacts introduced by the immobilization of reactants to solid supports are omitted.
本文介绍了一种从溶液相中选择性沉淀抗原的通用方法。该方法基于在生物素化载体蛋白存在下抗生物素蛋白与生物素标记抗原的快速定量结合。以癌胚抗原（CEA）作为模型抗原，在每管高达5000 ng的量下，生物素标记的CEA沉淀率大于97%。该方法应用于测定溶液中单克隆抗CEA抗体的表位特异性和亲和力。放射性标记的Fab片段非特异性沉淀不超过2%，放射性标记的IgG不超过6%。由于抗原 - 抗体结合在溶液中发生，避免了反应物固定在固相载体上所引入的假象。
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