Abelev G I, Karamova E R, Yazova A K, Gussev A I
Laboratory of Immunochemistry, Cancer Research Center, Moscow, U.S.S.R.
Mol Immunol. 1989 Jan;26(1):49-52. doi: 10.1016/0161-5890(89)90019-9.
The method for comparison of epitope specificity of different monoclonal antibodies to one antigen, using a panel of monoclonal antibodies to mouse and human alpha-fetoprotein is described. The method used exploits the special properties of electroendosmotic flow in nitrocellulose membranes under the conditions of anionic isotachophoresis. Electroendosmosis allows successive transfer of several immunoreagents to the dots of monoclonals previously bound to the nitrocellulose membrane. The inhibition of antigen binding to monoclonal dot, if the antigen is mixed with excess of another monoclonal antibody, demonstrates that both monoclonals are directed to the same epitope, and vice versa. The method requires neither purified monoclonals nor antigens, or radio labelling, and is performed semi-automatically. It was shown that each monoclonal antibody to mouse and human alpha-fetoprotein had its own epitope specificity.
描述了一种使用一组针对小鼠和人甲胎蛋白的单克隆抗体来比较不同单克隆抗体对一种抗原的表位特异性的方法。所使用的方法利用了在阴离子等速电泳条件下硝酸纤维素膜中电渗流的特殊性质。电渗作用允许将几种免疫试剂连续转移到先前结合在硝酸纤维素膜上的单克隆抗体点上。如果抗原与过量的另一种单克隆抗体混合，抗原与单克隆抗体点的结合受到抑制，这表明两种单克隆抗体都针对相同的表位，反之亦然。该方法既不需要纯化的单克隆抗体，也不需要抗原，也不需要放射性标记，并且是半自动进行的。结果表明，针对小鼠和人甲胎蛋白的每种单克隆抗体都有其自身的表位特异性。
