Beresten' S F, Rubikaĭte B I, Kiselev L L
Bioorg Khim. 1986 Mar;12(3):316-26.
A method for localization of antigenic determinants in a polypeptide chain of unknown primary structure was proposed. A protein is modified at NH2-terminal and epsilon-NH2-groups of lysine residues with maleic anhydride and then is subjected to partial enzymatic cleavage. Newly formed NH2-terminal groups are tagged with radioiodinated Bolton--Hunter's reagent. The labeled fragments of the antigen are then demaleylated. Comparison of the two longest labeled fragments, only one of which still binds monoclonal antibody, makes it possible to define the location of the antigenic determinant along the polypeptide chain. The method was tested on the bovine tryptophanyl-tRNA synthetase using earlier prepared monoclonal antibodies against this enzyme.
提出了一种在一级结构未知的多肽链中定位抗原决定簇的方法。用马来酸酐对蛋白质的氨基末端和赖氨酸残基的ε-氨基进行修饰，然后进行部分酶切。新形成的氨基末端用放射性碘化的博尔顿-亨特试剂进行标记。然后将抗原的标记片段进行脱马来酰化处理。比较两个最长的标记片段（其中只有一个仍能结合单克隆抗体），就有可能确定抗原决定簇在多肽链上的位置。使用先前制备的针对该酶的单克隆抗体，在牛色氨酰-tRNA合成酶上对该方法进行了测试。
