Brodsky F M
J Cell Biol. 1985 Dec;101(6):2047-54. doi: 10.1083/jcb.101.6.2047.
Three monoclonal antibodies that react with previously undefined antigenic determinants on the clathrin molecule have been produced and characterized. They were isolated from a fusion between myeloma cells and popliteal lymphocytes from SJL mice that had received footpad injections of human brain clathrin. This protocol was chosen to favor the production of antibodies to poorly immunogenic proteins and thereby increase the repertoire of anti-clathrin monoclonal antibodies. One antibody (X16) reacts preferentially with the heavier of the two clathrin light chains (LCa) when it is not associated with heavy chain. This specificity is different from that of the anti-LCa antibody, CVC.6, which has preferential reactivity with heavy chain-associated LCa. In addition, X16 and CVC.6 bound simultaneously to LCa, confirming that they react with different sites. The other two antibodies produced, X19 and X22, react with two different determinants on the clathrin heavy chain, based on immunoprecipitation, Western blot, and binding studies. Competitive binding studies with anti-clathrin monoclonal antibodies showed that they define a total of five distinct antigenic determinants on bovine clathrin.
已制备并鉴定出三种单克隆抗体，它们可与网格蛋白分子上先前未定义的抗原决定簇发生反应。这些抗体是从骨髓瘤细胞与接受了足垫注射人脑网格蛋白的SJL小鼠腘窝淋巴细胞的融合产物中分离得到的。选择该方案是为了有利于产生针对免疫原性较差的蛋白质的抗体，从而增加抗网格蛋白单克隆抗体的种类。一种抗体（X16）在不与重链结合时，优先与两条网格蛋白轻链中较重的那条（LCa）发生反应。这种特异性不同于抗LCa抗体CVC.6，后者优先与重链相关的LCa发生反应。此外，X16和CVC.6可同时结合到LCa上，证实它们与不同的位点发生反应。根据免疫沉淀、蛋白质印迹和结合研究，产生的另外两种抗体X19和X22与网格蛋白重链上的两个不同决定簇发生反应。与抗网格蛋白单克隆抗体的竞争性结合研究表明，它们总共定义了牛网格蛋白上五个不同的抗原决定簇。
