Skubitz K M, Snook R W
J Immunol. 1987 Sep 1;139(5):1631-9.
A variety of monoclonal antibodies has been used to study the roles of surface proteins in neutrophil function. Many monoclonal antibodies that bind to human neutrophils react with the oligosaccharide lacto-N-fucopentaose III. Sequential immunoprecipitation of radiolabeled proteins from extracts of neutrophils labeled at the cell surface with 125I, and partial proteolysis peptide mapping studies were used to compare the proteins recognized by several widely used monoclonal antibodies that react with human neutrophils. The monoclonal antibodies that react with lacto-N-fucopentaose III (CD15) immunoprecipitated five distinct neutrophil surface proteins. The data indicate that CD15 monoclonal antibodies react with a subset of the LFA-1/HMac-1/gp 150,95 glycoprotein family as well as with CR1 on human neutrophils. The CD15 antibodies studied differed in their avidities for these proteins. The molecules immunoprecipitated by the CD15 antibodies tested were more resistant to proteolysis than the homologous proteins immunoprecipitated by the other monoclonal antibodies studied that react directly with the alpha M (CD11) or beta (CD18) chains of the LFA-1/HMac-1/gp 150,95 glycoprotein family. Some of the differences in antibody reactivity and protease sensitivity of the membrane proteins recognized by these antibodies may be due to differences in glycosylation. The data suggest that the antibodies studied can detect differences in post-translational modification among copies of certain surface proteins.
多种单克隆抗体已被用于研究表面蛋白在中性粒细胞功能中的作用。许多与人中性粒细胞结合的单克隆抗体与寡糖乳糖-N-岩藻五糖III发生反应。从用125I在细胞表面标记的中性粒细胞提取物中对放射性标记蛋白进行连续免疫沉淀，并使用部分蛋白酶解肽图谱研究来比较几种广泛使用的与人中性粒细胞反应的单克隆抗体所识别的蛋白。与乳糖-N-岩藻五糖III（CD15）反应的单克隆抗体免疫沉淀了五种不同的中性粒细胞表面蛋白。数据表明，CD15单克隆抗体与LFA-1/HMac-1/gp 150,95糖蛋白家族的一个亚群以及人中性粒细胞上的CR1发生反应。所研究的CD15抗体对这些蛋白的亲和力不同。与所测试的CD15抗体免疫沉淀的分子相比，与其他直接与LFA-1/HMac-1/gp 150,95糖蛋白家族的αM（CD11）或β（CD18）链反应的单克隆抗体免疫沉淀的同源蛋白对蛋白酶解更具抗性。这些抗体识别的膜蛋白在抗体反应性和蛋白酶敏感性方面的一些差异可能归因于糖基化的差异。数据表明，所研究的抗体可以检测某些表面蛋白拷贝之间翻译后修饰的差异。
