Evaluation of platelet surface antigens: localization of the PlA1 alloantigen
Evaluation of platelet surface antigens: localization of the PlA1 alloantigen
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血小板表面抗原的评估：PlA1 同种异体抗原的定位
DOI：
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发表时间：
1982
影响因子：
6.5
通讯作者：
G. M. Schmidt
中科院分区：
文献类型：
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作者：
Robert McMillan;D. Mason;P. Tani;G. M. Schmidt
摘要描述了一种用于将抗原决定簇定位于特定血小板蛋白的方法。将血小板溶解、溶解并转移到硝酸纤维素纸上，在那里它们被固定。与放射性标记的特异性抗体孵育后，通过放射自显影术定位抗原决定簇。可以用这种方式研究大小在50 000至540 000之间的纯化蛋白质，并且仍然保留它们的抗原性。将溶解的固定化血小板蛋白与已知具有抗PlA1抗体的两名输血后紫癜患者的血清IgG反应。放射性与表观分子量为100 000道尔顿的主要蛋白条带相关。然而，当使用从PlA1（+）血小板洗脱的纯化抗PlA1抗体时，观察到至少6个额外的次要条带，分子量范围约为86 000至175 000。使用PlA1阴性血小板或PlA1（+）血小板蛋白减少时未观察到放射性条带。来自一名患者的抗体与来自血栓性血小板的蛋白质的反应导致所有放射性条带的强度显著降低。此外，观察到一个新的高分子量条带，该条带在含有PlA1（+）血小板的反应中不存在。这些数据表明，PlA1抗原主要但不完全定位于糖蛋白IIIa。这种方法相对简单，只需要最少量的抗体，通常可用于研究细胞抗原和其他复杂的蛋白质混合物。
Summary. An approach is described for localizing antigenic determinants to specific platelet proteins. Platelets are solubilized, electrophoresed and transferred to nitrocellulose paper where they are immobilized. After incubation with radiolabelled specific antibody, the antigenic determinants are localized by radioautography. Purified proteins ranging in size from 50 000 to 540 000 could be studied in this manner and still retain their antigenicity. Solubilized immobilized platelet proteins were reacted with serum IgG from two patients with post‐transfusion purpura known to have anti‐PlA1 antibodies. The radioactivity was associated with a major protein band with an apparent molecular weight of 100 000 daltons. However, when purified anti‐PlA1 antibody eluted from PlA1 (+) platelets was used, at least six additional minor bands were noted with molecular weights ranging from about 86 000 to 175 000. No radioactive bands were noted using PlA1 negative platelets or reduced PlA1 (+) platelet proteins. Reaction of the antibody from one patient with proteins from thrombasthenic platelets resulted in a marked reduction in the intensity of all radioactive bands. In addition, a new high molecular weight band was noted which was not present in reactions containing PlA1 (+) platelets. These data suggest that the PlA1 antigen is primarily but not entirely localized to glycoprotein IIIa. This approach is relatively simple, requires only minimal amounts of antibody and should be generally useful in the study of cellular antigens and other complex mixtures of proteins.