Quackenbush E J, Cruz T F, Moscarello M A, Letarte M
Biochem J. 1985 Jan 15;225(2):291-9. doi: 10.1042/bj2250291.
Monoclonal antibodies prepared against a non-T and non-B acute-lymphocytic-leukaemia cell line were tested for reactivity against human brain tissue. Several of the monoclonal antibodies were found to react specifically with brain fractions. Three antigens, 44H4, 44D7 and 44D10, were identified in white matter. Although 44D10 was absent from grey matter, the levels of 44H4 and 44D7 antigens present in grey matter were 2- and 4-fold higher respectively than in white matter. Fractionation of white matter indicated that all three antigens were absent from the multilamellar compact myelin, but associated with a membrane fraction of higher density. All three antigens, which required detergent for solubilization from the membranes, were purified by affinity to monoclonal antibodies and/or were analysed by immunoblotting. The 44H4 and 44D10 antigens were single polypeptide chains with Mr 94000 and 80000 respectively when resolved by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. Monoclonal antibody 44D7 reacted with a complex of a Mr greater than 120000 under non-reducing conditions in the presence of sodium dodecyl sulphate. This complex dissociated on reduction into four bands with Mr values of 80000, 57000, 47000 and 41000. The brain antigens are present on proteins similar to, or identical with, those isolated from acute-lymphocytic-leukaemia cells.
针对一种非T和非B急性淋巴细胞白血病细胞系制备的单克隆抗体，检测其与人脑组织的反应性。发现几种单克隆抗体能与脑部分馏物特异性反应。在白质中鉴定出三种抗原，即44H4、44D7和44D10。虽然灰质中不存在44D10，但灰质中存在的44H4和44D7抗原水平分别比白质高2倍和4倍。白质分级分离表明，所有三种抗原均不存在于多层紧密髓鞘中，而是与较高密度的膜部分相关。所有三种抗原都需要去污剂从膜上溶解下来，通过与单克隆抗体的亲和力进行纯化和/或通过免疫印迹分析。当通过十二烷基硫酸钠/聚丙烯酰胺凝胶电泳分离时，44H4和44D10抗原分别是分子量为94000和80000的单条多肽链。在十二烷基硫酸钠存在下的非还原条件下，单克隆抗体44D7与分子量大于120000的复合物反应。该复合物在还原时解离成四条带，分子量分别为80000、57000、47000和41000。脑抗原存在于与从急性淋巴细胞白血病细胞中分离出的蛋白质相似或相同的蛋白质上。
