抗牛脑胆碱乙酰转移酶的单克隆抗体及其免疫亲和纯化

Monoclonal antibodies to and immunoaffinity purification of choline acetyltransferase from bovine brain.

作者信息

Ishida I, Ichikawa T, Deguchi T

出版信息

J Neurochem. 1987 Sep;49(3):933-8. doi: 10.1111/j.1471-4159.1987.tb00983.x.

DOI:10.1111/j.1471-4159.1987.tb00983.x

Abstract

Three hybridomas producing monoclonal antibodies to bovine brain choline acetyltransferase (ChAT) have been established by fusion of the spleen cells from a mouse immunized with purified enzyme with myeloma NS-1 cells. All three clones produced IgGl antibodies that reacted with enzyme protein denatured with sodium dodecyl sulfate. By using one of the monoclonal antibodies, a rapid and efficient immunoaffinity purification procedure of bovine ChAT has been established. Immunoblot analysis and immunoaffinity purification indicated that bovine ChAT is a single 68-kilodalton protein. The monoclonal antibodies will offer us a good tool to isolate the cDNA clones encoding ChAT.

摘要

通过将用纯化酶免疫的小鼠脾细胞与骨髓瘤NS-1细胞融合，建立了三种产生抗牛脑胆碱乙酰转移酶（ChAT）单克隆抗体的杂交瘤。所有三个克隆都产生了IgG1抗体，这些抗体与用十二烷基硫酸钠变性的酶蛋白发生反应。通过使用其中一种单克隆抗体，建立了一种快速有效的牛ChAT免疫亲和纯化方法。免疫印迹分析和免疫亲和纯化表明，牛ChAT是一种单一的68千道尔顿蛋白。这些单克隆抗体将为我们提供一个分离编码ChAT的cDNA克隆的良好工具。