通过两步柱色谱法从鼠腹水纯化IgM单克隆抗体。

Purification of IgM monoclonal antibody from murine ascitic fluid by a two-step column chromatography procedure.

作者信息

Johnson E, Miribel L, Arnaud P, Tsang K Y

出版信息

Immunol Lett. 1987 Jan;14(2):159-65. doi: 10.1016/0165-2478(87)90096-4.

Abstract

Two IgM monoclonal antibodies (Mabs) were purified from murine ascitic fluid by a two-step procedure involving pseudoligand affinity chromatography on immobilized Cibacron Blue F3GA and gel filtration chromatography on ACA-22. The purified IgM from one ascites sample, H-1, was determined to be homogeneous by sodium dodecyl sulfate-polyacrylamide gel electrophoresis while ascites containing E3.1 IgM Mab, was purified to greater than 95% IgM. The retention of biological activity was confirmed by immunofluorescence staining of peripheral blood lymphocytes.

摘要

通过两步法从鼠腹水原液中纯化出两种IgM单克隆抗体（Mabs），该两步法包括在固定化的汽巴蓝F3GA上进行假配体亲和色谱以及在ACA - 22上进行凝胶过滤色谱。通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳确定，来自一个腹水样本H - 1的纯化IgM是均一的，而含有E3.1 IgM Mab的腹水被纯化至IgM含量大于95%。通过外周血淋巴细胞的免疫荧光染色证实了生物活性的保留。