摘要
本文采用两种方法纯化抗体。方法一,经DE-52阴离子层析纯化获IgG;方法二,采用Sepharose-4B-Protein A亲和层析抗体,用纯化的McAb 2H_6-E_3与蓖麻毒蛋白A链偶连形成免疫毒素。体外将免疫毒素加入培养的杜氏利什曼原虫前鞭毛体内培养,经计数测定,可见免疫毒素对前鞭毛体有一定程度的抑制杀伤作用。
McAb 2H6-E3, directed against surface membrane antigen from Leishmania donovani promastigotes, was purified on a sepharose-4B-protein A affinity column. The McAb conjugated SPDP with the molar ratio of McAb to SPDP being 1 : 6 to produce PDP-McAb. The PDP-McAb was purified by sephadex G-25. The PDP-McAb conjugated to RTA with the molar ratio of McAb to RTA being 1:3. The reaction lasted for 36-48 hours at the temperature of 12℃. The McAb-RTA conjugation, which may be called immunotoxin, was purified by sephadex G-150 and detected by SDS-PAGE. Promastigotes of Leishmania donovani Sichuan human isolate were grown at 26℃ in 199 medium supplemented with immunotoxin (100μg/ml), or RTA(100μg/ml), or McAb(100μg/ml). The promastigotes were observed and counted after 1.2.3.4.5 days of cultivation. The results revealed that the ricin A chain-immunotoxin can, to some extent, inhibit the growth of promastigotes.
出处
《地方病通报》
1991年第4期19-22,共4页
Endemic Diseases Bulletin
基金
国家自然科学基金资助课题
关键词
单克隆抗体
免疫毒素
利什曼原虫
Leishmania donovani
promastigote
McAb
Immunotoxin
Ricin
