通过离子交换和嗜硫吸附色谱法纯化针对Le(y)和Le(d)碳水化合物抗原的单克隆抗体。

Purification of monoclonal antibodies to Le(y) and Le(d) carbohydrate antigens by ion-exchange and thiophilic-adsorption chromatography.

作者信息

Rapoport E M, Zhigis L S, Vlasova E V, Piskarev V E, Bovin N V, Zubov V P

机构信息

Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, GSP.

出版信息

Bioseparation. 1995 Jun;5(3):141-6.

PMID:7655191

Abstract

The paper deals with convenient and fast methods for purification of monoclonal antibodies (MAbs) to carbohydrate antigens Le(y) and Le(d) from the cell culture and ascite fluid by ion-exchange chromatography on S-Sepharose and salt-promoted chromatography on a "thiophilic" adsorbent. One-step procedure on S-Sepharose of MAbs to Le(y) (IgG and IgM) provides significant purification (over 90% of contaminants were removed), while a purification factor for IgM to Le(d) is much lower. Highly purified IgM to Le(d) could be obtained by a two-step purification procedure including "thiophilic-adsorption" chromatography and gel-filtration (90-98% of contaminants from the cell culture and ascite fluid were removed). The preparations of IgG and IgM retain their initial antibody activity.

摘要

本文探讨了通过在S-Sepharose上进行离子交换色谱以及在“嗜硫”吸附剂上进行盐促进色谱，从细胞培养物和腹水液中纯化针对碳水化合物抗原Le(y)和Le(d)的单克隆抗体（MAb）的便捷快速方法。在S-Sepharose上对Le(y)的单克隆抗体（IgG和IgM）进行一步法可实现显著纯化（去除超过90%的污染物），而针对Le(d)的IgM的纯化因子则低得多。通过包括“嗜硫吸附”色谱和凝胶过滤的两步纯化程序可获得高度纯化的针对Le(d)的IgM（去除细胞培养物和腹水液中90 - 98%的污染物）。IgG和IgM制剂保留了其初始抗体活性。