用于人类治疗的抗CD4单克隆抗体的纯化及分析表征

Purification and analytical characterization of an anti-CD4 monoclonal antibody for human therapy.

作者信息

Guse A H, Milton A D, Schulze-Koops H, Müller B, Roth E, Simmer B, Wächter H, Weiss E, Emmrich F

机构信息

Max-Planck-Gesellschaft, Klinische Arbeitsgruppe für Rheumatologie/Immunologie, Universität Erlangen-Nürnberg, Germany.

出版信息

J Chromatogr A. 1994 Feb 11;661(1-2):13-23. doi: 10.1016/0021-9673(94)85173-5.

DOI:10.1016/0021-9673(94)85173-5

PMID:8136902

Abstract

A purification process for the monoclonal anti-CD4 antibody MAX.16H5 was developed on an analytical scale using (NH4)2SO4 precipitation, anion-exchange chromatography on MonoQ or Q-Sepharose, hydrophobic interaction chromatography on phenyl-Sepharose and gel filtration chromatography on Superdex 200. The purification schedule was scaled up and gram amounts of MAX.16H5 were produced on corresponding BioPilot columns. Studies of the identity, purity and possible contamination by a broad range of methods showed that the product was highly purified and free from contaminants such as mouse DNA, viruses, pyrogens and irritants. Overall, the analytical data confirm that the monoclonal antibody MAX.16H5 prepared by this protocol is suitable for human therapy.

摘要

使用硫酸铵沉淀、在MonoQ或Q-Sepharose上进行阴离子交换色谱、在苯基琼脂糖上进行疏水相互作用色谱以及在Superdex 200上进行凝胶过滤色谱，在分析规模上开发了单克隆抗CD4抗体MAX.16H5的纯化工艺。将纯化方案放大，并在相应的BioPilot柱上生产了克级量的MAX.16H5。通过多种方法对其身份、纯度和可能的污染物进行的研究表明，该产品高度纯化，不含小鼠DNA、病毒、热原和刺激物等污染物。总体而言，分析数据证实，通过该方案制备的单克隆抗体MAX.16H5适用于人类治疗。