cGMP 级 NY-ESO-1 的表达和纯化用于临床试验。

Expression and purification of cGMP grade NY-ESO-1 for clinical trials.

机构信息

Graduate Field of Microbiology, Cornell University, Ithaca, NY 14853, USA.

出版信息

Biotechnol Prog. 2011 Mar-Apr;27(2):435-41. doi: 10.1002/btpr.552. Epub 2011 Mar 1.

Abstract

NY-ESO-1 is a cancer testis antigen expressed in numerous cancers. Initial tests have shown its efficacy as a cancer vaccine, stimulating the body's own immune response against the invading tumor. To produce enough material for phase I clinical trials, a process using current good manufacturing practices to produce clinical grade material was developed and executed. His-tagged NY-ESO-1 was expressed in C41DE3 Escherichia coli under control of the T-7 promoter. NY-ESO-1 was produced in a 20 L fed-batch fermentation utilizing a pH-stat control scheme. The protein was then purified from inclusion bodies using a three-column process that achieved a yield of over 3.4 g and endotoxin below the detection limit of 0.005 EU/μg protein.

摘要

NY-ESO-1 是一种在多种癌症中表达的癌症睾丸抗原。初步测试表明，它作为癌症疫苗是有效的，能刺激机体自身的免疫反应来对抗入侵的肿瘤。为了生产足够用于 I 期临床试验的材料，开发并执行了使用现行良好生产规范生产临床级材料的过程。His 标记的 NY-ESO-1 在 C41DE3 大肠杆菌中受 T-7 启动子的控制下表达。利用 pH -stat 控制方案在 20L 分批补料发酵中生产 NY-ESO-1。然后，该蛋白从包涵体中通过三柱工艺进行纯化，产率超过 3.4g，内毒素低于 0.005EU/μg 蛋白的检测限。