Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology-KAIST, Republic of Korea.
J Biotechnol. 2013 May 20;165(2):102-8. doi: 10.1016/j.jbiotec.2013.03.007. Epub 2013 Mar 22.
Because of the lack of post-translational glycosylation, Escherichia coli is not a preferable host for immunoglobulin G (IgG) production. However, recent successes in the developments of aglycosylated IgG variants that do not require glycosylation for effector functions have increased the likelihood of using E. coli for IgG production. Here, we have developed a new E. coli host-vector system for enhanced production of recombinant IgG using: (i) a combination of SRP/Sec-dependent pathways for the efficient secretion of heavy and light chains in the periplasm; (ii) co-expression of periplasmic foldase (DsbC) for efficient assembly of IgG in the periplasm; and (iii) co-expression of Ffh for enhancing the SRP machinery. Finally, with engineered host-vector system, fed-batch cultivations were conducted at four different conditions, and under an optimized condition, up to 62 mg/L of active full-length IgG was produced during a 28-h cultivation.
由于缺乏翻译后糖基化，大肠杆菌不是生产免疫球蛋白 G（IgG）的理想宿主。然而，最近在开发不需要糖基化即可发挥效应功能的去糖基化 IgG 变体方面取得的成功，增加了使用大肠杆菌生产 IgG 的可能性。在这里，我们开发了一种新的大肠杆菌宿主载体系统，用于使用：（i）SRP/Sec 依赖性途径的组合，以有效地将重链和轻链分泌到周质中；（ii）共表达周质折叠酶（DsbC），以有效地在周质中组装 IgG；和（iii）共表达 Ffh 以增强 SRP 机制。最后，在工程化的宿主载体系统下，在四种不同条件下进行了分批补料培养，在优化条件下，在 28 小时的培养过程中生产了高达 62mg/L 的活性全长 IgG。
