在大肠杆菌中生产抗体片段。

Production of antibody fragments in Escherichia coli.

作者信息

Katsuda Tomohisa, Sonoda Hiroyuki, Kumada Yoichi, Yamaji Hideki

机构信息

Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, Kobe, Japan.

出版信息

Methods Mol Biol. 2012;907:305-24. doi: 10.1007/978-1-61779-974-7_18.

DOI:10.1007/978-1-61779-974-7_18

PMID:22907360

Abstract

Escherichia coli is a host widely used in the industrial production of recombinant proteins. However, the expression of heterologous proteins in E. coli often encounters the formation of inclusion bodies, which are insoluble and nonfunctional protein aggregates. For the successful production of antibody fragments, which includes single-chain variable fragments (scFvs), we describe here the modification of linker, signal, and Shine-Dalgarno (SD) sequences, the coexpression of cytoplasmic and periplasmic chaperones, and a method for fed-batch cultivation with exponential feed.

摘要

大肠杆菌是重组蛋白工业生产中广泛使用的宿主。然而，在大肠杆菌中表达异源蛋白常常会遇到包涵体的形成，包涵体是不溶性且无功能的蛋白质聚集体。为了成功生产包括单链可变片段（scFv）在内的抗体片段，我们在此描述了接头、信号和夏因-达尔加诺（SD）序列的修饰、细胞质和周质伴侣蛋白的共表达以及指数补料分批培养的方法。