Production of a soluble and active MBP‐scFv fusion: favorable effect of the leaky tolR strain

FEBS Letters

Baty, Daniel¹  Fieschi, Jacques¹  Chames, Patrick¹ 

[1] Institut de Biologie Structurale et de Microbiologie, Laboratoire d'Ingénierie et Dynamique des Systèmes Membranaires, CNRS-UPR 9027, 31 Chemin Joseph Aiguier, 13402 Marseille Cedex 20, France

关键词: Secretion;    Aggregation;    Inclusion body;    Antibody;    Anti-cortisol;    Steroid;    ABTS;    2;    2′-azino-bis[3-ethylbenzthiazoline sulfonate];    ELISA;    enzyme-linked immunosorbant assay;    Fv;    immunoglobulin-variable fragment;    scFv;    single-chain Fv;    IPTG;    isopropyl-β-d-thiogalactoside;    MBP;    maltose-binding protein;    MBP*;    MBP part of the degraded MBP-scFv fusion;   

DOI  :  10.1016/S0014-5793(97)00194-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The 6D6 anti-cortisol scFv was prepared as fusion protein with maltose-binding protein (MBP) to increase the amount of soluble product. This fusion was almost completely insoluble when produced in a wild-type strain of Escherichia coli. However, when MBP-scFv fusion was produced in a tolR leaky strain, it was secreted into the culture medium as an active, soluble protein. Production of recombinant proteins in the tolR strain greatly enhances the recovery of active protein and may be a useful system to produce MBP fusion proteins that would normally aggregate when produced in wild-type bacterial strains. © 1997 Federation of European Biochemical Societies

【 授权许可】

Unknown