Molecular chaperone effects on recombinant yield and binding characteristics of an ABA-specific scFv in <i>Escherichia coli</i>.

ABSTRACT:

Introduction

Recombinant single-chain variable fragments (scFvs) are promising antibody formats for cost-effective and scalable production. However, their soluble expression in Escherichia coli is often limited by misfolding and aggregation, particularly for scFvs targeting small molecule haptens such as abscisic acid (ABA). To address this bottleneck, molecular chaperones can be co-expressed to enhance folding efficiency and functional yield.

Methods

An ABA-specific scFv was expressed in E. coli BL21(DE3) using five different chaperone plasmids (pG-KJE8, pGro7, pKJE7, pG-Tf2, and pTf16). Soluble expression was quantified by His-tag ELISA, and protein identity was confirmed by SDS-PAGE and Western blot. Functional characterization included competitive ELISA for IC₅₀

SUBMITTER: Chen S 

PROVIDER: S-EPMC12547695 | biostudies-literature | 2025

REPOSITORIES: biostudies-literature