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Comprehensive engineering of Escherichia coli for enhanced expression of IgG antibodies

Paper ID Volume ID Publish Year Pages File Format Full-Text
31753 44835 2011 11 PDF Available
Title
Comprehensive engineering of Escherichia coli for enhanced expression of IgG antibodies
Abstract

The expression of IgG antibodies in Escherichia coli is of increasing interest for analytical and therapeutic applications. In this work, we describe a comprehensive and systematic approach to the development of a dicistronic expression system for enhanced IgG expression in E. coli encompassing: (i) random mutagenesis and high-throughput screening for the isolation of over-expressing strains using flow cytometry and (ii) optimization of translation initiation via the screening of libraries of synonymous codons in the 5′ region of the second cistron (heavy chain). The effects of different promoters and co-expression of molecular chaperones on full-length IgG production were also investigated. The optimized system resulted in reliable expression of fully assembled IgG at yields between 1 and 4 mg/L of shake flask culture for different antibodies.

Keywords
Escherichia coli; Strain optimization; IgG expression; Chemical mutagenesis; Fluorescence-activated cell sorting (FACS); Translational initiation region (TIR)IgG, immunoglobulin G; TIR, translational initiation region; PECS, periplasmic expression followe
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Publisher
Database: Elsevier - ScienceDirect
Journal: Metabolic Engineering - Volume 13, Issue 2, March 2011, Pages 241–251
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
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Price was $35.95
You save - $31
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