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Enhanced production of full-length immunoglobulin G via the signal recognition particle (SRP)-dependent pathway in Escherichia coli

Paper ID Volume ID Publish Year Pages File Format Full-Text
23522 43446 2013 7 PDF Available
Title
Enhanced production of full-length immunoglobulin G via the signal recognition particle (SRP)-dependent pathway in Escherichia coli
Abstract

•Full-length IgG were produced in E. coli host.•By combining SRP/Sec pathways, heavy and light chains could be produced much more efficiently.•By co-expression of DsbC and Ffh, production yield of IgG could be significantly improved.•By fed-batch cultivations, E. coli cells produce 62 mg/L of full-length IgG.

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.

Keywords
Escherichia coli; Immunoglobulin G; SRP pathway; Fed-batch cultivation
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Enhanced production of full-length immunoglobulin G via the signal recognition particle (SRP)-dependent pathway in Escherichia coli
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 165, Issue 2, 20 May 2013, Pages 102–108
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us