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Thermostable tag (TST) protein expression system: Engineering thermotolerant recombinant proteins and vaccines

Paper ID Volume ID Publish Year Pages File Format Full-Text
23882 43479 2011 9 PDF Available
Title
Thermostable tag (TST) protein expression system: Engineering thermotolerant recombinant proteins and vaccines
Abstract

Methods to increase temperature stability of vaccines and adjuvants are needed to reduce dependence on cold chain storage. We report herein creation and application of pVEX expression vectors to improve vaccine and adjuvant manufacture and thermostability. Defined media fermentation yields of 6 g/L thermostable toll-like receptor 5 agonist flagellin were obtained using an IPTG inducible pVEX-flagellin expression vector. Alternative pVEX vectors encoding Pyrococcus furiosus maltodextrin-binding protein (pfMBP) as a fusion partner improved Influenza hemagglutinin antigen vaccine solubility and thermostability. A pfMBP hemagglutinin HA2 domain fusion protein was a potent immunogen. Manufacturing processes that combined up to 5 g/L defined media fermentation yields with rapid, selective, thermostable pfMBP fusion protein purification were developed. The pVEX pfMBP-based thermostable tag (TST) platform is a generic protein engineering approach to enable high yield manufacture of thermostable recombinant protein vaccine components.

Keywords
A450, absorbance at 450 nm; A600, absorbance at 600 nm; EU, endotoxin units; HA, hemagglutinin; IMAC, immobilized metal affinity chromatography; IN, intranasal; MBP, maltose or maltodextrin-binding protein; pfMBP, Pyrococcus furiosus maltodextrin-binding
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Thermostable tag (TST) protein expression system: Engineering thermotolerant recombinant proteins and vaccines
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 151, Issue 3, 10 February 2011, Pages 242–250
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