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Enhancing expression of the classical swine fever virus glycoprotein E2 in yeast and its application to a blocking ELISA

Paper ID Volume ID Publish Year Pages File Format Full-Text
23273 43429 2014 6 PDF Available
Title
Enhancing expression of the classical swine fever virus glycoprotein E2 in yeast and its application to a blocking ELISA
Abstract

•Production of yeast-expressed CSFV E2 (yE2) is enhanced by optimizing the first codon.•N-terminal 190 residues of E2 are essential for an efficient neutralizing antibody response.•The blocking ELISA using yE2 and the anti-E2 MAb, 1B6, performed well when detecting antibody in infected pig sera.

Classical swine fever virus (CSFV) infection is a severe swine disease, often causing large economic losses. A Pichia pastoris yeast-expressed CSFV glycoprotein E2 (yE2) has been shown to induce a protective immune response against the virus. To improve the expression level of yE2, the first codon of E2 gene, Arg (CGG), which is the least used in P. pastoris, was optimized to the most favorite codon AGA. The yield of E2 protein was remarkably increased in the codon optimized strain (N342). Three truncated E2 subunits encoding the N-terminal 330 (N330), 301 (N301), and 190 (N190) residues, respectively, were also constructed. The immunogenicity of each recombinant E2 subunits was confirmed by immunization of pigs, and all immunized groups demonstrated high neutralizing antibody titers after boost immunization, which lasted for a long period of time. In addition, a monoclonal antibody (MAb), 1B6, specific to yE2, was generated and shown to recognize CSFV-infected cells. A panel of swine sera were tested by peroxidase-conjugated MAb 1B6-based blocking enzyme-linked immunosorbent assay (ELISA) using N330 as coated antigen, and the assay demonstrated high sensitivity and specificity. The recombinant yE2 subunits may provide potential subunit vaccine candidates and useful diagnostic reagents for CSFV with easy manipulation and low cost.

Keywords
Classical swine fever virus; Subunit marker vaccine; Codon optimization; Monoclonal antibody; Blocking ELISA
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 174, 20 March 2014, Pages 1–6
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