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Expression of Bovine Viral Diarrhea Virus Envelope Glycoprotein E2 in Yeast Pichia pastoris and its Application to an ELISA for Detection of BVDV Neutralizing Antibodies in Cattle.
- Source :
- Journal of Immunoassay & Immunochemistry; 2015, Vol. 36 Issue 6, p639-654, 16p
- Publication Year :
- 2015
-
Abstract
- The aim of this article is to express envelope glycoprotein E2 of bovine viral diarrhea virus (BVDV) in yeastPichia pastorisand its utility as a diagnostic antigen in ELISA. The BVDV E2 gene was cloned into the pPICZαA vector followed by integration into thePichia pastorisstrain X-33 genome for methanol-induced expression. SDS-PAGE and Western blot results showed that the recombinant BVDV E2 protein (72 kDa) was expressed and secreted into the medium at a concentration of 40 mg/L of culture under optimized conditions. An indirect ELISA was then developed by using the yeast-expressed E2 protein. Preliminary testing of 300 field cattle serum samples showed that the E2 ELISA showed a sensitivity of 91.07% and a specificity of 92.02% compared to the reference virus neutralization test. The concordance between the E2 ELISA and VNT was 91.67%. This study demonstrates feasibility of BVDV E2 protein expression in yeastPichia pastorisfor the first time and its efficacy as an antigen in ELISA for detecting BVDV neutralizing antibodies in cattle. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 15321819
- Volume :
- 36
- Issue :
- 6
- Database :
- Complementary Index
- Journal :
- Journal of Immunoassay & Immunochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 103144869
- Full Text :
- https://doi.org/10.1080/15321819.2015.1032305