Direct coating of culture medium from cells secreting classical swine fever virus E2 antigen on ELISA plates for detection of E2-specific antibodies

Ta-Chun Cheng; Chu-Hsiang Pan; Chien-Shu Chen; Kuo-Hsiang Chuang; Chih-Hung Chuang; Chien-Chaio Huang; Yu-Yi Chu; Ya-Chun Yang; Pei-Yu Chu; Chien-Han Kao; Yuan-Chin Hsieh; Tian-Lu Cheng

doi:10.1016/j.tvjl.2015.02.007

Direct coating of culture medium from cells secreting classical swine fever virus E2 antigen on ELISA plates for detection of E2-specific antibodies

Vet J. 2015 Jul;205(1):107-9. doi: 10.1016/j.tvjl.2015.02.007. Epub 2015 Feb 14.

Authors

Affiliations

¹ Graduate Institute of Pharmacognosy, Taipei Medical University, 252 Wu Hsing Street, Taipei 110, Taiwan.
² Division of Hog Cholera Research, Animal Health Research Institute, Council of Agriculture, 376 Chung-Cheng Road, New Taipei City 251, Taiwan.
³ School of Pharmacy, China Medical University, 91 Hsueh-Shih Road, Taichung 404, Taiwan.
⁴ Department of Biomedical and Environmental Biology, Kaohsiung Medical University, 100 Shih-Chuan 1st Road, Kaohsiung 807, Taiwan.
⁵ Institute of Bioinformatics and Biosignal Transduction, College of Bioscience and Biotechnology, National Cheng Kung University, 1 University Road, Tainan 701, Taiwan.
⁶ Graduate Institute of Medicine, Kaohsiung Medical University, 100 Shih-Chuan 1st Road, Kaohsiung 807, Taiwan.
⁷ Department of Biomedical and Environmental Biology, Kaohsiung Medical University, 100 Shih-Chuan 1st Road, Kaohsiung 807, Taiwan; Graduate Institute of Medicine, Kaohsiung Medical University, 100 Shih-Chuan 1st Road, Kaohsiung 807, Taiwan; Center for Biomarkers and Biotech Drugs, Kaohsiung Medical University, 100 Shih-Chuan 1st Road, Kaohsiung 807, Taiwan. Electronic address: tlcheng@kmu.edu.tw.

PMID: 25975854
DOI: 10.1016/j.tvjl.2015.02.007

Abstract

The envelope glycoprotein E2 of classical swine fever virus (CSFV) is widely used as a marker for measuring vaccine efficacy and antibody titer. The glycosylation profile of E2 may affect the immunogenicity of the vaccine and the timing of re-vaccination. In this study, a human embryonic kidney cell line was used to secrete fully-glycosylated CSFV E2, which was then coated onto ELISA plates without purification or adjustment. The resulting E2-secreting medium-direct-coating (E2-mDc) ELISA was successfully used to measure anti-E2 antibody titers in vaccinated and field pig sera samples. Compared with a virus neutralization test (as standard), the E2-mDc ELISA was found to be more accurate (90%) than a commercial CSFV antibody diagnostic kit (62%). In conclusion, the mammalian cell-secreted antigen can provide cheap, accurate and effective assays for vaccine efficacy and disease diagnoses.

Keywords: Classical swine fever virus; E2; Glycosylation; Mammalian expression; Medium-direct-coating ELISA.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antibodies, Viral / analysis*
Antibodies, Viral / immunology
Cell Line
Classical Swine Fever Virus / immunology*
Culture Media*
Enzyme-Linked Immunosorbent Assay / methods*
HEK293 Cells
Humans
Swine
Viral Envelope Proteins / immunology*
Viral Envelope Proteins / metabolism

Substances

Antibodies, Viral
Culture Media
Viral Envelope Proteins
glycoprotein E2, classical swine fever virus