title: Direct coating of culture medium from cells secreting classical swine fever virus E2 antigen on ELISA plates for detection of E2-specific antibodies.
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.
authors.affiliation: Graduate Institute of Pharmacognosy, Taipei Medical University, 252 Wu Hsing Street, Taipei 110, Taiwan.
authors.forename: Ta-Chun
authors.initials: TC
authors.lastname: Cheng
authors.affiliation: Division of Hog Cholera Research, Animal Health Research Institute, Council of Agriculture, 376 Chung-Cheng Road, New Taipei City 251, Taiwan.
authors.forename: Chu-Hsiang
authors.initials: CH
authors.lastname: Pan
authors.affiliation: School of Pharmacy, China Medical University, 91 Hsueh-Shih Road, Taichung 404, Taiwan.
authors.forename: Chien-Shu
authors.initials: CS
authors.lastname: Chen
authors.affiliation: Graduate Institute of Pharmacognosy, Taipei Medical University, 252 Wu Hsing Street, Taipei 110, Taiwan.
authors.forename: Kuo-Hsiang
authors.initials: KH
authors.lastname: Chuang
authors.affiliation: Department of Biomedical and Environmental Biology, Kaohsiung Medical University, 100 Shih-Chuan 1st Road, Kaohsiung 807, Taiwan.
authors.forename: Chih-Hung
authors.initials: CH
authors.lastname: Chuang
authors.affiliation: Department of Biomedical and Environmental Biology, Kaohsiung Medical University, 100 Shih-Chuan 1st Road, Kaohsiung 807, Taiwan.
authors.forename: Chien-Chaio
authors.initials: CC
authors.lastname: Huang
authors.affiliation: Institute of Bioinformatics and Biosignal Transduction, College of Bioscience and Biotechnology, National Cheng Kung University, 1 University Road, Tainan 701, Taiwan.
authors.forename: Yu-Yi
authors.initials: YY
authors.lastname: Chu
authors.affiliation: Department of Biomedical and Environmental Biology, Kaohsiung Medical University, 100 Shih-Chuan 1st Road, Kaohsiung 807, Taiwan.
authors.forename: Ya-Chun
authors.initials: YC
authors.lastname: Yang
authors.affiliation: Department of Biomedical and Environmental Biology, Kaohsiung Medical University, 100 Shih-Chuan 1st Road, Kaohsiung 807, Taiwan.
authors.forename: Pei-Yu
authors.initials: PY
authors.lastname: Chu
authors.affiliation: Graduate Institute of Medicine, Kaohsiung Medical University, 100 Shih-Chuan 1st Road, Kaohsiung 807, Taiwan.
authors.forename: Chien-Han
authors.initials: CH
authors.lastname: Kao
authors.affiliation: Graduate Institute of Medicine, Kaohsiung Medical University, 100 Shih-Chuan 1st Road, Kaohsiung 807, Taiwan.
authors.forename: Yuan-Chin
authors.initials: YC
authors.lastname: Hsieh
authors.affiliation: 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.
authors.forename: Tian-Lu
authors.initials: TL
authors.lastname: Cheng
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis* / Q000276:immunology; D002460:Cell Line; D006692:Classical Swine Fever Virus / Q000276:immunology*; D003470:Culture Media; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D013552:Swine; D014759:Viral Envelope Proteins / Q000276:immunology* / Q000378:metabolism
chemical_list: D000914:Antibodies, Viral; D003470:Culture Media; D014759:Viral Envelope Proteins; C087086:glycoprotein E2, classical swine fever virus
keywords: Classical swine fever virus; E2; Glycosylation; Mammalian expression; Medium-direct-coating ELISA