Enhanced expression of the Erns protein of classical swine fever virus in yeast and its application in an indirect enzyme-linked immunosorbent assay for antibody differentiation of infected from vaccinated animals
Enhanced expression of the Erns protein of classical swine fever virusin yeast and its application in an indirect enzyme-linkedimmunosorbent assay for antibody differentiation of infected fromvaccinated animals.pdf
[en] tClassical swine fever (CSF), caused by classical swine fever virus (CSFV), is a devastating disease of swineworldwide. Although a mandatory vaccination with the modified live vaccine C-strain has been imple-mented in China for decades, CSF remains a serious threat to the swine industry. To facilitate the controland eradication of CSF in China, the E2-based marker vaccine rAdV-SFV-E2, an adenovirus-delivered,alphavirus replicon-vectored vaccine, has been developed. Accordingly, an accompanying discriminatorytest that allows differentiating infected from vaccinated animals (DIVA) is required. Here, the enhancedexpression of Ernsprotein of CSFV was achieved in the methyltropic yeast Pichia pastoris by codon-optimization of the Ernsgene, and an indirect enzyme-linked immunosorbent assay (iELISA) based on theyeast-expressed Erns(yErns) was developed and evaluated. The optimized iELISA was able to detect CSFV-specific antibodies in the serum samples from the CSFV-infected pigs as early as 6 days post-infection,and discriminate the CSFV-infected pigs from those vaccinated with rAdV-SFV-E2. The iELISA was evalu-ated using a panel of swine sera, and showed comparable sensitivity (94.6%) and specificity (97.1%), andthe consistence rates with the virus neutralization test were 96.8% for CSFV-infected swine sera, 83.3%for C-strain-vaccinated swine sera, and 95.0% for field swine sera. In addition, the iELISA showed highersensitivity (90.4%) compared with PrioCHECK CSFV Erns(59.6%). Taken together, the yErns-based iELISA isspecific and sensitive, representing a promising DIVA test for E2-based marker vaccines against CSF.
Research Center/Unit :
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute (HVRI), Chinese Academy of Agricultural Sciences.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Luo, Yuzi ✱
Li, Lin ✱; Université de Liège - ULiège > Doct. sc. agro. & ingé. biol. (Paysage)
✱ These authors have contributed equally to this work.
Enhanced expression of the Erns protein of classical swine fever virus in yeast and its application in an indirect enzyme-linked immunosorbent assay for antibody differentiation of infected from vaccinated animals
Publication date :
21 May 2015
Journal title :
Journal of Virological Methods
ISSN :
0166-0934
eISSN :
1879-0984
Publisher :
Elsevier, Netherlands
Peer reviewed :
Peer Reviewed verified by ORBi
Name of the research project :
Development of Livestock Genetic Engineering Vaccine
Funders :
National 863Projects of China (No. 2011AA10A208) and the European Union’sSeventh Framework Programme LinkTADs (No. 613804).
Aebischer A., Müller M., Hofmann M.A. Two newly developed E(rns)-based ELISAs allow the differentiation of classical swine fever virus-infected from marker-vaccinated animals and the discrimination of pestivirus antibodies. Vet. Microbiol. 2013, 161:274-285.
Ahrens U., Kaden V., Drexler C., Visser N. Efficacy of the classical swine fever (CSF) marker vaccine Porcilis Pesti in pregnant sows. Vet. Microbiol. 2000, 77:83-97.
Cereghino J.L., Cregg J.M. Heterologous protein expression in the methylotrophic yeast Pichia pastoris. FEMS Microbiol. Rev. 2000, 24:45-66.
Cheng C.Y., Wu C.W., Lin G.J., Lee W.C., Chien M.S., Huang C. Enhancing expression of the classical swine fever virus glycoprotein E2 in yeast and its application to a blocking ELISA. J. Biotechnol. 2014, 174:1-6.
Clavijo A., Lin M., Riva J., Zhou E.M. Application of competitive enzyme-linked immunosorbent assay for the serologic diagnosis of classical swine fever virus infection. J. Vet. Diagn. Invest. 2001, 13:357-360.
Colijn E.O., Bloemraad M., Wensvoort G. An improved ELISA for the detection of serum antibodies directed against classical swine fever virus. Vet. Microbiol. 1997, 59:15-25.
Dewulf J., Laevens H., Koenen F., Mintiens K., de Kruif A. An E2 sub-unit marker vaccine does not prevent horizontal or vertical transmission of classical swine fever virus. Vaccine 2001, 20:86-91.
Edwards S., Fukusho A., Lefèvre P.C., Lipowski A., Pejsak Z., Roehe P., Westergaard J. Classical swine fever: the global situation. Vet. Microbiol. 2000, 73:103-119.
de Smit A.J. Evaluation of an ELISA for the detection of antibodies against Erns envelope protein of classical swine fever virus. Classical Swine Fever-Efficacy of Marker Vaccines and Laboratory Diagnosis 2000, 117-131. Faculteit Diergeneeskunde Thesis University, Utrecht.
Floegel-Niesmann G. Classical Swine Fever (CSF) marker vaccine: trial III. Evaluation of discriminatory ELISAs. Vet. Microbiol. 2001, 83:121-136.
Hohenblum H., Gasser B., Maurer M., Borth N., Mattanovich D. Effects of gene dosage, promoters, and substrates on unfolded protein stress of recombinant Pichia pastoris. Biotechnol. Bioeng. 2004, 85:367-375.
Huang C., Chien M.S., Hu C.M., Chen C.W., Hsieh P.C. Secreted expression of the classical swine fever virus glycoprotein E(rns) in yeast and application to a sandwich blocking ELISA. J. Virol. Methods 2006, 132:40-47.
König M., Lengsfeld T., Pauly T., Stark R., Thiel H.J. Classical swine fever virus: independent induction of protective immunity by two structural glycoproteins. J. Virol. 1995, 69:6479-6486.
Laemmli U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970, 227:680-685.
Li M., Wang Y.F., Wang Y., Gao H., Li N., Sun Y., Liang B.B., Qiu H.J. Immune responses induced by a BacMam virus expressing the E2 protein of classical swine fever virus in mice. Immunol. Lett. 2009, 125:145-150.
Luo Y., Li S., Sun Y., Qiu H.J. Classical swine fever in China: a minireview. Vet. Microbiol. 2014, 172:1-6.
Moormann R.J.M., Bouma A., Kramps J.A., Terpstra C., de Smit H.J. Development of a classical swine fever subunit marker vaccine and companion diagnostic test. Vet. Microbiol. 2000, 73:209-219.
Moser C., Ruggli N., Tratschin J.D., Hofmann M.A. Detection of antibodies against classical swine fever virus in swine sera by indirect ELISA using recombinant envelope glycoprotein E2. Vet. Microbiol. 1996, 51:41-53.
Outchkourov N.S., Stiekema W.J., Jongsma M.A. Optimization of the expression of equistatin in Pichia pastoris. Protein Expr. Purif. 2002, 24:18-24.
Qiu H.J., Shen R.X., Tong G.Z. The lapinized Chinese strain vaccine against classical swine fever virus: a retrospective review spanning half a century. Agric. Sci. China 2006, 5:1-14.
Reed L.J., Muench H. A simple method of estimating fifty percent endpoints. Am. J. Hyg. 1938, 27:493-497.
Reimann I., Depner K., Trapp S., Beer M. An avirulent chimeric Pestivirus with altered cell tropism protects pigs against lethal infection with classical swine fever virus. Virology 2004, 322:143-157.
Rümenapf T., Unger G., Strauss J.H., Thiel H.J. Processing of the envelope glycoproteins of pestiviruses. J. Virol. 1993, 67:3288-3294.
Schroeder S., von Rosen T., Blome S., Loeffen W., Haegeman A., Koenen F., Uttenthal A. Evaluation of classical swine fever virus antibody detection assays with an emphasis on the differentiation of infected from vaccinated animals. Rev. Sci. Tech. 2012, 31:997-1010.
Sinclair G., Choy F.Y. Synonymous codon usage bias and the expression of human glucocerebrosidase in the methylotrophic yeast, Pichia pastoris. Protein Expr. Purif. 2002, 26:96-105.
Sun Y., Li H.Y., Tian D.Y., Han Q.Y., Zhang X., Li N., Qiu H.J. A novel alphavirus replicon-vectored vaccine delivered by adenovirus induces sterile immunity against classical swine fever. Vaccine 2011, 29:8364-8372.
Sun Y., Tian D.Y., Li S., Meng Q.L., Zhao B.B., Li Y., Li D., Ling L.J., Liao Y.J., Qiu H.J. Comprehensive evaluation of the adenovirus/alphavirus-replicon chimeric vector-based vaccine rAdV-SFV-E2 against classical swine fever. Vaccine 2013, 31:538-544.
Tautz N., Elbers K., Stoll D., Meyers G., Thiel H.J. Serine protease of pestiviruses: determination of cleavage sites. J. Virol. 1997, 71:5415-5422.
van Gennip H.G., van Rijn P.A., Widjojoatmodjo M.N., de Smit A.J., Moormann R.J. Chimeric classical swine fever viruses containing envelope protein E(RNS) or E2 of bovine viral diarrhoea virus protect pigs against challenge with CSFV and induce a distinguishable antibody response. Vaccine 2000, 19:447-459.
