OBJECTIVE: To investigate the immunogenicity of different regions of glycoproteins in fish infectious hematopoietic necrosis virus (IHNV). METHODS: The glycoproteins was truncated and expressed according to the prediction of antigenicity and hydrophobicity by DNAStar 6.0 software. The protein was purified and used to produce antisera in rabbits. ELISA was performed to determine the antiserum titer and the immunogenicity of the protein was tested by indirect immunofluorescence assay (IFA). RESULTS: SDS-PAGE showed that protein with expected size 40 kDa was obtained. Purity of the obtained protein was as high as 90% as confirmed by high performance liquid chromatography (HPLC) analysis. The antisera against glycoprotein prepared in this study could react specifically with both natural glycoprotein of the IHNV-Sn1203 and the recombinant truncated glycoprotein in ELISA test, and the antisera titer against the natural glycoprotein and the recombinant glycoprotein was 1:20 000 and 1:80 000, respectively. IFA result showed that the antisera could recognize the glycoprotein on the surface of IHNV-Sn1203 and reference strain IHNV-WRAC. CONCLUSION: The antigenicity of the truncated glycoprotein produced in the study was as good as that of natural glycoprotein of IHNV.
OBJECTIVE: To investigate the immunogenicity of different regions of glycoproteins in fish infectious hematopoietic necrosis virus (IHNV). METHODS: The glycoproteins was truncated and expressed according to the prediction of antigenicity and hydrophobicity by DNAStar 6.0 software. The protein was purified and used to produce antisera in rabbits. ELISA was performed to determine the antiserum titer and the immunogenicity of the protein was tested by indirect immunofluorescence assay (IFA). RESULTS:SDS-PAGE showed that protein with expected size 40 kDa was obtained. Purity of the obtained protein was as high as 90% as confirmed by high performance liquid chromatography (HPLC) analysis. The antisera against glycoprotein prepared in this study could react specifically with both natural glycoprotein of the IHNV-Sn1203 and the recombinant truncated glycoprotein in ELISA test, and the antisera titer against the natural glycoprotein and the recombinant glycoprotein was 1:20 000 and 1:80 000, respectively. IFA result showed that the antisera could recognize the glycoprotein on the surface of IHNV-Sn1203 and reference strain IHNV-WRAC. CONCLUSION: The antigenicity of the truncated glycoprotein produced in the study was as good as that of natural glycoprotein of IHNV.