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Preparation of recombinant Epstein-Barr virus viral capsid antigen BALF4 protein and its application in nasopharyngeal carcinoma diagnosis
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Abstract:
Objective : To prepare recombinant BALF4 poptein of VCA (viral capsid antigen) in Epstein-Barr virus and analyze its application in detection of nasopharyngeal carcinoma (NPC). Methods: BALF4 gene was amplified from EBV DNA sequence by PCR and then inserted into pPICZαA pichia pastoris expression vector. Recombinant pPICZαA-BALF4 plasmid was transfected into GS115 yeast cells, and corresponding recombinant BALF4 protein was induced by methanol. The recombinant BALF4 protein was identified by SDS-PAGE and Western blotting analysis, and used as coating antigen for detection of EBV-IgA antibody in NPC patients by ELISA. Results: pPICZαA-BALF4 expression vector was succesffuly constructed, and recombinant BALF4 protein was effectively expressed GS115 yeast cells. The molecular weight of recombinant BALF4 protein is approximately 52 000, and the recombinant BALF4 protein showed good immunoreactivity as detected by Western blotting analysis. Recombinant BALF4 protein was used as coating antigen in detecting EBV-IgA antibody levels in serum samples collected from NPC patients and healthy controls, and the sensitivity and specificity of VCA-BALF4 in the test were 81% and 94%, respectively. Conclusion: The recombinant BALF4 protein has been succesffuly prepared and it has satisfying sensitivity and specificity in serum screening of NPC patients.
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Project Supported:
Project supported by the Special Scientific Foundation for Outstanding Young Teachers in Shanghai Higher Education Institutions(No.37-0111-08-003), and the Innovation Foundation of Shanghai University(No. 11-0111-09-08)
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