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J Virol Methods. 2001 Jan;91(1):75-84.

Diagnostic immunoassays for tick-borne encephalitis virus based on recombinant baculovirus protein expression.

Author information

1
Department of Microbiology, Medical School, University of Innsbruck, Fritz-Pregl Strasse 3/II, A-6020, Innsbruck, Austria. florentine.marx@uibk.ac.at

Abstract

The baculovirus expression system that utilizes Autographa californica nuclear polyhedrosis virus was used to express the highly antigenic envelope protein E of a tick-borne encephalitis (TBE) complex virus, as well as a C-terminally truncated form of protein E (Etr). The recombinant proteins were produced with a histidine-tag at their carboxy-terminus. Protein purification by nickel agarose chromatography resulted in high concentrations of pure Etr protein, but only poor yields of E protein. Therefore, Etr was used to develop a sensitive and specific enzyme-linked immunosorbent assay (ELISA), as well as an immunoblot assay to detect TBE virus-specific antibodies in sera from immunized human blood donors. Sera from non-vaccinated blood donors were used as controls. The data show that the recombinant TBE virus-specific Etr protein exhibits the antigenic epitopes and conformation necessary for specific antigen-antibody recognition. Thus, the baculovirus expression system provides a cheap and easy method to generate recombinant viral antigens for TBE virus-specific serodiagnosis.

PMID:
11164488
DOI:
10.1016/s0166-0934(00)00251-2
[Indexed for MEDLINE]

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