Diagnostic immunoassays for tick-borne encephalitis virus based on recombinant baculovirus protein expression.
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.
Marx
F
F
Department of Microbiology, Medical School, University of Innsbruck, Fritz-Pregl Strasse 3/II, A-6020, Innsbruck, Austria. florentine.marx@uibk.ac.at
Gritsun
T S
TS
Grubeck-Loebenstein
B
B
Gould
E A
EA
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000914:Antibodies, Viral / Q000097:blood; D015153:Blotting, Western; D003001:Cloning, Molecular; D004669:Encephalitis Viruses, Tick-Borne / Q000276:immunology / Q000302:isolation & purification*; D004675:Encephalitis, Tick-Borne / Q000175:diagnosis* / Q000821:virology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006801:Humans; D008875:Middle Aged; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012698:Serologic Tests; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology; D014765:Viral Vaccines / Q000008:administration & dosage
D000914:Antibodies, Viral; D010446:Peptide Fragments; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; D014765:Viral Vaccines; C065696:glycoprotein E, Flavivirus