Purification of a baculovirus-expressed hepatitis E virus structural protein and utility in an enzyme-linked immunosorbent assay.
We report on the purification of the full-length structural protein encoded by open reading frame 2 (ORF-2) of hepatitis E virus. The ORF-2 protein, expressed in Sf9 cells by using a recombinant baculovirus vector system, was successfully purified to homogeneity. Gel electrophoresis of the purified ORF-2 protein showed a single polypeptide of 75 kDa by Coomassie blue staining and by Western blot (immunoblot) analysis. We demonstrated that the partially purified ORF-2 protein could be used successfully in a sensitive and specific enzyme-linked immunosorbent assay for the detection of antibodies to hepatitis E virus.
He
J
J
Viral and Rickettsial Diseases Program, Naval Medical Research Institute, Bethesda, Maryland, USA.
Ching
W M
WM
Yarbough
P
P
Wang
H
H
Carl
M
M
D000818:Animals; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D015870:Gene Expression; D005822:Genetic Vectors; D006508:Hepatitis Antibodies / Q000097:blood; D016751:Hepatitis E / Q000175:diagnosis / Q000276:immunology; D016752:Hepatitis E virus / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D007075:Immunoglobulin M / Q000097:blood; D017924:Nucleopolyhedroviruses / Q000235:genetics; D016366:Open Reading Frames; D017510:Protein Folding; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D018411:Spodoptera; D015678:Viral Structural Proteins / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification
D006508:Hepatitis Antibodies; D007075:Immunoglobulin M; D011994:Recombinant Proteins; D015678:Viral Structural Proteins