Purification and diagnostic utility of a recombinant hepatitis E virus capsid protein expressed in insect larvae.
We report here the expression and purification of a truncated form of the hepatitis E virus ORF2 protein (ORF2delta111/deltaTM), from the fat bodies of Spodoptera litura larvae infected with a recombinant baculovirus. The purified protein migrated as a doublet of approximately 56 kDa on SDS-PAGE and was found to be glycosylated by staining with concanavalin A-linked horseradish peroxidase. The protein was used in a sensitive and specific enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to HEV. The results showed complete concordance with those obtained using a commercial kit for the detection of anti-HEV antibodies. Antigen expression in the insect larvae system presents a rapid and low-cost method that obviates the need for expensive tissue culture scale-ups or special equipment.
Sehgal
Deepak
D
Virology Group, International Center for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi 110 067, India. deepak@icgeb.res.in
Malik
Punjab Singh
PS
Jameel
Shahid
S
D000818:Animals; D016367:Baculoviridae; D036022:Capsid Proteins / Q000235:genetics / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay; D006031:Glycosylation; D016751:Hepatitis E / Q000175:diagnosis* / Q000276:immunology; D016752:Hepatitis E virus / Q000737:chemistry*; D006801:Humans; D007814:Larva / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification*; D018411:Spodoptera / Q000821:virology; D014764:Viral Proteins / Q000235:genetics / Q000302:isolation & purification*
D036022:Capsid Proteins; C400971:ORF2 protein, Hepatitis E virus; D011994:Recombinant Proteins; D014764:Viral Proteins