Purification and application of bacterially expressed chimeric protein E1E2 of hepatitis C virus.
E1 and E2 glycoproteins are structural components of hepatitis C virus (HCV) virion. They are involved in cellular receptors interaction, neutralising antibodies elicitation, and viral morphogenesis. They are considered as major candidates for anti-HCV vaccine. In this report, we first expressed tandem E1E2 as well as C-terminally truncated E1 fragment and C-terminally truncated E2 fragment, respectively, in Escherichia coli cells and the proteins were purified to homogenesis. All the purified proteins can react specifically with patient sera. Both purified chimeric protein E1E2 and protein E2 can interact with a putative cellular receptor CD81, while purified protein E1 cannot interact with CD81. The sera of rabbit immunized with the E1E2 inhibited the binding of E2 protein to the major extracellular loop of human CD81 and reacted with both proteins E1 and E2, respectively. Anti-E1 and E2 antibodies can be generated simultaneously in the rabbit immunized with the E1E2, and the titers of antibodies were 63 or 56% higher than the titers induced by E1 or E2 alone, respectively. The results suggest that E1 and E2 can enhance their immunogenicity each other in chimeric protein E1E2 and the E. coli-derived chimeric protein E1E2 and corresponding antisera can be used as an useful tools in anti-HCV vaccine research.
Xiang
Zhong-Hua
ZH
State Key Laboratory of Virology, College of Life Science, Wuhan University, Wuhan, Hubei Province 430072, PR China.
Cai
Wei-Jia
WJ
Zhao
Peng
P
Kong
Ling-Bao
LB
Ye
Lin-Bai
LB
Wu
Zheng-Hui
ZH
D000906:Antibodies / Q000276:immunology; D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D016174:Hepacivirus / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D060225:Tetraspanin 28; D014764:Viral Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*
D000906:Antibodies; D015703:Antigens, CD; C554798:CD81 protein, human; D011994:Recombinant Proteins; D060225:Tetraspanin 28; D014764:Viral Proteins