標題: Improving Antigenicity of the Recombinant Hepatitis C Virus Core Protein via Random Mutagenesis
作者: Huang, Chen-Ji
Peng, Hwei-Ling
Cheng, Chih-Yu
生物科技學系
Department of Biological Science and Technology
公開日期: 2011
摘要: In order to enhance the sensitivity of diagnosis, a recombinant clone containing domain I of HCV core (amino acid residues 1 to 123) was subjected to random mutagenesis. Five mutants with higher sensitivity were obtained by colony screening of 616 mutants using reverse ELISA. Sequence analysis of these mutants revealed alterations focusing on W(84), P(95), P(110), or V(129). The inclusion bodies of these recombinant proteins overexpressed in E. coli BL21(DE3) were subsequently dissolved using 6 M urea and then refolded by stepwise dialysis. Compared to the unfolded wild-type antigen, the refolded M3b antigen (W(84)S, P(110)S and V(129)L) exhibited an increase of 66% antigenicity with binding capacity of 0.96 and affinity of 113 mu M(-1). Moreover, the 33% decrease of the production demand suggests that M3b is a potential substitute for anti-HCV antibody detection.
URI: http://hdl.handle.net/11536/14820
http://dx.doi.org/10.1155/2011/359042
ISSN: 1110-7243
DOI: 10.1155/2011/359042
期刊: JOURNAL OF BIOMEDICINE AND BIOTECHNOLOGY
顯示於類別:期刊論文


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