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Journal of Biomedicine and Biotechnology
Volume 2011 (2011), Article ID 359042, 6 pages
Research Article

Improving Antigenicity of the Recombinant Hepatitis C Virus Core Protein via Random Mutagenesis

1Department of Biological Science and Technology, National Chiao Tung University, Hsinchu 30010, Taiwan
2Department of Marine Biotechnology, National Kaohsiung Marine University, Kaohsiung 81143, Taiwan

Received 14 March 2011; Accepted 10 May 2011

Academic Editor: Shahid Jameel

Copyright © 2011 Chen-Ji Huang et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


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 W84, P95, P110, or V129. 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 (W84S, P110S and V129L) exhibited an increase of 66% antigenicity with binding capacity of 0.96 and affinity of 113 μM−1. Moreover, the 33% decrease of the production demand suggests that M3b is a potential substitute for anti-HCV antibody detection.