A novel recombinant human interferon alpha2b with high antivirus activity by combinatorial mutagenesis.
- Author:
Guangrong ZHAO
1
;
Lei DU
;
Linqi ZHU
Author Information
1. Key Laboratory of Systems Bioengineering, Ministry of Education, Tianjin University, Tianjin 300072, China. grzhao@tju.edu.cn
- Publication Type:Journal Article
- MeSH:
Antiviral Agents;
pharmacology;
Base Sequence;
Combinatorial Chemistry Techniques;
Humans;
Interferon-alpha;
genetics;
pharmacology;
Molecular Sequence Data;
Mutagenesis, Site-Directed;
methods;
Recombinant Proteins;
genetics;
pharmacology
- From:
Journal of Biomedical Engineering
2011;28(2):347-351
- CountryChina
- Language:Chinese
-
Abstract:
In order to create a novel recombinant human interferon alpha2b (rh-IFN alpha2b) with higher biological activity, we subjected the rational designed sequence of rh-IFN alpha2b to direct evolution by strategy of the combinatorial mutagenesis. The amino acid residues at multiple sites of 52-53-55, 103-107, and 121-125 were simultaneously mutated. The resulted gene of the mutated rh-IFN a2b was cloned into the pET28a and expressed in E. coli BL21 Condon plus (RIL). The anti-virus activity of the novel interferon alpha2b was 9.3 x 10(7) IU/mg, 93 times higher than the wild type (1 x 10(6) IU/mg). The results showed that the multiple point mutation used in this study could effectively combine the site effects of rh-IFN alpha2b and increase its biological activity.
