Expression and renaturation of a novel human single-chain Fv antibody against SARS-CoV.
- Author:
Jin-Zhu DUAN
1
;
Cai QI
;
Wei HAN
;
Zhan-Hui WANG
;
Gang JIN
;
Xi-Yun YAN
Author Information
1. State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences (CAS), Beijing 100080, China.
- Publication Type:Journal Article
- MeSH:
Antibodies, Monoclonal;
biosynthesis;
genetics;
Antibodies, Viral;
immunology;
Escherichia coli;
genetics;
metabolism;
Humans;
Immunoglobulin Fragments;
biosynthesis;
genetics;
immunology;
Immunoglobulin Variable Region;
biosynthesis;
genetics;
immunology;
Inclusion Bodies;
genetics;
immunology;
Protein Renaturation;
Recombinant Proteins;
biosynthesis;
chemistry;
immunology;
SARS Virus;
immunology
- From:
Chinese Journal of Biotechnology
2005;21(5):692-697
- CountryChina
- Language:Chinese
-
Abstract:
A novel human ScFv H12 against SARS-CoV has been selected from a SARS immune library. In order to produce a large amount of ScFv H12, pET28a-H12 expression vector was constructed and ScFv H12 was expressed at yield about 30% of total proteins in E. coli . Here two different refolding procedures were used to refold ScFv H12 from inclusion body: gel filtration chromatography and dilution. The results showed that ScFv H12 could be efficiently refolded by both procedures. However, the refolding via gel filtration was 1.5 time more effective than that of dilution. The affinity of ScFv H12 to SARS-CoV virion was detected as Kd = 73.5 nmol/mL.
