title: [Expression and renaturation of a novel human single-chain Fv antibody against SARS-CoV].
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.
authors.affiliation: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences (CAS), Beijing 100080, China.
authors.forename: Jin-Zhu
authors.initials: JZ
authors.lastname: Duan
authors.forename: Cai
authors.initials: C
authors.lastname: Qi
authors.forename: Wei
authors.initials: W
authors.lastname: Han
authors.forename: Zhan-Hui
authors.initials: ZH
authors.lastname: Wang
authors.forename: Gang
authors.initials: G
authors.lastname: Jin
authors.forename: Xi-Yun
authors.initials: XY
authors.lastname: Yan
mesh_terms: D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics; D000914:Antibodies, Viral / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007128:Immunoglobulin Fragments / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D002479:Inclusion Bodies / Q000235:genetics / Q000276:immunology; D020673:Protein Renaturation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology; D045473:Severe acute respiratory syndrome-related coronavirus / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins; C055966:immunoglobulin Fv