Sequence analysis of a monoclonal antibody specific for the preS2 region of hepatitis B surface antigen, and the cloning, expression and characterisation of its single‐chain Fv construction

FEBS Letters

Passafiume, Maria¹  Vulliez-le Normand, Brigitte¹  Bentley, Graham A.¹  Riottot, Marie-Madeleine¹ 

[1] Unité d'Immunologie Structurale (CNRS URA 1961), Institut Pasteur, 25 rue du Dr. Roux, 75724 Paris cedex 15, France

关键词: Antibody engineering;    c-myc tag;    Hepatitis B virus;    PreS2;    Single chain Fv fragment;    CDR;    complementarity determining region;    FR;    framework region;    HBV;    hepatitis B virus;    HBsAg;    hepatitis B surface antigen;    r-HBsAg;    recombinant hepatitis B surface antigen particles;    scFv;    single-chain Fv fragment;    SDS-PAGE;    sodium dodecyl sulfate polyacrylamide gel electrophoresis;    VL;    light chain variable region;    VH;    heavy chain variable region;   

DOI  :  10.1016/S0014-5793(98)01581-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The nucleotide sequence of the monoclonal antibody F124, specific for the preS2 region of the surface antigen of hepatitis B virus, has been determined and an single-chain Fv fragment (scFv) recombinant construction has been cloned and expressed into the periplasmic region of Escherichia coli. The recombinant antibody fragment contains a (Gly₄Ser)₃ linker connecting the C-terminus of the heavy chain variable region (V_H) domain to the N-terminus of the light chain variable region (V_L) domain. A 23-residue peptide segment, containing a c-myc marker for immunochemical detection of the scFv and hexahistidine tag to facilitate its purification, was added C-terminal to the V_L domain. The scFv mimics the antibody in binding to the native antigen in the form of recombinant hepatitis B surface antigen (HBsAg) particles as well as to peptide fragments carrying the viral epitope.

【 授权许可】

Unknown