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.
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(L)) 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.
Passafiume
M
M
Unité d'Immunologie Structurale (CNRS URA 1961), Institut Pasteur, Paris, France.
Vulliez-le Normand
B
B
Riottot
M M
MM
Bentley
G A
GA
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D000918:Antibody Specificity; D001483:Base Sequence; D001667:Binding, Competitive; D003001:Cloning, Molecular; D018076:DNA, Complementary; D006514:Hepatitis B Surface Antigens / Q000276:immunology*; D007128:Immunoglobulin Fragments / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000276:immunology*
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D006514:Hepatitis B Surface Antigens; D007128:Immunoglobulin Fragments; D011498:Protein Precursors; C050807:presurface protein 2, hepatitis B surface antigen