Human recombinant antibodies specific for hepatitis C virus core and envelope E2 peptides from an immune phage display library.
Hepatitis C virus (HCV) is the aetiological agent responsible for most cases of non-A non-B hepatitis. Hepatitis C is a disease of clinical importance because of its high infection rate in blood donors and its persistence as chronic infections which may lead to cirrhosis and hepatocellular carcinoma in the long term. The variability of the HCV genome has posed difficulties in serological detection and vaccine design. The recent advance in phage technology offers a means of cloning human anti-HCV antibodies of a defined specificity that may have potential therapeutic use. We now report the generation of a phage display library using the V(H) genes of a HCV-infected patient and the V(L) genes of two non-immune individuals. From this library we were able to obtain specific IgG single-chain Fvs (scFvs) that recognize viral core and envelope proteins by selection on synthetic peptides derived from the core sequence PKARRPEGRTWAQPG and the envelope E2 sequence RPIDDFDQGWGPITY. The specificity of the scFvs was demonstrated by their specific reactions with homologous peptides in ELISA and the specific blocking of scFv binding by homologous peptides, in a dose-dependent manner, in inhibition ELISA. The binding of the anticore 4c2 to homologous peptide was blocked by HCV-positive human sera in an antibody-concentration-dependent manner, suggesting that the scFv recognizes a similar if not identical epitope to those of one or more of the polyclonal antibodies present in the sera.
Chan
S W
SW
Medical Research Council (MRC) Molecular Immunopathology Unit, MRC Centre, Cambridge, UK. swc21@cam.ac.uk
Bye
J M
JM
Jackson
P
P
Allain
J P
JP
D000595:Amino Acid Sequence; D000918:Antibody Specificity / Q000276:immunology*; D001435:Bacteriophages / Q000235:genetics; D003001:Cloning, Molecular; D005822:Genetic Vectors; D018937:Hepatitis C Antibodies / Q000235:genetics / Q000276:immunology*; D018936:Hepatitis C Antigens / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics; D008222:Lymphokines / Q000276:immunology; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology*; D012710:Serum Albumin, Bovine / Q000276:immunology; D012795:Sialoglycoproteins / Q000276:immunology; D014758:Viral Core Proteins / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000276:immunology*
C089422:Fv protein, human; D018937:Hepatitis C Antibodies; D018936:Hepatitis C Antigens; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D008222:Lymphokines; D010455:Peptides; D011993:Recombinant Fusion Proteins; D012795:Sialoglycoproteins; D014758:Viral Core Proteins; D014759:Viral Envelope Proteins; C069130:nucleocapsid protein, Hepatitis C virus; C091015:glycoprotein E2, Hepatitis C virus; D012710:Serum Albumin, Bovine