Human monoclonal recombinant Fabs specific for HCV antigens obtained by repertoire cloning in phage display combinatorial vectors.
Molecular cloning of the antibody repertoire in phage display combinatorial vectors is a powerful method enabling the dissection of the immunoresponse against a given pathogen. In this paper we describe the construction of a combinatorial library displayed on phage surface, containing the antibody repertoire of a patient with high serological response against hepatitis C virus (HCV) antigens. Following selection of the library against solid-phase-bound antigen, sixteen human antibody Fab fragments able to bind to HCV-specific antigens were generated and studied for binding characteristics. The majority of them appeared to have specificity for the HCV c33 peptide. All the clones reacting with the c33 peptide shared the same heavy-chain CDR3 sequence. This is the first report of molecular cloning in a combinatorial phage display vector of the antibody repertoire of an anti-HCV-positive patient.
Plaisant
P
P
Istituto di Microbiologia, Facoltà di Medicina, Università Cattolica del Sacro Cuore, Roma, Italy.
Burioni
R
R
Manzin
A
A
Solforosi
L
L
Candela
M
M
Gabrielli
A
A
Fadda
G
G
Clementi
M
M
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000276:immunology; D001435:Bacteriophages / Q000235:genetics*; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005822:Genetic Vectors; D018936:Hepatitis C Antigens / Q000276:immunology*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000235:genetics* / Q000276:immunology; D007135:Immunoglobulin Variable Region; D008875:Middle Aged; D008969:Molecular Sequence Data
D000911:Antibodies, Monoclonal; D018936:Hepatitis C Antigens; D007140:Immunoglobulin Fab Fragments; D007135:Immunoglobulin Variable Region