title: Recombinant human monoclonal antibodies against different conformational epitopes of the E2 envelope glycoprotein of hepatitis C virus that inhibit its interaction with CD81.
abstract: The antibody response to the envelope proteins of hepatitis C virus (HCV) may play an important role in controlling the infection. To allow molecular analyses of protective antibodies, we isolated human monoclonal antibodies to the E2 envelope glycoprotein of HCV from a combinatorial Fab library established from bone marrow of a chronically HCV-infected patient. Anti-E2 reactive clones were selected using recombinant E2 protein. The bone marrow donor carried HCV genotype 2b, and E2 used for selection was of genotype 1a. The antibody clones were expressed as Fab fragments in E. coli, and as Fab fragments and IgG1 in CHO cells. Seven different antibody clones were characterized, and shown to have high affinity for E2, genotype 1a. Three clones also had high affinity for E2 of genotype 1b. They all bind to conformation-dependent epitopes. Five clones compete for the same or overlapping binding sites, while two bind to one or two other epitopes of E2. Four clones corresponding to the different epitopes were tested as purified IgG1 for blocking the CD81-E2 interaction in vitro; all four were positive at 0.3-0.5 microg/ml. Thus, the present results suggest the existence of at least two conserved epitopes in E2 that mediate inhibition of the E2-CD81 interaction, of which one appeared immunodominant in this donor.
authors.affiliation: Karolinska Institute, Department of Medicine1 and Department of Laboratory Medicine2, Center for Molecular Medicine (L8:01), Karolinska Hospital, S-171 76 Stockholm, Sweden.
authors.forename: Tobias
authors.initials: T
authors.lastname: Allander
authors.affiliation: Karolinska Institute, Department of Medicine1 and Department of Laboratory Medicine2, Center for Molecular Medicine (L8:01), Karolinska Hospital, S-171 76 Stockholm, Sweden.
authors.forename: Katarina
authors.initials: K
authors.lastname: Drakenberg
authors.affiliation: Karolinska Institute, Department of Medicine1 and Department of Laboratory Medicine2, Center for Molecular Medicine (L8:01), Karolinska Hospital, S-171 76 Stockholm, Sweden.
authors.forename: Aster
authors.initials: A
authors.lastname: Beyene
authors.affiliation: Chiron-Biocine, IRIS, Siena, Italy3.
authors.forename: Domenico
authors.initials: D
authors.lastname: Rosa
authors.affiliation: Chiron-Biocine, IRIS, Siena, Italy3.
authors.forename: Sergio
authors.initials: S
authors.lastname: Abrignani
authors.affiliation: Chiron Corporation, Department of Virology, Emeryville, CA, USA4.
authors.forename: Michael
authors.initials: M
authors.lastname: Houghton
authors.affiliation: Lund University, Department of Clinical Microbiology, Malmö University Hospital, Malmö, Sweden5.
authors.forename: Anders
authors.initials: A
authors.lastname: Widell
authors.affiliation: Karolinska Institute, Department of Medicine1 and Department of Laboratory Medicine2, Center for Molecular Medicine (L8:01), Karolinska Hospital, S-171 76 Stockholm, Sweden.
authors.forename: Lena
authors.initials: L
authors.lastname: Grillner
authors.affiliation: Karolinska Institute, Department of Medicine1 and Department of Laboratory Medicine2, Center for Molecular Medicine (L8:01), Karolinska Hospital, S-171 76 Stockholm, Sweden.
authors.forename: Mats A A
authors.initials: MAA
authors.lastname: Persson
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000302:isolation & purification*; D015703:Antigens, CD / Q000378:metabolism*; D001853:Bone Marrow / Q000821:virology; D016466:CHO Cells; D006224:Cricetinae; D018604:Epitope Mapping; D005838:Genotype; D016174:Hepacivirus / Q000235:genetics / Q000276:immunology*; D018937:Hepatitis C Antibodies / Q000302:isolation & purification*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis; D007074:Immunoglobulin G / Q000096:biosynthesis; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D011487:Protein Conformation; D021281:Quantitative Structure-Activity Relationship; D011994:Recombinant Proteins / Q000276:immunology; D060225:Tetraspanin 28; D014019:Tissue Donors; D014759:Viral Envelope Proteins / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D015703:Antigens, CD; C554798:CD81 protein, human; D018937:Hepatitis C Antibodies; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D008565:Membrane Proteins; D011994:Recombinant Proteins; D060225:Tetraspanin 28; D014759:Viral Envelope Proteins; C091015:glycoprotein E2, Hepatitis C virus