title: The human antibody repertoire specific for rabies virus glycoprotein as selected from immune libraries.
abstract: Antibody phage display technology was used to identify human monoclonal antibodies that neutralize rabies virus (RV). A phage repertoire was constructed using antibody genes harvested from the blood of vaccinated donors. Selections using this repertoire and three different antigen formats of the RV glycoprotein (gp) resulted in the identification of 147 unique antibody fragments specific for the RV gp. Analysis of the DNA sequences of these antibodies demonstrated a large variation in the heavy- and light-chain germ-line gene usage, suggesting that a broad antibody repertoire was selected. The single-chain variable fragment (scFv) antibodies were tested in vitro for RV neutralization, resulting in 39 specificities that neutralize the virus. Of the scFv clones, 21 were converted into full-length human IgG(1) format. Analysis of viral escape variants and binding competition experiments indicated that the majority of the neutralizing antibodies are directed against antigenic site III of the RV gp. The obtained specificities expand the set of human anti-RV antibodies eligible for inclusion in an antibody cocktail aimed for use in rabies post-exposure prophylaxis.
authors.affiliation: Crucell Holland B.V., Leiden, The Netherlands.
authors.forename: R Arjen
authors.initials: RA
authors.lastname: Kramer
authors.forename: Wilfred E
authors.initials: WE
authors.lastname: Marissen
authors.forename: Jaap
authors.initials: J
authors.lastname: Goudsmit
authors.forename: Therese J
authors.initials: TJ
authors.lastname: Visser
authors.forename: Marieke
authors.initials: M
authors.lastname: Clijsters-Van der Horst
authors.forename: Arjen Q
authors.initials: AQ
authors.lastname: Bakker
authors.forename: Maureen
authors.initials: M
authors.lastname: de Jong
authors.forename: Mandy
authors.initials: M
authors.lastname: Jongeneelen
authors.forename: Sandra
authors.initials: S
authors.lastname: Thijsse
authors.forename: Harold H J
authors.initials: HH
authors.lastname: Backus
authors.forename: Amy B
authors.initials: AB
authors.lastname: Rice
authors.forename: William C
authors.initials: WC
authors.lastname: Weldon
authors.forename: Charles E
authors.initials: CE
authors.lastname: Rupprecht
authors.forename: Bernhard
authors.initials: B
authors.lastname: Dietzschold
authors.forename: Alexander B H
authors.initials: AB
authors.lastname: Bakker
authors.forename: John
authors.initials: J
authors.lastname: de Kruif
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000378:metabolism; D000914:Antibodies, Viral / Q000032:analysis* / Q000096:biosynthesis / Q000378:metabolism; D000956:Antigens, Viral / Q000276:immunology*; D006023:Glycoproteins / Q000276:immunology*; D006801:Humans; D007128:Immunoglobulin Fragments / Q000032:analysis / Q000096:biosynthesis; D007074:Immunoglobulin G / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism; D007135:Immunoglobulin Variable Region / Q000032:analysis / Q000096:biosynthesis; D008969:Molecular Sequence Data; D019151:Peptide Library; D010449:Peptide Mapping; D011820:Rabies virus / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D006023:Glycoproteins; D007128:Immunoglobulin Fragments; D007074:Immunoglobulin G; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D014759:Viral Envelope Proteins; C064951:glycoprotein G, Rabies virus