The human antibody repertoire specific for rabies virus glycoprotein as selected from immune libraries.
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.
Kramer
R Arjen
RA
Crucell Holland B.V., Leiden, The Netherlands.
Marissen
Wilfred E
WE
Goudsmit
Jaap
J
Visser
Therese J
TJ
Clijsters-Van der Horst
Marieke
M
Bakker
Arjen Q
AQ
de Jong
Maureen
M
Jongeneelen
Mandy
M
Thijsse
Sandra
S
Backus
Harold H J
HH
Rice
Amy B
AB
Weldon
William C
WC
Rupprecht
Charles E
CE
Dietzschold
Bernhard
B
Bakker
Alexander B H
AB
de Kruif
John
J
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*
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