Generation of an HFRS patient-derived neutralizing recombinant antibody to Hantaan virus G1 protein and definition of the neutralizing domain.
Hantaan virus (HTNV) in the Hantavirus genus, family Bunyaviridae, is the major cause of severe hemorrhagic fever with renal syndrome (HFRS). We prepared a combinatorial phage display library of human Fabs to HTNV from RNA extracted from the blood lymphocytes of a convalescent HFRS patient. We selected two G1 glycoprotein-specific clones and one nucleocapsid protein (N)-specific clone from the Fab library for further studies. The human Fab antibodies were converted to IgG form in baculovirus/insect cells system by using cassette vectors that we developed earlier. Characterization of the recombinant antibodies revealed that the two G1-specific IgGs, could bind to and neutralize HTNV but not Seoul virus (SEOV). The N-specific IgG did not neutralize either HTNV or SEOV. Sequence analysis revealed that the two G1-specific clones differed by only one predicted amino acid in their complementarity determining regions, CDR3. Epitope mapping studies were carried out with one of the two G1-specific clones and synthetic peptides representing portions of HTNV G1. Results indicated that the recombinant antibody recognizes the core amino acid sequence LTKTLVIGQ, which is found near the C-terminus of HTNV G1. These results are the first to define a neutralizing epitope on the G1 protein of HTNV using an antibody derived from an HFRS patient.
Liang
Mifang
M
Institute of Virology, Chinese Academy of Preventive Medicine, Beijing, China. liangmf@public3.bta.net.cn
Mahler
Michael
M
Koch
Joachim
J
Ji
Yan
Y
Li
Dexin
D
Schmaljohn
Connie
C
Bautz
Ekkehard K F
EK
D000914:Antibodies, Viral / Q000096:biosynthesis / Q000276:immunology*; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D018063:Hantaan virus / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D006480:Hemorrhagic Fever with Renal Syndrome / Q000821:virology*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000235:genetics; D009500:Neutralization Tests; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D017924:Nucleopolyhedroviruses / Q000235:genetics / Q000378:metabolism; D019151:Peptide Library; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism
D000914:Antibodies, Viral; D007140:Immunoglobulin Fab Fragments; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D019590:Nucleocapsid Proteins; D019151:Peptide Library; D011994:Recombinant Proteins