title: Generation of an HFRS patient-derived neutralizing recombinant antibody to Hantaan virus G1 protein and definition of the neutralizing domain.
abstract: 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.
authors.affiliation: Institute of Virology, Chinese Academy of Preventive Medicine, Beijing, China. liangmf@public3.bta.net.cn
authors.forename: Mifang
authors.initials: M
authors.lastname: Liang
authors.forename: Michael
authors.initials: M
authors.lastname: Mahler
authors.forename: Joachim
authors.initials: J
authors.lastname: Koch
authors.forename: Yan
authors.initials: Y
authors.lastname: Ji
authors.forename: Dexin
authors.initials: D
authors.lastname: Li
authors.forename: Connie
authors.initials: C
authors.lastname: Schmaljohn
authors.forename: Ekkehard K F
authors.initials: EK
authors.lastname: Bautz
mesh_terms: 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
chemical_list: D000914:Antibodies, Viral; D007140:Immunoglobulin Fab Fragments; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D019590:Nucleocapsid Proteins; D019151:Peptide Library; D011994:Recombinant Proteins