title: The neutralizing human recombinant antibodies to pathogenic Orthopoxviruses derived from a phage display immune library.
abstract: A panel of recombinant human antibodies to orthopoxviruses was isolated from a combinatorial phage display library of human scFv antibodies constructed from the Vh and Vl genes cloned from the peripheral blood lymphocytes of Vaccinia virus (VACV) immune donors. Plaque-reduction neutralization tests showed that seven selected phage-displaying scFv antibodies (pdAbs) neutralized both CPXV and VACV, and five of them neutralized Monkeypox virus (MPXV). Western blot analysis of VACV and CPXV proteins demonstrated that seven neutralizing antibodies recognized a 35 kDa protein. To identify this target protein, we produced a recombinant J3L protein of CPXV and showed that all the selected neutralizing antibodies recognized this protein. Neutralizing pdAb b9 was converted into fully human mAb b9 (fh b9), and scFv b9 displayed high binding affinities (K(d) of 0.7 and 3.2 nM). The fh b9 reduced VACV plaque formation in a dose-dependent manner.
authors.affiliation: Institute of Chemical Biology and Fundamental Medicine, Novosibirsk 630090, Russia. tikunova@niboch.nsc.ru
authors.forename: Nina
authors.initials: N
authors.lastname: Tikunova
authors.forename: Viktoriya
authors.initials: V
authors.lastname: Dubrovskaya
authors.forename: Vera
authors.initials: V
authors.lastname: Morozova
authors.forename: Tatiana
authors.initials: T
authors.lastname: Yun
authors.forename: Yana
authors.initials: Y
authors.lastname: Khlusevich
authors.forename: Nikolai
authors.initials: N
authors.lastname: Bormotov
authors.forename: Aleksandr
authors.initials: A
authors.lastname: Laman
authors.forename: Fedor
authors.initials: F
authors.lastname: Brovko
authors.forename: Aleksandr
authors.initials: A
authors.lastname: Shvalov
authors.forename: Eugeni
authors.initials: E
authors.lastname: Belanov
mesh_terms: D057134:Antibodies, Neutralizing / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D000915:Antibody Affinity; D015153:Blotting, Western; D006805:Human Experimentation; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D018155:Orthopoxvirus / Q000276:immunology*; D019151:Peptide Library; D011485:Protein Binding; D017422:Sequence Analysis, DNA; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D012900:Smallpox Vaccine / Q000008:administration & dosage / Q000276:immunology; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D019151:Peptide Library; D057127:Single-Chain Antibodies; D012900:Smallpox Vaccine; D014764:Viral Proteins