title: Characterization of a human antibody fragment Fab and its calcium phosphate nanoparticles that inhibit rabies virus infection with vaccine.
abstract: Recombinant antibody phage display technology has been used to mimic many aspects of the processes that govern the generation and selection of high-affinity natural human antibodies in the human immune system, especially for infectious disease prophylaxis. An anti-rabies virus immunized phage-display Fab library was constructed from peripheral blood lymphocytes from vaccinated volunteers. The immunized antibody library, with a diversity of 6.7×10(8), was used to select and produce antibodies that bound to rabies virus glycoprotein. After five rounds of immobilized fixed rabies virion panning, four unique DNA sequences were found in the higher binding clones, and only one, Fab094, showed neutralization activity. Fab094 components were analyzed by ELISA, immunoprecipitation and immunofluorescent staining. ELISA and immunofluorescence showed that Fab094 bound specifically to rabies virions. Immunoprecipitation and mass spectrometry showed that Fab094 reacted with rabies virus glycoprotein. To improve the penetration power of Fab094 antibodies, we developed Fab094 calcium phosphate nanoparticles (Fab094-CPNPs) and tested their efficacy. The rapid fluorescent focus inhibition test indicated that the neutralizing antibody titers of Fab094 and Fab094-CPNPs were reached at 200.17 IU/Kg and 246.12 IU/Kg, respectively. These findings were confirmed in vivo in a Kunming mouse challenge model. Our results demonstrate that human Fab094 and Fab094-CPNPs are efficacious candidate drugs to replace rabies immunoglobulin in post-exposure prophylaxis (PEP).
authors.affiliation: Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing, China.
authors.forename: Xinjian
authors.initials: X
authors.lastname: Liu
authors.forename: Hong
authors.initials: H
authors.lastname: Lin
authors.forename: Qi
authors.initials: Q
authors.lastname: Tang
authors.forename: Chen
authors.initials: C
authors.lastname: Li
authors.forename: Songtao
authors.initials: S
authors.lastname: Yang
authors.forename: Zhongcan
authors.initials: Z
authors.lastname: Wang
authors.forename: Changjun
authors.initials: C
authors.lastname: Wang
authors.forename: Qing
authors.initials: Q
authors.lastname: He
authors.forename: Brian
authors.initials: B
authors.lastname: Cao
authors.forename: Zhenqing
authors.initials: Z
authors.lastname: Feng
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Guan
authors.forename: Jin
authors.initials: J
authors.lastname: Zhu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000737:chemistry / Q000276:immunology*; D000941:Antigens / Q000276:immunology; D015153:Blotting, Western; D002130:Calcium Phosphates / Q000276:immunology*; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D006801:Humans; D055767:Immobilized Proteins / Q000276:immunology; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000276:immunology*; D047468:Immunoprecipitation; D053208:Kaplan-Meier Estimate; D013058:Mass Spectrometry; D051379:Mice; D008969:Molecular Sequence Data; D053758:Nanoparticles / Q000627:therapeutic use*; D009500:Neutralization Tests; D019151:Peptide Library; D011818:Rabies / Q000276:immunology / Q000517:prevention & control* / Q000821:virology; D011819:Rabies Vaccines / Q000276:immunology*; D011820:Rabies virus / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D000941:Antigens; D002130:Calcium Phosphates; D055767:Immobilized Proteins; D007140:Immunoglobulin Fab Fragments; D019151:Peptide Library; D011819:Rabies Vaccines; C020243:calcium phosphate