title: Generation and characterization of the human neutralizing antibody fragment Fab091 against rabies virus.
abstract: OBJECTIVE
  To transform the human anti-rabies virus glycoprotein (anti-RABVG) single-chain variable fragment (scFv) into a Fab fragment and to analyze its immunological activity.
  
  
  METHODS
  The Fab gene was amplified using overlap PCR and inserted into the vector pComb3XSS. The recombinant vector was then transformed into E coli Top10F' for expression and purification. The purified Fab was characterized using SDS-PAGE, Western blotting, indirect ELISA, competitive ELISA, and the fluorescent antibody virus neutralization test (FAVN), respectively, and examined in a Kunming mouse challenge model in vivo.
  
  
  RESULTS
  A recombinant vector was constructed. The Fab was expressed in soluble form in E coli Top10F'. Specific binding of the Fab to rabies virus was confirmed by indirect ELISA and immunoprecipitation (IP). The neutralizing antibody titer of Fab was 10.26 IU/mL. The mouse group treated with both vaccine and human rabies immunoglobulin (HRIG)/Fab091 (32 IU/kg) showed protection against rabies, compared with the control group (P<0.05, Logrank test).
  
  
  CONCLUSIONS
  The antibody fragment Fab was shown to be a neutralizing antibody against RABVG. It can be used together with other monoclonal antibodies for post-exposure prophylaxis of rabies virus in future studies.
authors.affiliation: Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Li
authors.forename: Feng
authors.initials: F
authors.lastname: Zhang
authors.forename: Hong
authors.initials: H
authors.lastname: Lin
authors.forename: Zhong-can
authors.initials: ZC
authors.lastname: Wang
authors.forename: Xin-jian
authors.initials: XJ
authors.lastname: Liu
authors.forename: Zhen-qing
authors.initials: ZQ
authors.lastname: Feng
authors.forename: Jin
authors.initials: J
authors.lastname: Zhu
authors.forename: Xiao-hong
authors.initials: XH
authors.lastname: Guan
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D000914:Antibodies, Viral / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D051379:Mice; D011818:Rabies / Q000517:prevention & control*; D011819:Rabies Vaccines / Q000008:administration & dosage*; D011820:Rabies virus / Q000235:genetics / Q000276:immunology*
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D007140:Immunoglobulin Fab Fragments; D011819:Rabies Vaccines