title: Recombinant human IgG antibodies against human cytomegalovirus.
abstract: OBJECTIVE
  To study the passive immunization with human monoclonal antibodies as for prophylaxis of human cytomegalovirus (HCMV) infection.
  
  
  METHODS
  Fab monoclonal antibodies to HCMV were recovered by repertoire cloning of mRNA from a HCMV infected individual. Antigen binding specificity, CDR sequence of V(H) and V(L) and neutralizing activity on HCMV AD169 stain were analyzed in vitro. The light and heavy chain Fd fragment genes of Fab antibodies were further cloned into a recombinant baculovirus expression vector pAC-kappa-Fc to express intact IgG. Secreted products were purified with affinity chromatography using protein G.
  
  
  RESULTS
  SDS-PAGE and Western blot confirmed the expression of the intact IgG. Immuno-blotting and -precipitation were used to identify HCMV proteins. One Fab monoclonal antibody recognized a conformational HCMV protein.
  
  
  CONCLUSIONS
  IgG antibodies can neutralize the HCMV AD169 strain efficiently at a titer of 2.5 microg/mL and may prove valuable for passive immunoprophylaxis against HCMV infection in humans.
authors.affiliation: Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Duan
authors.forename: Xiao-Fang
authors.initials: XF
authors.lastname: Wang
authors.forename: Shu-Yuan
authors.initials: SY
authors.lastname: Xiao
authors.forename: Shu-Yan
authors.initials: SY
authors.lastname: Gu
authors.forename: Mi-Fang
authors.initials: MF
authors.lastname: Liang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D003587:Cytomegalovirus / Q000276:immunology*; D004279:DNA, Viral / Q000235:genetics; D005814:Genes, Viral; D006801:Humans; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000276:immunology*; D047468:Immunoprecipitation; D007313:Insecta; D008969:Molecular Sequence Data; D019151:Peptide Library; D011994:Recombinant Proteins / Q000276:immunology*
chemical_list: D004279:DNA, Viral; D007074:Immunoglobulin G; D019151:Peptide Library; D011994:Recombinant Proteins