Recombinant human IgG antibodies against human cytomegalovirus.
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.
Duan
Tao
T
Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
Wang
Xiao-Fang
XF
Xiao
Shu-Yuan
SY
Gu
Shu-Yan
SY
Liang
Mi-Fang
MF
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*
D004279:DNA, Viral; D007074:Immunoglobulin G; D019151:Peptide Library; D011994:Recombinant Proteins