Recombinant human IgG antibodies against human cytomegalovirus.
- Author:
Tao DUAN
1
;
Xiao-Fang WANG
;
Shu-Yuan XIAO
;
Shu-Yan GU
;
Mi-Fang LIANG
Author Information
- Publication Type:Journal Article
- MeSH: Amino Acid Sequence; Animals; Base Sequence; Cell Line; Cloning, Molecular; Cytomegalovirus; immunology; DNA, Viral; genetics; Genes, Viral; Humans; Immunoblotting; Immunoglobulin G; immunology; Immunoprecipitation; Insecta; Molecular Sequence Data; Peptide Library; Recombinant Proteins; immunology
- From: Biomedical and Environmental Sciences 2008;21(5):372-380
- CountryChina
- Language:English
-
Abstract:
OBJECTIVETo study the passive immunization with human monoclonal antibodies as for prophylaxis of human cytomegalovirus (HCMV) infection.
METHODSFab 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.
RESULTSSDS-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.
CONCLUSIONIgG 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.
