Volume 21 Issue 5
Oct.  2008
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TAO DUAN, XIAO-FANG WANG, SHU-YUAN XIAO, SHU-YAN GU, MI-FANG LIANG. Recombinant Human IgG antibodies against Human Cytomegalovirus[J]. Biomedical and Environmental Sciences, 2008, 21(5): 372-380.
Citation: TAO DUAN, XIAO-FANG WANG, SHU-YUAN XIAO, SHU-YAN GU, MI-FANG LIANG. Recombinant Human IgG antibodies against Human Cytomegalovirus[J]. Biomedical and Environmental Sciences, 2008, 21(5): 372-380.

Recombinant Human IgG antibodies against Human Cytomegalovirus

  • Objective To study the passive immunization with human monoclonal antibodies as for prophylaxis of human eytomegalovirus (HCMV) infection. Methods Fab monochinal antibodies to HCMV were recovered by repertoire cloning of mRNA from a HCMV infected individual. Antigen binding specificity, CDR sequence of Vhand Vland 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-κ-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 antibodyrecognized a conformational HCMV protein. Conclusion IgG antibodies can neutralize the HCMV AD169 strain efficiently at a titer of 2.5 μg/mL and may prove valuable for passive immunoprophylaxis against HCMV infection in humans.
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    沈阳化工大学材料科学与工程学院 沈阳 110142

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Recombinant Human IgG antibodies against Human Cytomegalovirus

Abstract: Objective To study the passive immunization with human monoclonal antibodies as for prophylaxis of human eytomegalovirus (HCMV) infection. Methods Fab monochinal antibodies to HCMV were recovered by repertoire cloning of mRNA from a HCMV infected individual. Antigen binding specificity, CDR sequence of Vhand Vland 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-κ-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 antibodyrecognized a conformational HCMV protein. Conclusion IgG antibodies can neutralize the HCMV AD169 strain efficiently at a titer of 2.5 μg/mL and may prove valuable for passive immunoprophylaxis against HCMV infection in humans.

TAO DUAN, XIAO-FANG WANG, SHU-YUAN XIAO, SHU-YAN GU, MI-FANG LIANG. Recombinant Human IgG antibodies against Human Cytomegalovirus[J]. Biomedical and Environmental Sciences, 2008, 21(5): 372-380.
Citation: TAO DUAN, XIAO-FANG WANG, SHU-YUAN XIAO, SHU-YAN GU, MI-FANG LIANG. Recombinant Human IgG antibodies against Human Cytomegalovirus[J]. Biomedical and Environmental Sciences, 2008, 21(5): 372-380.

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