Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology · 2011
[Generation of recombinant human antibodies for EV71 virus].
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Sun Li-Na, Zhang Li, Zhang Fu-Shun, Li Chuan, Zhang Quan-Fu, Li De-Xin, Liang Mi-Fang
Аннотация
OBJECTIVE: To obtain recombinant human anti-EV71 antibodies from a EV71-associated hand-foot-and-mouth disease patient-derived antibody phage library.
METHODS: A combinatorial human scFv library to enterovirus 71 (EV71) virus was constructed using antibody genes harvested from the blood of EV71 virus patients. The library was panned and selected by using purified VP1 protein of EV71 virus with phage display. After that the specific antibody was converted to full human IgG antibody with recombinant baculovirus/insect cell system.
RESULTS: One unique human scFv antibody specific for EV71 virus VP1 protein was obtained by ELISA, IFA and analysis of the antibody DNA sequence. The specific anti-VP1 human scFv antibody was converted to full human IgG antibody with recombinant baculovirus/insect cell system. The full human IgG antibody was tested in vitro for EV71 virus neutralization, resulting in no neutralizing activity with EV71 A type and EV71 C4 subtype.
CONCLUSION: The obtained human anti-EV71 antibodies without neutralizing activity laid the foundation for diagnosis of human EV71-associated hand-foot-and-mouth disease.
METHODS: A combinatorial human scFv library to enterovirus 71 (EV71) virus was constructed using antibody genes harvested from the blood of EV71 virus patients. The library was panned and selected by using purified VP1 protein of EV71 virus with phage display. After that the specific antibody was converted to full human IgG antibody with recombinant baculovirus/insect cell system.
RESULTS: One unique human scFv antibody specific for EV71 virus VP1 protein was obtained by ELISA, IFA and analysis of the antibody DNA sequence. The specific anti-VP1 human scFv antibody was converted to full human IgG antibody with recombinant baculovirus/insect cell system. The full human IgG antibody was tested in vitro for EV71 virus neutralization, resulting in no neutralizing activity with EV71 A type and EV71 C4 subtype.
CONCLUSION: The obtained human anti-EV71 antibodies without neutralizing activity laid the foundation for diagnosis of human EV71-associated hand-foot-and-mouth disease.