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《Chinese Journal of Cellular and Molecular Immunology》 2011-09
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Preparation and identification of OmpW monoclonal antibodies

MA Zhen-ning1,ZHENG Lei1,LIN Hai-mei1,ZHENG Yun-quan2,TANG Feng-xiang2,SHI Xian-ai2,GUO Yang-hao1,21Institute of Pharmaceutical Biotechnology and Bioengineering,Fuzhou University;2Fujian Key Lab of Medical Instrumentation and Pharmaceutical Technology,Fuzhou 350002,China  
AIM: To prepare and characterize the mouse monoclonal antibodies against Vibrio parahaemolyticus OmpW.METHODS: The OmpW amino acid sequence from three diseased Vibrio was analyzed by Bioinformatics.Mice were immunized with r-OmpW which was highly expressed and purified in E.coli.Five Vibrio(Va,Vp,Vh,Vv,Van) were chosen as antigen for mAb selection.The characters of the anti-OmpW monoclonal antibodies were studied by Western blot,Flow Cytometry,indirect immunofluorescence.RESULTS: OmpW was testified a highly conservative membrane protein.Three clones of anti-OmpW mAb was obtained.The Ig subclass of the mAb secreted from fused cell S5C10 was IgG3,which of the titer was 4.6×104.The mAb could specifically recognize Vibrio parahaemolyticus,Vibrio alginolyticus,Vibrio harveyi,Vibrio anguillarum,Vibrio vulnificus,which could not react with Pseudomonas flurosecens,Aeromonas hydrophila,Aeromonas sobria,Aeromonas hydrophila,Escherichia coli.CONCLUSION: The mAb could specially recognize five diseased Vibrio,which is a useful tool for the further study of the diagnosis of Vibrio.
【Fund】: 福建省科技厅重点项目(2009N401010844);福建省科技厅重点项目(2010N0014);; 福建省教育厅科技项目(JA10024)
【CateGory Index】: R392.1
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