a rapid subtractive immunization method to prepare discriminatory monoclonal antibodies for food e. coli o157h7 contamination快速减去免疫方法准备歧视性的单克隆抗体对大肠杆菌o157h7污染食物.pdfVIP

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2017-09-07 发布于上海
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a rapid subtractive immunization method to prepare discriminatory monoclonal antibodies for food e. coli o157h7 contamination快速减去免疫方法准备歧视性的单克隆抗体对大肠杆菌o157h7污染食物.pdf

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a rapid subtractive immunization method to prepare discriminatory monoclonal antibodies for food e. coli o157h7 contamination快速减去免疫方法准备歧视性的单克隆抗体对大肠杆菌o157h7污染食物

A Rapid Subtractive Immunization Method to Prepare Discriminatory Monoclonal Antibodies for Food E. coli O157:H7 Contamination . . Min Jin , Jing Lang , Zhi-Qiang Shen, Zhao-Li Chen, Zhi-Gang Qiu, Xin-Wei Wang, Jun-Wen Li* Department of Environment and Health, Institute of Health and Environmental Medicine, Key Laboratory of Risk Assessment and Control for Environment and Food Safety, Tianjin, China Abstract To detect food E. coli O157:H7 contamination rapidly and accurately, it is essential to prepare high specific monoclonal antibodies (mAbs) against the pathogen. Cyclophosphamide (Cy)-mediated subtractive immunization strategy was performed in mice to generate mAbs that react with E. coli O157:H7, but not with other affiliated bacteria. Specificity of 19 mAbs was evaluated by ELISA and/or dot-immunogold filtration assay (DIGFA). Immunogloubin typing, affinity and binding antigens of 5 selected mAbs were also analysed. MAbs 1D8, 4A7, 5A2 were found to have high reactivity with E. coli O157:H7 and no cross-reactivity with 80 other strains of bacteria including Salmonella sp., Shigella sp., Proteus sp., Yersinia enterocolitica, Staphylococcus aureus, Klebsiella pneumoniae, Citrobacter freundii and other non-E. coli O157:H7 enteric bacteria. Their ascetic titers reached 1:106 with E. coli O157:H7 and affinity constants ranged from 1.57 61010 to 2.79 61010 L/ mol. The antigens recognized by them were different localized proteins. Furthermore, immune-colloidal gold probe coated with mAb 5A2 could specifically distinguish minced beef contaminated by E. coli O157:H7 from 84 other bacterial contaminations. The Cy-mediated subtractive immunization procedure coupled with hybridoma tec