| 恩诺沙星快速酶联免疫吸附法检测试剂盒的研制 |
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| 引用本文： | 陈雪岚,金晶,杨晓慧.恩诺沙星快速酶联免疫吸附法检测试剂盒的研制[J].食品科学,2010,31(8):16-19. |
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| 作者姓名： | 陈雪岚 金晶 杨晓慧 |
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| 作者单位： | 1.江西师范大学生命科学学院 2.北京中德大地食品安全技术开发有限责任公司 |
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| 基金项目： | 国家“863”计划项目(2007AA10Z438) |
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| 摘 要： | 目的：研制恩诺沙星酶联免疫吸附法检测试剂盒。方法：采用EDC-NHS 法制备ENR-M-BSA 免疫原和ENR-OVA 检测抗原，用ENR-M-BSA 免疫BALB/C 小鼠，采用甲基纤维素半固体培养基法筛选单克隆抗体并采用间接竞争ELISA 对其进行分析。结果：通过公式计算ENR-M-BSA 摩尔偶联比为15:1，ENR-OVA 摩尔偶联比为7.2:1。筛选出1 株特异性分泌株3E9，其腹水纯化后间接竞争ELISA 测其效价为107，分型试剂盒测试显示此抗体属于IgG1 型；检测限为1ng/mL，线性范围在1～100ng/mL 之间，半量抑制浓度(IC50 值)为10ng/mL，与4 种结构类似物交叉反应均小于1%。结论：此株单克隆抗体可以用来建立恩诺沙星ELISA 检测试剂盒。
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| 关 键 词： | 恩诺沙星 酶联免疫吸附法 单克隆抗体 |
| 收稿时间： | 2009-05-06 |
Development of Enrofloxacin ELISA Test Kit |
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| CHEN Xue-lan,JIN Jing,YANG Xiao-hui.Development of Enrofloxacin ELISA Test Kit[J].Food Science,2010,31(8):16-19. |
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| Authors: | CHEN Xue-lan JIN Jing YANG Xiao-hui |
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| Affiliation: | 1. School of Bioscience, Jiangxi Normal University, Nanchang 330022, China；
2. Beijing Zhongde-Dade Food Safety Technology Co. Ltd., Beijing 100081, China |
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| Abstract: | An enzyme-linked immunoabsorbence(ELISA) test kit was developed for enrofloxacin(ENR) detection.ENR-M-BSA and ENR-OVA antigens were prepared by EDC-NHS method.BALB/C mice were immunized with ENR-M-BSA for monoclonal antibody generation.A specific monoclonal antibody was screened using methylcellulose-based semi-solid medium and analyzed using indirect competition ELISA.The molar coupling ratios of ENR-conjugated BSA and ENR-conjugated OVA were calculated to be 15:1 and 7.2:1,respectively.The specific monocl... |
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| Keywords: | enrofloxacin(ENR) ELISA monoclonal antibody |
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