新霉素的直接竞争酶联免疫分析法

doi:10.7506/spkx1002-6630-201110050

食品科学 ›› 2011, Vol. 32 ›› Issue (10): 212-217.doi: 10.7506/spkx1002-6630-201110050

新霉素的直接竞争酶联免疫分析法

徐乃丰，胥传来，匡华，屈昌龙，许阳，彭池方*

江南大学食品学院

出版日期:2011-05-25 发布日期:2011-04-08

Direct Competitive Enzyme-linked Immunosorbent Assay (ELISA) for Neomycin

XU Nai-feng，XU Chuan-lai，KUANG Hua，QU Chang-long，XU Yang，PENG Chi-fang*

School of Food Science and Technology, Jiangnan University, Wuxi 214122, China

Online:2011-05-25 Published:2011-04-08

摘要/Abstract

摘要： 首先用碳二亚胺(EDC)法将新霉素(NEO)偶联于载体蛋白-卵清白蛋白(ovalbumin，OVA)，合成包被原OVA-NEO，SDS-PAGE 进行鉴定；用高碘酸钠法连接辣根过氧化物酶(horseradish peroxidase，HRP)，制备酶标抗原NEO-HRP并建立直接ELISA检测方法。通过一系列参数的优化，包括包被溶液、封闭溶液、竞争时间、抗体稀释液、pH值、反应温度、显色时间等，最终得到其IC20(抑制率为20%时的标准溶液质量浓度)＜1ng/mL、半数抑制量(IC50)为7.6ng/mL，线性方程为y =－0.2798x＋0.7456，R2=0.991。直接ELISA总耗时只需大约1h。

关键词: 新霉素, 间接竞争酶联免疫吸附试验, 直接竞争酶联免疫吸附试验

Abstract: The neomycin (NEO) was coupled to Ovalbumin (OVA) to form coating antigen NEO-OVA by the EDC methods, determined by SDS-PAGE, and then indirect competitive ELISA (ic-ELISA) method was established; The NEO was coupled to horseradish peroxidase (HRP) by NaIO4 method，making enzyme tracer NEO-HRP and direct competitive ELISA . Then, the direct ELISA was developed and optimized in many parameters, such as coating liquids, pH value, incubation time , incubation temperature and so on. At last, the IC20 value (20% inhibitory concentration) was less than 1 ng/mL and IC50 value (50% inhibitory concentration) was 7.6 ng/mL . The formation is y = -0.2798x + 0.74 56, R2= 0.991. The direct ELISA could run in only about 1 h while the indirect assay duration was at least 1.75 h.

Key words: neomycin, indirect competitive enzyme-linked immunosorbent assay, direct competitive enzyme-linked immunosorbent assay

中图分类号:

S859.84

徐乃丰，胥传来，匡华，屈昌龙，许阳，彭池方. 新霉素的直接竞争酶联免疫分析法[J]. 食品科学, 2011, 32(10): 212-217.

XU Nai-feng，XU Chuan-lai，KUANG Hua，QU Chang-long，XU Yang，PENG Chi-fang. Direct Competitive Enzyme-linked Immunosorbent Assay (ELISA) for Neomycin[J]. FOOD SCIENCE, 2011, 32(10): 212-217.

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新霉素的直接竞争酶联免疫分析法

Direct Competitive Enzyme-linked Immunosorbent Assay (ELISA) for Neomycin

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