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Position: Home > Articles > Preparation of an ELISA Method for Determining Buckwheat Allergen FOOD SCIENCE 2011,32 (20) 327-330

荞麦过敏原特异性抗体酶联免疫吸附法检测试剂制备

作  者:
张昕;崔晓东;王转花
单  位:
山西大学环境科学与工程研究中心;山西大学生物技术研究所
关键词:
苦荞;过敏蛋白;试剂盒;间接酶联免疫吸附法
摘  要:
以天然苦荞过敏蛋白包被聚苯乙烯微孔板,以鼠抗人IgE抗体标记辣根过氧化物酶,以间接酶联免疫吸附法(indirect enzyme-linked immunosorbent assay,IELISA)检测病人血清中的特异性抗体,分析并建立关键操作步骤及反应条件。结果表明:ELISA反应的最适工作条件为包被的抗原质量浓度100μg/mL、阳性血清稀释度1:50、酶标二抗稀释度1:1000,37℃时封闭的最佳时间为40min,包被抗原与阳性血清中抗体的最佳作用时间为40min,阳性血清与酶标二抗的最佳作用时间为40min。本方法具有良好的特异性、重复性和精密度。
译  名:
Preparation of an ELISA Method for Determining Buckwheat Allergen
作  者:
ZHANG Xin1,CUI Xiao-dong2,WANG Zhuan-hua2(1.Research Center for Environmental Science and Engineering,Shanxi University,Taiyuan 030006,China; 2.Institute of Biotechnology,Shanxi University,Taiyuan 030006,China)
关键词:
tartary buckwheat;allergen;kit;indirect enzyme-linked immunosorbent assay(IELISA)
摘  要:
An indirect enzyme-linked immunosorbent assay(IELISA) was proposed to detect specific antibodies in tartary buckwheat allergenic patients,sera using polystyrene micro-well plate coated with natural tartary buckwheat allergenic proteins and horseradish peroxidase(HRP)-labeled mouse anti-human IgE.Key operations and reaction conditions were optimized.The results showed that the optimal coating antigen concentration,positive serum dilution,HRP-labeled mouse anti-human IgE concentration,blocking time at 37 ℃,reaction time between coating antigen and antibodies in positive sera,and reaction time between positive sera and HRP-labeled mouse anti-human IgE were 100 μg/mL,1:50,1:1000,40 min,40 min and 40 min,respectively.This method proved highly specific,repeatable and sensitive.

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