中国食品学报2013，Vol.13Issue(7)：170-174,5.
绿豆过敏原多克隆抗体制备及间接竞争酶联免疫检测方法的建立
Preparation of Polyclonal Antibody Against Mung Bean Allergen and Development of Indirect Competitive ELISA
摘要
Abstract
Objective:To prepare polyclonal antibody against mung bean allergen and develop an indirect competitive ELISA method.Methods:Extracted mung bean total protein by method of alkali-soluble acid-extracted,the amount of total protein was measured by Coomassie Brilliant Blue G-250.SDS-PAGE were carried out to assess the molecular weight of the total protein.Then New Zealand white rabbits were immunized with the total protein of 1 mg per rabbit,primary immunization was carried out by intramuscular and subcutaneous injection with Freund's complete adjuvant.Booster immunization was immunized four times with Freund's incompleteadjuvant,respectively,after the primary immunization 14,28,32 and 46 days,immunization dose was 0.5 mg per rabbit.Antiserum were purified with Protein ASepharose 4B affinity chromatography.Finally,through the optimization of coating amount of mung bean protein and antibody dilution,developed an indirect competitive ELISA method.Results:The major mung bean protein molecular weight was 50,38,34 and 25 kDa.The titer of anti-mung bean polyclonal antibody was 32000.Determined mung bean protein coating quantity was 0.1 μg/well and antibody diluted with 1∶32 000.The IC50 and IC15 was (0.72±0.035) μg/mL,(0.72±0.035)μg/mL,separately.Conclusion:An rapid indirect competitive ELISA detection methods for mung bean allergens was established successfully,to achieve real-time monitoring for allergenic proteins in food.关键词
绿豆过敏原/多克隆抗体/间接竞争酶联免疫检测方法Key words
Mung bean allergen/ polyclonal antibody/ ic-ELISA引用本文复制引用
谈超,高爱华,张燕,王宵雪,徐小婧,王硕..绿豆过敏原多克隆抗体制备及间接竞争酶联免疫检测方法的建立[J].中国食品学报,2013,13(7):170-174,5.基金项目
国家科技支撑"十二五"食品致敏原分子检测技术研究(2011BAK10B03) （2011BAK10B03）
