玉米赤霉醇酶标抗原的制备及其直接和间接竞争ELISA检测方法的建立与比较  

Synthesis of Enzyme Labeled Antigens and Establishment of Direct Competitive ELISA and Indirect Competitive ELISA Methods for Zeranol

作　　者：刘媛[1] 刘贤进[1] 余向阳[1] 张存政[1] 张晓[1] 刘蓉蓉[1] 龚振明[2] 

机构地区：[1]江苏省农业科学院食品质量安全检测研究所,南京210014 [2]上海交通大学,上海200030

出　　处：《分析科学学报》2008年第2期141-144,共4页Journal of Analytical Science

基　　金：国家自然科学基金(No.30471155);上海市科技兴农重点攻关项目(沪农科攻字(2003)第9-4号)

摘　　要：采用活性酯法将半抗原玉米赤霉醇-16-羧丙基醚与辣根过氧化物酶连接,制备了三种结合比的酶标抗原。通过紫外吸收法和直接非竞争ELISA法对酶标抗原的偶联结果和抗原性进行鉴定。最终选择结合比为1.3∶1的酶标抗原建立直接竞争ELISA检测方法。并对建立的直接竞争ELISA(DC-ELISA)与间接竞争ELISA(IC-ELISA)方法在检出限、检测线性范围、检测时间和二抗的使用方面进行了比较。To produce a novel type of enzyme labeled antigen, zerenol-16-carboxy propyl ether was conjugated to horseradish peroxidase （HRP） by the activated ester method. Three enzyme labeled antigens with different conjugate ratios were identified by UV spectrometry and direct uncompetitive ELISA. The enzyme labeled antigen with conjugate ratio of 1.3：1 was finally chosen to establish direct competitive ELISA. The direct competitive ELISA and indirect competitive ELISA for zeranol were established and a comparison between these two methods in terms of detection limit, response range, detection time and use of commercial secondary antibodies was also made in this study.

关 键 词：玉米赤霉醇 酶标抗原 酶联免疫吸附试验(ELISA) 

分 类 号：Q503[生物学—生物化学]