摘要:
通过制备克伦特罗 (Clenbuterol, CL) 人工抗原获得高活性抗体, 用辣根过氧化物酶 (HRP) 标记CL抗原, 以鲁米诺 (luminol) 为发光底物, 建立直接竞争法检测分析盐酸克伦特罗, 并对包被液、包被条件、封闭蛋白、孵育时间、酶用量等参数进行优化, 成功研制出化学发光法检测CL试剂盒。该方法的线性检测范围0.18.1ng·mL-1, 线性相关系数r=0.996, IC50值为0.607ng·mL-1;理论检测限小于0.1ng·mL-1;批内变异系数6.76%8.31%;批间变异系数7.4%;回收率 (90±15) %, 与莱克多巴胺等其他β-激动剂不发生交叉反应。
Abstract:
A highly active antibody was produced by making manmade antigen of Clenbuterol.We established a direct competition method to detect Clenbuterol that used HRP as markers and luminol as chemiluminescent substrates, with which many parameters were optimized, including coating buffer, coating condition, blocking condition, incubating time, and enzyme consumption.Based on this method we developed a chemiluminescent detecting kit, whose linear range was 0.1-8.1ng·mL-1, the linear correlation coefficient was r=0.996, the IC50 was 0.607ng mL-1, the detection limit was under 0.1ng·mL-1, the within-run assay coefficient of variation was 6.76%-8.31%, the between-run assay coefficient of variation was 7.4%, the coefficient of recovery was between (90±15) %, and there was no cross reaction with otherβ-agonists.
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