CAJ | 학술논문

通过制备莱克多巴胺-牛血清白蛋白偶联物（Ractopamine-BSA, RAC-BSA）人工抗原，制备并获得高活性单克隆抗体，用辣根过氧化物酶（HRP）标记RAC抗原，以鲁米诺（luminol）为发光底物，建立直接竞争法检测分析莱克多巴胺，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RAC试剂盒。该方法的线性检测范围可达0.1～10.0 ng/mL，线性相关系数r=0.998，IC50值为0.763 ng/mL；理论检测限小于0.1 ng/mL；批内变异系数7.62%~8.65%；批间变异系数8.15%；回收率在85%±15%，与常见的克伦特罗等其他β-激动剂不发生交叉反应。
통과제비래극다파알-우혈청백단백우련물（Ractopamine-BSA, RAC-BSA）인공항원，제비병획득고활성단극륭항체，용랄근과양화물매（HRP）표기RAC항원，이로미낙（luminol）위발광저물，건립직접경쟁법검측분석래극다파알，병대포피액、포피조건、봉폐단백、부육시간、매용량등삼수진행우화，성공연제출화학발광법검측RAC시제합。해방법적선성검측범위가체0.1～10.0 ng/mL，선성상관계수r=0.998，IC50치위0.763 ng/mL；이론검측한소우0.1 ng/mL；비내변이계수7.62%~8.65%；비간변이계수8.15%；회수솔재85%±15%，여상견적극륜특라등기타β-격동제불발생교차반응。
A highly active monoclonal antibody was gained by making manmade antigen of Ractopamine and BSA conjugate. We es-tablished a direct competition method to detect Ractopamine that uses HRP as markers and luminol as chemiluminescent substrates. We optimized many parameters in this method, including coating buffer, coating condition, blocking condition, incubating time, and enzyme consumption. Based on this method we developed a chemiluminescent detecting kit. The linear range is 0.1~10.0 ng/mL, the linear correlation coefficient is r=0.998, the IC50 is 0.763 ng/mL, the detection limit is under 0.1 ng/mL, the within-run assay coeffi-cient of variation is 7.62%~8.65%, the between-run assay coefficient of variation is 8.15%, the coefficient of recovery is between 85%±15%, and there is no cross reaction with clenbuterol and other β-agonists.