CAJ | 학술논문

通过合成羧基化孔雀石绿,采用活泼酯法制备孔雀石绿免疫抗原CMG-BSA、检测抗原CMG-OVA.采用CMG-BSA免疫BALB/c小鼠,经融合、筛选和克隆化获得了一株稳定的抗孔雀石绿单克隆抗体,并建立了检测孔雀石绿的直接竞争ELISA方法.ELISA方法的最佳操作参数:孔雀石绿抗体稀释倍数1:16000(1mg/ml),酶标抗原(CMG-HRP)使用质量浓度0.2μg/ml.该方法具有良好的灵敏度和特异性,IC_(50)值0.3ng/ml,最低检测灵敏度达到0.05ng/ml,标准曲线方程为y=-20.888x+81.423(R~2=0.9813).
통과합성최기화공작석록,채용활발지법제비공작석록면역항원CMG-BSA、검측항원CMG-OVA.채용CMG-BSA면역BALB/c소서,경융합、사선화극륭화획득료일주은정적항공작석록단극륭항체,병건립료검측공작석록적직접경쟁ELISA방법.ELISA방법적최가조작삼수:공작석록항체희석배수1:16000(1mg/ml),매표항원(CMG-HRP)사용질량농도0.2μg/ml.해방법구유량호적령민도화특이성,IC_(50)치0.3ng/ml,최저검측령민도체도0.05ng/ml,표준곡선방정위y=-20.888x+81.423(R~2=0.9813).
Carboxyl-malachite green (CMG) was synthesized and immunogen was prepared by coupling CMG with carrier protein BSA through an ester activation method. BALB/c mice were subjected to injection of CMG-BSA and a hybridoma cell line secreting monoclonal antibody against MG was constructed after fusion, screening and cloning experiments. A direct competitive ELISA for detection MG was established with 0.2μg/ml CMG-HRP and 1:16000 of antibody dilution ratio. This method exhibited high sensitivity and excellent specificity for detecting MG. The standard regression equation of this method was y =- 20.888x + 81.423 (R~2 = 0.9813) with a detection limit for MG at the level of 0.05 ng/ml and IC_(50) was 0.3 ng/ml.