抗伏马菌素B_1单克隆抗体的制备及试剂盒的研制

目的为了能够快速检测粮谷类食品中伏马菌素B_1的污染水平,研制具有我国自主知识产权的针对伏马菌素B_1快速检测试剂盒。方法制备了能分泌抗伏马菌素B_1单克隆抗体的杂交瘤细胞株,建立了敏感、特异、快速的酶联免疫吸附试验检测方法,最终形成具有中国自主知识产权的伏马菌素B_1快速检测试剂盒。结果该试剂盒所用单克隆抗体的Ig亚类为IgG1,亲和常数为8·3×10-8mol/L。该抗体与其他结构类似物无明显交叉反应,具有较高的特异性。试剂盒对伏马菌素B_1最低检出限5ng/ml,标准曲线的线性范围(50~500ng/ml),回归方程^Y=-0·582X+1·793(r=0·99,P<0·05);对玉米作50ng/ml,200ng/ml和500ng/ml3个加标浓度的回收率在71·89%~112·95%之间;试剂盒在常温状态下有效期至少10个月;实验室内的变异系数和实验室间的变异系数均小于20%。

Development of monoclonal antibody against Fumonisin B1 and ELISA-kit of quantitative analysis for Fumonisin B1

OBJECTIVE: To detect the contamination level of fumonisin B1 in grain, and to develop rapid detection kit that possess patent of China. METHODS: Hybridoma cell line excreting monoclonal antibody against fumonisin B, was produced using B cell hybridoma technique and develop a rapid, sensitive, quantitative ELISA-kit for detection fumonisin B1. RESULTS: The monoclonal antibody used in the ELISA-kit were tested for subtype as IgG1 and its affinity constant was 8.3 x 10(-8) mol/L. The monoclonal antibody obtained in the present study was highly specific to fumonisin B1 , because no cross reactions between the monoclonal antibodies against fumonisin B1 with the analogues of fumonisin B, were found. The limited concentration of the ELISA-kit was 5 ng/ml, linear range was 50-500 ng/ml, the linear equation was Y = -0.582 x +1.793( = 0.99, P < 0.05). The recovery rate of maize on the level of 50n g/ml, 200 ng/ml and 500 ng/ml was among 71.89%-112.95%. The kit can be stored at normal tempertature in ten months at least. The coefficient of variant winthin-laboratory and between-laboratory was less than 20%.