摘要
目的 建立应用单克隆抗体的伏马菌素B1（fumonisin B1，FB1）间接竞争酶联免疫吸附检测方法。方法 利用B淋巴细胞杂交瘤技术制备抗FB1单克隆抗体，建立FB1间接竞争酶联免疫吸附方法。结果 建立稳定分泌抗FB1毒素单克隆抗体的杂交瘤细胞株，分泌的抗体为IgG1亚类，分子量150KD，腹水稀释度为1：2．0×10^8，亲和常数为6．72×10^9L／mol。FB1间接竞争酶联免疫吸附方法的最低检出浓度为5μg／L，校正曲线的线性范围50～500μg/L，线性方程Y=-0．582X＋1．793（r=0．99，P〈0．05）；与脱氧雪腐镰刀菌烯醇、玉米赤霉烯酮、T-2毒素均无交叉反应。回收率71．89％～112．95％，平均回收率100．23％。结论 所建立的检测方法具有快速、简便、灵敏的特点，可满足实际工作需要。
Objective To develop enzyme - linked immunosorbant assay for detection of FB1 which is based on monclonal antibodies. Methods To produce hybridoma cell lines excreting monoclonal antibodies against FB1 by using B cell hybridoma technique and develop enzyme - linked immunosorbant assay for detection of FB1. Results Hybridoma cell lines excreting monoclonal antibodies were developed and monoclonal antibodies against FB1 were prepared. Monoclonal antibodies produced by the hybridoma cells were tested for subtypes and designated as IgG1, the molecular weight was 150kD, the titer was 1 ： 2.0 108, the affinity constant was 6.72 × 10^9 L/tool. The limited concentration of detection was 5μg/L, linear range was50 500 μg/L, the linear equation was Y = - 0. 582x ＋ 1. 793, （ r = 0.99, P 〈 0.05 ）. There was no cross reaction with DON, ZEN, T-2 toxin. The recovery rate of spiked maize was among 71.89% - 112.95%, and mean recovery rate was 100. 23 %. Conclusion The detection method is simple, rapid, sensitive and can meet the demand of practical work.
出处
《中国公共卫生》
CAS
CSCD
北大核心
2006年第7期840-842,共3页
Chinese Journal of Public Health
基金
国家"十五"科技重大专项(2001BA804A20
2001BA804A3205)
关键词
伏马菌素B1
单克隆抗体
酶联免疫吸附试验
fumonisin B1
monoclonal antibody
enzyme-linked immunsorbant assay（ELISA）
