摘要
旨在制备伏马菌素B1单克隆抗体，并建立伏马菌素B，免疫学检测方法。以制备的免疫原FB1-BSA免疫小鼠，利用细胞融合技术建立能分泌抗FB，抗体的杂交瘤细胞株，体内诱生腹水的方法制备FB，单抗。基于制备的FB。单抗，建立间接竞争ELISA检测分析方法，并对检测方法性能进行了初步鉴定。结果表明，通过动物免疫、细胞融合筛选到1株分泌抗FB，抗体的杂交瘤细胞株3F6。建立的间接竞争ELISA检测分析方法检测范围40-600ng／mL之间，检测下限达到40ng／mL，半数抑制浓度IC50为136．81ng／mL，除与FBl特异性反应外，与同系物FB2及FB3交叉反应率分别为11．65％和7．85％，与黄曲霉毒素B1、黄曲霉毒素M1、β-玉米赤霉烯醇、呕吐毒素、T-2毒素、玉米赤酶烯酮、赭曲霉毒素A、玉米赤霉酮、α-玉米赤霉醇交叉反应率均低于0．2％，样品回收率在88．89％到115．45％之间，平均101．91％，变异系数为7．43％。本研究建立的ELISA方法与LC—MS—MS检测相同的样品时，二者的检测结果没有显著差异（P〉0．05）。本研究初步研发出灵敏度符合检测要求、特异较强、准确度较高、简便快捷的FBl免疫学检测方法。
The study was aimed at preparation of monoclonal antibodies of fumonisin B1 （ FB1 ） and establishing of immunoassay methods. The mice were immunized by irnmunogen FBl - BSA, and hybridoma cell line secreting monoclonal antibody against FB1 （FB1 mAb） was prepared by cell fusion technology, while FBl monoclonal antibody was prepared by in vivo induction. The indirectly competitive ELISA was established by the prepared FB1 mAb, with a preliminary evaluation. The results showed that one hybridoma cell line 3F6 could be obtained by animal immuni- zing and cell fusion. The indirectly competitive ELISA had a detection range between 40 ng/mL and 600 ng/mL, and the detection limit was 40 ng/mL; 50% inhibitory concentration （ICs0） was 136.81 ng/mL. The method has specific reaction with FB1 , the cross - reactivity ratio with FB2, FB3 and the ratio with other mycotoxins （ aflatoxin B1, aflatoxin M1, β- zearalenol, deoxynivalenol, T-2 toxin, zearalenone, ochratoxin A, zearalanone, zeranol） were 11.65% , 7.85% and less than 0.2% respectively. The recovery ranged from 88.89% to 115.45% , for an average as 101.91% ; and the CV was 7.43%. The same samples were assayed by LC -MS -MS and ELISA. There were no significant difference between the two methods （ P 〉 0.05 ）. In a conclusion, the sensitivity, specificity, accura- cy, simple and efficient FB1 immunological detection method was developed.
出处
《中国粮油学报》
EI
CAS
CSCD
北大核心
2015年第7期116-122,共7页
Journal of the Chinese Cereals and Oils Association
基金
公益性行业(农业)科研专项(201203040)
河南省农业科学院优秀青年科技基金(2013YQ29)
