中文摘要：本研究开发了一种灵敏的基于纳米抗体的间接竞争酶联免疫 (Nb-ELISA)法用于测定赭曲霉毒素A（OTA），能高抗基体干扰。研究证明了纳米抗体是可靠的检测食物基质中的霉菌毒素的有前途的免疫分析试剂，并且在生物传感器的开发中很有潜力。
外文摘要：A sensitive indirect competitive nanobody-based enzyme linked immunosorbent assay (Nb-ELISA) for ochratoxin A (OTA) with high resistance to cereal matrix interference was developed. Nanobodies against OTA (Nb15, Nb28, Nb32, Nb36) were expressed in E. coli cells and their thermal stabilities were compared with that of an OTA-specific monoclonal antibody 6H8. All nanobodies could still retain their antigen-binding activity after exposure to temperature 95 °C for 5 min or to 90 °C for 75 min. Nb28 that exhibited the highest sensitivity in ELISA was selected for further research. An indirect competitive ELISA based on Nb28 was developed for OTA, with an IC50 of 0.64 ng/mL and a linear range (IC20-IC80) of 0.27–1.47 ng/mL. Cereal samples were analyzed following a 2.5 fold dilution of sample extracts, showing the good resistance to matrix interference of the Nb-ELSIA. The recovery of spiked cereal samples (rice, oats, barley) ranged from 80% to 105% and the Nb-ELISA results of OTA content in naturally contamined samples were in good agreement with those determined by a commercial ELISA kit. The results indicated the reliablity of nanobody as a promising immunoassay reagent for detection of mycotoxins in food matrix and its potential in biosensor development.
外文关键词：Nanobody；Ochratoxin A；Immunoassay；Cereal；Matrix interference
作者：Liu, X；Tang, ZW；Duan, ZH；He, ZY；Shu, M；Wang, XX；Gee, SJ；Hammock, BD；Xu, Y
作者单位：Hainan Univ
期刊名称：TALANTA
期刊影响因子：4.035
出版年份：2017
出版刊次：3
点击下载：用高抗基体干扰的纳米抗体酶免疫测定法检测谷物中的赭曲霉毒素A