基于“抗体复制”策略的分子印迹微阵列芯片的构建及中药饮片中黄曲霉毒素B1的快速检测

The harmful residues in Chinese herbal pieces are common and mycotoxins need to be controlled strictly because of severe danger to human health. The immunoassays used in mycotoxin analysis for extensive screen and field test suffer from poor thermal and pH stability, short lifetime and high cost. A viable alternative would be the use of molecularly imprinted polymers (MIPs) endowed with high affinity for their target molecules. However, the problems like the toxicity and price of templates such as mycotoxins and decreasing selectivity with dummy templates still remains a challenge. From the results we obtained on the research of MIPs, we put forward the double imprinting strategy of “antibody imprinting – transcription imprinting” to obtain antibody replica with monoclonal antibody of AFB1 as the template. The trace AFB1 in Chinese herbal pieces could be detected fast via the interaction between the antibody replica and corresponding antigen AFB1. The antibody replica have considerable advantages over antibodies as they possess a higher chemical, physical, and thermal stability, low cost, and still keep the same specific selectivity as antibodies. The analysis methods with the antibody replica as the recognition elements are sensitive, accurate and stable. The absent of toxic templates and reuse of antibodies in the preparation of antibody replica also make the analysis green and economic.

中药饮片中外源性有害残留物存在普遍，真菌毒素因毒性极大痕量污染需严格控制。目前大量筛查及现场检测常用的免疫分析法存在抗体获取不易、性质不稳定、结果重现性差等不足，被称为“塑料抗体”的MIPs在真菌毒素的选择性富集及模拟生物传感检测方面优势凸显，但模板分子毒性大、价格贵，替代模板对目标分子的选择性低等问题难以突破。基于前期对MIPs认识及实践，本项目提出以AFB1的单克隆抗体为模板，通过“抗体印迹-转录印记”的策略，将AFB1的抗体进行复制，通过复制抗体与抗原AFB1之间的特异性结合，探讨二者之间相互作用的规律，以期实现中药饮片中AFB1的痕量分析。制备中不需使用有害分析物，且抗体可重复利用，绿色、经济；以复制抗体为识别元件，灵敏、准确、稳定性好。