高通量FcεRI-IgE电化学免疫感器检测食品中20种常见过敏原的研究

Food allergies can cause acute or chronic illnesses, and even life-threatening, seriously affecting the health of public life. Allergens in food are proteins from eight main groups - nuts and tree nuts, fish, shellfish, wheat, soy, eggs and milk - which trigger severe adverse reactions in the human body, involving IgE-type antibodies. Sensitive detection of allergens in a large variety of food matrices and appropriate labelling rules have become increasingly important considering the emergence of functional foods and food manufacturing technologies involving the use of allergen proteins. This project designed an electrochemical signal amplification system-FcεRI-IgE universal molecular linker and constructed a nanogold-FcεRI-IgE-allergen electrochemical immunosensor based on our previous results of biosensor research. The electrochemical multi-channel detection system was designed to solve the problem that the reproducibility of the immunosensor is poor and the standard curve is difficult to establish. In addition, on the basis of a large number of parallel detection data, it can also reduce external interference, improve the accuracy of results, and achieve high-throughput detection. This study is expected to provide a high-throughput, highly sensitive, and rapid detection solution for allergens in food.

食物过敏能够引起急性或慢性疾病,甚至威胁生命,严重地影响着公众的健康生活。鱼、蛋、奶等八大类食物是人们经常摄取的营养物质，在其富含的蛋白质中，含有多种致敏性的蛋白质。目前尚无治疗过敏的有效方法，所以对食品中潜在的过敏原进行检测和标识是避免食品过敏的必要手段。本项目在我们前期传感器研究的基础上，设计一种电化学信号放大系统-FcεRI-IgE通用分子接头，构建纳米金-FcεRI-IgE-过敏原电化学免疫传感器。结合设计的电化学多通道检测系统，解决免疫传感器重现性差、标准曲线建立难的问题。此外在大量平行检测数据的基础上还可以减少外界干扰、提高结果准确性，实现高通量检测。本研究有望为实现对多种食品过敏原同时进行高通量、高灵敏度的快速检测提供新的手段。

本项目基于HRP-NanoAu电化学信号放大系统偶联FcεRI-IgE通用分子接头，构建纳米金-FcεRI-IgE-过敏原电化学免疫传感器。通过构建SD大鼠过敏模型得到的8种常见食品过敏原（变应原）的抗血清IgE作为分子探针，分别测定其与8种蛋白类过敏原的联动变构常数Ka值。结果如下：来源于鸡蛋(Gallus domesticus)的卵类黏蛋白的Ka值为1.81×10-19 mol/L，卵清蛋白的Ka值为5.47×10-19 mol/L；来源于牛乳（Bos domesticus）的α-乳清蛋白的Ka值为2.51×10-19 mol/L，β-乳球蛋白的Ka值为3.93×10-18 mol/L，乳清蛋白的Ka值为1.25×10-19 mol/L，α-酪蛋白的Ka值为4.10×10-19 mol/L，β-酪蛋白的Ka值为1.02×10-18 mol/L，κ-酪蛋白的Ka值为1.59×10-19 mol/L。结果表明，该传感器在一定程度上模拟了体内带有FcεRI受体的肥大细胞在接触到变应原后所产生的超敏反应过程，并且通过这种方法测得的作用灵敏度与现有方法相比提高了近6个数量级。此外通过电化学多通道激励测试平台，设计了一套48通路的电信号检测平台实现高通量检测，为低剂量变应原导致的过敏反应提供了一个新的检测手段，同时为变应原诊断、食物变应原安全检测、消除过敏原工艺和效果评价等提供新的研究方法和思路。本项目共发表论文学术论文23篇，其中SCI收录论文12篇，EI收录4篇，中科院2022年JCR期刊分区：I区2篇，II区4篇；申请并受理专利1项；第1标注论文11篇；SCI总影响因子达到60.452。培养硕士研究生1名（获得学位），在读研究生7名。.