基于3D纸芯片的肥大细胞传感器检测食品过敏原蛋白新方法研究

Food allergy has become a serious threat to human health. Due to the lack of effective treatment methods, early warning and detection for food allergens serves as the only way to avoid allergic disease. Therefore, it is of both important theoretical and practical significance to establish a detection and evaluation method for the allergenicity of food allergens, thereby further perfecting the food allergen assessment strategies. In this project, we intend to investigate the immunological mechanisms during the sensitive stage of food allergy, and analyze the rules relating to the identification of different food allergen proteins by the mast cells. On this basis, microfluidic paper chips are prepared through the wax printing and screen printing technology, while the 3D cell culture system has been developed with biocompatible sodium alginate and multifunctional graphene oxide. Besides, TN-XXL plasmid containing the gene of fluorescent protein is constructed to transfect RBL cells. In the following parts, with the RBL-2H3 cells employed as a recognition component, a delicate photoelectric mast cell sensing platform has been built for the detection and evaluation of food allergen protein through the combination of electrochemical capacitance and Förster resonance energy transfer (FRET). It would thus improve and enrich the food allergen monitoring and early warning system. This project will offer a scientific and theoretical basis for the development of new methods in food safety detection and biosensing.

食品过敏严重威胁着人体健康，由于目前尚无有效治疗的手段，提前检测和发现食品过敏原是避免过敏病症发生的唯一途径。因此建立针对食品过敏原蛋白致敏性的检测评价方法，对完善食品过敏原评估策略具有重要的理论和实际意义。本项目拟采用细胞传感技术研究食物过敏发敏阶段的免疫学机制，探索肥大细胞识别不同食品过敏原蛋白的规律，在此基础上以RBL-2H3细胞作为识别元件，通过蜡打印和丝网印刷技术制备微流控纸芯片，采用亲生物性的海藻酸钠和多功能的氧化石墨烯建立3D细胞培养系统，同时合成TN-XXL荧光质粒并转染细胞，构建具有精细光电传导功能的肥大细胞传感平台。分别应用电化学电容法和荧光能量共振转移法（FRET），开发和建立一套基于细胞传感的食品过敏原蛋白检测评价新方法，完善和丰富食品过敏原监控和预警体系。为生物传感和食品安全检测新技术、新方法的开发提供科学依据和理论基础。

由食品过敏原引起的过敏疾病严重威胁着人体健康，由于目前尚无有效的治疗手段，而提前检测和发现食品过敏原是避免过敏病症发生的唯一途径，因此针对食品过敏原开展监测预警和建立快精准的检测评价方法，对保障食品安全具有重要的理论和实践意义。本项目采用细胞传感技术研发食品过敏原检测评价新方法，探索细胞识别不同食品过敏原蛋白的机制和规律，并对三大类四种具有代表性的过敏原蛋白（花生过敏原蛋白Ara h2、大豆球蛋白、β-伴大豆球蛋白和牛乳酪蛋白）完成了分离和提取纯化。通过丝网印刷和蜡打印技术制备了不同种类的一次性、低成本、便携式的微流控纸芯片，合成了高亲生物性和高导电性的纳米材料（如海藻酸钠-氧化石墨烯、石墨烯-碳纳米纤维-甲基丙烯酰化明胶等复合材料），建立体外3D培养系统，以RBL-2H3细胞作为识别元件，结合电化学技术构建了多种高灵敏的肥大细胞传感器，实现了对花生过敏原蛋白，牛乳过敏原蛋白等精确检测。为生物传感和食品安全检测新技术、新方法的开发提供科学依据和理论基础。