Zhang Yingming, Zhang Shikun, Ning Zhiyuan, Lin Xianfeng, Duan Nuo, Wang Zhouping, Wu Shijia
State Key Laboratory of Food Science and Resources, Jiangnan University, Wuxi 214122, China.
School of Food Science and Technology, Jiangnan University, Wuxi 214122, China.
J Agric Food Chem. 2024 Dec 18;72(50):28148-28156. doi: 10.1021/acs.jafc.4c10043. Epub 2024 Dec 4.
β-Conglycinin is the main allergen present in soybeans, and it is causing wide concern due to its notable allergenicity, heat, and digestive enzyme resistance. Screening for aptamers that both recognize β-conglycinin and inhibit the allergic reactions that it triggers is necessary. Conventional aptamer screening is labor-intensive, requires skilled personnel, and has limited reproducibility. To address these limitations, an automated device was developed to enhance the efficiency of aptamer selection in Capture-SELEX. The device achieves highly integrated, reproducible, and accurate contamination control. Using this device, a high-affinity and specific aptamer, β-5, was selected with a = 18.24 ± 2.42 nM for β-conglycinin, as confirmed by isothermal titration calorimetry and fluorescence polarization. Thermodynamic analysis revealed that enthalpy-driven binding and docking simulations clarified the recognition mechanism. Overall, this automated device enables high-efficiency aptamer generation for certain targets, with aptamer β-5 expected to play a vital role in the detection of β-conglycinin and the targeted inhibition of its allergic reaction.
β-伴大豆球蛋白是大豆中的主要过敏原，因其显著的致敏性、耐热性和抗消化酶性而备受关注。筛选既能识别β-伴大豆球蛋白又能抑制其引发的过敏反应的适体是必要的。传统的适体筛选工作强度大，需要技术熟练的人员，且重现性有限。为解决这些局限性，开发了一种自动化设备，以提高Capture-SELEX中适体选择的效率。该设备实现了高度集成、可重现且准确的污染控制。使用该设备，筛选出了一种高亲和力和特异性的适体β-5，等温滴定量热法和荧光偏振法证实其对β-伴大豆球蛋白的解离常数为18.24±2.42 nM。热力学分析表明，焓驱动的结合和对接模拟阐明了识别机制。总体而言，这种自动化设备能够高效生成针对特定靶标的适体，适体β-5有望在β-伴大豆球蛋白的检测及其过敏反应的靶向抑制中发挥重要作用。
