Development of an Automated Capture-SELEX Device for Efficient Screening of β-Conglycinin Aptamer
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 <i>K</i><sub>d</sub> = 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.
Authors
- Nuo Duan (ORCID: https://orcid.org/0000-0001-6493-4951)
- Zhiyuan Ning (ORCID: https://orcid.org/0009-0000-4718-1564)
- Zhouping Wang (ORCID: https://orcid.org/0000-0002-3868-8125)
- Xianfeng Lin (ORCID: https://orcid.org/0000-0002-7699-1395)
- Shijia Wu (ORCID: https://orcid.org/0000-0002-0736-6641)
- Yingming Zhang
- Shikun Zhang (ORCID: https://orcid.org/0000-0002-1321-1515)
Institutions
- Jiangnan University (CN)
- State Key Laboratory of Food Science and Technology (CN)
Publication Details
- Journal
- Journal of Agricultural and Food Chemistry
- Published
- 2024-12-04
- DOI
- https://doi.org/10.1021/acs.jafc.4c10043
- Citations
- 8
- Primary Topic
- Advanced biosensing and bioanalysis techniques
- Type
- article
- Field-Weighted Citation Impact
- 1.87
Funders
- National Key Research and Development Program of China
- Fundamental Research Funds for the Central Universities
- Collaborative Innovationcenter of Food Safety and Quality Control in Jiangsu Province
- Natural Science Foundation of Jiangsu Province for Distinguished Young Scholars