Zhang Yingming, Zhang Shikun, Ning Zhiyuan, Duan Mengxia, Lin Xianfeng, Duan Nuo, Wang Zhouping, Wu Shijia
State Key Laboratory of Food Science and Resources, Jiangnan University, Wuxi, China.
School of Food Science and Technology, Jiangnan University, Wuxi, China.
Crit Rev Anal Chem. 2024 Oct 8:1-21. doi: 10.1080/10408347.2024.2411244.
Aptamers, as short single-stranded nucleic acids, can bind to targets in a similar way to antibodies. Relying on the advantages of low cost, high stability, and flexibility, they are widely applied in biosensors, disease therapy, and synthetic biology. As an aptamer screening method, the Systematic Evolution of Ligands by Exponential Enrichment (SELEX) offers almost unlimited possibilities for functional aptamer generation. However, at present, the SELEX procedure has not reached a satisfactory level, and it still faces some challenges in practical application, such as the relatively blind initial library, laborious and time-consuming selection process, typically requires 9-20 rounds for screening, and the entire process generally extends over 2-3 months, and sub-optimal performance of aptamers obtained. In the past few years, researchers have made great efforts to address these obstacles. Hence, in this review, we first summarize the aptamer screening mechanism and the existing limitations of SELEX. Then analyze the principle and technical key points of the SELEX optimization screening strategy. By incorporating rational library design, novel screening awareness, and advanced screening equipment, the number of aptamer screening cycles is significantly reduced to <8 rounds, with some methods achieving single-round screenings. This has led to a considerable decrease in the overall screening time to <3 weeks, while simultaneously enhancing the performance of the aptamers. Finally, critically discuss the present challenges and future directions of aptamer screening. This review aims to provide a practical reference for designing suitable aptamer screening methods.
适体作为短单链核酸，能以与抗体相似的方式结合靶标。凭借低成本、高稳定性和灵活性等优势，它们在生物传感器、疾病治疗和合成生物学中得到广泛应用。作为一种适体筛选方法，指数富集配体系统进化技术（SELEX）为功能性适体的产生提供了几乎无限的可能性。然而，目前SELEX程序尚未达到令人满意的水平，在实际应用中仍面临一些挑战，如初始文库相对盲目、筛选过程费力且耗时，通常需要9至20轮筛选，整个过程一般持续2至3个月，以及所获得适体的性能欠佳。在过去几年中，研究人员已做出巨大努力来克服这些障碍。因此，在本综述中，我们首先总结适体筛选机制以及SELEX的现有局限性。然后分析SELEX优化筛选策略的原理和技术要点。通过结合合理的文库设计、新颖的筛选思路和先进的筛选设备，适体筛选循环次数显著减少至<8轮，有些方法甚至实现了单轮筛选。这使得总体筛选时间大幅缩短至<3周，同时提高了适体的性能。最后，批判性地讨论适体筛选当前面临的挑战和未来方向。本综述旨在为设计合适的适体筛选方法提供实用参考。
