Yao Lili, Feng Junmei, Zhou Yuefei, Gao Shengjie, Liu Shuai, Qu Hao, Mao Yu, Zheng Lei
School of Food and Biological Engineering, Hefei University of Technology, Hefei 230009, China.
Research (Wash D C). 2024 May 1;7:0372. doi: 10.34133/research.0372. eCollection 2024.
Circular aptamers are promising candidates for analytical and therapeutic applications due to their enhanced biological and structural stability. However, the process of circular aptamer selection remains a great challenge, as it requires multiple rounds of binding-separation-amplification that involves issues with nonspecific binding and amplification bias. Here, we develop a highly practical solution for reliable selection of circular aptamers in a single round based on magnetosome-like magnetic chain cross-linked graphene oxide (separation efficiency ≈ 10). High-affinity aptamer candidates can be rapidly selected from a preenriched circular DNA library, while low-affinity candidates are effectively adsorbed and separated by magnetosome-like magnetic chain cross-linked graphene oxide. With lipopolysaccharide as a representative model, the single-round selected lipopolysaccharide circular aptamer has been identified to have a high binding affinity with a value of low to nanomolar range. Using this method, circular aptamers for protein and small-molecule targets were also successfully generated. We envision that this approach will accelerate the discovery of various new circular aptamers and open up a new avenue for analytical and therapeutic studies.
由于具有增强的生物学和结构稳定性,环状适体在分析和治疗应用方面是很有前景的候选物。然而,环状适体的筛选过程仍然是一个巨大的挑战,因为它需要多轮的结合-分离-扩增,这涉及到非特异性结合和扩增偏差等问题。在此,我们基于类磁小体的磁链交联氧化石墨烯(分离效率≈10)开发了一种高度实用的解决方案,可在一轮中可靠地筛选环状适体。高亲和力的适体候选物可从预富集的环状DNA文库中快速筛选出来,而低亲和力的候选物则被类磁小体的磁链交联氧化石墨烯有效吸附和分离。以脂多糖作为代表性模型,已鉴定出单轮筛选的脂多糖环状适体具有高结合亲和力,解离常数在低至纳摩尔范围内。使用这种方法,还成功生成了针对蛋白质和小分子靶标的环状适体。我们设想这种方法将加速各种新型环状适体的发现,并为分析和治疗研究开辟一条新途径。