Productive screening of single aptamers with ddPCR
Productive screening of single aptamers with ddPCR
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使用 ddPCR 高效筛选单个适体
作者：
Jinyu Chen;Jinjun Wang;Zhaofeng Luo;Xiaona Fang;Lei He;Jianwei Zhu;Zahra Qurat ul ain;Jinlong He;Huan Ma;Haiyan Zhang;Minghou Liu;Liqun He
抗体现已广泛用于多种肿瘤的临床治疗。然而，存在与抗体治疗相关的严重问题，例如抗体与免疫系统的潜在相互作用以及长的生产周期。最近，已经发现适体在对某些蛋白质的亲和力和特异性方面具有与抗体相似的功能，并且由于其小尺寸而具有低免疫原性、易于化学合成和有效渗透到组织中而引起了广泛关注。然而，如何有效地获得高亲和力和高选择性的适体用于进一步的分析仍然是一个有待解决的问题。在此，报道了将连续流ddPCR技术与珠粒的流式细胞仪分选相结合的适体发现方法，使得我们获得了特异性结合PD-1的DNA适体，其亲和力比最佳报道方法高60倍以上。
Antibodies have now been widely used for clinical treatment of a number of tumors. However, there are serious problems associated with antibody therapy, such as potential interactions of antibodies with the immune system as well as long production cycles. Recently, aptamers have been found to function similar to antibodies in terms of affinity and specificity to certain proteins and are attracting much attention for their low immunogenicity, easy chemical synthesis, and efficient penetration into tissues due to their small size. However, how to access high affinity and selectivity aptamers efficiently for further analysis is still open to be resolved. Herein, an aptamer discovery method that combines the continuous flow ddPCR technology with cytometer sorting of beads is reported, such that we have obtained DNA aptamers binding specifically to PD-1 with an affinity of over 60-fold higher than that for the best-reported method.