Zhang Huimin, Yu Xiyuan, Liu Yilong, Lin Bingqian, Jiang Meng, Song Juan, Di Wen, Zhu Zhi, Yang Chaoyong
Institute of Molecular Medicine, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China.
MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, Key Laboratory for Chemical Biology of Fujian Province, State Key Laboratory of Physical Chemical of Solid Surfaces, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China.
Anal Chem. 2021 May 18;93(19):7235-7241. doi: 10.1021/acs.analchem.1c00330. Epub 2021 May 5.
Isolation and genetic analysis of circulating fetal cells from billions of maternal cells in peripheral blood are the cornerstone of fetal cell-based non-invasive prenatal testing. Inspired by the hierarchically multivalent architecture for enhanced capture of nature, an aptamer-based Hierarchically mUltivalent aNTibody mimic intERface (HUNTER) was designed with a tremendous avidity effect for highly efficient capture and non-destructive release of fetal cells. It was engineered by grafting Y-shaped DNA nanostructures to a linear polymer chain, creating a flexible polymer chain with bivalent aptamer side chains. This hierarchical arrangement of the aptamer ensures morphological complementarity, collective multiple-site interaction, and multivalent recognition between the aptamer and target cells. In combination with a deterministic lateral displacement (DLD)-patterned microdevice named as HUNTER-Chip, it achieves a binding affinity over 65-fold and a capture efficiency over 260%-fold due to the combination of hierarchically designed aptamers and frequent cell-ligand collision created by DLD. Moreover, a nuclease-assisted cell release strategy facilitates the release of fetal cells for gene analysis, such as fluorescence hybridization. With the advantages of high affinity, excellent capture efficiency, and compatible downstream analysis, the HUNTER-Chip holds great potential for non-invasive prenatal diagnosis.
从外周血数十亿母体细胞中分离循环胎儿细胞并进行基因分析，是基于胎儿细胞的无创产前检测的基石。受自然界增强捕获的分层多价结构启发，设计了一种基于适配体的分层多价抗体模拟界面（HUNTER），它对胎儿细胞具有高效捕获和无损释放的巨大亲和力效应。它是通过将Y形DNA纳米结构嫁接到线性聚合物链上构建而成，形成了具有二价适配体侧链的柔性聚合物链。适配体的这种分层排列确保了形态互补性、集体多位点相互作用以及适配体与靶细胞之间的多价识别。与名为HUNTER-Chip的确定性侧向位移（DLD）图案化微器件相结合，由于分层设计的适配体与DLD产生的频繁细胞-配体碰撞相结合，它实现了超过65倍的结合亲和力和超过260倍的捕获效率。此外，一种核酸酶辅助的细胞释放策略有助于释放胎儿细胞用于基因分析，如荧光杂交。凭借高亲和力、出色的捕获效率和兼容的下游分析等优点，HUNTER-Chip在无创产前诊断方面具有巨大潜力。
