中国科学院杭州医学研究所谭蔚泓团队报道了，在稀缺样品中发现针对低丰度细胞表面蛋白的DNA适体的有效策略。相关研究成果发表在2024年9月19日出版的《美国化学会杂志》。

靶向细胞表面蛋白（如适配体）的分子识别探针，在精确诊断和治疗中起着至关重要的作用。然而，在细胞表面原位选择针对低丰度蛋白质的适配体，特别是在稀缺样本中，仍然是一个尚未解决的挑战。

该文中，研究人员提出了一种单轮、单细胞适配体选择方法，通过采用称为DiDS选择的数字DNA测序策略来解决这一难题。这种方法结合了每个DNA模板的分子识别卡，从而减轻了多重PCR扩增带来的偏差，并确保了适配体候选者的准确鉴定。

通过DiDS筛选，成功获得了一系列针对细胞系、临床标本和神经元的高质量适配体。随后的靶标鉴定分析表明，来自DiDS选择的适配体对不同丰度水平的蛋白质具有识别能力。相比之下，多轮选择只富集了一种针对高丰度靶标的适配体。此外，利用富集的适配体池在单细胞水平上对细胞表面进行全面分析，揭示了每种细胞系的独特分子模式。

这种简化的方法有望在广泛的表达水平上，快速生成靶向细胞表面蛋白的特异性识别分子，并扩大其在细胞分析、特异性探针鉴定、生物标志物发现等方面的应用。

附：英文原文

Title: Efficient Strategy to Discover DNA Aptamers Against Low Abundance Cell Surface Proteins in Scarce Samples

Author: Xiaoqiu Wu, Yuqing Liu, Dengwei Zhang, Jingjing Yu, Mingxin Zhang, Shuwei Feng, Lifei Zhang, Ting Fu, Yamin Tan, Tao Bing, Weihong Tan

Issue&Volume: September 19, 2024

Abstract: Molecular recognition probes targeting cell surface proteins such as aptamers play crucial roles in precise diagnostics and therapy. However, the selection of aptamers against low-abundance proteins in situ on the cell surface, especially in scarce samples, remains an unmet challenge. In this study, we present a single-round, single-cell aptamer selection method by employing a digital DNA sequencing strategy, termed DiDS selection, to address this dilemma. This approach incorporates a molecular identification card for each DNA template, thereby mitigating biases introduced by multiple PCR amplifications and ensuring the accurate identification of aptamer candidates. Through DiDS selection, we successfully obtained a series of high-quality aptamers against cell lines, clinical specimens, and neurons. Subsequent analyses for target identification revealed that aptamers derived from DiDS selection exhibit recognition capabilities for proteins with varying abundance levels. In contrast, multiple rounds of selection resulted in the enrichment of only one aptamer targeting a high-abundance target. Moreover, the comprehensive profiling of cell surfaces at the single-cell level, utilizing an enriched aptamer pool, revealed unique molecular patterns for each cell line. This streamlined approach holds promise for the rapid generation of specific recognition molecules targeting cell surface proteins across a broad range of expression levels and expands its applications in cell profiling, specific probe identification, biomarker discovery, etc.

DOI: 10.1021/jacs.4c03129

Source: https://pubs.acs.org/doi/abs/10.1021/jacs.4c03129