癌细胞中血管内皮生长因子165（VEGF165）的过度表达通过促进肿瘤过度快速增殖和分裂而在促进肿瘤转移中发挥关键作用。因此，开发具有高灵敏度和抗干扰性的分析方法对于VEGF165的检测势在必行。多种类型的适体传感器已被设计用于 VEGF165 检测；然而，这些生物传感器的性能可能会受到未结合适体构象变化引起的非目标信号的影响。该论文展示了一种精确、灵敏的荧光生物传感器的创建，旨在通过使用 VEGF165 特异性分裂适体来检测 VEGF165。此外，该生物传感器采用切口酶辅助 DNA walker 与 CRISPR-Cas12a 耦合来实现双信号放大。 VEGF165 校准曲线显示检测限为 268 fM，并且具有 5 至 4000 nM 的宽线性范围。利用荧光生物传感器检测人血清和细胞匀浆样品中的 VEGF165，取得了良好的结果。该创新设计可作为概念验证，并展示出检测各种目标的巨大潜力。版权所有 © 2024 Elsevier B.V. 保留所有权利。

The overexpression of vascular endothelial growth factor 165 (VEGF165) in cancer cells plays a pivotal role in promoting tumor metastasis by facilitating their excessively rapid proliferation and division. Hence, the development of analytical methods possessing high sensitivity and resistance to interference is imperative for the detection of VEGF165. Various types of aptasensors have been devised for VEGF165 detection; however, the performance of these biosensors can be influenced by non-target signals caused by conformational changes in unbound aptamers. The paper shows the creation of a precise and sensitive fluorescence biosensor designed to detect VEGF165 by using a VEGF165-specific split aptamer. Additionally, this biosensor employs nicking enzyme-assisted DNA walker coupling with CRISPR-Cas12a to achieve dual-signal amplification. The VEGF165 calibration curve shows a detection limit of 268 fM and has a broad linear range from 5 to 4000 nM. The fluorometric biosensor was utilized to detect VEGF165 in human serum and cellular homogenate samples, yielding good outcomes. The innovative design serves as proof of concept and demonstrates significant potential in detecting various targets.Copyright © 2024 Elsevier B.V. All rights reserved.