在本文中，我们提出并制备了一种多功能微流控芯片，集成了离心分离区、水相双分相系统（ATPS）混合区和富集检测区。设计了自动高效的全血样中血管内皮生长因子165（VEGF165）的分离和定量分析方法，应用该微流控芯片实现。首先在离心分离区内将血样分为血细胞和血浆两部分，然后在ATPS混合区内，将血浆与含有Apt-Au NP纳米探针的PEG/KH2PO4水相双分相溶液混合。最后，在富集检测区内，将混合物分离在CN140上，并利用ZnO NP-anti VEGF165 纳米结构对其进行修饰。Apt-Au NPs捕获到的VEGF165分布在PEG相中，集中在CN140的前端，并与抗VEGF165结合形成夹心结构。通过荧光共振能量转移技术，在纳米探针上的罗丹明B和Au NPs之间完成对VEGF165的敏感检测。在优化的旋转程序下，毛细和离心力推动整个前处理和检测过程中的流体。该方法检测的线性范围为1 pg mL-1至50 ng mL-1，血液中VEGF165的检测限为0.22 pg mL-1，富集效率为983。这表明，基于该多功能微流控芯片的肿瘤标记物检测为临床癌症的早期筛查和预后提供了便利可靠的方法，并具有潜在的价值。

In this paper, a multifunctional microfluidic chip integrated with a centrifugal separation zone, aqueous two-phase system (ATPS) mixing zone and enrichment detection zone was proposed and fabricated. An automatic and efficient separation and quantitative analysis method for vascular endothelial growth factor 165 (VEGF165) in whole blood samples was established with the designed microfluidic chip. A blood sample was divided into blood cells and plasma in the centrifugation zone. In the ATPS mixing zone, plasma was mixed with PEG/KH2PO4 aqueous two-phase solution containing Apt-Au NP nanoprobes. In the enrichment detection zone, the mixture was separated on CN140 modified with a ZnO NP-anti VEGF165 nanostructure. The VEGF165 captured by Apt-Au NPs was distributed in the PEG phase, concentrated at the front of CN140 and combined with anti-VEGF165 to form a sandwich structure. The sensitive detection of VEGF165 was achieved through fluorescence resonance energy transfer between rhodamine B and Au NPs on the nanoprobe. Under the optimized rotation program, capillary and centrifugal forces propelled the fluid in the whole process of pretreatment and detection. The detection linear range was between 1 pg mL-1 and 50 ng mL-1, the detection limit of VEGF165 in blood was 0.22 pg mL-1 and the enrichment efficiency was 983. It was illustrated that a convenient and reliable way for detection of tumor markers based on the multifunctional microfluidic chip was provided and it has a potential value for early screening and prognosis of clinical cancer.