State Key Laboratory of Supramolecular Structure and Materials, College of Chemistry, Jilin University, Changchun, 130012, PR China; Harbin Medical University, Department Organic Chemistry, College of Pharmacy, Baojian Rd 157, Harbin, 150081, Heilongjiang, PR China.
Center for Supramolecular Chemical Biology, State Key Laboratory of Supramolecular Structure and Materials, School of Life Sciences, Jilin University, Changchun, 130012, PR China.
Biosens Bioelectron. 2024 Aug 15;258:116349. doi: 10.1016/j.bios.2024.116349. Epub 2024 Apr 29.
Detection of cancer-related exosomes in body fluids has become a revolutionary strategy for early cancer diagnosis and prognosis prediction. We have developed a two-step targeting detection method, termed PS-MIPs-NELISA SERS, for rapid and highly sensitive exosomes detection. In the first step, a phospholipid polar site imprinting strategy was employed using magnetic PS-MIPs (phospholipids-molecularly imprinted polymers) to selectively isolate and enrich all exosomes from urine samples. In the second step, a nanozyme-linked immunosorbent assay (NELISA) technique was utilized. We constructed Au/NaPMoO nanoparticles (NPs) with both surface-enhanced Raman scattering (SERS) property and peroxidase catalytic activity, followed by the immobilization of CD9 antibodies on the surface of Au/NaPMoO NPs. The Au/NaPMoO-CD9 antibody complexes were then used to recognize CD9 proteins on the surface of exosomes enriched by magnetic PS-MIPs. Lastly, the high sensitivity detection of exosomes was achieved indirectly via the SERS activity and peroxidase-like activity of Au/NaPMoO NPs. The quantity of exosomes in urine samples from pancreatic cancer patients obtained by the PS-MIPs-NELISA SERS technique showed a linear relationship with the SERS intensity in the range of 6.21 × 10-2.81 × 10 particles/mL, with a limit of detection (LOD) of 5.82 × 10 particles/mL. The SERS signal intensity of exosomes in urine samples from pancreatic cancer patients was higher than that of healthy volunteers. This bidirectional MIPs-NELISA-SERS approach enables noninvasive, highly sensitive, and rapid detection of cancer, facilitating the monitoring of disease progression during treatment and opening up a new avenue for rapid early cancer screening.
体液中癌症相关外泌体的检测已成为癌症早期诊断和预后预测的革命性策略。我们开发了一种两步靶向检测方法，称为 PS-MIPs-NELISA SERS，用于快速和高灵敏度的外泌体检测。在第一步中，使用磁性 PS-MIPs（磷脂分子印迹聚合物）采用磷脂极性印迹策略选择性地从尿液样本中分离和富集所有外泌体。在第二步中，采用纳米酶联免疫吸附测定（NELISA）技术。我们构建了具有表面增强拉曼散射（SERS）性质和过氧化物酶催化活性的 Au/NaPMoO 纳米粒子（NPs），随后将 CD9 抗体固定在 Au/NaPMoO NPs 表面。然后，Au/NaPMoO-CD9 抗体复合物用于识别通过磁性 PS-MIPs 富集的外泌体表面的 CD9 蛋白。最后，通过 Au/NaPMoO NPs 的 SERS 活性和过氧化物酶样活性间接实现对外泌体的高灵敏度检测。通过 PS-MIPs-NELISA SERS 技术获得的胰腺癌患者尿液样本中外泌体的数量与 SERS 强度呈线性关系，范围为 6.21×10-2.81×10 个/mL，检出限（LOD）为 5.82×10 个/mL。胰腺癌患者尿液中外泌体的 SERS 信号强度高于健康志愿者。这种双向 MIPs-NELISA-SERS 方法能够实现非侵入性、高灵敏度和快速的癌症检测，有助于监测治疗期间疾病的进展，并为快速早期癌症筛查开辟了新途径。
