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.

中文翻译：

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Au/Na7PMo11O39结合磷脂表位印迹提高外泌体SERS检测效率的双边努力


检测体液中与癌症相关的外泌体已成为早期癌症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、高灵敏度的外泌体检测。第一步，采用磷脂极性位点印迹策略，使用磁性 PS-MIP（磷脂分子印迹聚合物）选择性地分离和富集尿液样本中的所有外泌体。第二步，利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）特性和过氧化物酶催化活性的 Au/NaPMoO 纳米颗粒（NP），然后将 CD9 抗体固定在 Au/NaPMoO 纳米颗粒的表面。然后使用 Au/NaPMoO-CD9 抗体复合物识别磁性 PS-MIP 富集的外泌体表面上的 CD9 蛋白。最后，通过Au/NaPMoO NPs的SERS活性和类过氧化物酶活性间接实现了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颗粒/mL，检测限为（ LOD）为 5.82 × 10 颗粒/mL。胰腺癌患者尿液样本中外泌体的SERS信号强度高于健康志愿者。这种双向 MIPs-NELISA-SERS 方法能够实现无创、高灵敏度、快速检测癌症，有利于治疗期间疾病进展的监测，并为快速早期癌症筛查开辟了新途径。