Department of Oncology, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing, 400038, China; School of Medical and Life Sciences, Chengdu University of Traditional Chinese Medicine, Chengdu, 611137, China.
School of Medical and Life Sciences, Chengdu University of Traditional Chinese Medicine, Chengdu, 611137, China.
Anal Chim Acta. 2023 Apr 22;1251:340984. doi: 10.1016/j.aca.2023.340984. Epub 2023 Feb 17.
Programmed death ligand-1 (PD-L1) positive exosomes (P-Exo) have been widely used for tumor diagnosis. However, accurate and rapid quantification of P-Exo remains challenging due to the heterogeneity of clinical individuals and isolation techniques. In this study, the triple-helix molecular probe (THMP) coupled with high-affinity silica-based TiO magnetic beads was used to isolate exosomes and to analyze the relative abundance of P-Exo in total exosomes (T-Exo). By employing this strategy, the entire analysis was completed within 70 min and the detection limit for P-Exo was 880 particles μL. Additionally, the relative abundance of P-Exo in T-Exo (RA) was calculated from their fluorescence ratio, which could avoid errors due to differences in samples and separation methods, and identify 1.5 × 10 P-Exo from 5 × 10 T-Exo per microliter. RA values were not only effective in distinguishing healthy volunteers from breast cancer patients, but also highly positively correlated with the stage of breast carcinoma. Overall, this strategy opens a new avenue for rapid and quantitative analysis of P-Exo, providing an opportunity for precise diagnosis and prediction of treatment efficacy in cancer.
程序性死亡配体 1（PD-L1）阳性外泌体（P-Exo）已被广泛用于肿瘤诊断。然而，由于临床个体和分离技术的异质性，P-Exo 的准确和快速定量仍然具有挑战性。在本研究中，采用三联螺旋分子探针（THMP）与高亲和力硅基 TiO 磁珠结合，分离外泌体并分析总外泌体（T-Exo）中 P-Exo 的相对丰度。通过采用该策略，整个分析在 70 分钟内完成，P-Exo 的检测限为 880 个颗粒/μL。此外，通过其荧光比计算 T-Exo 中 P-Exo 的相对丰度（RA），可以避免因样品和分离方法的差异而导致的误差，并能够从 5×10^6 个 T-Exo 中鉴定出 1.5×10^5 个 P-Exo。RA 值不仅能够有效地区分健康志愿者和乳腺癌患者，而且与乳腺癌的分期高度正相关。总体而言，该策略为 P-Exo 的快速定量分析开辟了新途径，为癌症的精确诊断和治疗效果预测提供了机会。
