Cetinkaya Ahmet, Kızılelma Bilal, Farhangzad Nazila, Dener-Kılıç Elif, Ozcelikay-Akyıldız Goksu, Özkan Tülin, Unal Mehmet Altay, Sunguroglu Asuman, Ozkan Sibel A
Gülhane Faculty of Pharmacy, Department of Analytical Chemistry, University of Health Sciences, Ankara, Türkiye.
Faculty of Pharmacy, Department of Analytical Chemistry, Hacettepe University, Ankara, Türkiye.
Anal Bioanal Chem. 2025 Aug;417(20):4695-4708. doi: 10.1007/s00216-025-05987-w. Epub 2025 Jul 21.
Cancer stem cells (CSCs) have recently emerged as important variables affecting the development or invasiveness of solid tumors. Prominin 1 (PROM1, CD133) is a frequently used CSC biomarker in various tumors, including lung cancer, and CD133 overexpression is linked to chemoresistance, cancer recurrence, and a poor prognosis. Furthermore, CD133 has a significant impact on tumor growth and metastasis. In this work, the detection of CD133 using a single-use multi-walled carbon nanotube (MWCNT)-supported immunoplatform was achieved for the first time. The immunoplatform was designed using an anti-CD133 antibody and the CD133 protein. Differential pulse voltammetry (DPV) measurements were performed between -0.20 V and 0.8 V using a 5.0 mM [Fe(CN)] redox probe. The optimization parameters influencing the immunoassay response were thoroughly evaluated, and an analytical assessment of the single-use immunoplatform for CD133 detection created with anti-CD133 was performed. The designed immunosensor exhibited a rapid response time (30 min). In addition, the limit of detection (LOD) was determined for the immunosensors developed using both the commercial CD133 antibody and the 7D7 monoclonal antibody (MoAb; anti-CD133 antibodies produced by the Faculty of Medicine), with values of 0.135 and 0.052 µg mL obtained, respectively, in standard solutions. The determination of CD133 in commercial serum samples was used to test the applicability of the developed immunoplatform in real samples, and the results were satisfactory. This study provides a rapid, selective, and sensitive platform for determining the CD133 biomarker, which can be used for the early detection of lung cancer and monitoring of its development. Moreover, in a next step, it could be integrated into the design of an interesting single-use device with point-of-care (POC) application feasibility for measurement of CD133 levels.
癌症干细胞（CSCs）最近已成为影响实体瘤发展或侵袭性的重要变量。Prominin 1（PROM1，CD133）是包括肺癌在内的各种肿瘤中常用的癌症干细胞生物标志物，CD133的过表达与化疗耐药、癌症复发及不良预后相关。此外，CD133对肿瘤生长和转移有重大影响。在本研究中，首次实现了使用一次性多壁碳纳米管（MWCNT）支撑的免疫平台检测CD133。该免疫平台采用抗CD133抗体和CD133蛋白设计而成。使用5.0 mM [Fe(CN)]氧化还原探针在-0.20 V至0.8 V之间进行差分脉冲伏安法（DPV）测量。全面评估了影响免疫分析响应的优化参数，并对用抗CD133构建的用于检测CD133的一次性免疫平台进行了分析评估。所设计的免疫传感器显示出快速响应时间（30分钟）。此外，分别测定了使用商业CD133抗体和7D7单克隆抗体（MoAb；医学院生产的抗CD133抗体）开发的免疫传感器的检测限（LOD），在标准溶液中获得的值分别为0.135和0.052 µg/mL。通过测定商业血清样本中的CD133来测试所开发免疫平台在实际样本中的适用性，结果令人满意。本研究提供了一个用于测定CD133生物标志物的快速、选择性和灵敏的平台，可用于肺癌的早期检测及其发展监测。此外，下一步，它可以集成到一个有趣的一次性设备设计中，该设备具有即时检测（POC）应用可行性，用于测量CD133水平。
