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基于塑料抗体的新型电化学生物传感器的研制及其在癌症标志物 STEAP1 检测中的应用。

Development of a novel electrochemical biosensor based on plastic antibodies for detection of STEAP1 biomarker in cancer.

机构信息

BioMark Sensor Research/ISEP, School of Engineering, Polytechnic Institute, Porto, Portugal; CEB, Centre of Biological Engineering, Minho University, Braga, Portugal; CICS-UBI-Health Science Research Centre, University of Beira Interior, Av. Infante D. Henrique, 6200-506 Covilha, Portugal; Associate Laboratory i4HB-Institute for Health and Bioeconomy, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa, 2819-516 Caparica, Portugal.

BioMark Sensor Research/ISEP, School of Engineering, Polytechnic Institute, Porto, Portugal; CEB, Centre of Biological Engineering, Minho University, Braga, Portugal; Faculty of Engineering, Porto University, Portugal.

出版信息

Bioelectrochemistry. 2023 Aug;152:108461. doi: 10.1016/j.bioelechem.2023.108461. Epub 2023 May 9.

Abstract

STEAP1 is a cell surface protein of the STEAP family whose main function focuses on intercellular communication and cell growth. STEAP1 is considered a promising putative biomarker and a candidate target for prostate cancer treatment. For specific and selective detection of STEAP1, a molecularly imprinted polymers (MIP) was developed on a screen-printed electrode (C-SPE) whose surface was modified with a nanocomposite based on carbon nanotubes decorated with dendritic platinum nanoparticles (CNTs- PAH /Pt). Then, the MIPs were produced on the modified C-SPE by electropolymerization of a mixture of STEAP1 and a monomer (pyrrole-2-carboxylic acid). Then, the protein was removed from the polymeric network by enzymatic treatment with trypsin, which created the specific template cavities for further STEAP1 detection. Electrochemical techniques such as EIS and CV were used to follow the chemical modification steps of C-SPE. The analytical performance of the biosensor was evaluated by SWV in PBS buffer and in lysates of neoplastic prostate cancer cells (LNCaP) extracts. The MIP material showing a linear range from 130 pg/ml to 13 µg/ml. Overall, the biosensor exhibits essential properties such as selectivity, sensitivity and reproducibility for its application in medical and clinical research diagnosis and/or prognosis of prostate cancer.

摘要

STEAP1 是 STEAP 家族的细胞表面蛋白,其主要功能集中在细胞间通讯和细胞生长上。STEAP1 被认为是一种很有前途的潜在生物标志物和前列腺癌治疗的候选靶点。为了特异性和选择性地检测 STEAP1,在修饰有基于碳纳米管的纳米复合材料的丝网印刷电极 (C-SPE) 上开发了一种分子印迹聚合物 (MIP),该复合材料表面修饰有树枝状铂纳米粒子的聚电解质 (CNTs-PAH/Pt)。然后,通过将 STEAP1 和单体 (吡咯-2-羧酸) 的混合物电聚合在修饰后的 C-SPE 上生成 MIPs。然后,通过用胰蛋白酶进行酶处理从聚合物网络中去除蛋白质,这为进一步检测 STEAP1 创建了特定的模板腔。电化学技术,如 EIS 和 CV,用于跟踪 C-SPE 的化学修饰步骤。通过在 PBS 缓冲液和肿瘤前列腺癌细胞 (LNCaP) 提取物的裂解物中进行 SWV 来评估生物传感器的分析性能。MIP 材料在 130pg/ml 至 13μg/ml 的线性范围内表现出线性范围。总的来说,该生物传感器表现出选择性、灵敏度和重现性等基本特性,可用于医学和临床研究诊断和/或前列腺癌的预后。

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