Pãun Angela Gabriela, Popescu Simona, Ungureanu Alisa Ioana, Trusca Roxana, Popp Alina, Dumitriu Cristina, Buica George-Octavian
Faculty of Applied Chemistry and Materials Science, National University of Science and Technology Politehnica Bucharest, 313 Splaiul Independentei, Sector 6, 060042 Bucharest, Romania.
National Institute for Mother and Child Health "Alessandrescu-Rusescu", 120 Lacul Tei Boulevard, Sector 2, 020395 Bucharest, Romania.
Biosensors (Basel). 2025 Jan 13;15(1):42. doi: 10.3390/bios15010042.
A novel electrochemical detection method utilizing a cost-effective hybrid-modified electrode has been established. A glassy carbon (GC) modified electrode was tested for its ability to measure electrochemical tTG antibody levels, which are essential for diagnosing and monitoring Celiac disease (CD). Tissue transglutaminase protein biomolecules are immobilized on a quantum dots-polypyrrole nanocomposite in the improved electrode. Initial, quantum dots (QDs) were obtained from Bombyx mori silk fibroin and embedded in polypyrrole film. Using carbodiimide coupling, a polyamidoamine (PAMAM) dendrimer was linked with GQDs-polypyrrole film to improve sensor sensitivity. The tissue transglutaminase (tTG) antigen was cross-linked onto PAMAM using N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC)-N-hydroxy succinimide (NHS) chemistry to develop a nanoprobe that can detect human serum anti-tTG antibodies. The physicochemical characteristics of the synthesized nanocomposite were examined by FTIR, UV-visible, FE-SEM, EDX, and electrochemical studies. The novel electrode measures anti-tissue antibody levels in real time using human blood serum samples. The modified electrode has great repeatability and an 8.7 U/mL detection limit. Serum samples from healthy people and CD patients were compared to standard ELISA kit assays. SPSS and Excel were used for statistical analysis. The improved electrode and detection system can identify anti-tissue antibodies up to 80 U/mL.
一种利用具有成本效益的混合修饰电极的新型电化学检测方法已经建立。对玻碳(GC)修饰电极测量电化学组织转谷氨酰胺酶抗体水平的能力进行了测试,该抗体水平对于乳糜泻(CD)的诊断和监测至关重要。在改进的电极中,组织转谷氨酰胺酶蛋白质生物分子固定在量子点 - 聚吡咯纳米复合材料上。首先,从家蚕丝素蛋白中获得量子点(QDs)并将其嵌入聚吡咯膜中。使用碳二亚胺偶联,将聚酰胺胺(PAMAM)树枝状大分子与石墨烯量子点 - 聚吡咯膜连接以提高传感器灵敏度。使用N - (3 - 二甲基氨基丙基) - N' - 乙基碳二亚胺盐酸盐(EDC) - N - 羟基琥珀酰亚胺(NHS)化学方法将组织转谷氨酰胺酶(tTG)抗原交联到PAMAM上,以开发一种能够检测人血清抗tTG抗体的纳米探针。通过傅里叶变换红外光谱(FTIR)、紫外 - 可见光谱、场发射扫描电子显微镜(FE - SEM)、能谱分析(EDX)和电化学研究对合成的纳米复合材料的物理化学特性进行了研究。该新型电极使用人血清样本实时测量抗组织抗体水平。修饰电极具有良好的重复性和8.7 U/mL的检测限。将健康人和CD患者的血清样本与标准酶联免疫吸附测定试剂盒检测结果进行了比较。使用SPSS和Excel进行统计分析。改进后的电极和检测系统能够识别高达80 U/mL的抗组织抗体。