Mobed Ahmad, Kohansal Fereshteh, Dolati Sanam, Hasanzadeh Mohammad
Infectious and Tropical Diseases Research Center, Tabriz University of Medical Sciences Tabriz Iran.
Physical Medicine and Rehabilitation Research Center, Aging Research Institute, Tabriz University of Medical Sciences Tabriz Iran
RSC Adv. 2022 Oct 5;12(44):28473-28488. doi: 10.1039/d2ra04984h. eCollection 2022 Oct 4.
Hyaluronic Acid (HA) is a non-sulfated glycosaminoglycan, which is a potential biomarker that could be evaluated in the diagnosis of some cancers. For the first time, a novel label-free electrochemical immunosensor was developed based on modified ITO-PET (indium tin oxide-polyethylene terephthalate) electrodes for the sensitive recognition of hyaluronic acid (HA) in real samples. A disposable ITO-coated PET electrode was modified with gold nanoparticles (AuNPs) to construct a suitable substrate for the efficient immobilization of biotinylated antibodies of HA. Importantly, the encapsulation of biotinylated antibody of HA in KCC1-NH-CS was performed successfully, which was another innovative part of this bio-device construction. For determining the immobilization steps and optimization of the biosensor, electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) techniques were used. Furthermore, the morphological characterization of each ITO electrode surface was performed by field emission scanning electron microscopy (FESEM). Specific binding of gold nanoparticles supported CTAB to ITO-PET and its bioconjugation with the biotinylated antibody of HA was studied using the electroanalysis of the sensor performance. For the better performance of the antibody to generate an immunocomplex with HA (antigen), its encapsulation was performed, which led to the excellent behavior of the immunosensor. The proposed HA immunosensor indicated excellent reproducibility, high selectivity, and long-term stability. The HA electrochemical immunosensor performed perfectly with a wide determination range (0.078 to 160 ng mL) and a low limit of quantification (0.078 ng mL) in human plasma samples. It is recommended that the designed biosensor can be used as a diagnostic tool in clinical bioassays in the near future.
透明质酸（HA）是一种非硫酸化糖胺聚糖，它是一种潜在的生物标志物，可用于某些癌症的诊断评估。首次基于修饰的ITO-PET（氧化铟锡-聚对苯二甲酸乙二酯）电极开发了一种新型无标记电化学免疫传感器，用于灵敏识别实际样品中的透明质酸（HA）。用金纳米颗粒（AuNPs）修饰一次性ITO涂层PET电极，以构建适合高效固定HA生物素化抗体的基质。重要的是，成功地将HA生物素化抗体封装在KCC1-NH-CS中，这是该生物装置构建的另一个创新部分。为了确定固定步骤并优化生物传感器，使用了电化学阻抗谱（EIS）和循环伏安法（CV）技术。此外，通过场发射扫描电子显微镜（FESEM）对每个ITO电极表面进行形态表征。利用传感器性能的电分析研究了金纳米颗粒负载的CTAB与ITO-PET的特异性结合及其与HA生物素化抗体的生物共轭。为了使抗体更好地与HA（抗原）生成免疫复合物，对其进行了封装，这导致免疫传感器具有优异的性能。所提出的HA免疫传感器具有出色的重现性、高选择性和长期稳定性。该HA电化学免疫传感器在人血浆样品中表现出色，测定范围宽（0.078至160 ng/mL），定量下限低（0.078 ng/mL）。建议在不久的将来，所设计的生物传感器可作为临床生物测定中的诊断工具。
