Aye Nang Noon Shean, Maraming Pornsuda, Tippayawat Patcharaporn, Daduang Sakda, Techasen Anchalee, Sithithaworn Paiboon, Rujanakraikarn Roengrut, Jearanaikoon Nichada, Phatthanakun Rungrueang, Supruangnet Ratchadaporn, Photongkam Pat, Daduang Jureerut
Biomedical Sciences Program, Graduate School, Khon Kaen University, Khon Kaen 40002, Thailand.
Centre for Research and Development of Medical Diagnostic Laboratories, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand.
ACS Omega. 2025 Mar 28;10(13):13621-13633. doi: 10.1021/acsomega.5c00633. eCollection 2025 Apr 8.
Bioconjugates in electrochemical biosensors can significantly enhance the detection process and sensitivity. In this study, we synthesized a monoclonal antibody-conjugated nanocomposite of graphene oxide and Nafion (GO-Nf-mAb) for application in an electrochemical biosensor as a novel all-in-one bioreceptor. The incorporation of Nafion (Nf) improved the stability, dispersity, and antibody immobilization on the GO surface, thereby increasing the sensitivity of the biosensor. The impact of Nafion on GO stability and antibody conjugation was thoroughly investigated and compared to that of Nafion-free conjugation via various characterization techniques, including X-ray photoelectron spectroscopy (XPS) and synchrotron radiation near-edge X-ray absorption fine structure. The presence of Nafion during monoclonal antibody (mAb) conjugation resulted in an increased peak intensity of the NH band in XPS analysis and the highest intensity of C=O groups in O-K edge analysis, indicating a greater yield of the antibody. This innovative electrochemical biosensor exhibited a low detection limit of 1.68 ng mL in spiked urine, a wide linear range, and high reproducibility, outperforming conventional detection methods for (OV) antigen detection. Our developed electrochemical biosensor introduces a novel and straightforward fabrication process using an all-in-one bioconjugate that serves as a bioreceptor, transducer, and blocking reagent simultaneously. Overall, this study offers a new insight on Nafion application in bioconjugation, and the GO-Nf-mAb conjugate-based electrochemical biosensor promises high sensitivity and a hassle-free immobilization process for OV antigen quantification.
生物共轭物在电化学生物传感器中可显著增强检测过程和灵敏度。在本研究中，我们合成了一种氧化石墨烯与全氟磺酸（GO-Nf-mAb）共轭的单克隆抗体纳米复合材料，用于电化学生物传感器，作为一种新型的一体化生物受体。全氟磺酸（Nf）的加入提高了氧化石墨烯表面的稳定性、分散性和抗体固定能力，从而提高了生物传感器的灵敏度。通过各种表征技术，包括X射线光电子能谱（XPS）和同步辐射近边X射线吸收精细结构，深入研究了全氟磺酸对氧化石墨烯稳定性和抗体共轭的影响，并与无全氟磺酸共轭进行了比较。在单克隆抗体（mAb）共轭过程中全氟磺酸的存在导致XPS分析中NH带的峰强度增加，以及O-K边分析中C=O基团的强度最高，表明抗体的产量更高。这种创新的电化学生物传感器在加标尿液中的检测限低至1.68 ng/mL，线性范围宽且重现性高，优于传统的（OV）抗原检测方法。我们开发的电化学生物传感器引入了一种新颖且简单的制造工艺，使用一种一体化生物共轭物同时作为生物受体、换能器和封闭试剂。总体而言，本研究为全氟磺酸在生物共轭中的应用提供了新的见解，基于GO-Nf-mAb共轭物的电化学生物传感器有望实现高灵敏度和用于OV抗原定量的无麻烦固定过程。
