Department of Emergency Medicine, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung 83301, Taiwan.
Department of Engineering Science, National Cheng Kung University, Tainan 70101, Taiwan.
Biosensors (Basel). 2024 Nov 10;14(11):545. doi: 10.3390/bios14110545.
Rheumatoid arthritis (RA) is a chronic autoimmune disorder that causes extensive damage to multiple organs and tissues and has no known cure. This study introduces a microfluidic detection platform that combines a microfluidic reaction chip with a micro-spectrometer to accurately detect the anti-cyclic citrullinated peptide antibody (anti-CCP Ab) biomarker, commonly associated with arthritis. The surface of the microfluidic reaction chip is functionalized using streptavidin to enable the subsequent immobilization of biotinylated-labeled cyclic citrullinated peptide (biotin-CCP) molecules through a streptavidin-biotin reaction. The modified chip is then exposed to anti-CCP Ab, second antibody conjugated with horseradish peroxidase (HRP) (2nd Ab-HRP), 3,3',5,5'-tetramethylbenzidine (TMB), and a stop solution. Finally, the concentration of the anti-CCP Ab biomarker is determined by analyzing the optical density (OD) of the colorimetric reaction product at 450 nm using a micro-spectrometer. The detection platform demonstrated a strong correlation (R = 0.9966) between OD and anti-CCP Ab concentration. This was based on seven control samples with anti-CCP Ab concentrations ranging from 0.625 to 100 ng/mL. Moreover, for 30 artificial serum samples with unknown anti-CCP Ab concentrations, the biosensor achieves a correlation coefficient of (R = 0.9650). The proposed microfluidic detection platform offers a fast and effective method for accurately identifying and quantifying the anti-CCP Ab biomarker. Thus, it offers a valuable tool for the early diagnosis and monitoring of RA and its progression in point-of-care settings.
类风湿性关节炎（RA）是一种慢性自身免疫性疾病，可导致多个器官和组织广泛受损，目前尚无已知的治愈方法。本研究介绍了一种微流控检测平台，该平台将微流控反应芯片与微光谱仪相结合，可准确检测与关节炎相关的抗环瓜氨酸肽抗体（anti-CCP Ab）生物标志物。微流控反应芯片的表面通过链霉亲和素进行功能化，使随后通过链霉亲和素-生物素反应固定生物素化标记的环瓜氨酸肽（biotin-CCP）分子。然后将修饰后的芯片暴露于抗-CCP Ab、辣根过氧化物酶（HRP）（2nd Ab-HRP）、3,3',5,5'-四甲基联苯胺（TMB）和终止溶液中。最后，通过使用微光谱仪分析比色反应产物在 450nm 处的光密度（OD）来确定抗-CCP Ab 生物标志物的浓度。该检测平台在七个具有 0.625 至 100ng/mL 抗-CCP Ab 浓度的对照样本之间显示出 OD 和抗-CCP Ab 浓度之间的强相关性（R = 0.9966）。此外，对于 30 个未知抗-CCP Ab 浓度的人工血清样本，生物传感器的相关系数为（R = 0.9650）。该微流控检测平台提供了一种快速有效的方法，可以准确识别和定量抗-CCP Ab 生物标志物。因此，它为 RA 的早期诊断和监测以及在即时护理环境中监测其进展提供了有价值的工具。
