Rapid Microfluidic Biosensor for Point-of-Care Determination of Rheumatoid Arthritis via Anti-Cyclic Citrullinated Peptide Antibody Detection
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-cyc...
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MDPI AG
2024-11-01
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| Series: | Biosensors |
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| author | Wei-Yu Tai To-Lin Chen Hsing-Meng Wang Lung-Ming Fu |
| author_facet | Wei-Yu Tai To-Lin Chen Hsing-Meng Wang Lung-Ming Fu |
| author_sort | Wei-Yu Tai |
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| description | 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<sup>2</sup> = 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<sup>2</sup> = 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. |
| format | Article |
| id | doaj-art-8dc95b9fcfc84bd1a3e784cb4979b32c |
| institution | OA Journals |
| issn | 2079-6374 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | MDPI AG |
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| series | Biosensors |
| spelling | doaj-art-8dc95b9fcfc84bd1a3e784cb4979b32c2025-08-20T02:28:02ZengMDPI AGBiosensors2079-63742024-11-01141154510.3390/bios14110545Rapid Microfluidic Biosensor for Point-of-Care Determination of Rheumatoid Arthritis via Anti-Cyclic Citrullinated Peptide Antibody DetectionWei-Yu Tai0To-Lin Chen1Hsing-Meng Wang2Lung-Ming Fu3Department of Emergency Medicine, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung 83301, TaiwanDepartment of Engineering Science, National Cheng Kung University, Tainan 70101, TaiwanDepartment of Engineering Science, National Cheng Kung University, Tainan 70101, TaiwanDepartment of Engineering Science, National Cheng Kung University, Tainan 70101, TaiwanRheumatoid 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<sup>2</sup> = 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<sup>2</sup> = 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.https://www.mdpi.com/2079-6374/14/11/545microfluidicsmicrochipELISARAanti-CCP antibody |
| spellingShingle | Wei-Yu Tai To-Lin Chen Hsing-Meng Wang Lung-Ming Fu Rapid Microfluidic Biosensor for Point-of-Care Determination of Rheumatoid Arthritis via Anti-Cyclic Citrullinated Peptide Antibody Detection Biosensors microfluidics microchip ELISA RA anti-CCP antibody |
| title | Rapid Microfluidic Biosensor for Point-of-Care Determination of Rheumatoid Arthritis via Anti-Cyclic Citrullinated Peptide Antibody Detection |
| title_full | Rapid Microfluidic Biosensor for Point-of-Care Determination of Rheumatoid Arthritis via Anti-Cyclic Citrullinated Peptide Antibody Detection |
| title_fullStr | Rapid Microfluidic Biosensor for Point-of-Care Determination of Rheumatoid Arthritis via Anti-Cyclic Citrullinated Peptide Antibody Detection |
| title_full_unstemmed | Rapid Microfluidic Biosensor for Point-of-Care Determination of Rheumatoid Arthritis via Anti-Cyclic Citrullinated Peptide Antibody Detection |
| title_short | Rapid Microfluidic Biosensor for Point-of-Care Determination of Rheumatoid Arthritis via Anti-Cyclic Citrullinated Peptide Antibody Detection |
| title_sort | rapid microfluidic biosensor for point of care determination of rheumatoid arthritis via anti cyclic citrullinated peptide antibody detection |
| topic | microfluidics microchip ELISA RA anti-CCP antibody |
| url | https://www.mdpi.com/2079-6374/14/11/545 |
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