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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Main Authors: Wei-Yu Tai, To-Lin Chen, Hsing-Meng Wang, Lung-Ming Fu
Format: Article
Language:English
Published: MDPI AG 2024-11-01
Series:Biosensors
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Online Access:https://www.mdpi.com/2079-6374/14/11/545
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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
collection DOAJ
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.
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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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