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Microfluidic immunosensor design for the quantification of interleukin-6 in human serum samples
Germán A Messina1, Nancy V Panini, Noelia A Martinez
1INQUISAL, Department of Chemistry, National University of San Luis, CONICET, Chacabuco y Pedernera, D5700BWS, San Luis, Argentina.
Insights
This study presents a novel microfluidic immunosensor for detecting Interleukin-6 (IL-6), a key inflammatory cytokine. The developed biosensor offers faster and more sensitive detection of IL-6 in human serum samples.
Area of Science:
- Biomedical Engineering
- Immunosensing Technology
- Cytokine Detection
Background:
- Interleukin-6 (IL-6) is a critical inflammatory cytokine involved in fever and acute phase responses.
- Accurate and rapid detection of IL-6 is essential for diagnosing and managing inflammatory conditions.
- Existing methods for IL-6 detection can be time-consuming and require large sample volumes.
Purpose of the Study:
- To develop and validate an integrated microfluidic immunosensor for sensitive electrochemical detection of IL-6.
- To assess the performance of the microfluidic immunosensor in terms of limit of detection (LOD) and reproducibility.
- To compare the developed immunosensor with traditional IL-6 sensing methods.
Main Methods:
- Utilized a microfluidic biochip with a sandwich immunoassay format employing anti-IL-6 monoclonal antibodies.
- Immobilized antibodies on a 3-aminopropyl-modified controlled-pore glass (APCPG) packet within the microfluidic system.
- Employed electrochemical detection of an enzymatic product (p-aminophenol) generated by alkaline phosphatase.
Main Results:
- Achieved a limit of detection (LOD) of 0.41 pg/mL for electrochemical detection of IL-6.
- Demonstrated high reproducibility with intra- and inter-assay coefficients of variation below 6.5%.
- The microfluidic immunosensor required smaller sample volumes and offered faster detection compared to traditional methods.
Conclusions:
- The integrated microfluidic immunosensor provides a sensitive, reproducible, and rapid method for IL-6 detection in human serum.
- This technology holds promise for improved diagnostics of IL-6-related inflammatory diseases.
- The microfluidic platform offers advantages in sample volume reduction and assay speed.
Abstract:
Interleukin-6 (IL-6), an inflammatory cytokine, is one of the most important mediators of fever, the acute phase response, and inflammatory conditions. Described here is an integrated microfluidic immunosensor capable of detecting the concentration of IL-6 in human serum samples by use of an electrochemical method in a microfluidic biochip format. The detection of IL-6 was carried out using a sandwich immunoassay method based on the use of anti-IL-6 monoclonal antibodies, immobilized on a 3-aminopropyl-modified controlled-pore glass (APCPG) packet in a central channel (CC) of the microfluidic system. The IL-6 in the serum sample is allowed to react immunologically with the immobilized anti-IL-6 and biotin-labeled second antibodies specific to IL-6. After washing, the streptavidin-alkaline phosphatase conjugate is added. p-Aminophenyl phosphate is converted to p-aminophenol by alkaline phosphatase, and the electroactive product is quantified on a gold electrode at 0.10 V. For electrochemical detection and enzyme immunoassay, the LOD was 0.41 and 1.56 pg mL(-1), respectively. Reproducibility assays employed repetitive standards of IL-6, and the intra- and inter-assay coefficients of variation were below 6.5%. Compared with the traditional IL-6 sensing method, the integrated microfluidic immunosensor required smaller amounts of sample to perform faster detection.
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