Integrated Bioluminescent Immunoassays for High-Throughput Sampling and Continuous Monitoring of Cytokines

Immunoassays show great potential for the detection of low levels of cytokines, due to their high sensitivity and excellent specificity. There is a particular demand for biosensors that enable both high-throughput screening and continuous monitoring of clinically relevant cytokines such as interleuk...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2023-06, Vol.95 (23), p.8922-8931
Main Authors: van Aalen, Eva A., Rosier, Bas J. H. M., Jansen, Tom, Wouters, Simone F. A., Vermathen, Robin T., van der Veer, Harmen J., Yeste Lozano, José, Mughal, Sheeza, Fernández-Costa, Juan M., Ramón-Azcón, Javier, den Toonder, Jaap M. J., Merkx, Maarten
Format: Article
Language:English
Subjects:
Bioluminescence
Biosensors
Breast cancer
Breast carcinoma
Chemistry
Cytokines
Digital cameras
Endotoxins
High-throughput screening
Humans
Immunoassay
Immunoassay - methods
Immunoassays
Immunologic Tests
Interleukin 6
Interleukins
Low concentrations
Microfluidic devices
Microfluidics
Monitoring
Protein G
Three dimensional models
Tumor Necrosis Factor-alpha
Tumor necrosis factor-α
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Summary:Immunoassays show great potential for the detection of low levels of cytokines, due to their high sensitivity and excellent specificity. There is a particular demand for biosensors that enable both high-throughput screening and continuous monitoring of clinically relevant cytokines such as interleukin-6 (IL-6) and tumor necrosis factor-α (TNFα). To this end, we here introduce a novel bioluminescent immunoassay based on the ratiometric plug-and-play immunodiagnostics (RAPPID) platform, with an improved intrinsic signal-to-background and an >80-fold increase in the luminescent signal. The new dRAPPID assay, comprising a dimeric protein G adapter connected via a semiflexible linker, was applied to detect the secretion of IL-6 by breast carcinoma cells upon TNFα stimulation and the production of low concentrations of IL-6 (∼18 pM) in an endotoxin-stimulated human 3D muscle tissue model. Moreover, we integrated the dRAPPID assay in a newly developed microfluidic device for the simultaneous and continuous monitoring of changes in IL-6 and TNFα in the low-nanomolar range. The luminescence-based read-out and the homogeneous nature of the dRAPPID platform allowed for detection with a simple measurement setup, consisting of a digital camera and a light-sealed box. This permits the usage of the continuous dRAPPID monitoring chip at the point of need, without the requirement for complex or expensive detection techniques.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.3c00745