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• W2800785622 abstract "Purpose: Osteoarthritis is a highly debilitation disease affecting an increasing part of the population. The disease development is characterized by a change in homeostasis leading to progressive degradation of the joint. Early measurement of biomarkers in the joint to detect these changes could lead to new sensitive diagnostic methods and improve treatment efficacy. However, no sufficiently sensitive method exists that can measure multiple biomarkers simultaneously in the small volume of the joint to capture the complexity of the disease. In addition, current existing assays are end point measurements, which limits the opportunities for quality control. This is particularly relevant in multiplex and could lead to failure to detect spotting errors, cross-reactivity or heterophilic antibodies, resulting in false positives or negatives. Previously, we have proposed a new method to measure biomarkers in the synovial fluid and serum. This method combines real-time interaction measurements via surface plasmon resonance array imaging (SPRi) with an enhancement cascade to sequentially increase the signal. This has resulted in a large improvement in sensitivity of 40,000 times. In a proof of concept study we have shown we can measure 4 cytokines simultaneously (IL-1β, IL-6, TNF-α and IFN-γ) in multiplex in concentrations as low as 50 fg/ml and with a dynamic detection range of 7 logarithms. In addition, this method was shown to be accurate in synovial fluid with good recovery of spikes compared to buffer controls. In this work, we will present this method of powerful quality control opportunities essential for reliable research and clinical application. Methods: The IBIS MX96 was used in combination with custom sensors, allowing for free choice of antibody pairs. We subsequently implemented an antibody sandwich assay with a biotinylated gold nanoparticle signal enhancement cascade to quantitatively measure potential biomarkers with both high sensitivity and in a large dynamic concentration range. The real time nature of SPRi has allowed us to accurately fit the interaction curves of the cascade and thereby determine the underlying kinetics. To achieve this we have applied a 1-1 langmuir fit in combination with heterogeneity of binding sites. Using the determined kinetics of individual cascade components and their underlying relation for IL-1β, IL-6, TNF-α and IFN-γ, we subsequently predicted the enhancement cascade depending on biomarker concentration. Results: We have achieved accurate fits for all the components of the enhancement cascade for the 4 cytokines tested over a concentration range of 7 logs. The subsequent characterization of the underlying relation allowed us to accurately predict the enhancement cascade with small residuals in comparison to actual measured values. This enables calibration free measurements, which saves time and reagents. In addition, this allows for many interesting opportunities to improve quality control both pre- and post- measurement. Pre- measurement, the kinetics enable the detection and elimination of spotting irregularities, which is a major quality issue in all multiplex assays based on antibody technology. Post-measurement, the prediction of the cascade can be used to assess cross-reactivity and heterophilic antibodies. When any step in this cascade deviates from the expected curve we can determine if a confounding event has occurred. This can be analyzed fully automatic and can assist in determining confounders and avoid false positives or negatives. Conclusions: We have developed a nanoparticle enhancement cascade for SPRi to measure biomarker panels for the early diagnosis of osteoarthritis. Our extensive quality control opportunities further improve this technology, making it highly robust for both research and clinical application. This can lead to exciting new possibilities in early diagnosis, prognosis and treatment efficacy monitoring in osteoarthritis." @default.
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• W2800785622 date "2018-04-01" @default.
• W2800785622 modified "2023-09-28" @default.
• W2800785622 title "A new method for multiplex measurements of biomarkers in synovial fluid aspirates with high sensitivity and extensive quality control" @default.
• W2800785622 doi "https://doi.org/10.1016/j.joca.2018.02.413" @default.
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