A ten-minute, single step, label-free, sample-to-answer assay for qualitative detection of cytokines in serum at femtomo
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A ten-minute, single step, label-free, sample-to-answer assay for qualitative detection of cytokines in serum at femtomolar levels Pengfei Xie 1 & Naixin Song 2 & Wen Shen 2 & Mark Allen 2 & Mehdi Javanmard 1 Accepted: 22 September 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Label-free electronic affinity based immuno-sensing is an attractive candidate as a platform technology for analyzing biomarkers due to the ease of miniaturization and minimal use of reagents. Electronic based sensing approaches, however, have lagged behind their optical counterparts in terms of detection limit, selectivity, and reliability. Also, the matrix dependent nature of electronic sensing modalities makes difficult the analysis of biomarkers in high salt concentration samples such as serum due to charge screening. We present a novel sensing platform, the micro-well sensor, that works by functionalizing nanoscale volume wells with antibodies and monitoring the impedance change inside the wells due binding of target protein. This detection modality is advantageous to many label-free electronic sensors in that signal power scales with increase in salt concentration, thus improving the sensitivity of the platform. We demonstrate rapid label-free qualitative detection of cytokines within ten minutes at femtoMolar concentrations and a dynamic range of 3 orders of magnitude in serum samples. We describe the design, fabrication, and characterization of the micro-well sensor in serum samples using inflammatory protein biomarkers. Keywords Label-free biosensors . Cytokine quantifications . Impedance detection . Protein detection . Proteomics . Biomarker detection . Diagnostics
1 Introduction Affinity based biosensors detect proteins and nucleic acids through immobilization of probe molecules on a transducer substrate to attain specificity in detection of the target macromolecule of interest. In the context of protein detection, protein array technologies provide a valuable platform for functional proteomic analysis (Ray et al. 2010). A protein microarray provides a multi-functional platform enabling comprehensive and high throughput studies, which can be widely used in biomarker validation studies (Hudson et al. 2007), study of protein–protein interactions (Schweitzer et al. 2003), protein–DNA interactions (Kersten et al. 2004), and detection of various antigens and antibodies (Chamritski et al. 2007). Among affinity-based biosensing technologies, Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10544-020-00525-0) contains supplementary material, which is available to authorized users. * Mehdi Javanmard [email protected] 1
Rutgers University, New Brunswick, NJ, USA
2
University of Pennsylvania, Philadelphia, PA, USA
there are two major detection strategies, label-based and labelfree. Both approaches have their merits and disadvantages. Label-based technologies typically achieve lower detection limits and higher selectivity, yet require multiple st
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