A microfluidic multiplex proteomic immunoassay device for translational research

ObjectiveMicrofluidic technology has the potential to miniaturize and automate complex laboratory procedures. The objective of this study was to assess a microfluidic immunoassay device, Simple Plex, which simultaneously measured IL-1β, TNF-α, IL-6, and IL-10 in serum samples. This assessment is important to understanding the potentials of this microfluidic device as a valuable tool in translational research efforts.MethodsWe studied the operational characteristics of Simple Plex, and compared to other immunoassay systems including bead-based (i.e., Bio-Plex® from Bio-Rad) and planar micro-spot based (i.e., Multi-Array from Meso Scale Discovery) multiplex assays. We determined imprecisions for each of the Simple Plex assays and evaluated the ability of Simple Plex to detect IL-1β, TNF-α, IL-6, and IL-10 in serum samples.ResultsSimple Plex assays required 25 µL serum, and 1.5 h to run 16 samples per cartridge per instrument. Assay imprecisions, evaluated by measurement of 6 replicates in duplicate from a serum pool using three different cartridges, were less than 10 % for all 4 cytokine protein biomarkers, comparable to the imprecisions of traditional ELISAs. The Simple Plex assays were able to detect 32, 95, 97, and 100 % [i.e., percentages of the results within the respective analytical measurement ranges (AMRs)] of IL-1β, TNF-α, IL-6, and IL-10, respectively, in 66 serum samples.ConclusionsSimple Plex is a microfluidic multiplex immunoassay device that offers miniaturized, and automated analysis of protein biomarkers. Microfluidic devices such as Simple Plex represent a promising platform to be used in translational research to measure protein biomarkers in real clinical samples.

[1]  D. Chan,et al.  Integrated analyses of proteins and their glycans in a magnetic bead-based multiplex assay format. , 2013, Clinical chemistry.

[2]  S. Hwang,et al.  Microchip‐based multiplex electro‐immunosensing system for the detection of cancer biomarkers , 2008, Electrophoresis.

[3]  Ferdousi Chowdhury,et al.  Validation and comparison of two multiplex technologies, Luminex and Mesoscale Discovery, for human cytokine profiling. , 2009, Journal of immunological methods.

[4]  Peter K Sorger,et al.  Microfluidics closes in on point-of-care assays , 2008, Nature Biotechnology.

[5]  Steven A Carr,et al.  Protein biomarker discovery and validation: the long and uncertain path to clinical utility , 2006, Nature Biotechnology.

[6]  D. Chan,et al.  Validation of a multiplex immunoassay for serum angiogenic factors as biomarkers for aggressive prostate cancer. , 2012, Clinica chimica acta; international journal of clinical chemistry.

[7]  Samuel K Sia,et al.  Commercialization of microfluidic point-of-care diagnostic devices. , 2012, Lab on a chip.

[8]  A. Manz,et al.  Miniaturized total chemical analysis systems: A novel concept for chemical sensing , 1990 .

[9]  G. Whitesides.  The origins and the future of microfluidics , 2006, Nature.

[10]  Wei Cao,et al.  A highly efficient and versatile microchip capillary electrophoresis method for DNA separation using gold nanoparticle as a tag. , 2011, Journal of separation science.

[11]  Lori J Sokoll,et al.  Glycoproteomics for prostate cancer detection: changes in serum PSA glycosylation patterns. , 2009, Journal of proteome research.

[12]  Pingbo Zhang,et al.  Multiplex assays for biomarker research and clinical application: Translational science coming of age , 2010, Proteomics. Clinical applications.

[13]  Jin-Woo Choi,et al.  Disposable smart lab on a chip for point-of-care clinical diagnostics , 2004, Proceedings of the IEEE.

[14]  V. Srinivasan,et al.  Heterogeneous immunoassays using magnetic beads on a digital microfluidic platform. , 2008, Lab on a chip.

[15]  Martin Pumera,et al.  Nanomaterials as electrochemical detectors in microfluidics and CE: Fundamentals, designs, and applications , 2009, Electrophoresis.

[16]  A. Roda,et al.  Recent advancements in chemical luminescence-based lab-on-chip and microfluidic platforms for bioanalysis. , 2014, Journal of pharmaceutical and biomedical analysis.

[17]  Vijay Srinivasan,et al.  Development of a digital microfluidic platform for point of care testing. , 2008, Lab on a chip.

[18]  Samuel K Sia,et al.  Microfluidics and point-of-care testing. , 2008, Lab on a chip.

[19]  C T Lim,et al.  Bead-based microfluidic immunoassays: the next generation. , 2007, Biosensors & bioelectronics.

[20]  A. Helmicki,et al.  An integrated microfluidic biochemical detection system for protein analysis with magnetic bead-based sampling capabilities. , 2002, Lab on a chip.

[21]  James F Rusling,et al.  Measurement of biomarker proteins for point-of-care early detection and monitoring of cancer. , 2010, The Analyst.

[22]  Agustina Gómez-Hens,et al.  Strategies to improve the analytical features of microfluidic methods using nanomaterials , 2014 .

[23]  Mariana Medina-Sánchez,et al.  Nanomaterials and lab-on-a-chip technologies. , 2012, Lab on a chip.

[24]  Henry Rodriguez,et al.  The journey to regulation of protein-based multiplex quantitative assays. , 2011, Clinical chemistry.

[25]  P. Mitchell.  Microfluidics—downsizing large-scale biology , 2001, Nature Biotechnology.

[26]  Qin Fu,et al.  Comparison of multiplex immunoassay platforms. , 2010, Clinical chemistry.

[27]  S. Maerkl,et al.  A Microfluidic Platform for High-Throughput Multiplexed Protein Quantitation , 2014, PloS one.