Abstract
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.
Highlights
The technology of microfluidics is one that manipulates small volumes of fluid and flow that has the potential to miniaturize complex laboratory procedures [1, 2]
Assay imprecisions were evaluated on 6 replicates of duplicate measurements from a pooled serum sample on three different cartridges for Interleukin-1 beta (IL-1β), Tumor necrosis factor-α (TNF-α), IL-6, and IL-10, which were found to be 6.2, 8.0, 6.2 and 9.1 %, respectively
0.21–2000 a Analytical measurement range (AMR) of the Simple Plex assays were provided by the manufacturer lower limit of the AMR, which was 0.21 pg/mL (Table 2)
Summary
The technology of microfluidics is one that manipulates small volumes of fluid and flow that has the potential to miniaturize complex laboratory procedures [1, 2]. Microfluidic technology has been widely used in point-of-care (POC) devices for clinical diagnostics (e.g., iSTAT) [3,4,5,6,7] Since these devices generally require small sample volumes there is much interest in applying microfluidic technology to areas outside of the traditional realms of POC diagnostics and into areas in translational research efforts such as the quantitative measurement of multiple protein biomarkers (multiplexing) [8,9,10,11]. Microfluidics allows for separate incubation chambers for every analyte; that is, each incubation chamber is limited to one antibody pair to react with its respective analyte This prevents the potential issue of cross-reactivity (i.e., antigen cross-reacting to other antibody pairs) [13]; second, the consumption of reagent and sample volume is low due to miniaturized design, and researchers are able to conserve precious
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