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. 2018 Jun 19;90(12):7569-7577.
doi: 10.1021/acs.analchem.8b01198. Epub 2018 May 31.

Automated 3D-Printed Microfluidic Array for Rapid Nanomaterial-Enhanced Detection of Multiple Proteins

Karteek Kadimisetty, Spundana Malla, Ketki S Bhalerao, Islam M Mosa  1 , Snehasis Bhakta, Norman H Lee  2 , James F Rusling  3   4
Affiliations

Automated 3D-Printed Microfluidic Array for Rapid Nanomaterial-Enhanced Detection of Multiple Proteins

Karteek Kadimisetty et al. Anal Chem. .

Abstract

We report here the fabrication and validation of a novel 3D-printed, automated immunoarray to detect multiple proteins with ultralow detection limits. This low cost, miniature immunoarray employs electrochemiluminescent (ECL) detection measured with a CCD camera and employs touch-screen control of a micropump to facilitate automated use. The miniaturized array features prefilled reservoirs to deliver sample and reagents to a paper-thin pyrolytic graphite microwell detection chip to complete sandwich immunoassays. The detection chip achieves high sensitivity by using single-wall carbon nanotube-antibody conjugates in the microwells and employing massively labeled antibody-decorated RuBPY-silica nanoparticles to generate ECL. The total cost of an array is $0.65, and an eight-protein assay can be done in duplicate for $0.14 per protein with limits of detection (LOD) as low as 78-110 fg mL-1 in diluted serum. The electronic control system costs $210 in components. Utility of the automated immunoarray was demonstrated by detecting an eight-protein prostate cancer biomarker panel in human serum samples in 25 min. The system is well suited to future clinical and point-of-care diagnostic testing and could be used in resource-limited environments.

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Figures

Figure 1.
Figure 1.
Representations of 3D-printed immunoarray: (A) 3D-printed microfluidic array with chambers to hold sample, wash buffers, detection nanoparticles, and coreactant for ECL generation. Array is shown on left without detection chip and on right bonded to pyrolytic graphite sheet (PGS) microwell detection chip with reagent and sample chambers filled with dye solutions for visualization; (B) representative disposable PGS chip with heat transferred microwells printed using hydrophobic toner ink. Inset illustrates sandwich immunoassay on a single-wall carbon nanotube forest (SWCNT) in one microwell.
Figure 2.
Figure 2.
Immunoassay controller with touch-screen user interface for immunoarray. A microfluidic array connected to a micropump is shown with dye-filled reagent chambers and graphite detection chip. Inset figures show multiple immunoassay steps along with messages to inform the user.
Figure 3.
Figure 3.
Recolorized CCD images for five arrays showing increase in ECL light with increase in concentration for all eight proteins on a single array with an acquisition time of 180 s at 1.0 V Ag/AgCl in the presence of 500 mM TrPA.
Figure 4.
Figure 4.
Calibration data for multiplexed detection in undiluted calf serum from ECL responses at 1.0 V vs Ag/AgCl. (A−H) are multiplex assay calibration curves for IGF-1, PSA, PF-4, CD-14, VEGF-D, GOLM-1, PSMA, and IGFBP-3. Standard deviations for n = 4.
Figure 5.
Figure 5.
Statistical summary of patient sample results using clustered multiple variable graphs for each protein. (A,B) Box-and-whisker plots show data point distribution classified as cancer-free and prostate cancer samples for each biomarker. The plots present lower, upper quartile, and median values along with minimum and maximum ranges.
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References

    1. Hanash SM; Baik CS; Kallioniemi O Nat. Rev. Clin. Oncol 2011, 8, 142−150. - PubMed
    1. National Cancer Institute. Cancer Screening and Early Detection Research https://www.cancer.gov/research/areas/screening (accessed February 15, 2018).
    1. Cohen JD; Li L; Wang Q; Thoburn C; Afsari B; Danilova L; Douville C; Javed AA; Wong F; Mattox A; et al. Science 2018, 359, 926. - PMC - PubMed
    1. Giljohann DA; Mirkin CA Nature 2009, 462, 461−464. - PMC - PubMed
    1. Kulasingam V; Diamandis EP Nat. Clin. Pract. Oncol 2008, 5, 588−599. - PubMed

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