µFBI: A Microfluidic Bead-Based Immunoassay for Multiplexed Detection of Proteins from a µL Sample Volume

Yu, Xiaobo; Hartmann, Michael; Wang, Quan; Poetz, Oliver; Schneiderhan‐Marra, Nicole; Stoll, Dieter; Kazmaier, Cornelia; Joos, Thomas

doi:10.1371/journal.pone.0013125

Cited by 27 publications

(27 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Another device recirculated fluid over a small number of beads immobilized behind a filter 11 and performed detection using fluorescence microscopy. Beads have also been confined in a capillary tube, collected at the completion of the assay, and analyzed off-chip using flow cytometry 10 . In the absence of a permanent mechanical trap PDMS valves 47 and magnetic trapping have also been utilized in conjunction with fluorescence imaging 48 .…”

Section: Resultsmentioning

confidence: 99%

Development and validation of a microfluidic immunoassay capable of multiplexing parallel samples in microliter volumes

et al. 2015

View full text Add to dashboard Cite

Immunoassays are widely utilized due to their ability to quantify a vast assortment of biomolecules relevant to biological research and clinical diagnostics. Recently, immunoassay capabilities have been improved by the development of multiplex assays that simultaneously measure multiple analytes in a single sample. However, these assays are hindered by high costs of reagents and relatively large sample requirements. For example, in vitro screening systems currently dedicate individual wells to each time point of interest and this limitation is amplified in screening studies when the investigation of many experimental conditions is necessary; resulting in large volumes for analysis, a correspondingly high cost and a limited temporal experimental design. Microfluidics based immunoassays have been developed in order to overcome these drawbacks. Together, previous studies have demonstrated on-chip assays with either a large dynamic range, high performance sensitivity, and/or the ability to process samples in parallel on a single chip. In this report, we develop a multiplex immunoassay possessing all of these parallel characteristics using commercially available reagents, which allows the analytes of interest to be easily changed. The device presented can measure 6 proteins in 32 samples simultaneously using only 4.2 µL of sample volume. High quality standard curves are generated for all 6 analytes included in the analysis, and spiked samples are quantified throughout the working range of the assay. In addition, we demonstrate a strong correlation (R2=0.8999) between in vitro supernatant measurements using our device and those obtained from a bench-top multiplex immunoassay. Finally, we describe cytokine secretion in an in vitro inflammatory hippocampus culture system, establishing proof-of-concept of the ability to use this platform as an in vitro screening tool. The low-volume, multiplexing abilities of the microdevice described in this report could be broadly applied to numerous situations where sample volumes and costs are limiting.

show abstract

Section: Resultsmentioning

confidence: 99%

Development and validation of a microfluidic immunoassay capable of multiplexing parallel samples in microliter volumes

et al. 2015

View full text Add to dashboard Cite

show abstract

“…By use of bead-based assays in microfluidics, multiplexed protein detection of TNF-α, CXCL2, IL-6, and IL-1b has been reported [111] with the sensitivity of 1000 pg/mL with the sample volume of 4.7 nL. A capillary-based system (~100 μm diameter), in which the antibody-coupled microbeads were captured on a 0.5 μm filter, was developed to detect protein expression of receptor tyrosine kinase in breast cancer tissues [112]. Our group has developed a microfluidic platform for immobilization of single-layer microbead array for multiplexed high-throughput analysis of biomolecules [30].…”

Section: Immunoassaysmentioning

confidence: 99%

Microfluidics for DNA and Protein Analysis with Multiplex Microbead-Based Assays

Yue

Yang

2016

Microfluidic Methods for Molecular Biology

View full text Add to dashboard Cite

Early screening and diagnosis of diseases require the development of sensitive, reliable, and inexpensive high-throughput assays. There is a growing need for innovative diagnostic technologies that provide accurate detection of a broad range of diseases and enhance laboratory productivity. Developments in micro-and nanotechnology have advanced the "lab-on-a-chip" concept towards a new generation of point-of-care diagnostic devices that enable parallel detection of multiple analytes in small-volume samples with high sensitivity in a short time. These features fulfill some of the important criteria of bioanalysis used for biochemical studies, environmental analyses, and clinical diagnostics. However, the multiplexing capability of microfluidic-based assay is limited compared to conventional flow cytometric assays, particularly for encoded microbead-based assays, which are capable of simultaneously analyzing multiple analyte. It is possible to incorporate the microbead-based assays into microfluidic devices in order to retain all the advantages of the microdevice and significantly improve multiplexing capability. In this chapter, we summarize the latest development in microbeadbased assays and discuss the integration of the microbead technology with microfluidic devices. Applications of the integrated approach in multiplexed protein and DNA analysis are growing rapidly in the areas of biomarker research, cancer screening, and disease diagnosis. The prospects for future development and commercialization of these microbead-based microfluidic devices are also discussed.

show abstract

“…In this approach antibody-coated beads were captured on a 0.5 μm filter while washing and incubation steps were done by manipulating the flow across the beads. Quantitative analysis of the beads was performed using a flow cytometer or collecting the beads in a microwell plate [85] (Fig. 6).…”

Section: Bioanalytical Analysis Technologiesmentioning

confidence: 99%

Particles and microfluidics merged: perspectives of highly sensitive diagnostic detection

et al. 2011

View full text Add to dashboard Cite

There is a growing need for diagnostic technologies that provide laboratories with solutions that improve quality, enhance laboratory system productivity, and provide accurate detection of a broad range of infectious diseases and cancers. Recent advances in micro- and nanoscience and engineering, in particular in the areas of particles and microfluidic technologies, have advanced the “lab-on-a-chip” concept towards the development of a new generation of point-of-care diagnostic devices that could significantly enhance test sensitivity and speed. In this review, we will discuss many of the recent advances in microfluidics and particle technologies with an eye towards merging these two technologies for application in medical diagnostics. Although the potential diagnostic applications are virtually unlimited, the most important applications are foreseen in the areas of biomarker research, cancer diagnosis, and detection of infectious microorganisms.

show abstract

µFBI: A Microfluidic Bead-Based Immunoassay for Multiplexed Detection of Proteins from a µL Sample Volume

Cited by 27 publications

References 35 publications

Development and validation of a microfluidic immunoassay capable of multiplexing parallel samples in microliter volumes

Development and validation of a microfluidic immunoassay capable of multiplexing parallel samples in microliter volumes

Microfluidics for DNA and Protein Analysis with Multiplex Microbead-Based Assays

Particles and microfluidics merged: perspectives of highly sensitive diagnostic detection

Contact Info

Product

Resources

About