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qFlow Cytometry-Based Receptoromic Screening: A High-Throughput Quantification Approach Informing Biomarker Selection and Nanosensor Development

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Biomedical Nanotechnology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1570))

Abstract

Nanosensor-based detection of biomarkers can improve medical diagnosis; however, a critical factor in nanosensor development is deciding which biomarker to target, as most diseases present several biomarkers. Biomarker-targeting decisions can be informed via an understanding of biomarker expression. Currently, immunohistochemistry (IHC) is the accepted standard for profiling biomarker expression. While IHC provides a relative mapping of biomarker expression, it does not provide cell-by-cell readouts of biomarker expression or absolute biomarker quantification. Flow cytometry overcomes both these IHC challenges by offering biomarker expression on a cell-by-cell basis, and when combined with calibration standards, providing quantitation of biomarker concentrations: this is known as qFlow cytometry. Here, we outline the key components for applying qFlow cytometry to detect biomarkers within the angiogenic vascular endothelial growth factor receptor family. The key aspects of the qFlow cytometry methodology include: antibody specificity testing, immunofluorescent cell labeling, saturation analysis, fluorescent microsphere calibration, and quantitative analysis of both ensemble and cell-by-cell data. Together, these methods enable high-throughput quantification of biomarker expression.

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Acknowledgments

We would like to thank Dr. Barbara Pilas for her advice and help with flow cytometry. We would also like to thank Spencer B. Mamer and Ali Ansari for their help with editing. Finally, we would like to thank the American Heart Association Grant #16SDG26940002, American Cancer Society Illinois Division Basic Research Grant #282802, and National Science Foundation CBET Grant #1512598 for funding support.

Author information

Authors and Affiliations

  1. Bioengineering Department, University of Illinois at Urbana-Champaign, Urbana, IL, USA

    Si Chen, Jared Weddell & Princess I. Imoukhuede

  2. Chicago College of Osteopathic Medicine, Downers Grove, IL, USA

    Pavan Gupta

  3. Indiana University School of Medicine, Indianapolis, IN, USA

    Grace Conard

  4. Bioengineering Department, California Institute of Technology, Pasadena, CA, USA

    James Parkin

Authors
  1. Si Chen
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  2. Jared Weddell
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  3. Pavan Gupta
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  4. Grace Conard
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  6. Princess I. Imoukhuede
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Editor information

Editors and Affiliations

  1. Department of Chemistry and International Institute for Nanotechnology, Northwestern University, Evanston, Illinois, USA

    Sarah Hurst Petrosko

  2. Department of Biomedical Engineering, University of Delaware, Newark, Delaware, USA

    Emily S. Day

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Chen, S., Weddell, J., Gupta, P., Conard, G., Parkin, J., Imoukhuede, P.I. (2017). qFlow Cytometry-Based Receptoromic Screening: A High-Throughput Quantification Approach Informing Biomarker Selection and Nanosensor Development. In: Petrosko, S., Day, E. (eds) Biomedical Nanotechnology. Methods in Molecular Biology, vol 1570. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6840-4_8

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  • DOI: https://doi.org/10.1007/978-1-4939-6840-4_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6838-1

  • Online ISBN: 978-1-4939-6840-4

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