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Flow-Based Combinatorial Antibody Profiling: An Integrated Approach to Cell Characterization

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Flow Cytometry Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 699))

Abstract

BD FACS™ CAP (CAP = combinatorial antibody profile) is a screening tool for rapid characterization of human cell surface protein expression profiles using semi-automated high-throughput flow cytometry. The current configuration consists of 229 directly conjugated antibodies arrayed in a 96-well plate as three-color cocktails, which enables the characterization of each of the 229 individual surface markers. Each individual cell type of interest is analyzed on the 96-well screening plates and the data are acquired on a flow cytometer equipped with a high-throughput sampler. The expression level of each marker for each cell type is then calculated using semiautomated custom flow cytometry software. The process of characterizing these surface markers in a highly efficient manner using BD FACS™ CAP is enabled by automated liquid handling for staining, automated flow cytometry for data acquisition, and standardized algorithms for automated data analysis.

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Acknowledgments

We would like to acknowledge the following individuals for important contributions to the development of the FACS™ CAP technology: Keith Deluca, Megan Gottlieb, Errol Strain, Julie Leonard, Dylan Wilson, Stacy Xu, Jamal Sirriyah, John Dunne, Sharon Presnell, David Hodl, Mary Meyer, and William Busa. For the construction of the antibody plates, we would like to acknowledge the excellent technical assistance provided by Christine Chuang.

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Authors and Affiliations

  1. BD Biosciences, San Diego, CA, USA

    Shane Bruckner & Amitabh Gaur

  2. BD Technologies, Research Triangle Park, NC, USA

    Ling Wang, Ruiling Yuan & Perry Haaland

Authors
  1. Shane Bruckner
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  2. Ling Wang
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  3. Ruiling Yuan
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  4. Perry Haaland
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  5. Amitabh Gaur
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Corresponding author

Correspondence to Amitabh Gaur .

Editor information

Editors and Affiliations

  1. Medical Center, Flow Cytometry Core Facility, George Washington University, I Street NW 2300, Washington, 20037, District of Columbia, USA

    Teresa S. Hawley

  2. Medical Center, Dept. Anatomy & Regenerative Biology, George Washington University, I Street NW 2300, Washington, 20037, District of Columbia, USA

    Robert G. Hawley

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© 2011 Springer Science+Business Media, LLC

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Bruckner, S., Wang, L., Yuan, R., Haaland, P., Gaur, A. (2011). Flow-Based Combinatorial Antibody Profiling: An Integrated Approach to Cell Characterization. In: Hawley, T., Hawley, R. (eds) Flow Cytometry Protocols. Methods in Molecular Biology, vol 699. Humana Press. https://doi.org/10.1007/978-1-61737-950-5_6

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  • DOI: https://doi.org/10.1007/978-1-61737-950-5_6

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61737-949-9

  • Online ISBN: 978-1-61737-950-5

  • eBook Packages: Springer Protocols

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