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Development of multiplexed protein profiling and detection using near infrared detection of reverse-phase protein microarrays

Clinical Proteomics volume 1, pages 81–89 (2004)Cite this article

Abstract

Protein microarrays have been recently employed for signal pathway profiling and high-throughput protein expression analysis. Reversephase arrays, where the array consists of immobilized analytes and lysates has especially shown promise in low abundance analyte detection and signal pathway profiling using phospho-specific antibodies. A limitation to current reverse phase array methodology is the inability to multiplex proteomic-based endpoints as each array can only report one analyte endpoint. In this study, we report on the use of a dual dye based approach that can effectively double the number of endpoints observed per array allowing, for example, both phosphospecific and total protein levels to be measured and analyzed at once. The method utilizes antibody bound dyes that emit in the infrared spectral region as a means of sensitive and specific detection.

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Affiliations

  1. Office of Cell and Gene Therapy, Center for Biologics Evaluation and Research, Food and Drug Administration, NCI/FDA Clinical Proteomics Program, 20892, Bethesda, Maryland

    Valerie S. Calvert & Emanuel F. Petricoin III

  2. LI-COR Biosciences, 68504, Lincoln, NE

    Yihui Tang, Vince Boveia, Amy Schutz-Geschwender & D. Michael Olive

  3. Laboratory of Pathology, Center for Cancer Research, National Cancer Institute, NCI/FDA Clinical Proteomics Program, 20892, Bethesda, Maryland

    Julie Wulfkuhle & Lance A. Liotta

Authors
  1. Valerie S. Calvert
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  2. Yihui Tang
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  3. Vince Boveia
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  4. Julie Wulfkuhle
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  5. Amy Schutz-Geschwender
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  6. D. Michael Olive
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  7. Lance A. Liotta
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  8. Emanuel F. Petricoin III
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Corresponding author

Correspondence to Emanuel F. Petricoin III.

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Cite this article

Calvert, V.S., Tang, Y., Boveia, V. et al. Development of multiplexed protein profiling and detection using near infrared detection of reverse-phase protein microarrays. Clin Proteom 1, 81–89 (2004). https://doi.org/10.1385/CP:1:1:081

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  • DOI: https://doi.org/10.1385/CP:1:1:081

Key Words

  • Proteomics
  • signal profiling
  • networks
  • protein microarrays

Clinical Proteomics

ISSN: 1559-0275