Integration of Lyoplate Based Flow Cytometry and Computational Analysis for Standardized Immunological Biomarker Discovery

نویسندگان

  • Federica Villanova
  • Paola Di Meglio
  • Margaret Inokuma
  • Nima Aghaeepour
  • Esperanza Perucha
  • Jennifer Mollon
  • Laurel Nomura
  • Maria Hernandez-Fuentes
  • Andrew Cope
  • A. Toby Prevost
  • Susanne Heck
  • Vernon Maino
  • Graham Lord
  • Ryan R. Brinkman
  • Frank O. Nestle
چکیده

Discovery of novel immune biomarkers for monitoring of disease prognosis and response to therapy in immune-mediated inflammatory diseases is an important unmet clinical need. Here, we establish a novel framework for immunological biomarker discovery, comparing a conventional (liquid) flow cytometry platform (CFP) and a unique lyoplate-based flow cytometry platform (LFP) in combination with advanced computational data analysis. We demonstrate that LFP had higher sensitivity compared to CFP, with increased detection of cytokines (IFN-γ and IL-10) and activation markers (Foxp3 and CD25). Fluorescent intensity of cells stained with lyophilized antibodies was increased compared to cells stained with liquid antibodies. LFP, using a plate loader, allowed medium-throughput processing of samples with comparable intra- and inter-assay variability between platforms. Automated computational analysis identified novel immunophenotypes that were not detected with manual analysis. Our results establish a new flow cytometry platform for standardized and rapid immunological biomarker discovery with wide application to immune-mediated diseases.

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عنوان ژورنال:

دوره 8  شماره 

صفحات  -

تاریخ انتشار 2013