Integration of lyoplate based flow cytometry and computational analysis for standardized immunological biomarker discovery

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

Research output: Contribution to journalArticlepeer-review

13 Citations (Scopus)

Abstract

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.
Original languageEnglish
Article numbere65485
Number of pages8
JournalPLoS ONE
Volume8
Issue number7
DOIs
Publication statusPublished - 3 Jul 2013

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