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PLoS One. 2013 Jul 3;8(7):e65485. doi: 10.1371/journal.pone.0065485. Print 2013.

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

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1
St John's Institute of Dermatology, King's College London, London, United Kingdom.

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.

PMID:
23843942
PMCID:
PMC3701052
DOI:
10.1371/journal.pone.0065485
[Indexed for MEDLINE]
Free PMC Article

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