title: Integration of lyoplate based flow cytometry and computational analysis for standardized immunological biomarker discovery.
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.
authors.affiliation: St John's Institute of Dermatology, King's College London, London, United Kingdom.
authors.forename: Federica
authors.initials: F
authors.lastname: Villanova
authors.forename: Paola
authors.initials: P
authors.lastname: Di Meglio
authors.forename: Margaret
authors.initials: M
authors.lastname: Inokuma
authors.forename: Nima
authors.initials: N
authors.lastname: Aghaeepour
authors.forename: Esperanza
authors.initials: E
authors.lastname: Perucha
authors.forename: Jennifer
authors.initials: J
authors.lastname: Mollon
authors.forename: Laurel
authors.initials: L
authors.lastname: Nomura
authors.forename: Maria
authors.initials: M
authors.lastname: Hernandez-Fuentes
authors.forename: Andrew
authors.initials: A
authors.lastname: Cope
authors.forename: A Toby
authors.initials: AT
authors.lastname: Prevost
authors.forename: Susanne
authors.initials: S
authors.lastname: Heck
authors.forename: Vernon
authors.initials: V
authors.lastname: Maino
authors.forename: Graham
authors.initials: G
authors.lastname: Lord
authors.forename: Ryan R
authors.initials: RR
authors.lastname: Brinkman
authors.forename: Frank O
authors.initials: FO
authors.lastname: Nestle
mesh_terms: D000328:Adult; D000906:Antibodies; D015415:Biomarkers; D019295:Computational Biology / Q000379:methods*; D005260:Female; D005434:Flow Cytometry / Q000379:methods* / Q000592:standards; D006801:Humans; D016130:Immunophenotyping / Q000379:methods* / Q000592:standards; D007249:Inflammation / Q000276:immunology* / Q000378:metabolism*; D007371:Interferon-gamma / Q000378:metabolism; D016753:Interleukin-10 / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D055815:Young Adult
chemical_list: D000906:Antibodies; D015415:Biomarkers; D016753:Interleukin-10; D007371:Interferon-gamma