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dc.contributor.authorBringeland, Gerd Hagaen_US
dc.contributor.authorBader, Luciusen_US
dc.contributor.authorBlaser, Nelloen_US
dc.contributor.authorBudzinski, Lisaen_US
dc.contributor.authorSchulz, Axel R.en_US
dc.contributor.authorMei, Henrik E.en_US
dc.contributor.authorMyhr, Kjell-Mortenen_US
dc.contributor.authorVedeler, Christian A.en_US
dc.contributor.authorGavasso, Soniaen_US
dc.PublishedBringeland GH, Bader L, Blaser N, Budzinski L, Schulz AR, Mei HE, Myhr KM, Vedeler CA, Gavasso S. Optimization of receptor occupancy assays in mass cytometry: Standardization across channels with QSC beads. Cytometry Part A. 2019;95(3):314-322eng
dc.description.abstractReceptor occupancy, the ratio between amount of drug bound and amount of total receptor on single cells, is a biomarker for treatment response to therapeutic monoclonal antibodies. Receptor occupancy is traditionally measured by flow cytometry. However, spectral overlap in flow cytometry limits the number of markers that can be measured simultaneously. This restricts receptor occupancy assays to the analysis of major cell types, although rare cell populations are of potential therapeutic relevance. We therefore developed a receptor occupancy assay suitable for mass cytometry. Measuring more markers than currently available in flow cytometry allows simultaneous receptor occupancy assessment and high‐parameter immune phenotyping in whole blood, which should yield new insights into disease activity and therapeutic effects. However, varying sensitivity across the mass cytometer detection range may lead to misinterpretation of the receptor occupancy when drug and receptor are detected in different channels. In this report, we describe a method for optimization of mass cytometry receptor occupancy measurements by using antibody‐binding quantum simply cellular (QSC) beads for standardization across channels with different sensitivities. We evaluated the method in a mass cytometry‐based receptor occupancy assay for natalizumab, a therapeutic antibody used in multiple sclerosis treatment that binds to α4‐integrin, which is expressed on leukocyte cell surfaces. Peripheral blood leukocytes from a treated patient were stained with a panel containing metal‐conjugated antibodies for detection of natalizumab and α4‐integrin. QSC beads with known antibody binding capacity were stained with the same metal‐conjugated antibodies and were used to standardize the signal intensity in the leukocyte sample before calculating receptor occupancy. We found that QSC bead standardization across channels corrected for sensitivity differences for detection of drug and receptor and generated more accurate results than observed without standardization.en_US
dc.rightsAttribution CC BY-NC-NDeng
dc.titleOptimization of receptor occupancy assays in mass cytometry: Standardization across channels with QSC beadsen_US
dc.typePeer reviewed
dc.typeJournal article
dc.rights.holderCopyright 2019 The Authors
dc.source.journalCytometry Part A

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