Experiment Overview
| Repository ID: | FR-FCM-ZYRR | Experiment name: | Natalizumab RO with QSC beads: triplicate experiment | MIFlowCyt score: | 70.86% |
| Primary researcher: | Gerd Haga Bringeland | PI/manager: | Gerd Haga Bringeland | Uploaded by: | Gerd Haga Bringeland |
| Experiment dates: | 2018-05-01 - 2018-07-31 | Dataset uploaded: | Nov 2018 | Last updated: | Mar 2019 |
| Keywords: | [mass cytometry] [CyTOF] [Standardization] [quantitative analysis] [Receptor occupancy] [biomarkers] [precision medicine] [natalizumab] [multiple sclerosis] [optimization] [QSC beads] | Manuscripts: | [30688025] |
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| Organizations: |
University of Bergen, Dept of Clinical Medicine, Bergen, (Norway)
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| Purpose: | Demonstrate a method for standardization across channels in mass cytometry receptor occupancy (RO) assays, where drug and receptor are detected in mass cytometry channels with different sensitivity. We exemplify this in a RO assay of the MS-therapeutic antibody natalizumab. | ||||
| Conclusion: | RO is the ratio between bound drug and total target receptor. Estimation of RO is inaccurate when drug and receptor are detected in mass cytometry channels with different sensitivity. Accurate values are obtained by standardizing with QSC beads coated with known numbers of the antibodies used to detect drug and receptor in the sample. | ||||
| Comments: | None | ||||
| Funding: | This research was supported in part by Norwegian Novartis' research grant for MS 2017 | ||||
| Quality control: | Negative controls: - Unstained PBL sample (preexisting metal isotopes in the detection range of the mass cytometer) - Mass-minus-one controls stained with the whole panel minus antibody for detection of drug or receptor (spillover) - Untreated healthy donor PBL (specificity of anti-IgG4). Positive control: in vitro drug saturated patient sample with expected RO of 100%. Competitivity: equal detection of receptor in the abcence and precence of drug. | ||||
Experiment variables
| Individuals | |
|---|---|
| · HD | c01_D_2 PBL+_02_0_HD.fcs · c01_D_2 PBL-_02_0_HD.fcs · c01_D_1 PBL+_01_1_HD.fcs · c01_D_1 PBL-_01_0_HD.fcs · c01_D_3 PBL+_02_0_HD.fcs · c01_D_3 PBL-_02_0_HD.fcs · c01_Unstained_01_0_HD.fcs |
| · Patient | c02_D_2 PBL+_02_0_Pt.fcs · c02_D_2 PBL-_02_0_Pt.fcs · c02_D_1 PBL+_01_1_Pt.fcs · c02_D_1 PBL-_01_0_Pt.fcs · c02_D_3 PBL+_02_0_Pt.fcs · c02_D_3 PBL-_02_0_Pt.fcs · c02_Unstained_01_0_Pt.fcs |
| · QSC beads | D_1 CD_B1_01.FCS · D_1 CD_B2_01.FCS · D_1 CD_B3_01.FCS · D_1 CD_B4_01.FCS · D_1 CD_B5_01.FCS · D_1 IgG_B1_2_01.FCS · D_1 IgG_B2_02.FCS · D_1 IgG_B3_02.FCS · D_1 IgG_B4_02.FCS · D_1 IgG_B5_02.FCS · D2_CD_B1_006_0.fcs · D2_CD_B2_01_1.fcs · D2_CD_B3_01_0.fcs · D2_CD_B4_01_0.fcs · D2_CD_B5_02_0.fcs · D2_IgG_B1_01_1.fcs · D2_IgG_B2_01_1.fcs · D2_IgG_B3_01_1.fcs · D2_IgG_B4_01_0.fcs · D2_IgG_B5_01_0.fcs · D3_CD_B1_01_0.fcs · D3_CD_B2_01_0.fcs · D3_CD_B3_01_0.fcs · D3_CD_B4_01_0.fcs · D3_CD_B5_01_0.fcs · D3_IgG_B1_01_1.fcs · D3_IgG_B2_01_0.fcs · D3_IgG_B3_01_0.fcs · D3_IgG_B4_01_0.fcs · D3_IgG_B5_01_0.fcs |
| Timepoints | |
|---|---|
| · D_1 | c01_D_1 PBL+_01_1_HD.fcs · c01_D_1 PBL-_01_0_HD.fcs · c02_D_1 PBL+_01_1_Pt.fcs · c02_D_1 PBL-_01_0_Pt.fcs · D_1 CD_B1_01.FCS · D_1 CD_B2_01.FCS · D_1 CD_B3_01.FCS · D_1 CD_B4_01.FCS · D_1 CD_B5_01.FCS · D_1 IgG_B1_2_01.FCS · D_1 IgG_B2_02.FCS · D_1 IgG_B3_02.FCS · D_1 IgG_B4_02.FCS · D_1 IgG_B5_02.FCS |
| · D_2 | c01_D_2 PBL+_02_0_HD.fcs · c01_D_2 PBL-_02_0_HD.fcs · c02_D_2 PBL+_02_0_Pt.fcs · c02_D_2 PBL-_02_0_Pt.fcs · D2_CD_B1_006_0.fcs · D2_CD_B2_01_1.fcs · D2_CD_B3_01_0.fcs · D2_CD_B4_01_0.fcs · D2_CD_B5_02_0.fcs · D2_IgG_B1_01_1.fcs · D2_IgG_B2_01_1.fcs · D2_IgG_B3_01_1.fcs · D2_IgG_B4_01_0.fcs · D2_IgG_B5_01_0.fcs |
| · D_3 | c01_D_3 PBL+_02_0_HD.fcs · c01_D_3 PBL-_02_0_HD.fcs · c01_Unstained_01_0_HD.fcs · c02_D_3 PBL+_02_0_Pt.fcs · c02_D_3 PBL-_02_0_Pt.fcs · c02_Unstained_01_0_Pt.fcs · D3_CD_B1_01_0.fcs · D3_CD_B2_01_0.fcs · D3_CD_B3_01_0.fcs · D3_CD_B4_01_0.fcs · D3_CD_B5_01_0.fcs · D3_IgG_B1_01_1.fcs · D3_IgG_B2_01_0.fcs · D3_IgG_B3_01_0.fcs · D3_IgG_B4_01_0.fcs · D3_IgG_B5_01_0.fcs |
| Doses | |
|---|---|
| · + | c01_D_2 PBL+_02_0_HD.fcs · c02_D_2 PBL+_02_0_Pt.fcs · c01_D_1 PBL+_01_1_HD.fcs · c02_D_1 PBL+_01_1_Pt.fcs · c01_D_3 PBL+_02_0_HD.fcs · c02_D_3 PBL+_02_0_Pt.fcs |
| · - in vitro natalizumab | c01_D_2 PBL-_02_0_HD.fcs · c02_D_2 PBL-_02_0_Pt.fcs · c01_D_1 PBL-_01_0_HD.fcs · c02_D_1 PBL-_01_0_Pt.fcs · c01_D_3 PBL-_02_0_HD.fcs · c02_D_3 PBL-_02_0_Pt.fcs |
| Sample Type | |
|---|---|
| · Unstained | c01_Unstained_01_0_HD.fcs · c02_Unstained_01_0_Pt.fcs |
