Assay performance: Linking repeatability, reproducibility and accuracy
Here we present results from the award-winning* Maxpar Direct Immune Profiling System, developed for use on the Helios™ mass cytometer, a CyTOF® system, and designed for comprehensive single-tube immune monitoring studies. The system consists of the Maxpar® Direct™ Immune Profiling Assay™ for processing PBMC or whole blood samples and Maxpar Pathsetter™ software for automated five-minute data analysis.
Take a look at the results using healthy human peripheral blood mononuclear cell (PBMC) samples to see the intra-assay repeatability, inter-site reproducibility and accuracy of the dry panel format.
To see the whole blood data from the same performance studies, download the full white paper, Deep Immune Profiling with the Maxpar Direct Immune Profiling System (FLDM-400247), or see the peer-reviewed publication:
Intra-assay repeatability refers to the precision of the assay within the run, demonstrating the reliability of the results obtained with the same lot and instrument.
Figure 1. Workflow for assessing the intra-assay repeatability of the Maxpar Direct Immune Profiling System with PBMC
Figure 2. Repeatability of cell frequency measurements from samples stained by three technicians in triplicate. The Y-axis is the measured percentage of total single live cells. All populations with a frequency ≥5% had %CV of the mean <9%. For details, see the Intra-Assay Repeatability section of the immune profiling white paper (FLDM-400247).
In the multi-site study presented here, the variability in immune cell population frequencies from single-donor PBMC samples was measured across five independent testing sites.
Figure 3. Workflow for assessing inter-site reproducibility from five different sites for cryopreserved PBMC samples isolated from a single donor
Figure 4. The inter-site reproducibility of cell frequency measurements at multiple sites for PBMC processed and analyzed with the Maxpar Direct Immune Profiling System. For all populations ≥5% in frequency, the %CV of the mean was <20%. For details, see the Inter-Site Reproducibility section of the immune profiling white paper (FLDM-400247).
Accuracy: Liquid Panel vs. Dry Panel
In order to test the accuracy of the dry format reagents, staining was performed using tubes from the Maxpar Direct Immune Profiling Assay (dry format) and a panel containing the same antibodies in liquid form in parallel.
Figure 5. The workflow for assessing accuracy of the liquid vs. the dry antibody panel. Single PBMC and whole blood donor samples were independently stained and processed by three technicians using both panel formats.
Figure 6. Accuracy of the liquid vs. the dry antibody panel. The coefficient of determination (R2) for the average frequency of each population was >0.99 for the two methods. For details, see the Inter-Site Reproducibility section of the immune profiling white paper (FLDM-400247).