Figure 1

Diagram illustrating the process used for the selection and evaluation of markers for the NGF MM MRD panel. Left and lower right panels show the sequential steps followed to select those markers providing the best resolution between aPCs and nPC, including principal component 1 (PC1) vs PC2 —automatic population separator (APS1)— plots illustrating the described comparisons for steps 1, 2 and 4, respectively. (right; a) An APS1 plot corresponding to the simultaneous evaluation of 31 normal/reactive BM samples vs all 63 MM patients studied at diagnosis, in which the resolution power of the EuroFlow diagnostic PCD antibody panel combination is illustrated. Please note that PC from five samples (highlighted by the black arrows) showed suboptimal separation in the overall comparison; nonetheless, when individually compared vs the normal/reactive reference PC pool (b–f), these cases also showed sufficient phenotypic discrimination from nPC. Markers contributing to PC1 (and their percentage contribution) for each panel on the right include: (a) CD19(20%), CD56(17%), CD81(13%), CD45(11%), CD117(10%), CD27(8%), CyIgλ(5%), CD38(5%), CyIgκ(4%), β2 micro(3%), CD138(3%), CD28(1%); (b) CyIgκ(27%), CyIgλ(13%), β2 micro(12%), CD38(11%), CD56(10%), CD138(7%), CD28(16%), CD45(4%), CD27(4%), CD117(4%), CD19(2%), CD81(0%); (c) CyIgκ(20%), CD45(16%), CD56(11%), CD28(10%), CD19(10%), CD27(8%), CD117(6%), CD81(6%), CD38(6%), CyIgλ(4%), CD138(3%), β2 micro(0%); (d), CD19(29%), β2 micro(13%), CyIgκ(13%), CD56(13%), CD45(11%), CD38(5%), CD27(4%), CyIgλ(3%), CD28(3%), CD138(2%), CD117(2%), CD81(2%); (e) CD19(23%), β2 micro(13%), CD45(12%), CyIgλ(11%), CD81(9%), CD56(7%), CyIgκ(6%), CD38(6%), CD28(4%), CD138(4%), CD27(4%), CD117(1%); (f) CD19(20%), CD117(19%), CD81(14%), CD45(13%), CD56(12%), CyIgλ(9%), β2 micro(6%), CD38(3%), CD27(2%), CD28(1%), CD138(1%), CyIgκ(0%). In all PC1 vs PC2 plots, solid circles represent median values for the 12 fluorescence-associated parameters evaluated, inner (dotted) and outer (solid) lines represent the first and second standard deviations for individual PC. nPC populations are depicted in green while aPC are shown as red dots, circles and lines, respectively.