The present invention recognizes that current clinical
laboratory testing methods for multiparametric
single cell analysis are limited to analysis of intact live cells, and are insufficient for identification of signaling activation profile defining certain
cell types, including but not limited to neoplastic and immunologically activated
cell subsets. One aspect of the present invention generally relates to
marker selection in panels to include proteins routinely assessed in standard FCM, while preferably also incorporating markers for surface
receptor proteins within activated signaling cascades. A further aspect of the present invention generally relates to
panel design for the following indications in neoplastic and non-neoplastic clinical applications as examples of the technology: (a) identification of CML
progenitor cell subsets in the setting of
disease recurrence
after treatment discontinuation or relapse due to
treatment resistance, and (b) characterization of activated basophils to predict the severity of an
allergic response. Another aspect of the present invention generally relates to methods to measure levels of surface and IC biomarkers in separate or combined assays for robust characterization of each or select cell compartment, and
data analysis based on results from each or all method(s) used for optimal detection of the markers. A further aspect of the present invention generally relates to the identification and profiling of
cell subpopulations based on analysis of surface markers including those associated with lineage and maturation of cell types and
receptor proteins, and the corresponding IC phosphoproteins including those in activated signaling cascades to predict certain
disease states or
response to treatment.