Patent ID: 7507548
Filing Date: 2009-03-24
Classification: G01N,Y10S,Y10T

Abstract:
1. A method for the identification of aberrant phenotypes expressed by neoplastic cells comprising the steps of: a) separately staining one or more normal/reactive samples and one neoplastic sample with overlapping multiple combinations of monoclonal antibodies, each monoclonal antibody in each combination being conjugated to a different fluorochrome and each combination having in common at least three fluorochrome conjugated monoclonal antibodies that are specific for tumor cells of interest contained in said neoplastic samples; b) sequentially measuring at least two light scatter emissions and the at least four fluorescence emissions of each stained cell from the normal/reactive samples and the neoplastic sample, using flow cytometry; c) storing two independent list mode data files, one containing measurement information on the specific light scatter and fluorescence characteristics of each cell analyzed from the normal/reactive samples in step b) and the other containing measurement information on the specific light scatter and fluorescence characteristics of each cell analyzed from the neoplastic sample in step b); d) creating new data files from step c) by merging, at known proportions cellular events from the data file containing measurement information about the cells present in the neoplastic sample from step b) into the data file containing measurement information on the cells present in the normal/reactive samples from step b); e) defining in a multidimensional space generated by the flow cytometric measurements of light scatter and fluorescence emissions from preestablished standards, those areas occupied by events corresponding to normal cells and those areas corresponding to empty spaces in normal/reactive samples and that may be occupied by tumor cells in neoplastic samples; f) sequentially identifying in the data files containing measurement information about the cells present in the neoplastic sample and merged as described in step d), those events corresponding to neoplastic cells as those populations of events contained in the neoplastic sample which fall into the empty spaces identified in the multidimensional space generated by the flow cytometric measurements of light scatter and fluorescence emissions from pre-established standards in step e); and g) establishing and identifying the phenotypic aberrations displayed by the neoplastic cells as compared to their normal counterpart, as those combinations of flow cytometric measurements of light scatter and fluorescence emissions from the events corresponding to the neoplastic cells contained in the merged data file from step d), that provide their identification and distinction from those events corresponding to cells from normal/reactive samples contained in the same merged data file from step d).