MAGNETIC ENRICHMENT OF CIRCULATING CELLS, FRAGMENTS AND DEBRIS FOR ENABLING HTS PROTEOMICS AND GENOMICS IN DISEASE DETECTION

摘要

The methods and reagents described in this invention are used to analyze circulating tumor cells, clusters, fragments, and debris. Analysis is performed with a number of platforms, including flow cytometry, CellSpotter<SUP>TM</SUP> fluorescent microscopy imaging andmass spectrometric profiling. In addition to analyzing and enumerating morphologicaly classified cells by imaging, the presence of damaged cells and derived debris in magnetically enriched specimens has been shown to be an important "smoking gun" indicator of demised tumor cells. Described herein are improved methods to screen, diagnose and monitor disease based on intact circulating rare cells and associated fragments or debris. The present invention incorporates proteomic and genomic profiling of cellular debris in cancer cell analysis, either complementary to or independent of image cytometry. The technology provides diagnosis and prognostic tools in diseases such as cancer that incorporate not only detection and quantitation of intact circulating cells but also of cellular fragments and debris that bear generic cytoskeletal and/or tumor-specific markers. The presence of such markers reflects immune function in early disease and/or therapeutic effectiveness in later stages.