Patent ID: 8951720
Filing Date: 2015-02-10
Classification: G01N

Abstract:
1. A method for identifying co-expressed differentiator marker proteins comprising: a. providing a first sample from a cancer patient comprising malignant tissue from the gingivo-buccal area of the oral cavity of said cancer patient; b. providing a second sample comprising non malignant tissue from the gingivo-buccal area of the oral cavity of said cancer patient; c. micro-dissecting transformed epithelial cells from frozen sections of the malignant tissue and the non transformed epithelial layer from frozen sections of the non malignant tissue; d. separating proteins of lysates prepared from the micro-dissected transformed and non transformed epithelial cells from (c) by two dimensional gel electrophoresis and staining with silver; e. generating micro-dissected epithelial cell protein profiles from the samples in c) and d); f. comparing the micro-dissected epithelial cell protein profiles generated in step (e) and generating a short list of protein spots commonly expressed in both normal and transformed epithelium; g. punching out the short listed protein spots from a silver stained two dimensional gel of micro-dissected transformed epithelial cells; h. destaining and drying the protein spots and digesting obtained protein with trypsin; i. analyzing eluted dried peptides by mass spectroscopy to obtain identities of proteins; j. comparing the expression of each short-listed protein spot in non malignant and transformed epithelial cells to identity a set of co expressed differentiator marker proteins for the transformed epithelium from the group consisting of lactate dehydrogenase, alpha enolase, prohibitin, cathepsin D, apolipoprotein A-I, tumor protein translationally controlled-1, a stratifin (SFN) family protein, 14-3-3σ, tropomyosin and 14-3-3ζ (YWHAZ); and k. identifying key differentiator protein spots using Linear Discriminator Analysis.