Method for Detection of L523S Expression in Biological Samples

Abstract

The present invention discloses methods for differentiating between normal or reactive cells and malignant cells in biological samples comprising: exposing the biological sample to an antibody to L523S protein and detecting the presence of the antibody bound to L523S protein within the malignant cells, in embodiments, such methods may further comprise: exposing the biological sample to an antibody to a second protein that is a marker of cell lineage and detecting the presence of the antibody to the second protein within cells corresponding to a particular cell lineage and/or may comprise: exposing the biological sample to an antibody to a third protein which is a marker of cells that are normal or reactive and detecting the presence of the antibody to the third protein within cells that are normal or reactive.

1 . A method for differentiating between normal or reactive cells and malignant cells in biological samples comprising:
 (a) exposing a the biological sample to an antibody to L523S protein and detecting the presence of the antibody bound to L523S protein within malignant cells;   (b) exposing the biological sample to an antibody to a second protein that is a marker of cell lineage and detecting the presence of the antibody to the second protein within cells corresponding to a particular cell lineage; and   (c) exposing the biological sample to an antibody to a third protein that is a marker of cells that are normal or reactive and detecting the presence of the antibody to the third protein within cells that are normal or reactive.   
   
   
       2 . (canceled) 
   
   
       3 . The method of  claim 1 , wherein the second protein is a marker of mesothelial cell lineage. 
   
   
       4 . The method of  claim 2 , wherein the second protein is calretinin. 
   
   
       5 . (canceled) 
   
   
       6 . The method of  claim 1 , wherein the third protein is desmin. 
   
   
       7 . The method of  claim 1 , wherein at least one detecting step comprises detecting an immunohistochemical stain or flow cytometric evaluation. 
   
   
       8 . The method of  claim 1 , wherein the type of malignant cells is chosen from melanoma, mesothelioma, lung adenocarcinoma, spindle cell carcinoma, squamous cell carcinoma, colonic adenocarcinoma, ovarian carcinoma, gastric carcinoma, pancreatic carcinoma, asctrocytoma, Hodgkin's lymphoma, lymphoma, malignant fibrous histocytoma, Ewing's sarcoma, and carcinoid. 
   
   
       9 . (canceled) 
   
   
       10 . The method of  claim 1 , wherein the biological sample is a tissue biopsy, biological fluid, cell monolayer, or cell pellet derived from a biological fluid. 
   
   
       11 . The method according to  claim 10 , wherein the tissue biopsy is obtained from a pigmented skin lesion. 
   
   
       12 - 13 . (canceled) 
   
   
       14 . The method according to  claim 10 , wherein the biological fluid is lung pleural effusion. 
   
   
       15 . The method of  claim 1 , wherein the step of detecting the presence of the antibody bound to L523S protein within the malignant cells comprises the sub-steps of:
 further exposing the biological sample to a labeled secondary antibody that reacts with the antibody bound to L523S protein; and   detecting the presence of the secondary antibody bound to the antibody bound to L523S protein.   
   
   
       16 . The method of  claim 1 , wherein the step of detecting the presence of the antibody bound to L523S protein within the malignant cells comprises the sub-steps of:
 further exposing the biological sample to a labeled reagent comprising an enzyme-conjugated polymer backbone that carries secondary antibody molecules that react with the antibody bound to L523S protein; and   detecting the presence of the labeled reagent.   
   
   
       17 . The method of  claim 10 , wherein the biological sample is a tissue biopsy, and wherein step (a) is performed on a first portion of the sample, step (b) is performed on a second portion of the sample, and step (c) is performed on a third portion of the sample. 
   
   
       18 - 26 . (canceled) 
   
   
       27 . The method of  claim 17 , wherein the first and second portions are different microtomed sections from the same tissue biopsy. 
   
   
       28 . The method of  claim 17 , wherein the first, second, and third portions are different microtomed sections from the same tissue biopsy. 
   
   
       29 - 38 . (canceled) 
   
   
       39 . The method of  claim 1 , wherein the step of detecting the presence of the antibody bound to the second protein within cells corresponding to a particular cell lineage comprises the sub-steps of:
 further exposing the biological sample to a labeled secondary antibody that reacts with the antibody bound to the second protein; and   detecting the presence of the secondary antibody bound to the antibody bound to the second protein.   
   
   
       40 . The method of  claim 1 , wherein the step of detecting the presence of the antibody bound to the second protein within cells corresponding to a particular cell lineage comprises the sub-steps of:
 further exposing the biological sample to a labeled reagent comprising an enzyme-conjugated polymer backbone that carries secondary antibody molecules that react with the antibody bound to the second protein; and   detecting the presence of the labeled reagent.   
   
   
       41 . The method of  claim 1 , wherein the step of detecting the presence of the antibody bound to the third protein within cells which are normal or reactive comprises the sub-steps of:
 further exposing the biological sample to a labeled secondary antibody that reacts with the antibody bound to the third protein; and   detecting the presence of the secondary antibody bound to the antibody bound to the third protein.   
   
   
       42 . The method of any one of  claim 1 , wherein the step of detecting the presence of the antibody bound to the third protein within cells which are normal or reactive comprises the sub-steps of:
 further exposing the biological sample to a labeled reagent comprising an enzyme-conjugated polymer backbone that carries secondary antibody molecules that react with the antibody bound to the third protein; and   detecting the presence of the labeled reagent.   
   
   
       43 . A panel for differentiating between normal or reactive cells and malignant cells in biological samples, wherein said panel comprises an antibody to L523S protein, an antibody to a second protein that is a marker of cell lineage, and an antibody to a third protein which is a marker of cells that are normal or reactive. 
   
   
       44 . The panel of  claim 43 , wherein the second protein is calretinin. 
   
   
       45 . The panel of  claim 43 , wherein the third protein is desmin. 
   
   
       46 . The panel of  claim 43 , wherein said panel consists of an antibody to L523S, an antibody to calretinin, and an antibody to desmin. 
   
   
       47 . The panel of  claim 43 , wherein at least one of the antibodies is monoclonal. 
   
   
       48 . The panel of  claim 43 , wherein said panel is used for the detection of melanoma. 
   
   
       49 . The panel of  claim 43 , wherein said panel comprises at least one labeled antibody. 
   
   
       50 . A kit for the detection of melanoma, comprising a panel of antibodies as defined in  claim 43 . 
   
   
       51 . The kit of claim  34 , comprising an antibody to L523S, an antibody to calretinin, and an antibody to desmin.