Kits for multiparametric phospho analysis

Abstract

As disclosed herein, the present invention provides for kits and a composition for diagnosis, prognosis, drug discovery, drug development, and patient stratification. The kits can comprise a plurality of binding elements for cell surface markers, and a plurality of binding elements for state-specific intracellular markers. The kits can further comprise a plurality of modulators directed for the particular cell function or signaling pathways. The kits can further include fixatives, permeabilizing agent, buffers, containers, instructions, and software for data analysis/compilation.

1 - 35 . (canceled) 
     
     
         36 . A kit comprising:
 (i) at least one binding element specific for a cell surface marker selected from the group consisting of CD7, CD11b, CD14, CD 33, CD34, and CD45; and   (ii) at least one binding element specific for an intracellular marker selected from the group consisting of cleaved PARP, cytochrome C, p-ATM, p-ATR, p-CHK1, p-CHK2, p-H2AX, p-p53, cleaved Caspase 3 and cleaved Caspase 8.   
     
     
         37 . The kit of  claim 36  comprising at least two binding elements specific for a cell surface marker. 
     
     
         38 . The kit of  claim 36  comprising at least three binding elements specific for a cell surface marker. 
     
     
         39 . The kit of  claim 37  wherein the cell surface markers are selected from the group consisting of CD11b, CD34, and CD45. 
     
     
         40 . The kit of  claim 39  wherein the cell surface markers comprise CD34 and CD45. 
     
     
         41 . The kit of  claim 36  wherein one or more of the binding elements comprises a detection element. 
     
     
         42 . The kit of  claim 41  wherein the detection element comprises a fluorophore. 
     
     
         43 . The kit of  claim 41  wherein the detection element is detectable by mass spectrometry. 
     
     
         44 . The kit of  claim 36  further comprising at least one modulator selected from the group consisting of Etoposide, Ara-C, Daunorubicin, Gemtuzumab, and Staurosporin. 
     
     
         45 . The kit of  claim 44  wherein the modulator comprises etoposide, Ara-c, daunorubicin, or a combination thereof. 
     
     
         46 . The kit of  claim 44  wherein the modulator comprises etoposide. 
     
     
         47 . The kit of  claim 44  wherein the modulator comprises Ara-c, daunorubicin, or a combination thereof. 
     
     
         48 . The kit of  claim 36  wherein the intracellular marker comprises cleaved PARP. 
     
     
         49 . The kit of  claim 36  wherein the binding elements comprise antibodies. 
     
     
         50 . The kit of  claim 36  further comprising control cells. 
     
     
         51 . The kit of  claim 50  wherein the control cells are fixed/preserved. 
     
     
         52 . The kit of  claim 50  wherein the control cells are lyophilized. 
     
     
         53 . The kit of  claim 36  further comprising calibration beads for calibration of a fluorescence detector. 
     
     
         54 . The kit of  claim 36  further comprising instructions. 
     
     
         55 . The kit of  claim 54  wherein the instructions comprise printed instructions. 
     
     
         56 . The kit of  claim 36  further comprising a reagent selected from the group consisting of a fixative, a permeabilizing agent, or a buffer. 
     
     
         57 . The kit of  claim 36  wherein the components are packaged together in a single container. 
     
     
         58 . The kit of  claim 36  further comprising at least one dye selected from the group consisting of Annexin-V, Yo-Pro, propidium iodide (PI), Amine Aqua, DRAQ5, and XIAP. 
     
     
         59 . The kit of  claim 36  wherein the dye comprises Amine Aqua. 
     
     
         60 . A kit comprising:
 (i) at least one binding element specific for a cell surface marker selected from the group consisting of CD7, CD11b, CD14, CD15, CD33, CD34, and CD45; and   (ii) at least one state-specific one binding element specific for a the protein selected from the group consisting of STAT1, STAT3, STAT5, S6, Erk, Akt, ATM, ATR, Chk1, Chk2, 53BP1, PARP, H2AX, Caspase 3, Caspase 8, CRKL, Histone H3, Cyclin B1, Cyclin D1, Cyclin E, Cyclin A, p15, p16, p21, PLCγ2, p53, SLP-76, and CREB.   
     
     
         61 . The kit of  claim 60  comprising at least two binding elements specific for at least two cell surface markers selected from the group consisting of CD7, CD11b, CD14, CD15, CD33, CD34, and CD45. 
     
     
         62 . The kit of  claim 60  comprising at least three binding elements specific for at least three cell surface markers selected from the group consisting of CD7, CD11b, CD14, CD15, CD33, CD34, and CD45. 
     
     
         63 . The kit of  claim 60  comprising at least two state-specific binding elements specific for at least two proteins selected from the group consisting of STAT1, STAT3, STAT5, S6, Erk, Akt, ATM, ATR, Chk1, Chk2, 53BP1, PARP, H2AX, Caspase 3, Caspase 8, CRKL, Histone H3, Cyclin B1, Cyclin D1, Cyclin E, Cyclin A, p15, p16, p21, PLCg2, p53, SLP-76, and CREB. 
     
     
         64 . The kit of  claim 60  comprising at least three state-specific binding elements specific for at least three proteins selected from the group consisting of STAT1, STAT3, STAT5, S6, Erk, Akt, ATM, ATR, Chk1, Chk2, 53BP1, PARP, H2AX, Caspase 3, Caspase 8, CRKL, Histone H3, Cyclin B1, Cyclin D1, Cyclin E, Cyclin A, p15, p16, p21, PLCg2, p53, SLP-76, and CREB. 
     
     
         65 . A kit comprising
 (i) at least one binding element specific for a cell surface marker selected from the group consisting of CD20, CD19, CD79a, IgM, IgD, CD5, CD3, CD38, kappa light chain, and lambda light chain; and   (ii) at least one binding element specific for an intracellular marker selected from the group consisting of p-Lyn, p-Syk, p-BLNK, p-PLCg2, p-Akt, p-Erk, p-s6, p-Stat5, p-Stat1, p-Stat3, cleaved PARP, and cleaved caspase 3.   
     
     
         66 . The kit of  claim 65  comprising at least two binding elements specific for a cell surface marker. 
     
     
         67 . The kit of  claim 65  comprising at least three binding elements specific for a cell surface marker. 
     
     
         68 . The kit of  claim 66  wherein the cell surface markers are selected from the group consisting of CD20, CD19, CD3, and C5. 
     
     
         69 . The kit of  claim 67  wherein the cell surface markers are selected from the group consisting of CD20, CD19, CD3, and C5. 
     
     
         70 . The kit of  claim 65  wherein one or more of the binding elements comprises a detection element. 
     
     
         71 . The kit of  claim 70  wherein the detection element comprises a fluorophore. 
     
     
         72 . The kit of  claim 70  wherein the detection element is detectable by mass spectrometry. 
     
     
         73 . The kit of  claim 65  further comprising at least one modulator selected from the group consisting of anti-m, anti-g, fludarabine (F-ara-a), and bendamustine. 
     
     
         74 . The kit of  claim 73  wherein the modulator comprises anti-μ. 
     
     
         75 . The kit of  claim 65  wherein the at least one binding element is specific for an intracellular marker selected from the group consisting of p-Syk, p-Akt, p-Erk, p-s6, and cleaved PARD. 
     
     
         76 . The kit of  claim 65  wherein the binding elements comprise antibodies. 
     
     
         77 . The kit of  claim 65  further comprising control cells. 
     
     
         78 . The kit of  claim 77  wherein the control cells are fixed/preserved. 
     
     
         79 . The kit of  claim 78  wherein the control cells are lyophilized. 
     
     
         80 . The kit of  claim 65  further comprising calibration beads for calibration of a fluorescence detector. 
     
     
         81 . The kit of  claim 65  further comprising instructions. 
     
     
         82 . The kit of  claim 81  wherein the instructions comprise printed instructions. 
     
     
         83 . The kit of  claim 65  further comprising a reagent selected from the group consisting of a fixative, a permeabilizing agent, and a buffer. 
     
     
         84 . The kit of  claim 65  wherein the components are packaged together in a single container. 
     
     
         85 . The kit of  claim 65  further comprising at least one dye selected from the group consisting of Annexin-V, Yo-Pro, propidium iodide (PI), Amine Aqua, DRAQ5, and XIAP. 
     
     
         86 . The kit of  claim 65  wherein the dye comprises Amine Aqua.