Claims
- 1. A method for detecting protein modification of a target protein in a sample, which method comprises:
a) contacting a sample containing or suspected of containing a target protein with a capture molecule immobilized on a solid support, said capture molecule is capable of specifically binding to said target protein, whereby said target protein is immobilized on said solid support; and b) assessing modification status and/or identity of said immobilized target protein.
- 2. The method of claim 1, wherein a plurality of target proteins is contacted with the immobilized capture molecule simultaneously.
- 3. The method of claim 2, wherein the plurality of target proteins comprises a group of structurally and/or functionally related proteins.
- 4. The method of claim 1, wherein a target protein is contacted with a plurality of immobilized capture molecules simultaneously.
- 5. The method of claim 1, wherein a plurality of target proteins is contacted with a plurality of immobilized capture molecules simultaneously.
- 6. The method of claim 1, wherein the capture molecule is capable of specifically binding to both a modified and an unmodified forms of a target protein.
- 7. The method of claim 1, wherein the capture molecule is capable of specifically binding to a modified form of the target protein but is not capable of specifically binding to an unmodified form of the target protein.
- 8. The method of claim 1, wherein the capture molecule is an antibody.
- 9. The method of claim 1, wherein the solid support is selected from the group consisting of silicon, plastic, nylon, glass, ceramic, photoresist, rubber or polymer support.
- 10. The method of claim 1, wherein the solid support comprises a flat support, a set of sticks, or a set of beads.
- 11. The method of claim 10, wherein the flat support comprises a slide, a chip, a filter, or a membrane.
- 12. The method of claim 1, wherein the protein modification is selected from the group consisting of phosphorylation, acetylation, methylation, ADP-ribosylation, addition of a polypeptide side chain, addition of a hydrophobic group, and addition of a carbohydrate.
- 13. The method of claim 12, wherein the phosphorylation is on an amino acid residue selected from the group consisting of tyrosine, serine, threonine and histidine.
- 14. The method of claim 12, wherein the addition of a polypeptide side chain is the addition of ubiquitin.
- 15. The method of claim 12, wherein the addition of a hydrophobic group is the addition of a fatty acid, addition of an isoprenoid, or addition of a glycosyl-phosphatidyl inositol anchor.
- 16. The method of claim 15, wherein the fatty acid is myristate or palmitate.
- 17. The method of claim 15, wherein the isoprenoid is famesyl or genranylgenranyl.
- 18. The method of claim 12, wherein the carbohydrate group comprises glycosyl.
- 19. The method of claim 6, wherein the modification status of the immobilized target protein is assessed by contacting the immobilized target protein with a detection molecule that is capable of specifically binding to the modified target protein but is not capable of specifically binding to the unmodified target protein itself.
- 20. The method of claim 6, wherein the modification status of the immobilized target protein is determined by a physical or chemical means.
- 21. The method of claim 20, wherein the physical or chemical means comprises chemical or radioisotopic label of the protein modification moiety.
- 22. The method of claim 20, wherein the physical or chemical means is selected from the group consisting of chromatographic, electrophoretic, protein sequencing, mass spectrometry and NMR means for detecting the protein modification moiety.
- 23. The method of claim 7, wherein the identity of the immobilized target protein is assessed by contacting the immobilized target protein with a detection molecule that is capable of specifically binding to the unmodified target protein itself but is not capable of specifically binding to the modified target protein.
- 24. The method of claim 1, wherein the sample is a biological sample.
- 25. The method of claim 1, wherein the target protein is involved in a biological pathway, belongs to a group of proteins with identical or similar biological function, expressed in a stage of cell cycle, expressed in a cell type, expressed in a tissue type, expressed in an organ type, expressed in a developmental stage, a protein whose expression and/or activity is altered in a disease or disorder type or stage, or a protein whose expression and/or activity is altered by drug or other treatments.
- 26. A method for identifying biologically distinguishable marker(s) associated with a biosample, which comprises:
1) assessing protein modification profile of a biosample through the method of claim 1;2) assessing protein modification profile of a comparable control biosample through the method of claim 1; and 3) comparing the protein modification profile obtained in step 1) with the protein modification profile obtained in step 2) to identify biologically distinguishable protein modification profile marker(s) associated with said biosample.
- 27. A kit for detecting protein modification, which kit comprises:
a) a capture molecule immobilized on a solid support, said capture molecule is capable of specifically binding to a target protein; and b) means for assessing modification status and/or identity of said target protein.
- 28. The kit of claim 27, wherein a plurality of capture molecules is immobilized on the solid support, each of said capture molecules is capable of specifically binding to a member protein of a group of structurally and/or functionally related target proteins.
- 29. The kit of claim 27, wherein the capture molecule is capable of specifically binding to both a modified and an unmodified forms of a target protein.
- 30. The kit of claim 27, wherein the capture molecule is capable of specifically binding to a modified form of the target protein but is not capable of specifically binding to an unmodified form of the target protein.
- 31. The kit of claim 27, wherein the capture molecule is an antibody.
- 32. The kit of claim 27, wherein the protein modification is selected from the group consisting of phosphorylation, acetylation, methylation, ADP-ribosylation, addition of a polypeptide side chain, addition of a hydrophobic group, and addition of a carbohydrate.
- 33. The kit of claim 29, wherein the modification status of the immobilized target protein is assessed by contacting the immobilized target protein with a detection molecule that is capable of specifically binding to the modified target protein but is not capable of specifically binding to the unmodified target protein itself.
- 34. The kit of claim 29, wherein the modification status of the immobilized target protein is determined by a physical or chemical means.
- 35. The kit of claim 30, wherein the identity of the immobilized target protein is assessed by contacting the immobilized target protein with a detection molecule that is capable of specifically binding to the unmodified target protein itself but is not capable of specifically binding to the modified target protein.
- 36. The kit of claim 27, further comprising:
a) instructions for using the kit; b) reagents and buffers; and/or c) a container(s) for the kit contents.
- 37. An array of protein capture molecules, which array comprises:
a) a solid support; and b) a plurality of capture molecules immobilized on said solid support, wherein each of said molecules is capable of specifically binding to both a modified and an unmodified form of a member protein of a plurality of target proteins.
- 38. The array of claim 37, wherein the plurality of target proteins comprises a group of structurally and/or functionally related proteins.
- 39. The array of claim 38, wherein the modified and unmodified forms of the same target protein have different biological activities.
- 40. The array of claim 38, wherein the modified and unmodified forms of the same target protein represent different physiological conditions or biological statuses.
- 41. The array of claim 40, wherein the array is used to identify pathway activation.
- 42. The array of claim 40, wherein the array is used to identify activation of a group of structurally and/or functionally related protein.
- 43. The array of claim 40, wherein the array is used to generate a modification profile correlated to a physiological condition, drug treatment and disease.
- 44. The array of claim 40, wherein the array is used to identify a physiological or pathological status.
- 45. The array of claim 40, wherein the array is used to record biological perturbation caused by drug and other treatment.
- 46. An array of protein capture molecules, which array comprises:
a) a solid support; and b) a plurality of capture molecules immobilized on said solid support, wherein each of said molecules is capable of specifically binding to an epitope generated by a specific modification moiety of a modified protein.
- 47. The array of claim 46, wherein the modified protein is Rb.
- 48. An array of enzyme substrates, which array comprises:
a) a solid support; and b) a plurality of substrates immobilized on said solid support, wherein each of said substrates is a substrate of a member enzyme of a group of structurally and/or functionally related enzymes.
- 49. The array of claim 48, wherein at least one of the member enzymes catalyzes a protein modification reaction.
- 50. A kit for detecting enzymatic activity, which kit comprises:
a) the array of claim 48; and b) means for assessing activity of each of the member enzymes.
- 51. A method for detecting enzymatic activity in a sample, which method comprises:
a) contacting a sample containing or suspected of containing a group of structurally and/or functionally related target enzymes with a plurality of substrates immobilized on a solid support, wherein each of said substrates is a substrate of a member enzyme of said group of target enzymes under conditions suitable for said target enzymes to catalyze enzymatic reactions involving said immobilized substrates; and b) assessing enzymatic activities of said target enzymes.
- 52. The method of claim 51, wherein at least one of the target enzymes catalyzes a protein modification reaction.
Parent Case Info
[0001] This application claims the benefit of the priority date of the U.S. Provisional Patent Application Serial No. 60/158,560, filed Oct. 8, 1999 under 35 U.S.C. §119(e). The content of the above-referenced application is herein incorporated by reference in its entirety.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60158560 |
Oct 1999 |
US |
Divisions (1)
|
Number |
Date |
Country |
Parent |
09678644 |
Oct 2000 |
US |
Child |
10356442 |
Jan 2003 |
US |