Patent ID: 8030083
Filing Date: 2011-10-04
Classification: B01J,G01N,Y10T

Abstract:
1. A method for analyzing a structural affinity relationship between plural kinds of proteins and plural kinds of test compounds, comprising the steps of: (a) using a detection compound-immobilized carrier to purify plural kinds of 1st isotope-labeled proteins bound to the detection compound immobilized on the detection compound-immobilized carrier, from a 1st group of proteins labeled with the 1st isotope; (b) using the detection compound-immobilized carrier to purify plural kinds of proteins, which may or may not be labeled with a 2nd isotope, bound to the detection compound immobilized on the detection compound-immobilized carrier, from a 2nd group of proteins, which may or may not be labeled with the 2nd isotope, wherein the 2nd group of proteins is brought into contact with a 1st test compound before using the detection compound-immobilized carrier; (c) using the detection compound-immobilized carrier to purify plural kinds of proteins, which may or may not be labeled with the 2nd isotope, bound to the detection compound immobilized on the detection compound-immobilized carrier, from a 3rd group of proteins, which may or may not be labeled with the 2nd isotope, wherein the 3rd group of proteins is brought into contact with a 2nd test compound before using the detection compound-immobilized carrier; wherein the detection compound is the same for steps (a) to (c), wherein the 1st test compound is the same compound as the detection compound, wherein the 1st and 2nd isotopes are different from each other, wherein the 1st and 2nd test compounds are different from each other, wherein the 1st group of proteins does not have contact with the 1st or 2nd test compound, wherein the 2nd group of proteins does not have contact with the 2nd test compound, and wherein the 3rd group of proteins does not have contact with the 1st test compound before using the detection-compound-immobilized carrier; (d) mixing the plural kinds of proteins purified in step (a) together with the plural kinds of proteins purified in step (b) to prepare a 1st protein mixture; (e) analyzing the plural kinds of proteins in the 1st protein mixture prepared in step (d) with mass spectrometry; (f) identifying each of the plural kinds of proteins analyzed in step (e) based on information obtained by the mass spectrometry performed in step (e); (g) mixing the plural kinds of proteins purified in step (a) together with the plural kinds of proteins purified in step (c) to prepare a 2nd protein mixture; (h) analyzing the plural kinds of proteins in the 2nd protein mixture prepared in step (g) with mass spectrometry; (i) identifying each of the plural kinds of proteins analyzed in step (h) based on information obtained by the mass spectrometry performed in step (h); and (j) obtaining, regarding each identified protein, an intensity ratio between a peak derived from one of the plural kinds of proteins purified in step (a) and a peak derived from one of the plural kinds of proteins purified in step (b) and an intensity ratio between a peak derived from one of the plural kinds of proteins purified in step (a) and a peak derived from one of the plural kinds of proteins purified in step (c), thereby quantitating affinity ratios of the 1st and 2nd test compounds to each identified protein.