Method for analysis of compound-binding ability of protein

a technology of compound binding ability and protein, which is applied in the field of protein binding ability analysis, can solve the problems that methods cannot be used to study the binding ability of substances, and inhibitors cannot fully function as inhibitors

Inactive Publication Date: 2015-04-16
EISIA R&D MANAGEMENT CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

Enables the ranking of binding abilities of multiple proteins to a compound and the analysis of binding abilities in complex mixtures, providing valuable information on the interactions between proteins and compounds without the need for prior purification or identification of proteins.

Problems solved by technology

For example, when an inhibitor against an enzyme is developed, if the binding ability of the inhibitor to the enzyme is lower than the binding ability of the original substrate of the enzyme to the enzyme, the inhibitor cannot fully function as an inhibitor.
Therefore, these methods cannot be used for studying the binding ability of a substance to each of proteins contained in a mixture of plural kinds of unknown proteins.

Method used

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  • Method for analysis of compound-binding ability of protein

Examples

Experimental program
Comparison scheme
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first embodiment

(1) First Embodiment

[0093]A first embodiment of the present invention comprises the following steps for analyzing a binding ability of protein to a compound, wherein the group of proteins as the target of analysis are isotope-labeled before fractionation:[0094](a) step of fractionating a first group of isotope-labeled proteins into plural fractions using a carrier having the compound immobilized thereon;[0095](b) step of fractionating a second group of proteins into one or plural fractions using a carrier having the compound immobilized thereon;[0096](c) step of adding a certain amount of the one fraction obtained in step (b), or a certain amount of a mixture of all the fractions or a mixture of plural contiguous fractions among the fractions obtained in step (b), to each of the fractions obtained in step (a);[0097](d) step of analyzing the fractions obtained in step (c) with mass spectrometry; and[0098](e) step of, based on the mass spectrometry information, obtaining, regarding ea...

second embodiment

(2) Second Embodiment

[0162]A second embodiment of the present invention comprises the following steps for analyzing a binding ability of protein to a compound, wherein samples for the internal standard fraction are isotope-labeled before fractionation:[0163](a) step of fractionating a first group of proteins into plural fractions using a carrier having the compound immobilized thereon;[0164](b) step of fractionating a second group of isotope-labeled proteins into one or plural fractions using a carrier having the compound immobilized thereon;[0165](c) step of adding a certain amount of the one fraction obtained in step (b), or a certain amount of a mixture of all the fractions or a mixture of plural contiguous fractions among the fractions obtained in step (b), to each of the fractions obtained in step (a);[0166](d) step of analyzing the fractions obtained in step (c) with mass spectrometry; and[0167](e) step of, based on the mass spectrometry information, obtaining, regarding each ...

third embodiment

(3) Third Embodiment

[0169]A third embodiment of the present invention is directed to a method for analyzing a binding ability of protein to a compound, comprising the following steps, wherein test protein samples are labeled after fractionation:[0170](a) step of fractionating a first group of proteins into plural fractions using a carrier having the compound immobilized thereon;[0171](b) step of fractionating a second group of proteins into one or plural fractions using a carrier having the compound immobilized thereon;[0172](c) step of labeling the fractions obtained in step (a);[0173](d) step of adding a certain amount of the one fraction obtained in step (b), or a certain amount of a mixture of all the fractions or a mixture of plural contiguous fractions among the fractions obtained in step (b), to each of the fractions labeled in step (c);[0174](e) step of analyzing the fractions obtained in step (d) with mass spectrometry; and[0175](f) step of, based on the mass spectrometry i...

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Abstract

A method for analyzing a binding ability of protein to a compound, involves fractionating first and second groups of proteins into plural fractions using a carrier having the compound immobilized thereon; combining fractions, analyzing the combined fractions with mass spectrometry; based on the mass spectrometry information, obtaining, regarding each fraction, an intensity ratio between peaks derived from a protein in each of the groups of fractions; and comparing degrees of the binding ability of the plural kinds of proteins to the compound.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application is a division of application Ser. No. 11 / 989,565, filed Jan. 28, 2008, which was a U.S. National Phase Application of PCT International Application No. PCT / JP2006 / 315545, filed on Jul. 31, 2006, which claims priority to Japanese Patent Application No. 2005-222097, filed Jul. 29, 2005, all of which are hereby incorporated herein by reference.TECHNICAL FIELD[0002]The present invention relates to a method for analyzing a binding ability of protein to a compound using mass spectrometry.BACKGROUND OF THE INVENTION[0003]Analysis of protein-protein binding gives us a lot of useful information for studying functions of each protein. Analysis of binding between a compound available for a drug and protein gives us important information for revealing influences of such a compound on living organisms.[0004]For analyzing the binding between a substance and protein, not only the specificity of the protein which binds to the substance, ...

Claims

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Application Information

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Patent Type & AuthorityApplications(United States)
IPC IPC(8): G01N33/68G01N33/58G01N33/543
CPCG01N33/6848G01N33/54306G01N33/58G01N2458/15G01N2560/00G01N30/72G01N30/84G01N2030/027G01N2030/8411H01J49/0409Y10T436/24
InventorODA, YOSHIYAKATAYAMA, HIROYUKI
OwnerEISIA R&D MANAGEMENT CO LTD