Systems and methods for analysis of protein post-translational modification
First Claim
Patent Images
1. A method for identifying modified amino acids within a protein, comprising:
- (i) providing one or more samples and an affinity capture reagent for isolating, from said samples, those proteins post-translationally modified by a moiety of interest;
(ii) processing said samples to chemically modify at least one of the C-terminal carboxyl, the N-terminal amine and amino acid side chains of polypeptides in said samples so as to increase the specificity of said affinity capture reagent for those proteins post-translationally modified by said moiety of interest;
(iii) isolating said proteins post-translationally modified by said moiety of interest from said samples using said affinity capture reagent;
(iv) eluting said proteins bound to said affinity capture reagent by manipulating the oxidation state of said affinity capture reagent; and
, (v) determining the identity of said proteins eluted in (iv) by mass spectroscopy.
2 Assignments
0 Petitions
Accused Products
Abstract
The invention relates to a method for the detection and identification of amino acid modifications, such as phosphorylation, using a combination of affinity capture and mass-spectroscopy.
-
Citations
31 Claims
-
1. A method for identifying modified amino acids within a protein, comprising:
-
(i) providing one or more samples and an affinity capture reagent for isolating, from said samples, those proteins post-translationally modified by a moiety of interest;
(ii) processing said samples to chemically modify at least one of the C-terminal carboxyl, the N-terminal amine and amino acid side chains of polypeptides in said samples so as to increase the specificity of said affinity capture reagent for those proteins post-translationally modified by said moiety of interest;
(iii) isolating said proteins post-translationally modified by said moiety of interest from said samples using said affinity capture reagent;
(iv) eluting said proteins bound to said affinity capture reagent by manipulating the oxidation state of said affinity capture reagent; and
,(v) determining the identity of said proteins eluted in (iv) by mass spectroscopy. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 23, 24, 25, 26, 27, 28, 29, 30, 31)
-
-
16. A method for analyzing a phosphoproteome, comprising:
-
(i) providing one or more protein sample(s);
(ii) chemically modifying the side chains of glutamic acid and aspartic acid residues of polypeptides in said protein sample(s) to neutral derivatives;
(ii) isolating phosphorylated proteins from said protein sample(s) by using immobilized metal affinity chromatography;
(iii) eluting said phosphorylated proteins from said affinity capture reagent by manipulating the oxidation state of said reagent; and
,(iv) determining the identity of said phosphorylated p roteins eluted in (iii) by m ass spectroscopy. - View Dependent Claims (17, 18, 19, 20, 21, 22)
-
Specification