Proteome epitope tags and methods of use thereof in protein modification analysis
AssigneeMillipore Corporation
InventorsFrank D. Lee, Xun Meng, David Livingston
Life of this patent
Estimating the term…
Abstract
Disclosed are methods for reliably detecting the presence of proteins, especially proteins with various post-translational modifications (phosphorylation, glycosylation, methylation, acetylation, etc.) in a sample by the use of one or more capture agents that recognize and interact with recognition sequences uniquely characteristic of a set of proteins (Proteome Epitope Tags, or PETs) in the sample. Arrays comprising these capture agents or PETs are also provided.
Claims10
1-125. (canceled)
A method for detecting the presence or absence of a post-translational modification at a location on a target protein within a sample, comprising:
- (1) subjecting the target protein to a fragmentation treatment to produce a peptide fragment comprising (i) at least one potential post-translational modification site and, separate therefrom, (ii) a proteome epitope tag (PET) comprising an amino acid sequence unique to the peptide fragment within the sample;
- (2) contacting the peptide fragment with a capture agent specific for the PET, separate from the post-translational modification on the peptide fragment, to bind the fragment at the PET, to the capture agent;
- (3) detecting, on the peptide fragment bound to the capture agent, the presence or absence of the post-translational modification by using a secondary capture agent specific for the post-translational modification separate from the PET on the peptide fragment, wherein the secondary capture agent is labeled with a detectable moiety.
A method for detecting the presence or absence of a post-translational modification at a location on a target protein within a sample, comprising:
- (1) subjecting the target protein to a fragmentation treatment to produce a peptide fragment comprising (i) at least one potential post-translational modification site and, separate therefrom, (ii) a proteome epitope tag (PET) comprising an amino acid sequence unique to the peptide fragment within the sample;
- (2) contacting the peptide fragment with first and second capture agents respectively specific for the PET and the post-translational modification, one of which is immobilized and the other of which is labeled with a detectable moiety, to bind the fragment at the PET and at the post-translational modification to form a complex;
- and
- (3) detecting, the presence of the detectable moiety in said complex as an indication of the presence of said post-translational modification at a location on a target protein.
Dependencies are inferred from the claim text.