Method for detecting protein modifications using specific antibodies
AssigneeMerck Patent GmbH
InventorsFrank Jeahrling, Friedhelm Bladt, Jessica Witzgall
Life of this patent
Estimating the term…
Abstract
Method, kit and composition for analyzing analytes for modifications using modification site specific antibodies to bind an analyte with his specific modification sites of interest to different dyes simultaneously with an antibody which is specific to the non-modificated analyte binding to another dye to determine the concentration of the analyte for quantification of the modified analyte in the identical sample.
Claims14
A method for detecting modification sites of a protein or polypeptide in an analyte of a sample to be analyzed comprising said protein or polypeptide, wherein the modification site is selected from the group consisting of phosphorylation, autophosphorylation, methylation, hydroxylation, glycosylation, ubiquination, acetylation, prenylation, amidation or N-terminal methionine modification sites, comprising
- (a) contacting the sample with a first capture antibody that is specific for or binds to a first modification site and is conjugated or associated with a dye serving as a first detection marker,
- (b) contacting the sample with a second antibody that is specific for or binds to said protein or polypeptide at a protein- or polypeptide-specific site or epitope, which protein- or polypeptide-specific antibody binding site or epitope is different from the modification site in (a), and which second antibody is conjugated to or associated with a second detection marker which is distinguishable from said dye of step (a),
- (c) contacting the sample with a third capture antibody that is specific for or binds to a second modification site on said protein or polypeptide that is different from the first modification site in (a) and which second modification site is different from the protein- or polypeptide-specific site or epitope, and which third capture antibody is conjugated or associated with a dye serving as a third detection marker which is distinguishable from the detection markers of step (a) and step (b),
- (d) separating unbound protein or polypeptide from said antibodies, and
- (e) detecting the presence of bound detection markers.
Dependencies are inferred from the claim text.