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US 2017/0362624 A1Published application
Labeling and detection of post translationally modified proteins
AssigneeLife Technologies Corporation
InventorsBrian AGNEW, Kyle GEE, Tamara NYBERG
Life of this patent
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Abstract
Provided in certain embodiments are new methods for forming azido modified biomolecule conjugates of reporter molecules, carrier molecules or solid support. In other embodiments are provided methods for enzymatically labeling a biomolecules with an azide group.
Claims13
-16. (canceled)
A method of detecting an azido modified biomoelcule, comprising:
- a) forming an azide-alkyne cycloaddition reaction mixture comprising:
- a reporter molecule that comprises a terminal alkyne moiety:
- an azido modified biomoelcule;
- copper ions;
- at least one reducing agent;
- and
- a copper chelator;
- b) incubating the azide-alkyne cycloaddition reaction mixture for a sufficient amount of time to form a biomoelcule-reporter molecule conjugate;
- c) separating the biomoelcule-reporter molecule conjugate by size and/or weight of the glycoprotein-reporter molecule conjugate to form a separated biomoelcule-reporter molecule conjugate;
- d) illuminating the separated biomoelcule-reporter molecule conjugate with an appropriate wavelength to form an illuminated biomoelcule-reporter molecule conjugate;
- e) observing the illuminated biomolecule-reporter molecule conjugate wherein the glycoprotein is detected.
8 more elements
-39. (canceled)
A kit comprising:
- UDP-GalNAz;
- a GalT enzyme;
- an azide reactive reporter molecule, carrier molecule or solid support.
A method of forming a glycoprotein conjugate, wherein the method comprises:
- a) contacting the glycoprotein with an endonuclease enzyme to form a deglycosylated glycoprotein;
- b) contacting the deglycosylated glycoprotein with an azido modified carbohydrate in the presence of a galactosyltransferase enzyme to form an azido-modified glycoprotein;
- and
- c) contacting the azido-modified glycoprotein with a reporter molecule, carrier molecule or solid support that comprises an azide reactive moiety to form the glycoprotein conjugate.
1 more element
Dependencies are inferred from the claim text.