High-Throughput Method for Quantifying Sialylation of Glycoproteins
AssigneeMassachusetts Institute of Technology
InventorsDaniel I. C. Wang, Lam Raga Anggara Markely
Life of this patent
Estimating the term…
Abstract
Provided herein methods and kits for detecting and/or quantifying sialic acid content of glycosylated molecules that does not require purification of the glycosylated molecule of interest or purification of the labeled product. The methods and kits provided herein are fast and suitable for high-throughput use.
Claims31
A method for detecting sialylation of one or more glycosylated molecules in a sample comprising;
- a) reducing a sample by contacting the sample with a reducing agent, wherein the sample comprises one or more glycosylated molecules;
- b) contacting the reduced sample with an agent capable of removing terminal sialic acid residues from the one or more glycosylated molecules, generating freed sialic acid residues;
- c) detectably labeling the freed sialic acid residues with a labeling agent, and
- d) detecting the detectably labeled freed sialic acid residues and optionally quantifying sialylation of the one or more glycosylated molecules,
- thereby detecting sialylation of one or more glycosylated molecules.
A method for detecting sialylation of one or more glycosylated molecules in a sample comprising;
- a) providing at least one reagent selected from the group consisting of:
- a reducing agent, a reagent suitable for removing terminal sialic acid residues from a glycoprotein, and a labeling reagent suitable for detectably labeling free sialic acid residues, and
- b) providing instructions for using the at least one reagent to detect sialylation of one or more glycosylated molecules in a sample and optionally quantify sialylation of one or more glycosylated molecules in a sample.
A kit for detecting sialylation of one or more glycosylated molecules in a sample comprising;
- a) one or more reagents selected from the group consisting of:
- a reducing agent, a reagent suitable for removing a terminal sialic acid residue from a glycoprotein, and a labeling reagent suitable for detectably labeling free sialic acid residues, and
- b) instructions describing how to use the one or more reagents to detect sialylation of one or more glycosylated molecules in a sample and optionally how to quantify sialylation of one or more glycosylated molecules in a sample.
Dependencies are inferred from the claim text.