Hydrophobic interaction chromatography-coupled native mass spectrometry for antibody analysis
AssigneeRegeneron Pharmaceuticals, Inc.
InventorsYuetian Yan, Shunhai Wang
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Abstract
The present invention provides rapid, sensitive high-throughput methods and systems for characterizing peptides or proteins using hydrophobic interaction chromatography-coupled native mass spectrometry to improve manufacturing process of biopharmaceutical products, such as identifying impurities during antibody purification, monitoring post-translational modification variants during production, or characterizing drug-to-antibody ratio of antibody-drug conjugates. The separation profiles of the peptides or proteins are generated and compared to identify or qualify the peptides or proteins.
Claims13
A method for determining glycosylation profiles for at least one peptide or protein in a sample, comprising:
- contacting the sample to a solid surface, wherein the solid surface comprises a hydrophobic group and the solid surface is included in a chromatography column;
- washing the solid surface using a mobile phase to produce at least one eluent, wherein the eluent comprises the at least one peptide or protein;
- determining the glycosylation profile of the at least one peptide or protein using a mass spectrometer;
- and
- determining the O-glycosylation variants in the sample;
- wherein the mass spectrometer is coupled online to the chromatography column.
A method for determining intact masses of at least on peptide or protein in a sample, comprising:
- contacting the sample to a solid surface, wherein the solid surface comprises a hydrophobic group and the solid surface is included in a chromatography column;
- washing the solid surface using a mobile phase to produce at least one eluent, wherein the eluent comprises the at least one peptide or protein;
- determining the intact masses of the at least one peptide or protein in the at least one eluent using a mass spectrometer under native conditions;
- and
- identifying or quantifying a drug location, a positional isomer or a degraded payload of the antibody-drug conjugate based on the at least one separation profile or a comparison with another separation profile;
- wherein the mass spectrometer is coupled online to the chromatography column.
Dependencies are inferred from the claim text.