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US 12,699,101 B2Granted
Maximizing hydrophobic peptide recovery using a mass spectrometry compatible surfactant
AssigneeRegeneron Pharmaceuticals, Inc.
InventorsSong Nie, Reid O'Brien Johnson, Tyler Greer, Xiaojing Zheng
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Abstract
The present invention generally pertains to methods of characterizing peptides or proteins of interest. In particular, the present invention pertains to the use of the nonionic surfactant DDM to improve recovery of hydrophobic peptides in pharmacokinetic, reduced peptide mapping, or other biotherapeutic protein analyses.
Claims18
A method for peptide mapping analysis, comprising:
- (a) subjecting a sample including a protein of interest to conditions suitable for denaturing to form a denatured sample;
- (b) contacting said denatured sample to at least one alkylating agent under conditions suitable for alkylation to form an alkylated sample;
- (c) contacting said alkylated sample to N-dodecyl-β-D-maltoside (DDM) to form a mixture;
- (d) contacting said mixture to at least one digestive enzyme under conditions suitable for digestion to form a peptide digest;
- and
- (e) subjecting said peptide digest to liquid chromatography-mass spectrometry for a peptide mapping analysis;
- wherein the protein of interest is an antibody having a peptide from a complementarity-determining region (CDR) of the antibody, and wherein the sample with the DDM exhibits an increased signal for the peptide from the CDR relative to a signal for the peptide from the CDR in a sample without DDM.
4 more elements
Dependencies are inferred from the claim text.