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US 12,669,508 B2Granted
Methods to prevent disulfide scrambling for MS-based proteomics
AssigneeRegeneron Pharmaceuticals, Inc.
InventorsAndrew Kleinberg, Yuan Mao, Ning Li
Life of this patent
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Abstract
The present invention generally pertains to methods of preventing disulfide scrambling in non-reducing liquid chromatography-mass spectrometry analysis of a protein of interest. In particular, the present invention pertains to the addition of maleimide to a non-reducing liquid chromatography-mass spectrometry analysis of a protein to prevent disulfide scrambling.
Claims20
A method for performing a non-reduced peptide mapping of a protein of interest in a sample, said method comprising:
- a. contacting said sample to a NEM analog to form an alkylated protein of interest, wherein the NEM analog is maleimide or N-hydroxy maleimide;
- b. contacting said alkylated protein of interest to at least one digestive enzyme to form a peptide digest;
- and
- c. analyzing said peptide digest using liquid chromatography-mass spectrometry to obtain said non-reduced peptide mapping of said protein of interest.
1 more element
A method for characterizing a protein of interest in a sample, said method comprising:
- a. contacting said sample to a NEM analog to form an alkylated protein of interest, wherein the NEM analog is maleimide or N-hydroxy maleimide;
- b. contacting said alkylated protein of interest to at least one digestive enzyme to form a peptide digest;
- and
- c. analyzing said peptide digest using liquid chromatography-mass spectrometry to obtain a non-reduced peptide mapping to characterize said protein of interest.
1 more element
Dependencies are inferred from the claim text.