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US 2017/0362294 A1Published application
Methods for Protein and Peptide Reduction
AssigneeDH Technologies Development Pte. Ltd.
InventorYves Le Blanc
Life of this patent
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Abstract
Methods and systems are provided herein for on-line preparation of a sample for mass spectrometry. In accordance with various aspects of applicant's teachings, the methods and systems can provide for the reduction of a polypeptide, for example, on a liquid chromatography column and can reduce or eliminate the need to incubate the reducing agent with the polypeptide and/or expose the reduced polypeptide to an alkylating agent.
Claims20
A method for on-line preparation of a sample for mass spectrometry analysis comprising:
- a. introducing a sample comprising a polypeptide having at least one disulfide bond to a liquid chromatography column, wherein the liquid chromatography column is configured to fluidly couple to an ion source of a mass spectrometer;
- b. retaining the polypeptide on the liquid chromatography column;
- c. flowing a solution comprising a reducing agent through the liquid chromatography column so as to cleave one or more of the at least one disulfide bonds of the retained polypeptide while the reducing solution is flowed therethrough;
- d. flowing an eluent through the liquid chromatography column so as to elute the reduced polypeptide from the liquid chromatography column;
- and
- e. delivering the eluate containing the reduced polypeptide to the ion source.
3 more elements
A method of preparing a sample for mass spectrometry analysis comprising:
- a. introducing a sample comprising a polypeptide to a liquid chromatography column;
- b. retaining the polypeptide on the liquid chromatography column;
- c. reducing the polypeptide by a reducing agent;
- wherein the polypeptide comprises one or more disulfide bonds capable of being reduced;
- and wherein reducing the polypeptide comprises:
- i. flowing a solution comprising the reducing agent through the liquid chromatography column at a first flow rate;
- and
- ii. flowing an eluent so as to elute the reduced polypeptide from the liquid chromatography column at a second flow rate;
- wherein the second flow rate is greater than the first flow rate;
- and
- d. delivering the eluate from a liquid chromatography column comprising the reduced polypeptide to a mass spectrometer.
8 more elements
Dependencies are inferred from the claim text.