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US 11,740,246 B2Granted
Dual-column LC-MS system and methods of use thereof
AssigneeRegeneron Pharmaceuticals, Inc.
InventorShunhai Wang
Life of this patent
Estimating the term…
Abstract
Methods for achieving complete sequence coverage of monoclonal antibodies by trypsin digestion and dual-column LC-MS system are provided. The disclosed method improves upon current techniques for standard peptide mapping.
Claims20
A method for determining the sequence of a protein comprising:
- loading the peptide fragments onto a first high performance liquid chromatography column comprising material for separating non-polar peptides under conditions in which a portion of the peptide fragments are retained on the first column and a portion of the peptide fragments are not retained on the first column, wherein the peptides not retained on the first column flow directly into a second high performance liquid chromatography column comprising material for separating polar peptides, wherein at least the portion of the peptide fragments not retained on the first column are retained on the second column;
- eluting the portion of the peptide fragments retained on the first column;
- eluting the portion of the peptide fragments retained on the second column;
- and
- analyzing the portion of the peptide fragments retained on the first column and the portion of the peptide fragments retained on the second column to determine the amino acid sequence of the portion of the peptide fragments retained on the first column and the portion of the peptide fragments retained on the second column.
2 more elements
A dual column liquid chromatography system for determining the sequence of a protein comprising:
- a first column in fluid communication with a first multi-port switching diverter valve, wherein the first column comprises material for separating non-polar peptides;
- a second column in fluid communication with the first multi-port switching diverter valve, wherein the second column comprises material for separating polar peptides;
- a second multi-port switching diverter value in fluid communication with the first multi-port switching diverter valve;
- and
- a computer in electronic communication with the dual column chromatography system for controlling fluid flow through the system by configuring the ports of the first multi-port switching diverter valve and the second multi-port switching diverter valve in multiple configurations.
2 more elements
Dependencies are inferred from the claim text.