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公开(公告)号：US20230243841A1
公开(公告)日：2023-08-03
申请号：US17976330
申请日：2022-10-28
Applicant: Regeneron Pharmaceuticals, Inc.
Inventor： Andrew Kleinberg , Yuan Mao , Ning Li
IPC: G01N33/68
CPC classification number: G01N33/6848 , G01N2560/00
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.
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公开(公告)号：US20230027480A1
公开(公告)日：2023-01-26
申请号：US17716154
申请日：2022-04-08
Applicant: Regeneron Pharmaceuticals, Inc.
Inventor： Xuefei Zhong , Yuan Mao
Abstract: The present invention generally pertains to methods of quantitating therapeutic proteins co-administered to a subject using LC-MRM-MS. In particular, the present invention pertains to the use of dual enzymatic digestion to generate unique surrogate peptides allowing for the accurate quantitation of co-administered therapeutic proteins using LC-MRM-MS.
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公开(公告)号：US20210231676A1
公开(公告)日：2021-07-29
申请号：US17159136
申请日：2021-01-26
Applicant: Regeneron Pharmaceuticals, Inc.
Inventor： Yuan Mao , Andrew Kleinberg
Abstract: Disclosed are methods of quantifying multiple quality attributes, such as post translational modifications, of multiple samples in a single mass spectrometry (MS) run, including contacting two or more samples with a digesting solution under conditions sufficient to digest samples, wherein each sample is digested separately and the digesting solution is a Tris-free buffer solution; contacting each of the two or more digested samples with a specific Tandem Mass Tag (TMT) labeling reagent under conditions sufficient to label peptides within each of the digested samples with the specific TMT labeling reagent; quenching labeling of peptides within each of the two or more digested samples; combining equal volumes of the two or more labeled, digested samples into a single combined sample solution; and analyzing the single combined sample solution by targeted mass spectral analysis, thereby allowing multiple quality attributes of the two or more samples to be quantified in a single mass spectrometry (MS) run.
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公开(公告)号：US12135328B2
公开(公告)日：2024-11-05
申请号：US18123753
申请日：2023-03-20
Applicant: Regeneron Pharmaceuticals, Inc.
Inventor： Yuan Mao , Andrew Kleinberg
Abstract: Disclosed are methods of quantifying multiple quality attributes, such as post translational modifications, of multiple samples in a single mass spectrometry (MS) run, including contacting two or more samples with a digesting solution under conditions sufficient to digest samples, wherein each sample is digested separately and the digesting solution is a Tris-free buffer solution; contacting each of the two or more digested samples with a specific Tandem Mass Tag (TMT) labeling reagent under conditions sufficient to label peptides within each of the digested samples with the specific TMT labeling reagent; quenching labeling of peptides within each of the two or more digested samples; combining equal volumes of the two or more labeled, digested samples into a single combined sample solution; and analyzing the single combined sample solution by targeted mass spectral analysis, thereby allowing multiple quality attributes of the two or more samples to be quantified in a single mass spectrometry (MS) run.
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公开(公告)号：US20230417759A1
公开(公告)日：2023-12-28
申请号：US18203854
申请日：2023-05-31
Applicant: Regeneron Pharmaceuticals, Inc.
Inventor： Yuan Mao , Andrew Kleinberg , Yunlong Zhao , Lili Guo
CPC classification number: G01N33/6848 , G01N33/6857 , G01N30/34 , G01N30/7233
Abstract: A method of enhancing a mass spectral signal is disclosed. The method can include contacting a sample to a separation column under conditions that permit sample components to bind to the substrate; applying a first mobile gradient to the separation column, wherein the first mobile phase gradient comprises trifluoroacetic acid (TFA) and a small molecule additive (e.g., an amino acid) or formic acid (FA) and a small molecule additive (e.g., an amino acid); applying a second mobile gradient to the separation column, wherein the second mobile phase gradient comprises TFA in acetonitrile (ACN) and a small molecule additive (e.g., an amino acid) or formic acid (FA) in ACN and a small molecule additive (e.g., an amino acid); and performing mass spectrometric analysis on eluted sample components.
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Patent Agency Ranking
公开(公告)号：US20210223259A1
公开(公告)日：2021-07-22
申请号：US17143890
申请日：2021-01-07
Applicant: Regeneron Pharmaceuticals, Inc.
Inventor： Yuan Mao , Andrew Kleinberg , Yunlong Zhao , Lili Guo
IPC: G01N33/68
Abstract: A method of enhancing a mass spectral signal is disclosed. The method can include contacting a sample to a separation column under conditions that permit sample components to bind to the substrate; applying a first mobile gradient to the separation column, wherein the first mobile phase gradient comprises trifluoroacetic acid (TFA) and a small molecule additive (e.g., an amino acid) or formic acid (FA) and a small molecule additive (e.g., an amino acid); applying a second mobile gradient to the separation column, wherein the second mobile phase gradient comprises TFA in acetonitrile (ACN) and a small molecule additive (e.g., an amino acid) or formic acid (FA) in ACN and a small molecule additive (e.g., an amino acid); and performing mass spectrometric analysis on eluted sample components.
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公开(公告)号：US20250067750A1
公开(公告)日：2025-02-27
申请号：US18787598
申请日：2024-07-29
Applicant: Regeneron Pharmaceuticals, Inc.
Inventor： Abu Nahiyaan Navid , Yunlong Zhao , Yuan Mao , Ning Li
Abstract: The present invention generally pertains to methods of characterizing non-consensus glycosylation sites of a protein. In particular, the present invention pertains to the use of high-throughput automated processes through digestion, enrichment of glycopeptides by liquid chromatography-mass spectrometry for identifying identification of non-consensus glycosylation sites.
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8.发明公开公开(公告)号：US20230349916A1
公开(公告)日：2023-11-02
申请号：US18123753
申请日：2023-03-20
Applicant: REGENERON PHARMACEUTICALS, INC.
Inventor： Yuan Mao , Andrew Kleinberg
CPC classification number: G01N33/6848 , G01N30/7233 , G01N2030/027
Abstract: Disclosed are methods of quantifying multiple quality attributes, such as post translational modifications, of multiple samples in a single mass spectrometry (MS) run, including contacting two or more samples with a digesting solution under conditions sufficient to digest samples, wherein each sample is digested separately and the digesting solution is a Tris-free buffer solution; contacting each of the two or more digested samples with a specific Tandem Mass Tag (TMT) labeling reagent under conditions sufficient to label peptides within each of the digested samples with the specific TMT labeling reagent; quenching labeling of peptides within each of the two or more digested samples; combining equal volumes of the two or more labeled, digested samples into a single combined sample solution; and analyzing the single combined sample solution by targeted mass spectral analysis, thereby allowing multiple quality attributes of the two or more samples to be quantified in a single mass spectrometry (MS) run.
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公开(公告)号：US20210278414A1
公开(公告)日：2021-09-09
申请号：US17327889
申请日：2021-05-24
Applicant: Regeneron Pharmaceuticals, Inc.
Inventor： Yuan Mao
Abstract: A method for determining an amino acid sequence of a polypeptide, including comprising: contacting a first sample containing the polypeptide with a first protease (e.g., Trypsin) to produce a first set of digested peptide fragments; fragmenting the first set of digested peptide fragments to produce a first set of fragmented peptide ions; determining masses of the first set of fragmented peptide ions; contacting a second sample containing the polypeptide with a second protease (e.g., Tryp-N); fragmenting the second set of digested peptide fragments to produce a second set of fragmented peptide ions; selecting pairs of peptide ions from the first and the second set of fragmented peptide ions that differ in mass by a mass of an arginine amino acid residue or a lysine amino acid residue; assigning an ion type (either N-terminal peptide ion or C-terminal peptide ion) to the selected pairs of the peptide ions from two sets of fragmented peptide ions; selecting a mass ladder of the same-type peptide ions in either set of fragmented peptide ions with incremental mass by the mass of amino acid residue(s), and assembling the identified amino acid residues from the mass ladder to determine the amino acid sequence of the polypeptide of interest.
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公开(公告)号：US12000840B2
公开(公告)日：2024-06-04
申请号：US17327889
申请日：2021-05-24
Applicant: Regeneron Pharmaceuticals, Inc.
Inventor： Yuan Mao
CPC classification number: G01N33/6851 , C07K16/065 , G16B20/00 , G16B40/10 , H01J49/305
Abstract: A method for determining an amino acid sequence of a polypeptide, including comprising: contacting a first sample containing the polypeptide with a first protease (e.g., Trypsin) to produce a first set of digested peptide fragments; fragmenting the first set of digested peptide fragments to produce a first set of fragmented peptide ions; determining masses of the first set of fragmented peptide ions; contacting a second sample containing the polypeptide with a second protease (e.g., Tryp-N); fragmenting the second set of digested peptide fragments to produce a second set of fragmented peptide ions; selecting pairs of peptide ions from the first and the second set of fragmented peptide ions that differ in mass by a mass of an arginine amino acid residue or a lysine amino acid residue; assigning an ion type (either N-terminal peptide ion or C-terminal peptide ion) to the selected pairs of the peptide ions from two sets of fragmented peptide ions; selecting a mass ladder of the same-type peptide ions in either set of fragmented peptide ions with incremental mass by the mass of amino acid residue(s), and assembling the identified amino acid residues from the mass ladder to determine the amino acid sequence of the polypeptide of interest.
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