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WO2024220916A1 - Procédés de détermination de teneur relative en glycane non apparié - Google Patents

Procédés de détermination de teneur relative en glycane non apparié Download PDF

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Publication number
WO2024220916A1
WO2024220916A1 PCT/US2024/025561 US2024025561W WO2024220916A1 WO 2024220916 A1 WO2024220916 A1 WO 2024220916A1 US 2024025561 W US2024025561 W US 2024025561W WO 2024220916 A1 WO2024220916 A1 WO 2024220916A1
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WO
WIPO (PCT)
Prior art keywords
unpaired
glycan
content
antibody
relative
Prior art date
Application number
PCT/US2024/025561
Other languages
English (en)
Inventor
Esme Candish
Alla Polozova
Original Assignee
Amgen Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Amgen Inc. filed Critical Amgen Inc.
Publication of WO2024220916A1 publication Critical patent/WO2024220916A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6854—Immunoglobulins
    • G01N33/6857—Antibody fragments
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
    • G01N33/6848—Methods of protein analysis involving mass spectrometry
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/40—Immunoglobulins specific features characterized by post-translational modification
    • C07K2317/41—Glycosylation, sialylation, or fucosylation
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/52—Constant or Fc region; Isotype
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
    • C07K2317/732—Antibody-dependent cellular cytotoxicity [ADCC]

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Analytical Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Pathology (AREA)
  • General Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Bioinformatics & Computational Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Biophysics (AREA)
  • Peptides Or Proteins (AREA)

Abstract

L'invention concerne des procédés de détermination de la teneur relative en glycane non apparié d'une composition d'anticorps IgG. Dans des modes de réalisation donnés à titre d'exemple, le procédé comprend le traitement de la composition d'anticorps IgG avec deux enzymes pour former un mélange de fragments Fab et de fragments Fc, une enzyme clivant une chaîne lourde d'anticorps au niveau d'un site N-terminal vers les liaisons disulfure de région charnière et l'autre enzyme clivant la liaison β1,4 entre les résidus GlcNAc centraux pour former des fragments Fc comprenant chacun une paire de structures de glycane noyau, (b) la séparation des fragments Fab des fragments Fc, et (c) la quantification de l'abondance de (i) fragments Fc afucosylés appariés, (ii) fragments Fc afucosylés non appariés, (iii) fragments Fc à teneur élevée en mannose appariés et/ou (iv) fragments Fc à teneur élevée en mannose non appariés, pour déterminer la teneur relative en glycane afucosylé (AF) non apparié et la teneur relative en glycane à teneur élevée en mannose (HM) non apparié.
PCT/US2024/025561 2023-04-20 2024-04-19 Procédés de détermination de teneur relative en glycane non apparié WO2024220916A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US202363576448P 2023-04-20 2023-04-20
US63/576,448 2023-04-20

Publications (1)

Publication Number Publication Date
WO2024220916A1 true WO2024220916A1 (fr) 2024-10-24

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ID=91128285

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2024/025561 WO2024220916A1 (fr) 2023-04-20 2024-04-19 Procédés de détermination de teneur relative en glycane non apparié

Country Status (1)

Country Link
WO (1) WO2024220916A1 (fr)

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