WO2025094054 - METHOD FOR PURIFYING SMALL MULTI-DOMAIN PROTEINS

National phase entry:
Publication Number WO/2025/094054
Publication Date 08.05.2025
International Application No. PCT/IB2024/060657
International Filing Date 29.10.2024
Title **
[English] METHOD FOR PURIFYING SMALL MULTI-DOMAIN PROTEINS
[French] PROCÉDÉ DE PURIFICATION DE PETITES PROTÉINES À DOMAINES MULTIPLES
Applicants **
IMMUNOCORE LIMITED
Inventors
PATEL, Viren
JOSEPH, Erin
Priority Data
63/594,987   01.11.2023   US
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Quotation for National Phase entry

Country StagesTotal
China Filing, Examination, Granting4283
EPO Filing, Examination, Granting64169
Japan Filing, Examination, Granting4211
South Korea Filing, Examination, Granting6174
USA Filing, Examination, Granting18740
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Total: 97,577

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Abstract[English] The present disclosure provides a method for purifying a T cell engaging peptide-major histocompatibility complex binding domain (TCE-pMHC) molecule produced by a host cell, the method comprising: isolating a supernatant of a host cell culture comprising the TCE-pMHC molecule to form Product (a), subjecting Product (a) to affinity chromatography wherein the TCE-pMHC molecule is eluted from the affinity chromatography at a pH of about 3.0 to about 4.0 to form Product (b); neutralizing Product (b) by adding MES buffer with a pH of less than 7.0 to form Product (c), wherein Product (c) has a pH of about 5.0 to about 7.0; and subjecting Product (c) to anion exchange chromatography to form Product (d), wherein the pMHC binding domain and the T cell engaging immune effector domain are capable of binding to a pMHC complex and a T cell respectively.[French] La présente divulgation concerne un procédé de purification d'une molécule à domaine de liaison de complexe majeur d'histocompatibilité (TCE-pMHC) de peptide d'activation des lymphocytes T produite par une cellule hôte, le procédé comprenant les étapes consistant à : isoler un surnageant d'une culture de cellules hôtes comprenant la molécule TCE-pMHC pour former un produit (a), soumettre le produit (a) à une chromatographie d'affinité, la molécule TCE-pMHC étant éluée à partir de la chromatographie d'affinité à un pH d'environ 3,0 à environ 4,0 pour former un produit (b) ; un produit neutralisant (b) par ajout d'un tampon MES ayant un pH inférieur à 7,0 pour former un produit (c), le produit (c) ayant un pH d'environ 5,0 à environ 7,0 ; et soumettre le produit (c) à une chromatographie par échange d'anions pour former un produit (d), le domaine de liaison pMHC et le domaine effecteur immunitaire d'activation des lymphocytes T étant capables de se lier à un complexe pMHC et à un lymphocyte T respectivement.