A Method Of Purifying A Binding Protein Produced By A Host Cell

Abstract: A method of purifying a binding protein produced by a host cell, comprising: (a) producing in a host cell a binding protein comprising four polypeptide chains that fonn three antigen binding sites that specifically bind one or more antigen targets or target proteins, wherein a first polypeptide chain comprises a structure represented by the formula: VLz-LJ-Vu-Lz-CL [1] and a second polypeptide chain comprises a structure represented by the fommla: VH1-L3-VH2-L4-CHt-hinge-CH2-Cm [II] and a third polypeptide chain comprises a structure represented by the formula: [III] and a fourth polypeptide chain comprises a structure represented by the fonnula: V13-CL [IV] Vu is a first immunoglobulin light chain variable domain; Vu is a second immwJOglobuli.tl light chai.t1 variable domain; VLJ is a third immunoglobulin light chain variable domain; VHt is a first immunoglobulin heavy chain variable domain; VH2 is a second illllllunoglobulinheavy chain variable domain; VHJ is a thi.t·d immunoglobulin heavy chain variable domain; CL is an immunoglobulin light chain constant domain; CH.1 is an immunoglobulin CRt heavy chain constant domain; Cm is an immunoglobulin CH2 heavy chain constant domain; Cm is an immunoglobulin Cm heavy chain constant domain; hinge is an immunoglobulin hinge region connecting the Cm and Cm domains; and L 1, Lz, L3 and L4 are amino acid linkers; wherein the polypeptide of fonnula 1 and the polypeptide of fonnula II form a cross-over light chainheavy chain pair; wherein only one of the Cm domain of the second polypeptide chain and the CH3 domain of the third polypeptide chain comprises amino acid substitutions at positions corresponding to positions 435 and 436 of humanlgGl or lgG4 according to EU Index, wherein the amino acid substitutions are H435R and Y436F; (b) contacting the binding protein produced in (a) with Protein A; and (c) eluti.t1g the binding protein from Protein A under cooditious sllitable for isolating the binding pruldu away from binding proteins comprising either 0 or 2 Cm domains comprising the amino acid substitutions are H435R and Y436F, thereby purifying the binding protein.

Patent Information

Application #

Filing Date

08 June 2023

Publication Number

32/2024

Publication Type

INA

Invention Field

BIOTECHNOLOGY

Status

Email

Parent Application

Applicants

SANOFI

54 RUE LA BOETIE, PARIS FRANCE 75008

Inventors

1. YANG, Zhi-Yong

C/O SANOFI 55 CORPORATE DRIVE, MAIL CODE 55A-505A , BRIDGEWATER, NJ UNITED STATES OF AMERICA 08807

2. NABEL, Gary, J.

C/O SANOFI 55 CORPORATE DRIVE, MAIL CODE 55A-505A BRIDGEWATER, NJ UNITED STATES OF AMERICA 08807

3. WU, Lan

C/O SANOFI 55 CORPORATE DRIVE, MAIL CODE 55A-505A BRIDGEWATER, NJ UNITED STATES OF AMERICA 08807

4. SEUNG, Edward

C/O SANOFI 55 CORPORATE DRIVE, MAIL CODE 55A-505A BRIDGEWATER, NJ UNITED STATES OF AMERICA 08807