US 12,327,614 B2
Multi-domain proteins with increased native state colloidal stability
Ralph Adams, Slough (GB); James Heads, Slough (GB); Sebastian Kelm, Slough (GB); and Alastair David Griffiths Lawson, Slough (GB)
Assigned to UCB BIOPHARMA SRL, Brussels (BE)
Appl. No. 16/972,458
Filed by UCB Biopharma SRL, Brussels (BE)
PCT Filed Jun. 5, 2019, PCT No. PCT/EP2019/064635
§ 371(c)(1), (2) Date Dec. 4, 2020,
PCT Pub. No. WO2019/234094, PCT Pub. Date Dec. 12, 2019.
Claims priority of application No. 1809341 (GB), filed on Jun. 7, 2018.
Prior Publication US 2021/0166780 A1, Jun. 3, 2021
Int. Cl. G16B 15/20 (2019.01); C07K 16/00 (2006.01); G16B 15/00 (2019.01); G16B 30/00 (2019.01); G16B 40/00 (2019.01); G16C 20/50 (2019.01); G16B 5/00 (2019.01)
CPC G16B 15/20 (2019.02) [C07K 16/00 (2013.01); G16B 15/00 (2019.02); G16B 30/00 (2019.02); G16B 40/00 (2019.02); G16C 20/50 (2019.02); C07K 2317/52 (2013.01); C07K 2317/92 (2013.01); C07K 2317/94 (2013.01); G16B 5/00 (2019.02)] 18 Claims
OG exemplary drawing
 
1. A method of producing an IgG4 antibody with an improved colloidal stability comprising:
calculating for each domain of said IgG4 antibody the total net charge at a given pH;
modifying the DNA sequence encoding the IgG4 antibody to introduce one or more modifications to the amino acid residues of the constant region of said IgG4 antibody to minimize the charge sign difference between the domains, wherein said one or more modification is, or each are, a substitution of a charged amino acid by a polar, non-charged amino acid, wherein said one or more modifications is, or are, selected from the list consisting of residues at positions R133, E137, D203, R214, E356, R409, E419 of Fc domain; and
producing the IgG4 antibody with improved colloidal stability at the given pH.