Granted Patent
B2
US 12,637,521 · App. 18/644,757 · Granted May 26, 2026
Low-viscosity antigen binding proteins and methods of making them
Inventors:
Joon Hoi Huh (Culver City, CA); Riki Stevenson (Ventura, CA); Pavel Bondarenko (Thousand Oaks, CA); Andrew Nichols (Calabasas, CA); Da Ren (Thousand Oaks, CA); Neeraj Jagdish Agrawal (Natick, MA); Richard Smith (Belmont, CA)
Assignee:
AMGEN INC.
C07K16/40C07K16/2869C07K2317/21C07K2317/24C07K2317/52C07K2317/55C07K2317/90C07K2317/92C07K2317/94G01N11/14
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Abstract
The present invention concerns a method for preparing antigen binding proteins specific for PCSK9 with reduced viscosity. The method proceeds by replacing residues in high viscosity variable domain subfamilies with residues in correlating low viscosity subfamilies. The method further comprises substituting residues in the Fc domain with residues associated with low viscosity and adding charged residues to the C-terminus of the Fc domain. The present invention further concerns antigen binding proteins produced by this method.
Claims (16)
1 . A method of reducing the viscosity of an antibody of the VH3|3-33 germline subfamily, which comprises making viscosity- reducing substitutions 1X 4 , 17X 5 , and 85X 6 in the
VH3 amino acid sequence of the antibody wherein X 4 is selected from D and E, X 5 is selected from G, A, V, I, L, and M, and W, and X 6 is selected from G, A, V, I, L, and M; and
wherein the amino acids are numbered according to the Aho numbering system.
2 . The method of claim 1 , wherein the viscosity-reducing substitutions comprise 1E, 17G, and 85A.
3 . The method of claim 1 , wherein the antibody comprises a heavy chain amino acid sequence comprising SEQ ID NO: 160, except for substitutions 1E, 17G, and 85A, and a light chain amino acid sequence comprising SEQ ID NO: 158.
4 . The method of claim 1 , wherein the antibody comprises a heavy chain amino acid sequence comprising SEQ ID NO: 144, except for substitutions 1E, 17G, and 85A, and a light chain amino acid sequence comprising SEQ ID NO: 142.
5 . The method of claim 1 , wherein the antibody comprises a heavy chain amino acid sequence comprising SEQ ID NO: 128, except for substitutions 1E, 17G, and 85A, and a light chain amino acid sequence comprising SEQ ID NO: 126.
6 . The method of claim 1 , wherein the antibody comprises a heavy chain amino acid sequence comprising SEQ ID NO: 120, except for substitutions 1E, 17G, and 85A, and a light chain amino acid sequence comprising SEQ ID NO: 118.
7 . An antibody of the VH3|3-33 germline subfamily, which comprises
a VH3 sequence comprising viscosity- reducing substitutions 1X 4 , 17X 5 , and 85X 6 , wherein X 4 is selected from D and E, X 5 is selected from G, A, V, I, L, and M, and X 6 is selected from G, A, V, I, L, and M;
wherein the amino acids in subparagraphs a, b, and e are numbered according to the Aho numbering system.
8 . The antibody of claim 7 , wherein the viscosity-reducing substitutions comprise 1E, 17G, and 85A.
9 . The antibody of claim 7 , comprising a heavy chain amino acid sequence comprising SEQ ID NO: 160, except for substitutions 1E, 17G, and 85A, and a light chain amino acid sequence comprising SEQ ID NO: 158.
10 . The antibody of claim 7 , comprising a heavy chain amino acid sequence comprising SEQ ID NO: 144, except for substitutions 1E, 17G, and 85A, and a light chain amino acid sequence comprising SEQ ID NO: 142.
11 . The antibody of claim 7 , comprising a heavy chain amino acid sequence comprising SEQ ID NO: 128, except for substitutions 1E, 17G, and 85A, and a light chain amino acid sequence comprising SEQ ID NO: 126.
12 . The antibody of claim 7 , comprising a heavy chain amino acid sequence comprising SEQ ID NO: 120, except for substitutions 1E, 17G, and 85A, and a light chain amino acid sequence comprising SEQ ID NO: 118.
Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Jul 17, 2024
From: HUH, JOON; STEVENSON, RIKI; BONDARENKO, PAVEL; NICHOLS, ANDREW; REN, DA; AGRAWAL, NEERAJ JAGDISH; SMITH, RICHARD
To: AMGEN INC.
Reel/Frame 068014/0804 →
Continuity (6)
Division
16338292
Provisional Application
62546469
· Aug 16, 2017
Provisional Application
62430773
· Dec 6, 2016
Provisional Application
62401770
· Sep 29, 2016
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