	US 12,325,733 B2
	Binding domain
Anna Bulek, London (GB); Martin Pulé, London (GB); Shaun Cordoba, London (GB); Simon Thomas, London (GB); Shimobi Onuoha, London (GB); Mathieu Ferrari, London (GB); and Vania Baldan, London (GB)
Assigned to AUTOLUS LIMITED, London (GB)
Appl. No. 17/290,188
Filed by AUTOLUS LIMITED, London (GB)
PCT Filed Oct. 31, 2019, PCT No. PCT/GB2019/053100 § 371(c)(1), (2) Date Apr. 29, 2021, PCT Pub. No. WO2020/089644, PCT Pub. Date May 7, 2020.
Claims priority of application No. 1817822 (GB), filed on Oct. 31, 2018.
Prior Publication US 2022/0041718 A1, Feb. 10, 2022
Int. Cl. A61P 35/00 (2006.01); A61K 47/68 (2017.01); C07K 14/705 (2006.01); C07K 14/725 (2006.01); C07K 16/28 (2006.01); G01N 33/574 (2006.01)

CPC C07K 14/7051 (2013.01) [A61K 47/6849 (2017.08); C07K 14/70517 (2013.01); C07K 16/2809 (2013.01); G01N 33/57407 (2013.01); G01N 33/57426 (2013.01); C07K 2317/56 (2013.01); C07K 2319/03 (2013.01)]

15 Claims

1. A variant antigen-binding domain which comprises mutations in the VH domain compared to a reference antibody having a VH domain with the sequence shown in SEQ ID NO: 1 and a VL domain with the sequence shown in SEQ ID NO: 2, wherein the variant antigen-binding domain comprises the following mutation combinations:

T28K, Y32F, A100N, Y102F in the VH domain;

T28K, Y32F, A100N, Y102L, N103M in the VH domain and N35R in the VL domain;

T28K, Y32F, A100N, Y102L, N103W in the VH domain and N35K in the VL domain;

T28K, Y32F, A100N, Y102L, N103W in the VH domain and N35R in the VL domain;

T28K, Y32F, A100N, N103E in the VH domain;

V2K, T28K, Y32F, A100N in the VH domain;

N103E, T28K, T32F, A100N in the VH domain and N35M in the VL domain;

T28K, Y32F, A100N, N103F in the VH domain and N35K in the VL domain;

T28K, Y32F, A100N, A107S in the VH domain;

G31S, T28K, Y32F, A100N in the VH domain;

V2R, T28K, Y32F, A100N in the VH domain;

T28K, Y32F, A100N, N103M in the VH domain and N35R in the VL domain;

N103W, T28K, Y32F, A100N in the VH domain and N35F in the VL domain;

T28K, G31R, Y32F, A100N in the VH domain;

N103F, T28K, T32F, A100N in the VH domain and N35F in the VL domain;

N103F, T28K, T32F, A100N in the VH domain and N35M in the VL domain;

N103M, T28K, T32F, A100N in the VH domain and N32F in the VL domain;

T28K, Y32F, A100N, N103S in the VH domain;

T28K, Y32F, A100N, N103Q in the VH domain;

Y27F, T28K, Y32F, A100N in the VH domain;

N103F, T28K, Y32F, A100N in the VH domain and N35Y in the VL domain;

N103W, T28K, Y32F, A100N in the VH domain and N35M in the VL domain;

N103L, T28K, Y32F, A100N in the VH domain and N35F in the VL domain;

N103L, T28K, Y32F, A100N in the VH domain and N35K in the VL domain;

T28K, Y32F, A100N, N103W in the VH domain;

T28K, Y32F, A100N, N103F in the VH domain;

N103L, T28K, Y32F, A100N in the VH domain and N35Y in the VL domain;

N103W, T28K, Y32F, A100N in the VH domain and N35K in the VL domain;

N103L, T28K, Y32F, A100N in the VH domain and N35R in the VL domain;

N103L, T28K, Y32F, A100N in the VH domain and N35M in the VL domain;

N103S, T28K, Y32F, A100N in the VH domain and N35Y in the VL domain;

N103S, T28K, Y32F, A100N in the VH domain and N35F in the VL domain;

T28K, Y32F, A100N, N103L in the VH domain;

T28K, Y32F, A100N, N103M in the VH domain and N35Y in the VL domain;

N103S, T28K, Y32F, A100N in the VH domain and N35R in the VL domain;

T28K, Y32F, R98K, A100N in the VH domain;

N103S, T28K, Y32F, A100N in the VH domain and N35K in the VL domain;

T28K, Y32F, A100N in the VH domain and N35F in the VL domain;

N103W, T28K, T32F, A100N in the VH domain and N35R in the VL domain;

T28K, Y32F, A100N, N103M in the VH domain;

N103S, T28K, Y32F, A100N in the VH domain and N35M in the VL domain;

T28K, Y32F, A100N in the VH domain and N35R in the VL domain;

T28K, Y32F, A100N, N103Y in the VH domain;

T28K, Y32F, A100N, N103A in the VH domain;

T28K, Y32F, A100N, N103H in the VH domain;

T28K, Y32F, A100N, N103M in the VH domain and N35M in the VL domain;

T28K, Y32F, A100N in the VH domain and N35K in the VL domain;

T28K, Y32F, A100N in the VH domain and R55K in the VL domain;

T28K, Y32F, A100N in the VH domain;

Y27W, T28K, Y32F, A100N in the VH domain;

T28K, Y32F, A100N, Y102F in the VH domain and N35R in the VL domain;

T28K, Y32F, A100N, Y102F, N103M in the VH domain and N35R in the VL domain;

T28K, Y32F, A100N, Y102F, N103M in the VH domain and N35F in the VL domain;

Y27M, T28K, Y32F, A100N in the VH domain;

T28K, G31K, Y32F, A100N in the VH domain;

T28K, Y32F, A100N, Y102F, N103M in the VH domain and N35K in the VL domain; or

Y27N, T28K, V32F, A100N in the VH domain;

and wherein the variant antigen-binding domain displays an increased affinity for TRBC2 over the reference antibody.