Anti-LAG-3 antibodies and uses thereof

1. An isolated monoclonal antibody or antigen-binding fragment thereof comprising a heavy chain variable region comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, and a light chain variable region comprising a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3, having the polypeptide sequences of:

a. SEQ ID NOs:35, 36, 37, 86, 87, and 88, respectively;

b. SEQ ID NOs:38, 39, 40, 89, 90, and 91, respectively;

c. SEQ ID NOs:41, 42, 137, 138, 93, and 94, respectively;

d. SEQ ID NOs:139, 140, 141, 142, 99, and 143, respectively;

e. SEQ ID NOs:144, 145, 146, 147, 148, and 149, respectively;

f. SEQ ID NOs:65, 66, 67, 116, 117, and 118, respectively;

g. SEQ ID NOs:68, 69, 70, 119, 120, and 121, respectively;

h. SEQ ID NOs:71, 72, 73, 122, 123, and 124, respectively;

i. SEQ ID NOs:74, 75, 76, 125, 126, and 127, respectively;

j. SEQ ID NOs:77, 78, 79, 128, 129, and 130, respectively; or

k. SEQ ID NOs:80, 150, 151, 131, 132, and 133, respectively;

l. SEQ ID NOs:68, 69, 70, 119, 120, and 121, respectively;

m. SEQ ID NOs:71, 72, 73, 122, 123, and 124, respectively;

n. SEQ ID NOs:74, 75, 76, 125, 126, and 127, respectively;

o. SEQ ID NOs:77, 78, 79, 128, 129, and 130, respectively;

p. SEQ ID NOs: 80, 81, 82, 131, 132, and 133, respectively; or

q. SEQ ID NOs: 83, 84, 85, 134, 135, and 136, respectively;

wherein the antibody or antigen-binding fragment thereof specifically binds LAG-3.

2. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1 , comprising:

a. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:1, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:2;

b. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:3, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:4;

c. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:5, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:6;

d. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:7, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:8;

e. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:9, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:10;

f. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:11, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:12;

g. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:13, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:14;

h. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:15, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:16;

i. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:17, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:18;

j. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:19, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:20;

k. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:21, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:22;

l. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:23, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:24;

m. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:25, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:26;

n. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:27, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:28;

o. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:29, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:30;

p. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:31, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:32;

q. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:33, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:34;

r. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:195, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:208;

s. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:196, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:206;

t. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:197, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:205;

u. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:197, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:206;

v. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:197, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:207;

w. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:198, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:205;

x. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:199, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:209;

y. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:200, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:209;

z. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:201, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:209;

aa. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:202, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:209;

bb. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:203, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:209;

cc. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:204, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:209;

dd. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:199, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:210;

ee. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:200, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:210;

ff. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:201, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:210;

gg. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:202, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:210;

hh. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:203, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:210; or

ii. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:204, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:210.

3. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1 , comprising:

a. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:1, and a light chain variable region having the polypeptide sequence of SEQ ID NO:2;

b. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:3, and a light chain variable region having the polypeptide sequence of SEQ ID NO:4;

c. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:5, and a light chain variable region having the polypeptide sequence of SEQ ID NO:6;

d. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:7, and a light chain variable region having the polypeptide sequence of SEQ ID NO:8;

e. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:9, and a light chain variable region having the polypeptide sequence of SEQ ID NO:10;

f. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:11, and a light chain variable region having the polypeptide sequence of SEQ ID NO:12;

g. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:13, and a light chain variable region having the polypeptide sequence of SEQ ID NO:14;

h. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:15, and a light chain variable region having the polypeptide sequence of SEQ ID NO:16;

i. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:17, and a light chain variable region having the polypeptide sequence of SEQ ID NO:18;

j. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:19, and a light chain variable region having the polypeptide sequence of SEQ ID NO:20;

k. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:21, and a light chain variable region having the polypeptide sequence of SEQ ID NO:22;

l. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:23, and a light chain variable region having the polypeptide sequence of SEQ ID NO:24;

m. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:25, and a light chain variable region having the polypeptide sequence of SEQ ID NO:26;

n. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:27, and a light chain variable region having the polypeptide sequence of SEQ ID NO:28;

o. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:29, and a light chain variable region having the polypeptide sequence of SEQ ID NO:30;

p. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:31, and a light chain variable region having the polypeptide sequence of SEQ ID NO:32;

q. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:33, and a light chain variable region having the polypeptide sequence of SEQ ID NO:34;

r. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:195, and a light chain variable region having the polypeptide sequence of SEQ ID NO:208;

s. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:196, and a light chain variable region having the polypeptide sequence of SEQ ID NO:206;

t. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:197, and a light chain variable region having the polypeptide sequence of SEQ ID NO:205;

u. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:197, and a light chain variable region having the polypeptide sequence of SEQ ID NO:206;

v. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:197, and a light chain variable region having the polypeptide sequence of SEQ ID NO:207;

w. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:198, and a light chain variable region having the polypeptide sequence of SEQ ID NO:205;

x. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:199, and a light chain variable region having the polypeptide sequence of SEQ ID NO:209;

y. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:200, and a light chain variable region having the polypeptide sequence of SEQ ID NO:209;

z. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:201, and a light chain variable region having the polypeptide sequence of SEQ ID NO:209;

aa. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:202, and a light chain variable region having the polypeptide sequence of SEQ ID NO:209;

bb. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:203, and a light chain variable region having the polypeptide sequence of SEQ ID NO:209;

cc. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:204, and a light chain variable region having the polypeptide sequence of SEQ ID NO:209;

dd. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:199, and a light chain variable region having the polypeptide sequence of SEQ ID NO:210;

ee. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:200, and a light chain variable region having the polypeptide sequence of SEQ ID NO:210;

ff. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:201, and a light chain variable region having the polypeptide sequence of SEQ ID NO:210;

gg. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:202, and a light chain variable region having the polypeptide sequence of SEQ ID NO:210;

hh. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:203, and a light chain variable region having the polypeptide sequence of SEQ ID NO:210; or

ii. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:204, and a light chain variable region having the polypeptide sequence of SEQ ID NO:210.

4. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1 , wherein the isolated monoclonal antibody or antigen-binding fragment thereof specifically binds to an epitope comprising an amino acid sequence selected from the group consisting of SEQ ID NO:192, SEQ ID NO:193, SEQ ID NO:158, SEQ ID NO:180, and SEQ ID NO:194.

5. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1 , wherein the monoclonal antibody or antigen-binding fragment thereof inhibits LAG-3 activity and/or is capable of blocking binding to LAG-3 to MHC class II molecules.

6. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1 , wherein the antibody or antigen-binding fragment thereof is chimeric and/or human or humanized.

7. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1 , wherein the monoclonal antibody or antigen-binding fragment thereof specifically binds human LAG-3.

8. A pharmaceutical composition comprising the isolated monoclonal antibody or antigen-binding fragment thereof of claim 1 and a pharmaceutically acceptable carrier.

9. A method of producing a pharmaceutical composition comprising

the monoclonal antibody or antigen-binding fragment of claim 1 , comprising combining the monoclonal antibody or antigen-binding fragment with a pharmaceutically acceptable carrier to obtain the pharmaceutical composition.

10. A bispecific antibody comprising the monoclonal antibody or antigen-binding fragment thereof of claim 1 .

11. An isolated nucleic acid encoding the monoclonal antibody or antigen-binding fragment thereof of claim 1 .

12. A vector comprising the isolated nucleic acid of claim 11 .

13. A host cell comprising the vector of claim 12 .

14. A method of producing the monoclonal antibody or antigen-binding fragment thereof of claim 1 , comprising culturing a cell comprising a nucleic acid encoding the monoclonal antibody or antigen-binding fragment under conditions to produce the monoclonal antibody or antigen-binding fragment, and recovering the antibody or antigen-binding fragment from the cell or culture.

15. A method of determining a level of LAG-3 in a subject, the method comprising:

a. obtaining a sample from the subject;

b. contacting the sample with an isolated monoclonal antibody or antigen-binding fragment thereof of claim 1 ; and

c. determining a level of LAG-3 in the subject.

16. The method of claim 15 , wherein the sample is a tissue sample or a blood sample.

17. The method of claim 16 , wherein the tissue sample is a cancer tissue sample.