Anti-lag-3 antibody polypeptide
The present disclosure provides anti-LAG-3 heavy-chain antibody or the antigen-binding domain thereof, isolated polynucleotides encoding the same, pharmaceutical compositions comprising the same, and the uses thereof.
1. An isolated single domain antibody polypeptide comprising a heavy chain variable domain that specifically binds to LAG-3, wherein the heavy chain variable domain comprises CDR1, CDR2 and CDR3, wherein
the CDR1 comprises the amino acid sequence of GLTLSQYTMG (SEQ ID NO: 1),
the CDR2 comprises the amino acid sequence of AIHWTSSVTDYADSVX 1 G (SEQ ID NO:33), and
the CDR3 comprises the amino acid sequence of TX 2 YYTHRGX 3 FDY (SEQ ID NO:34),
wherein X 1 is K, Y, M, D, or R, X 2 is H or W, and X 3 is S or P.
2. The antibody polypeptide of claim 1 , wherein:
the CDR1 comprises the sequence of SEQ ID NO: 1,
the CDR2 comprises a sequence selected from SEQ ID NOs: 2, 4, 8, 9, and 10, and
the CDR3 comprises a sequence selected from SEQ ID NOs: 3, 5, 6, and 7.
3. The antibody polypeptide of claim 1 , comprising:
a) a heavy chain variable region comprising CDR1 comprising the amino acid sequence of SEQ ID NO: 1, CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and CDR3 comprising the amino acid sequence of SEQ ID NO: 3;
b) a heavy chain variable region comprising CDR1 comprising the amino acid sequence of SEQ ID NO: 1, CDR2 comprising the amino acid sequence of SEQ ID NO: 4, and CDR3 comprising the amino acid sequence of SEQ ID NO: 5;
c) a heavy chain variable region comprising CDR1 comprising the amino acid sequence of SEQ ID NO: 1, CDR2 comprising the amino acid sequence of SEQ ID NO: 4, and CDR3 comprising the amino acid sequence of SEQ ID NO: 6;
d) a heavy chain variable region comprising CDR1 comprising the amino acid sequence of SEQ ID NO: 1, CDR2 comprising the amino acid sequence of SEQ ID NO: 4, and CDR3 comprising the amino acid sequence of SEQ ID NO: 7;
e) a heavy chain variable region comprising CDR1 comprising the amino acid sequence of SEQ ID NO: 1, CDR2 comprising the amino acid sequence of SEQ ID NO: 8, and CDR3 comprising the amino acid sequence of SEQ ID NO: 7;
f) a heavy chain variable region comprising CDR1 comprising the amino acid sequence of SEQ ID NO: 1, CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and CDR3 comprising the amino acid sequence of SEQ ID NO: 5;
g) a heavy chain variable region comprising CDR1 comprising the amino acid sequence of SEQ ID NO: 1, CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and CDR3 comprising the amino acid sequence of SEQ ID NO: 6;
h) a heavy chain variable region comprising CDR1 comprising the amino acid sequence of SEQ ID NO: 1, CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and CDR3 comprising the amino acid sequence of SEQ ID NO: 7;
i) a heavy chain variable region comprising CDR1 comprising the amino acid sequence of SEQ ID NO: 1, CDR2 comprising the amino acid sequence of SEQ ID NO: 10, and CDR3 comprising the amino acid sequence of SEQ ID NO: 5; or
j) a heavy chain variable region comprising CDR1 comprising the amino acid sequence of SEQ ID NO: 1, CDR2 comprising the amino acid sequence of SEQ ID NO: 4, and CDR3 comprising the amino acid sequence of SEQ ID NO: 3.
4. The antibody polypeptide of claim 1 , comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, and SEQ ID NO: 31.
5. The antibody polypeptide of claim 1 , wherein the heavy chain variable domain is derived from a VHH domain.
6. The antibody polypeptide of claim 1 , further comprising an immunoglobulin constant region, optionally a constant region of human Ig, or optionally an Fc region of human IgG.
7. The antibody polypeptide of claim 1 , which is of camelid origin or is humanized.
8. The antibody polypeptide of claim 1 , which is a nanobody.
9. The antibody polypeptide of claim 1 , capable of specifically binding to human LAG-3 at a K D value of no more than 5×10 −9 , 2×10 −10 , 2.5×10 −12 M as measured by surface plasmon resonance (SPR).
10. The antibody polypeptide of claim 1 , capable of specifically binding to human LAG-3 expressed on a cell surface at a K D value of no more than 10 −9 , 5×10 −10 , 6×10 −11 M as measured by flow cytometry.
11. The antibody polypeptide of claim 1 , capable of specifically binding to cynomolgus monkey LAG-3, and/or mouse LAG-3.
12. The antibody polypeptide of claim 1 , linked to one or more conjugate moieties.
13. The antibody polypeptide of claim 12 , wherein the conjugate moiety selected from the group consisting of a clearance-modifying agent, a chemotherapeutic agent, a toxin, a radioactive isotope, a lanthanide, a luminescent label, a fluorescent label, an enzyme-substrate label, a DNA-alkylators, a topoisomerase inhibitor, a tubulin-binders, or an anticancer drug.
14. A pharmaceutical composition comprising the antibody polypeptide of claim 1 , and a pharmaceutically acceptable carrier.
15. An isolated polynucleotide encoding the antibody polypeptide of claim 1 .
16. The isolated polynucleotide of claim 15 , comprising a nucleotide sequence selecting from a group consisting of SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, and SEQ ID NO: 32.
17. A vector comprising the isolated polynucleotide of claim 15 .
18. A host cell comprising the vector of claim 17 .
19. A method of expressing the antibody polypeptide of claim 1 , comprising culturing the host cell of claim 18 under the condition at which the vector of claim 17 is expressed.
20. A method of detecting presence or amount of LAG-3 in a sample, comprising contacting the sample with the antibody polypeptide of claim 1 , and determining the presence or the amount of LAG-3 in the sample.
21. A method of diagnosing a Lag-3 related disease or condition in a subject, comprising: a) contacting a sample obtained from the subject with the antibody polypeptide of claim 1 ; b) determining presence or amount of LAG-3 in the sample; and c) correlating the presence or the amount of LAG-3 to existence or status of the LAG-3 related disease or condition in the subject.
22. A kit comprising the antibody polypeptide of claim 1 , useful in detecting LAG-3.