Antigen binding constructs to CD4
Antigen binding constructs that bind to CD4, for example antibodies, including antibody fragments (such as scFv, minibodies, and cys-diabodies) that bind to CD4, are described herein. Methods of use are described herein.
1. An antigen binding construct that comprises:
a HCDR1 having the amino acid sequence KASGYSFANYWMH (SEQ ID NO: 45);
a HCDR2 having the amino acid sequence EWIGALYPGNVDTT (SEQ ID NO: 46);
a HCDR3 having the amino acid sequence TRMGTTLEAPLDY (SEQ ID NO: 47);
a LCDR1 having the amino acid sequence SARSSVSYLY (SEQ ID NO:48);
a LCDR2 having the amino acid sequence YDTSNLAS (SEQ ID NO:49); and
a LCDR3 having the amino acid sequence QQWSDYPLT (SEQ ID NO:50).
2. The antigen binding construct of claim 1 , further comprising at least one VL framework residue selected from the group consisting of:
a Glutamic acid at position 1 of the sequence in SEQ ID NO: 4;
a Leucine at position 13 of the sequence in SEQ ID NO: 4;
an Arginine at position 18 of the sequence in SEQ ID NO: 4;
a Threonine at position 20 of the sequence in SEQ ID NO: 4;
a Leucine at position 21 of the sequence in SEQ ID NO: 4;
a Serine at position 22 of the sequence in SEQ ID NO: 4;
a Glutamine at position 41 of the sequence in SEQ ID NO: 4;
an Alanine at position 42 of the sequence in SEQ ID NO: 4;
an Isoleucine at position 57 of the sequence in SEQ ID NO: 4;
an Aspartic acid at position 59 of the sequence in SEQ ID NO: 4;
a Phenylalanine at position 70 of the sequence in SEQ ID NO: 4;
a Threonine at position 71 of the sequence in SEQ ID NO: 4;
a Leucine at position 77 of the sequence in SEQ ID NO: 4;
a Proline at position 79 of the sequence in SEQ ID NO: 4;
a Valine at position 84 of the sequence in SEQ ID NO: 4; and
a Glycine at position 99 of the sequence in SEQ ID NO: 4.
3. The antigen binding construct of claim 1 , further comprising at least one VH framework residue selected from the group consisting of:
a Glutamine at position 1 of the sequence in SEQ ID NO: 2;
a Valine at position 5 of the sequence in SEQ ID NO: 2;
an Alanine at position 9 of the sequence in SEQ ID NO: 2;
a Valine at position 11 of the sequence in SEQ ID NO: 2;
a Lysine at position 13 of the sequence in SEQ ID NO: 2;
an Arginine at position 44 of the sequence in SEQ ID NO: 2;
a Glutamine at position 65 of the sequence in SEQ ID NO: 2;
a Glycine at position 66 of the sequence in SEQ ID NO: 2;
an Arginine at position 67 of the sequence in SEQ ID NO: 2;
a Valine at position 68 of the sequence in SEQ ID NO: 2;
a Threonine at position 69 of the sequence of SEQ ID NO: 2;
an Arginine at position 87 of the sequence in SEQ ID NO: 2; and
a Serine at position 88 of the sequence in SEQ ID NO: 2.
4. The antigen binding construct of claim 1 , further comprising a detectable marker.
5. The antigen binding construct of claim 1 , further comprising a variable heavy domain (VH), wherein the VH comprises the amino acid sequence in SEQ ID NO: 2.
6. The antigen binding construct of claim 1 , further comprising a variable light domain (VL), wherein the VL comprises the amino acid sequence in SEQ ID NO: 4.
7. The antigen binding construct of claim 1 , wherein the antigen binding construct is a minibody.
8. The antigen binding construct of claim 7 , further comprising the amino acid sequence in SEQ ID NO: 5, 6, 7, or 8.
9. A nucleic acid sequence encoding the antigen binding construct of claim 7 , wherein the nucleic acid sequence comprises the nucleic acid sequence in SEQ ID NO: 9 or 10.
10. The antigen binding construct of claim 1 , wherein the antigen binding construct is a humanized cys-diabody.
11. The antigen binding construct of claim 10 comprising the amino acid sequence in SEQ ID NO: 11, 12, 13, 14, 15, 16, 17, or 18.
12. The antigen binding construct of claim 1 , wherein the antigen binding construct comprises a polypeptide that comprises a single-chain variable fragment (scFv) comprising a variable heavy domain (VH) linked to a variable light domain (VL).
13. The antigen binding construct of claim 1 , further comprising a hinge region comprising the amino acid sequence in SEQ ID NO: 34, 35, 36, or 37.
14. An antigen binding construct that comprises:
a HCDR1 of the HCDR1 in SEQ ID NO: 2;
a HCDR2 of the HCDR2 in SEQ ID NO: 2;
a HCDR3 of the HCDR3 in SEQ ID NO: 2;
a LCDR1 of the LCDR1 in SEQ ID NO: 4;
a LCDR2 of the LCDR2 in SEQ ID NO: 4;
a LCDR3 of the LCDR3 in SEQ ID NO: 4; and
at least one of:
a) a VH comprising at least one of: E1Q, Q5V, T9A, V10E, L11V, A12K, R13K, Q19K, M20V, K38R, R40A, Q46E, T59, N61, K65Q, D66G, K67R, A68V, K69T, L70I, A72R, V73D, T87R, N88S, S91T, T115L, or L116V;
b) a VL comprising at least one of: Q1E, I10T, M11L, A13L, K18R, V19A, A20T, M21L, T22S, S41Q, S42A, V57I, V59D, I62S, S69D, Y70F, S71T, M77L, A79P, A82, T84V, A99G, or L106I;
c) the VH of a) and the VL of b);
d) any one of a), b), and c), wherein the VH is at least 80% identical to SEQ ID NO: 2;
e) any one of a), b), and c), wherein the VL is at least 80% identical to SEQ ID NO: 4;
f) any one of a), b), and c), wherein the VH is at least 80% identical to SEQ ID NO: 2; and any one of a), b), and c), wherein the VL is at least 80% identical to SEQ ID NO: 4;
g) a VH with T59, N61, or both, wherein the VH is a humanized VH;
h) a VL with A82, wherein the VL is a humanized VL;
i) a VH according to g) and a VL according to h);
j) a VH with Q46E;
k) a VH with Q46E, wherein
i) the VH is at least 80% identical to SEQ ID NO: 2;
ii) the VL is at least 80% identical to SEQ ID NO: 4; or
iii) the VH is at least 80% identical to SEQ ID NO: 2 and the VL is at least 80% identical to SEQ ID NO: 4;
l. A VH with at least one of T115L, L116V;
m) a VL with at least one of S41Q, A99G, or L106I;
n) a VH with at least one of T115L, L116V and a VL with at least one of S41Q, A99G, or L106I; or
o) l), m), or n), wherein
i) the VH is at least 80% identical to SEQ ID NO: 2;
ii) the VL is at least 80% identical to SEQ ID NO: 4; or
iii) the VH is at least 80% identical to SEQ ID NO: 2 and the VL is at least 80% identical to SEQ ID NO: 4.
15. The antigen binding construct of claim 1 , comprising:
a variable heavy domain (VH), wherein the VH is at least 90% identical to SEQ ID NO: 2; and/or
a variable light domain (VL), wherein the VL is at least 90% identical to SEQ ID NO: 4.
16. The antigen binding construct of claim 1 , wherein the antigen binding construct binds to CD4.
17. The antigen binding construct of claim 1 , wherein the antigen binding construct is a minibody, and wherein the minibody is covalently linked to a detectable marker.
18. The antigen binding construct of claim 17 , wherein the detectable marker is 18 F or 89 Zr.
19. The antigen binding construct of claim 1 , wherein the antigen binding construct comprises a VH with T59, N61, or both, wherein the VH is a humanized VH; and wherein the antigen binding construct comprises a VL with A82, wherein the VL is a humanized VL.