Anti-transferrin receptor (TFR) antibody, encoding nucleic acid thereof and method of use thereof to deliver an oligonucleotide to a subject
Aspects of the disclosure relate to antibodies that bind to transferrin receptor (e.g., transferrin receptor 1) and complexes comprising the antibody covalently linked to a molecular payload. Methods of using the antibodies are also provided.
1 . An antibody that binds to human transferrin receptor (TfR), wherein the antibody comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 76 and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 75.
2 . The antibody of claim 1 , wherein the antibody comprises a heavy chain comprising an N-terminal pyroglutamate.
3 . The antibody of claim 1 , wherein the antibody is not covalently linked to a molecular payload.
4 . A composition comprising the antibody of claim 1 and a pharmaceutically acceptable carrier.
5 . The composition of claim 4 , wherein the antibody comprises a heavy chain comprising an N-terminal pyroglutamate.
6 . The composition of claim 4 , wherein the antibody is not covalently linked to a molecular payload.
7 . The composition of claim 4 , wherein the composition is suitable for intravenous administration.
8 . The composition of claim 4 , wherein the composition is suitable for intravenous infusion.
9 . The composition of claim 4 , wherein the composition is a sterile injectable solution.
10 . The composition of claim 4 , wherein the composition is an aqueous solution.
11 . The composition of claim 4 , wherein the composition is lyophilized.
12 . The composition of claim 4 , wherein the composition is present in a container.
13 . The composition of claim 4 , wherein the composition is present in a column.
14 . The composition of claim 4 , wherein the composition is suitable for administration to a subject.
15 . The composition of claim 4 , wherein the composition is present in a cell in vitro.
16 . The composition of claim 4 , wherein the antibody is bound to a solid support.
17 . A nucleic acid comprising nucleotide sequences encoding a heavy chain and/or a light chain of the antibody of claim 1 .
18 . The nucleic acid of claim 17 , comprising a nucleotide sequence encoding the heavy chain of the antibody.
19 . The nucleic acid of claim 17 , comprising a nucleotide sequence encoding the light chain of the antibody.
20 . The nucleic acid of claim 17 , comprising a nucleotide sequence encoding the heavy chain and the light chain of the antibody.
21 . The nucleic acid of claim 18 , wherein the nucleotide sequence is operably linked to a promoter.
22 . The nucleic acid of claim 19 , wherein the nucleotide sequence is operably linked to a promoter.
23 . The nucleic acid of claim 20 , wherein the nucleotide sequence is operably linked to a promoter.
24 . A vector comprising the nucleic acid of claim 17 .
25 . The vector of claim 24 , wherein the vector is an expression vector.
26 . A cell comprising the vector of claim 24 .
27 . A method of producing the antibody of claim 1 , wherein the method comprises culturing a cell comprising a nucleic acid sequence encoding a heavy chain of the antibody and a nucleic acid sequence encoding a light chain of the antibody.
28 . The method of claim 27 , wherein the nucleic acid sequence encoding the heavy chain and the nucleic acid sequence encoding the light chain are on the same vector.
29 . The method of claim 27 , wherein the nucleic acid sequence encoding the heavy chain and the nucleic acid sequence encoding the light chain are on different vectors.
30 . The method of claim 27 , wherein the nucleic acid sequence encoding the heavy chain is operably linked to a promoter.
31 . The method of claim 27 , wherein the nucleic acid sequence encoding the light chain is operably linked to a promoter.
32 . The method of claim 27 , wherein the nucleic acid sequence encoding the heavy chain and the nucleic acid sequence encoding the light chain are operably linked to a promoter.
33 . The method of claim 27 , wherein the cell is a CHO cell.
34 . The method of claim 33 , wherein the CHO cell is a dhfr-CHO cell.
35 . The method of claim 33 , wherein the CHO cell is a CHO-S cell.
36 . The method of claim 33 , wherein the CHO cell is a CHO-K1 cell.
37 . The method of claim 27 , further comprising isolating the antibody.
38 . The method of claim 37 , wherein the antibody is isolated via affinity chromatography.
39 . The method of claim 37 , wherein the antibody is isolated by affinity chromatography with a Protein A or Protein G coupled matrix.
40 . A complex comprising the antibody of claim 1 covalently linked to a molecular payload.
41 . The complex of claim 40 , wherein the antibody comprises a heavy chain comprising an N-terminal pyroglutamate.
42 . The complex of claim 40 , wherein the molecular payload comprises an oligonucleotide.
43 . The complex of claim 42 , wherein the oligonucleotide is single stranded.
44 . The complex of claim 42 , wherein the oligonucleotide is double stranded.
45 . The complex of claim 42 , wherein the oligonucleotide comprises an SiRNA.
46 . The complex of claim 42 , wherein the oligonucleotide comprises one or more modified nucleosides.
47 . The complex of claim 46 , wherein the one or more modified nucleosides comprise 2′-modified nucleosides.
48 . The complex of claim 42 , wherein the oligonucleotide comprises one or more modified internucleoside linkages.
49 . The complex of claim 42 , wherein the oligonucleotide comprises one or more phosphorothioate internucleoside linkages.
50 . The complex of claim 40 , wherein the antibody is covalently linked to the molecular payload via a linker.
51 . The complex of claim 50 , wherein the linker comprises a cleavable linker.
52 . The complex of claim 50 , wherein the linker comprises a valine-citrulline sequence.
53 . The complex of claim 42 , comprising a structure of:
wherein n is 3 and m is 4, and wherein L1 comprises a spacer that is a substituted or unsubstituted aliphatic, substituted or unsubstituted heteroaliphatic, substituted or unsubstituted carbocyclylene, substituted or unsubstituted heterocyclylene, substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene, —O—, —N(R A )—, —S—, —C(═O)—, —C(═O)O—, —C(═O)NR A —, —NR A C(═O)—, —NR A C(═O) R A —, —C(═O)R A —, —NR A C(═O)O—, —NR A C(═O)N(R A )—, —OC(═O)—, —OC(═O)O—, —OC(═O)N(R A )—, —S(O) 2 NR A —, —NR A S(O) 2 —, or a combination thereof, wherein each R A is independently hydrogen or substituted or unsubstituted alkyl.
54 . A method of delivering an oligonucleotide to a subject, comprising administering to the subject the complex of claim 42 .
55 . The method of claim 54 , wherein the oligonucleotide is delivered to a muscle cell of the subject.
56 . The method of claim 54 , wherein the oligonucleotide is single stranded.
57 . The method of claim 54 , wherein the oligonucleotide is double stranded.
58 . The method of claim 54 , wherein the oligonucleotide is an siRNA.
59 . The method of claim 54 , wherein the subject is human.
60 . The method of claim 59 , wherein the subject has a muscle disorder.
61 . The method of claim 60 , wherein the muscle disorder is a muscular dystrophy.
62 . The method of claim 61 , wherein the muscular dystrophy is DMD.
63 . The method of claim 61 , wherein the muscular dystrophy is DM1.
64 . The method of claim 61 , wherein the muscular dystrophy is FSHD.