DESIGN AND DEVELOPMENT OF MASKED THERAPEUTIC ANTIBODIES TO LIMIT OFF-TARGET EFFECTS: APPLICATION TO ANTI-EGFR ANTIBODIES
In one embodiment, a masked monoclonal antibody (mAb) is provided, the mAb, encoded by a nucleic acid sequence or an amino acid sequence molecule comprising a signal sequence, a masking epitope sequence, a linker sequence that is cleavable by a protease specific to a target tissue; and an antibody or a functional fragment thereof. In another embodiment, a cross-masked mAb heterodimer complex is provided, comprising a first masked mAb, comprising a first signal sequence, a first masking epitope sequence, a first linker that is cleavable by a protease specific to a target tissue, and a first antibody or fragment thereof; and a second masked mAb, comprising a second signal sequence, a second masking epitope sequence, a second linker that is cleavable by a protease specific to a target tissue, and a second antibody or fragment thereof.
1 . A recombinant nucleic acid molecule comprising:
(a) a signal nucleic acid sequence;
(b) a nucleic acid sequence which encodes a masking epitope;
(c) a nucleic acid sequence which encodes a linker that is cleavable by a protease specific to a target tissue; and
(d) a nucleic acid sequence which encodes an antibody or an antigen binding fragment thereof.
2 . The recombinant nucleic acid molecule of claim 1 , wherein the masking epitope comprises an epitope specific to an antigenic recognition site of the antibody or functional fragment thereof.
3 . The recombinant nucleic acid molecule of claim 2 , wherein the antigenic recognition site is one or more complementary determining regions (CDRs).
4 . The recombinant nucleic acid molecule of claim 1 , wherein the protease is matrix metalloproteinase-9 (MMP-9).
5 . The recombinant nucleic acid molecule of claim 1 , wherein the antibody or antigen binding fragment thereof is specific to EGFR.
6 . The recombinant nucleic acid molecule of claim 1 , wherein the antibody or antigen binding fragment thereof is an IgG.
7 . The recombinant nucleic acid molecule of claim 1 , wherein the antibody or antigen binding fragment thereof is an scFv.
8 . The recombinant nucleic acid molecule of claim 1 , comprising SEQ ID NO:1 or SEQ ID NO:2.
9 . A masked monoclonal antibody (mAb) which comprises an amino acid sequence, the amino acid sequence comprising:
(a) a masking epitope sequence;
(b) a linker sequence that is cleavable by a protease specific to a target tissue; and
(c) an antibody sequence or an antigen binding fragment thereof.
10 . The masked mAb of claim 9 , wherein the nucleic acid molecule which encodes the masking epitope sequence comprises a nucleic acid molecule which encodes an epitope specific to an antigenic recognition site of the antibody or functional fragment thereof.
11 . The masked mAb of claim 10 , wherein the antigenic recognition site is one or more complementary determining regions (CDRs).
12 . The masked mAb of claim 9 , wherein the protease is matrix metalloproteinase-9 (MMP-9).
13 . The masked mAb of claim 9 , wherein the linker sequence comprises SEQ ID NO:3.
14 . The masked mAb of claim 9 , wherein the linker sequence comprises SEQ ID NO:4.
15 . The masked mAb of claim 9 , wherein the antibody or antigen binding fragment thereof is specific to EGFR.
16 . The masked mAb of claim 9 , wherein the antibody or antigen binding fragment thereof comprises an IgG.
17 . The masked mAb of claim 9 , wherein the antibody sequence or antigen binding fragment thereof is an scFv.
18 . The masked mAb of claim 9 , wherein the amino acid sequence of the masked mAb comprises SEQ ID NO:5 or SEQ ID NO:6.
19 . A masked monoclonal antibody (mAb) comprising:
(a) a masking epitope amino acid sequence;
(b) a linker sequence comprising SEQ ID NO:3; and
(c) an antibody amino acid sequence or an antigen binding fragment thereof.
20 . The masked mAb of claim 19 , wherein the linker sequence further comprises SEQ ID NO:4.