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 masked monoclonal antibody (mAb), encoded by a nucleic acid sequence or an amino acid sequence molecule comprising:
(a) a signal sequence;
(b) a masking epitope sequence;
(c) a linker sequence that is cleavable by a protease specific to a target tissue; and
(d) an antibody or a functional fragment thereof.
2 . The masked mAb of claim 1 , wherein the masking epitope sequence encodes an epitope specific to an antigenic recognition site of the antibody or functional fragment thereof.
3 . The masked mAb of claim 2 , wherein the antigenic recognition site is one or more complementary determining regions (CDRs).
4 . The masked mAb of claim 1 , wherein the protease is matrix metalloproteinase-9 (MMP-9).
5 . The masked mAb of claim 1 , wherein the antibody or functional fragment thereof is specific to EGFR.
6 . The masked mAb of claim 1 , wherein the antibody or functional fragment thereof is an IgG sequence.
7 . The masked mAb of claim 1 , wherein the antibody or functional fragment thereof is an scFv sequence.
8 . The masked mAb of claim 1 , wherein the masked mAb is a nucleic acid sequence consisting of SEQ ID NO:1.
9 . The masked mAb of claim 1 , wherein the masked mAb is a nucleic acid sequence consisting of SEQ ID NO:2.
10 . The masked mAb of claim 1 , wherein the masked mAb is an amino acid sequence consisting of SEQ ID NO:5.
11 . The masked mAb of claim 1 , wherein the masked mAb is an amino acid sequence consisting of SEQ ID NO:6.
12 . A cross-masked monoclonal antibody (mAb) heterodimer complex, 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.
13 . The cross-masked mAb heterodimer complex of claim 12 , wherein the first and second masking epitope sequences encode an epitope specific to an antigenic recognition site of the antibody or functional fragment thereof.
14 . The cross-masked mAb heterodimer complex of claim 12 , wherein the protease is matrix metalloproteinase-9 (MMP-9).
15 . The cross-masked mAb heterodimer complex of claim 12 , wherein the first and/or second antibody or functional fragment thereof is specific to EGFR.
16 . The cross-masked mAb heterodimer complex of claim 12 , wherein the first and second antibody or functional fragment thereof are an IgG or scFv sequence.
17 . The cross-masked mAb heterodimer complex of claim 12 , wherein the first masked mAb is encoded by a nucleic acid sequence consisting of SEQ ID NO:1 and the second masked mAb is encoded by a nucleic acid sequence consisting of SEQ ID NO:2.
18 . The cross-masked mAb heterodimer complex of claim 12 , wherein the first masked mAb is encoded by an amino acid sequence consisting of SEQ ID NO:5 and the second masked mAb is encoded by an amino acid sequence consisting of SEQ ID NO:6.
19 . The cross-masked mAb heterodimer complex of claim 12 , wherein the first and second masked mAbs form the cross-masked mAb heterodimer complex when first and second antigen recognition sites are occluded by the first and second masking epitopes.
20 . A cross-masked monoclonal antibody (mAb) heterodimer complex, comprising:
a first masked mAb encoded by a nucleic acid sequence of an amino acid sequence of SEQ ID NO:1 or SEQ ID NO:5, respectively, and a second masked mAb encoded by a nucleic acid sequence of an amino acid sequence of SEQ ID NO:2 or SEQ ID NO:6, respectively.