Anti-CD20 antibody and uses thereof
Provided herein are anti-CD20 antibodies and uses thereof for treatment and diagnosis. Also provided are CD20 antigens for the production of anti-CD20 antibodies and methods of generating anti-CD20 antibodies using the CD20 antigens.
1 . A method of treatment comprising
a. isolating T-cells from a subject,
b. transfecting the T-cells with a vector comprising a nucleic acid encoding a chimeric antigen receptor (CAR) comprising
(i) an antibody or an antigen binding portion thereof that specifically binds to a canine CD20 cyclic peptide having the sequence of SEQ ID NO: 20, wherein the cysteine at position 7 of SEQ ID NO: 20 forms a disulfide bond with the cysteine at position 23 of SEQ ID NO: 20, wherein the antibody or the antigen binding portion comprises a heavy chain variable domain (VH) complementarity determining region (CDR) 1 of SEQ ID NO: 40, a VH CDR2 of SEQ ID NO: 42, a VH CDR3 of SEQ ID NO: 44, and a light chain variable domain (VL) CDR1 of SEQ ID NO: 46, a VL CDR2 of LVS, and a VL CDR3 of SEQ ID NO: 50;
(ii) a transmembrane portion; and
(iii) a cytoplasmic signaling portion, and
c. administering the transfected T-cells to the subject.
2 . The method of claim 1 , wherein the subject is a canine subject.
3 . The method of claim 1 , wherein the antibody or the antigen binding portion thereof binds to the canine CD20 cyclic peptide at a higher affinity than a linear peptide having the sequence of SEQ ID NO: 21.
4 . The method of claim 1 , wherein the antibody or the antigen binding portion thereof of the CAR comprises a heavy chain variable domain (VH) comprising SEQ ID NO: 4, or is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 4.
5 . The method of claim 1 , wherein the antibody or the antigen binding portion thereof of the CAR comprises a light chain variable domain (VL) comprising SEQ ID NO: 6, or is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 6.
6 . The method of claim 1 , wherein the antibody or the antigen binding portion thereof of the CAR is a Fab fragment, a F(ab′)2 fragment, or a single chain variable fragment (scFv).
7 . The method of claim 6 , wherein the antibody or the antigen binding portion thereof of the CAR is a scFv.
8 . The method of claim 1 , wherein the antibody or the antigen binding portion thereof of the CAR comprises SEQ ID NO: 8, or is at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 8.
9 . The method of claim 1 , wherein the transmembrane portion of the CAR comprises a CD28 transmembrane domain.
10 . The method of claim 1 , wherein the cytoplasmic signaling portion of the CAR comprises a CD28 costimulatory domain, a CD3ζ-chain, a 4-1BBL costimulatory domain, a PD1-DNR domain, a CD34 domain, or any combination thereof.
11 . The method of claim 1 , wherein the CAR comprises a CD8 leader sequence.
12 . The method of claim 1 , wherein the subject suffers from B cell lymphoma or an autoimmune disease.
13 . The method of claim 12 , wherein the autoimmune disease is selected from the group consisting of rheumatoid arthritis, systemic lupus erythematosus (SLE), Sjogren's syndrome, vasculitis, multiple sclerosis, Graves' disease, idiopathic thrombocytopenia, dermatomyositis, immune mediated thrombocytopenia, polymyocytosis, pemphigus, immune mediated hemolytic anemia and bullous pemphigoid.
14 . A chimeric antigen receptor comprising
(i) an antibody or an antigen binding portion thereof that specifically binds to a canine CD20 cyclic peptide having the sequence of SEQ ID NO: 20, wherein the cysteine at position 7 of SEQ ID NO: 20 forms a disulfide bond with the cysteine at position 23 of SEQ ID NO: 20, wherein the antibody or the antigen binding portion comprises a heavy chain variable domain (V H ) complementarity determining region (CDR) 1 of SEQ ID NO: 40, a V H CDR2 of SEQ ID NO: 42, a V H CDR3 of SEQ ID NO: 44, and a light chain variable domain (V L ) CDR1 of SEQ ID NO: 46, a V L CDR2 of LVS, and a V L CDR3 of SEQ ID NO: 50;
(ii) a transmembrane portion; and
(iii) a cytoplasmic signaling portion.
15 . The chimeric antigen receptor of claim 14 , wherein the chimeric antigen receptor comprises a CD8 leader sequence, a CD28 costimulatory domain, a CD3-chain, a 4-1BBL costimulatory domain, a PD1-DNR domain, a CD34 domain, or any combination thereof.
16 . A cell expressing a chimeric antigen receptor of claim 15 , wherein the cell is a T cell or natural killer (NK) cell.
17 . A method of treating a condition mediated by B-cells in a subject in need thereof comprising administering to the subject an effective amount of the cell of claim 16 wherein the condition mediated by B-cells is a B cell lymphoma or an immune mediated disease.
18 . The method of claim 17 , wherein the immune mediated disease is an autoimmune disease.
19 . The autoimmune disease of claim 18 , wherein the autoimmune disease is selected from the group consisting of rheumatoid arthritis, systemic lupus erythematosus (SLE), Sjogren's syndrome, vasculitis, multiple sclerosis, Graves' disease, idiopathic thrombocytopenia, dermatomyositis, immune mediated thrombocytopenia, polymyocytosis, pemphigus, immune mediated hemolytic anemia and bullous pemphigoid.