Humanized antibody or fragment thereof specific for CD45R0
The present invention provides a humanized antibody or fragment thereof specific for the antigen CD45R0, wherein said antibody or fragment thereof comprises a humanized heavy chain variable domain comprising a CDR1 region of SEQ ID NO:1, a CDR2 region of SEQ ID NO:2, and a CDR3 region of SEQ ID NO:3, and a humanized light chain variable domain comprising a CDR1 region of SEQ ID NO:4, a CDR2 region of SEQ ID NO:5, and a CDR3 region of SEQ ID NO:6. It is also provided the use of the present antibody or fragment thereof for enrichment or depletion of CD45R0-expressing cells from a sample comprising CD45R0-expressing cells.
1. A humanized antibody or fragment thereof specific for the antigen CD45R0, wherein said antibody or fragment thereof comprises a humanized heavy chain variable domain comprising a CDR1 region of SEQ ID NO:1, a CDR2 region of SEQ ID NO:2, and a CDR3 region of SEQ ID NO:3, and a humanized light chain variable domain comprising a CDR1 region of SEQ ID NO:4, a CDR2 region of SEQ ID NO:5, and a CDR3 region of SEQ ID NO:6;
wherein said humanized heavy chain variable domain has artificial framework regions FR1, FR2, FR3, and FR4, and wherein said humanized light chain variable domain has artificial framework regions FR1, FR2, FR3, and FR4;
wherein said artificial framework regions FR1, FR2, FR3, and FR4 of said humanized heavy chain variable domain are amino acid sequences having an identity of at least 70% to the artificial heavy chain variable sequence framework regions FR1, FR2, FR3, and FR4 of SEQ ID NO:19, and wherein said artificial framework regions FR1, FR2, FR3, and FR4 of said humanized light chain variable domain are amino acid sequences having an identity of at least 70% to the light chain variable sequence framework regions FR1, FR2, FR3, and FR4 of SEQ ID NO:20;
and wherein said artificial framework regions FR1, FR2, FR3, and FR4 of said humanized heavy and light chain variable domains are amino acid sequences comprising within the heavy chain variable domain amino acid F at position 25, V at position 39, G at position 40, I at position 42, L at position 53, T at position 54, H at position 55, Y at position 66, K at position 80, S at position 82, and V at position 87; and within the light chain variable domain amino acid S at position 25, L at position 39, H at position 40, N at position 66, R at position 67, and F at position 68 according to the IMGT® nomenclature;
and wherein said antibodies or fragments thereof bind specifically to CD45R0 with a relative affinity of at least 20% for the CD45R0 antigen when compared to the murine CD45R0 antibody comprising the heavy chain variable domain set forth SEQ ID NO:13 and the light chain variable domain set forth in SEQ ID NO:14.
2. The antibody or fragment thereof according to claim 1 , wherein said antibody or fragment thereof comprises within the heavy chain variable domain amino acid Q at position 1, I at position 2, F at position 25, V at position 39, G at position 40, I at position 42, L at position 53, T at position 54, H at position 55, Y at position 66, R at position 72, K at position 80, S at position 82, and V at position 87, and within the light chain variable domain amino acid V at position 2, V at position 3, T at position 7, S at position 22, S at position 25, L at position 39, H at position 40, N at position 66, R at position 67, F at position 68, D at position 74, and G at position 80 according to the IMGT® nomenclature.
3. A humanized antibody or fragment thereof specific for the antigen CD45R0, wherein said antibody or fragment thereof comprises a humanized heavy chain variable domain comprising a CDR1 region of SEQ ID NO:1, a CDR2 region of SEQ ID NO:2, and a CDR3 region of SEQ ID NO:3, and a humanized light chain variable domain comprising a CDR1 region of SEQ ID NO:4, a CDR2 region of SEQ ID NO:5, and a CDR3 region of SEQ ID NO:6;
wherein said humanized heavy chain variable domain has natural framework regions FR1, FR2, and FR3, and wherein said humanized light chain variable domain has natural framework regions FR1, FR2, FR3;
wherein said natural framework regions FR1, FR2, FR3, and FR4 of said humanized heavy chain variable domain are amino acid sequences having an identity of at least 70% to the natural heavy chain variable sequence framework regions FR1, FR2, and FR3 encoded by one allele of the IGHV2-5 gene, and wherein said natural framework regions FR1, FR2, and FR3 of said humanized light chain variable domain are amino acid sequences having an identity of at least 70% to the light chain variable sequence framework regions FR1, FR2, and FR3 encoded by one allele of the IGKV2-30 gene; and
wherein said natural framework regions FR1, FR2, FR3, and FR4 of said humanized light chain variable domain are amino acid sequences comprising within the heavy chain variable domain amino acid T at position 54, amino acid H at position 55, Y at position 66, and S at position 82, and within the light chain variable domain amino acid T at position 7, H at position 40, Y at position 42, L at position 52, and F at position 68 according to the IMGT® nomenclature,
and wherein said antibodies or fragments thereof bind specifically to CD45R0 with a relative affinity of at least 20% for the CD45R0 antigen when compared to the murine CD45R0 antibody comprising the heavy chain variable domain set forth SEQ ID NO:13 and the light chain variable domain set forth in SEQ ID NO:14.
4. The antibody or fragment thereof according to claim 3 , wherein said IGHV2-5 allele is IGHV2-5*01, and wherein said IGKV2-30 allele is IGKV2-30*01.