JPWO2021113776A5 - - Google Patents

Description

In some of any such embodiments, the target antibody or antigen-binding fragment thereof is an scFv and the VH and VL regions are joined by a flexible linker. In some of any such embodiments, the flexible linker comprises the amino acid sequence set forth in SEQ ID NO:14 and the scFv comprises the amino acid sequence set forth in SEQ ID NO:27.
[Invention 1001]
An anti-idiotypic antibody or antigen-binding fragment thereof that binds to an anti-BCMA target antibody or antigen-binding fragment thereof,
CDR-H1 containing the amino acid sequence shown in SEQ ID NO:74 or 104, CDR-H2 containing the amino acid sequence shown in SEQ ID NO:75 or 105, and amino acid sequence shown in SEQ ID NO:76 or 106. a VH region, including CDR-H3,
CDR-L1 containing the amino acid sequence shown in SEQ ID NO:81 or 111, CDR-L2 containing the amino acid sequence shown in SEQ ID NO:82 or 112, and amino acid sequence shown in SEQ ID NO:83 or 113. The VL region, including CDR-L3, and
anti-idiotypic antibodies or antigen-binding fragments thereof, including:
[Present invention 1002]
An anti-idiotypic antibody or antigen-binding fragment thereof that binds to an anti-BCMA target antibody or antigen-binding fragment thereof,
A VH region containing CDR-H1, CDR-H2, and CDR-H3 containing the amino acid sequences of CDR-H1, CDR-H2, and CDR-H3 contained within the amino acid sequence of SEQ ID NO: 84 or 114, respectively; ,
A VL region containing CDR-L1, CDR-L2, and CDR-L3 containing the amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 contained within the amino acid sequence of SEQ ID NO:85 or 115, respectively.
anti-idiotypic antibodies or antigen-binding fragments thereof, including:
[Present invention 1003]
An anti-idiotypic antibody or antigen-binding fragment thereof that binds to an anti-BCMA target antibody or antigen-binding fragment thereof,
a VH region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:84 or 114;
a VL region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:85 or 115;
anti-idiotypic antibodies or antigen-binding fragments thereof, including:
[Present invention 1004]
An anti-idiotypic antibody or antigen-binding fragment thereof that binds to an anti-BCMA target antibody or antigen-binding fragment thereof,
A VH region comprising the amino acid sequence shown in SEQ ID NO:84 or 114,
A VL region containing the amino acid sequence shown in SEQ ID NO:85 or 115 and
anti-idiotypic antibodies or antigen-binding fragments thereof, including:
[Present invention 1005]
The VH region is CDR-H1 containing the amino acid sequence shown in SEQ ID NO:74 or 104, CDR-H2 containing the amino acid sequence shown in SEQ ID NO:75 or 105, and CDR-H2 containing the amino acid sequence shown in SEQ ID NO:76 or 106. contains CDR-H3 containing the amino acid sequence
The VL region is CDR-L1 containing the amino acid sequence shown in SEQ ID NO: 81 or 111, CDR-L2 containing the amino acid sequence shown in SEQ ID NO: 82 or 112, and CDR-L2 containing the amino acid sequence shown in SEQ ID NO: 83 or 113. Contains CDR-L3 containing the amino acid sequence
The anti-idiotypic antibody or antigen-binding fragment thereof according to any of the inventions 1002 to 1004.
[Present invention 1006]
The VH region comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:84, and the VL region comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:85. contains an amino acid sequence with 90% sequence identity, or
The VH region comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:114, and the VL region comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:115. comprising an amino acid sequence with 90% sequence identity,
The anti-idiotypic antibody or antigen-binding fragment thereof according to any of the inventions 1001 to 1003 and 1005.
[Present invention 1007]
The VH region contains the amino acid sequence shown in SEQ ID NO:84, and the VL region contains the amino acid sequence shown in SEQ ID NO:85, or
The VH region contains the amino acid sequence shown in SEQ ID NO:114, and the VL region contains the amino acid sequence shown in SEQ ID NO:115,
The anti-idiotype antibody or antigen-binding fragment thereof according to any one of the present invention 1001 to 1006.
[Present invention 1008]
CDR-H1 whose VH region contains the amino acid sequence shown in SEQ ID NO:74, CDR-H2 which contains the amino acid sequence shown in SEQ ID NO:75, and CDR whose VH region contains the amino acid sequence shown in SEQ ID NO:76. - Contains H3, and
CDR-L1 whose VL region contains the amino acid sequence shown in SEQ ID NO:81, CDR-L2 which contains the amino acid sequence shown in SEQ ID NO:82, and CDR whose VL region contains the amino acid sequence shown in SEQ ID NO:83. -Including L3,
The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of the present inventions 1001 to 1007.
[Present invention 1009]
The anti-idiotypic antibody of any one of the present invention 1001 to 1008 or its Antigen-binding fragment.
[Present invention 1010]
CDR-H1 whose VH region contains the amino acid sequence shown in SEQ ID NO:104, CDR-H2 which contains the amino acid sequence shown in SEQ ID NO:105, and CDR whose VH region contains the amino acid sequence shown in SEQ ID NO:106. - Contains H3, and
CDR-L1 whose VL region contains the amino acid sequence shown in SEQ ID NO:111, CDR-L2 which contains the amino acid sequence shown in SEQ ID NO:112, and CDR whose VL region contains the amino acid sequence shown in SEQ ID NO:113. -Including L3,
The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of the present inventions 1001 to 1007.
[Present invention 1011]
The anti-idiotype antibody according to any one of the present invention 1001 to 1007 and 1010, wherein the VH region contains the amino acid sequence shown in SEQ ID NO: 114, and the VL region contains the amino acid sequence shown in SEQ ID NO: 115. or antigen-binding fragment thereof.
[Invention 1012]
The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of the present invention 1001 to 1011, wherein the anti-BCMA target antibody or antigen-binding fragment thereof binds to human BCMA.
[Present invention 1013]
The present invention 1001 to 1012, wherein the anti-BCMA target antibody or antigen-binding fragment thereof comprises a VH region comprising the amino acid sequence shown in SEQ ID NO:25 and a VL region comprising the amino acid sequence shown in SEQ ID NO:26. any anti-idiotypic antibody or antigen-binding fragment thereof.
[Present invention 1014]
The anti-idiotypic antibody or antigen-binding fragment thereof of the present invention 1013, wherein the anti-BCMA target antibody or antigen-binding fragment thereof is a single chain fragment.
[Present invention 1015]
1014. The anti-idiotypic antibody or antigen-binding fragment thereof of the invention 1014, wherein the single chain fragment comprises a flexible linker disposed between the VH and VL regions.
[Invention 1016]
The anti-idiotypic antibody or antigen-binding fragment thereof of the present invention 1015, wherein the flexible linker comprises the amino acid sequence shown in SEQ ID NO:14.
[Invention 1017]
Inventions 1014-1016, wherein the single chain fragment of the anti-BCMA target antibody or antigen binding fragment thereof is a single chain variable region fragment (scFv), optionally said scFv comprising the amino acid sequence set forth in SEQ ID NO:27. any anti-idiotypic antibody or antigen-binding fragment thereof.
[Invention 1018]
The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of the present inventions 1001 to 1017, which binds to an epitope within a CDR of the target antibody or antigen-binding fragment or an epitope comprising all or a portion of the CDR.
[Invention 1019]
the anti-BCMA targeting antibody or antigen-binding fragment is present within or included in the antigen-binding domain of the extracellular portion of the CAR; and/or
the anti-idiotypic antibody or antigen-binding fragment binds to an anti-BCMA target antibody or antigen-binding fragment contained within or contained in the antigen-binding domain of the extracellular portion of the CAR;
The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of the present invention 1001 to 1018.
[Invention 1020]
the scFv is present within or included in the extracellular portion of the CAR, and/or
the anti-idiotypic antibody or antigen-binding fragment binds to an scFv contained within or contained in the extracellular portion of the CAR;
The anti-idiotype antibody or antigen-binding fragment thereof of the present invention 1017 or the present invention 1018.
[Invention 1021]
The anti-idiotypic antibody or antigen-binding fragment thereof of the invention 1019 or the invention 1020, wherein the CAR further comprises a transmembrane domain linked to the antigen-binding domain via a spacer.
[Invention 1022]
The anti-idiotypic antibody or antigen-binding fragment thereof of the present invention 1021, wherein the spacer is an immunoglobulin spacer, and optionally the spacer comprises the amino acid sequence shown in SEQ ID NO:50.
[Invention 1023]
The anti-idiotypic antibody or antigen-binding fragment thereof of the invention 1021 or the invention 1022, wherein the transmembrane domain comprises a transmembrane portion of CD28, optionally human CD28.
[Invention 1024]
The anti-idiotype antibody or antigen-binding fragment thereof according to any of the invention 1021-1023, which does not bind to an epitope in the spacer domain of CAR.
[Invention 1025]
The anti-idiotypic antibody or antigen-binding fragment thereof of any of the invention 1001-1024, which does not bind or specifically bind to CD28, optionally human CD28, or a portion thereof.
[Invention 1026]
The anti-idiotypic antibody or antigen-binding fragment thereof of any of the invention 1001-1025, which does not bind to an epitope in the Fc domain, optionally in the human IgG1 Fc domain.
[Invention 1027]
The anti-idiotypic antibody or antigen-binding fragment thereof according to any of the inventions 1001 to 1026, which specifically binds to an anti-BCMA target antibody or antigen-binding fragment thereof.
[Invention 1028]
Anti-idiotype antibodies are 100nM, 50nM, 40nM, 30nM, 25nM, 20nM, 19nM, 18nM, 17nM, 16nM, 15nM, 14nM, 13nM, 12nM, 11nM, 10nM, 9nM, 8nM, 7nM, 6nM, 5nM, 4nM, 3nM, 2nM or 1nM, or about 100nM, 50nM, 40nM, 30nM, 25nM, 20nM, 19nM, 18nM, 17nM, 16nM, 15nM, 14nM, 13nM, 12nM, 11nM, 10nM, 9nM, 8nM, 7nM, 6nM, 5nM, 4nM, 3nM, 2nM or 1nM, or 100nM, 50nM, 40nM, 30nM, 25nM, 20nM, 19nM, 18nM, 17nM, 16nM, 15nM, 14nM, 13nM, 12nM, 11nM, 10nM, 9nM, 8nM, 7nM, 6nM, 5nM , 4nM, 3nM, 2nM or less than 1nM, or about 100nM, 50nM, 40nM, 30nM, 25nM, 20nM, 19nM, 18nM, 17nM, 16nM, 15nM, 14nM, 13nM, 12nM, 11nM, 10nM, 9nM, 8nM, 7nM, The anti-idiotypic antibody or antigen-binding fragment thereof of any of the inventions 1001 to 1027, which has a dissociation constant for the anti-BCMA target antibody or antigen-binding fragment thereof that is less than 6nM, 5nM, 4nM, 3nM, 2nM or 1nM.
[Invention 1029]
Any anti-idiotypic antibody of the present invention 1001 to 1028 or its antigen that does not cross-react with another anti-BCMA antibody and, optionally, with another anti-BCMA antibody contained in the extracellular antigen-binding domain of another CAR. Combined fragment.
[Invention 1030]
Said another anti-BCMA antibody,
(i) a VH region comprising an amino acid sequence having less than 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:25 and/or 90% to the amino acid sequence set forth in SEQ ID NO:26; comprises a VL region comprising an amino acid sequence with sequence identity of less than or equal to
(ii) comprises an scFv with less than 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:13; and/or
(iii) CDR-H1 having 1, 2, 3 or 4 or more amino acid residues that differ from the amino acid sequence of CDR-H1 shown in SEQ ID NO:15 and/or shown in SEQ ID NO:16; CDR-H2 having 1, 2, 3 or 4 or more amino acid residues different from the amino acid sequence of CDR-H2 shown in SEQ ID NO:17, and/or 1 different from the amino acid sequence of CDR-H3 shown in SEQ ID NO:17. , CDR-H3 having 2, 3 or 4 or more amino acid residues, and/or 1, 2, 3 or 4 or more amino acid residues different from the amino acid sequence of CDR-L1 as shown in SEQ ID NO:22. and/or CDR-L2 having 1, 2, 3 or 4 or more amino acid residues different from the amino acid sequence of CDR-L2 shown in SEQ ID NO:23, and/or comprises a CDR-L3 having 1, 2, 3 or 4 or more amino acid residues different from the amino acid sequence of CDR-L3 shown in SEQ ID NO:24, and/or
(iv) binds to an epitope of human BCMA that is not the same epitope of human BCMA to which the anti-BCMA targeting antibody or antigen-binding fragment thereof binds;
The anti-idiotypic antibody or antigen-binding fragment thereof of the present invention 1029.
[Present invention 1031]
The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of the present invention 1001 to 1030, which is an agonist of CAR, including an anti-BCMA target antibody or antigen-binding fragment thereof.
[Invention 1032]
When in soluble form, the anti-idiotypic antibody or antigen-binding fragment thereof of the invention 1031 is an agonist of said CAR.
[Present invention 1033]
The anti-idiotypic antibody or antigen-binding fragment thereof of the invention 1031 or the invention 1032, which is an agonist of said CAR when immobilized on a support or stationary phase.
[Present invention 1034]
The anti-idiotypic antibody or antigen-binding fragment thereof of the present invention 1033, wherein the support is a plate.
[Invention 1035]
The anti-idiotypic antibody or antigen-binding fragment thereof of the present invention 1033, wherein the support is a bead.
[Invention 1036]
The anti-idiotype antibody or anti-idiotype antibody of any of the present invention 1001 to 1035, wherein the binding of the anti-idiotype antibody or antigen-binding fragment thereof to the anti-BCMA target antibody or antigen-binding fragment thereof is not blocked by human BCMA or human BCMA-Fc. its antigen-binding fragment.
[Present invention 1037]
An anti-idiotype antibody or antigen-binding fragment thereof that binds to a first anti-BCMA target antibody or antigen-binding fragment thereof and a second anti-BCMA target antibody or antigen-binding fragment thereof,
CDR-H1 containing the amino acid sequence shown in SEQ ID NO:28 or 52, CDR-H2 containing the amino acid sequence shown in SEQ ID NO:29 or 53, and amino acid sequence shown in SEQ ID NO:30 or 54. a VH region, including CDR-H3,
CDR-L1 containing the amino acid sequence shown in SEQ ID NO:35 or 59, CDR-L2 containing the amino acid sequence shown in SEQ ID NO:36 or 60, and the amino acid sequence shown in SEQ ID NO:37 or 61. The VL region, including CDR-L3, and
anti-idiotypic antibodies or antigen-binding fragments thereof, including:
[Invention 1038]
An anti-idiotype antibody or antigen-binding fragment thereof that binds to a first anti-BCMA target antibody or antigen-binding fragment thereof and a second anti-BCMA target antibody or antigen-binding fragment thereof,
A VH region containing CDR-H1, CDR-H2, and CDR-H3 containing the amino acid sequences of CDR-H1, CDR-H2, and CDR-H3 contained within the amino acid sequence of SEQ ID NO: 38 or 62, respectively; ,
A VL region containing CDR-L1, CDR-L2, and CDR-L3 containing the amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 contained within the amino acid sequence of SEQ ID NO: 39 or 63, respectively.
anti-idiotypic antibodies or antigen-binding fragments thereof, including:
[Invention 1039]
An anti-idiotype antibody or antigen-binding fragment thereof that binds to a first anti-BCMA target antibody or antigen-binding fragment thereof and a second anti-BCMA target antibody or antigen-binding fragment thereof,
a VH region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:38 or 62;
a VL region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:39 or 63;
anti-idiotypic antibodies or antigen-binding fragments thereof, including:
[Invention 1040]
An anti-idiotype antibody or antigen-binding fragment thereof that binds to a first anti-BCMA target antibody or antigen-binding fragment thereof and a second anti-BCMA target antibody or antigen-binding fragment thereof,
A VH region comprising the amino acid sequence shown in SEQ ID NO:38 or 62,
A VL region containing the amino acid sequence shown in SEQ ID NO:39 or 63 and
anti-idiotypic antibodies or antigen-binding fragments thereof, including:
[Present invention 1041]
The VH region is CDR-H1 containing the amino acid sequence shown in SEQ ID NO:28 or 52, CDR-H2 containing the amino acid sequence shown in SEQ ID NO:29 or 53, and CDR-H2 containing the amino acid sequence shown in SEQ ID NO:30 or 54. contains CDR-H3 containing the amino acid sequence
The VL region is CDR-L1 containing the amino acid sequence shown in SEQ ID NO: 35 or 59, CDR-L2 containing the amino acid sequence shown in SEQ ID NO: 36 or 60, and CDR-L2 containing the amino acid sequence shown in SEQ ID NO: 37 or 61. Contains CDR-L3 containing the amino acid sequence
The anti-idiotype antibody or antigen-binding fragment thereof according to any one of the present invention 1038 to 1040.
[Present invention 1042]
the VH region comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:38 or 62, and
the VL region comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:39 or 63;
The anti-idiotypic antibody or antigen-binding fragment thereof according to any of the inventions 1037, 1038, and 1041.
[Invention 1043]
the VH region contains the amino acid sequence shown in SEQ ID NO:38 or 62, and
the VL region comprises the amino acid sequence shown in SEQ ID NO:39 or 63,
The anti-idiotypic antibody or antigen-binding fragment thereof according to any of the inventions 1037 to 1039, 1041, and 1042.
[Present invention 1044]
The VH region comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:38, and the VL region comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:39. contains an amino acid sequence with 90% sequence identity, or
The VH region comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:62, and the VL region comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:63. comprising an amino acid sequence with 90% sequence identity,
The anti-idiotypic antibody or antigen-binding fragment thereof according to any of the inventions 1037 to 1039, 1041, and 1042.
[Invention 1045]
The VH region contains the amino acid sequence shown in SEQ ID NO:38, and the VL region contains the amino acid sequence shown in SEQ ID NO:39, or
The VH region contains the amino acid sequence shown in SEQ ID NO:62, and the VL region contains the amino acid sequence shown in SEQ ID NO:63,
The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of the present invention 1037 to 1044.
[Invention 1046]
CDR-H1 whose VH region contains the amino acid sequence shown in SEQ ID NO:28, CDR-H2 which contains the amino acid sequence shown in SEQ ID NO:29, and CDR whose VH region contains the amino acid sequence shown in SEQ ID NO:30. - Contains H3, and
CDR-L1 whose VL region contains the amino acid sequence shown in SEQ ID NO:35, CDR-L2 which contains the amino acid sequence shown in SEQ ID NO:36, and CDR whose VL region contains the amino acid sequence shown in SEQ ID NO:37. -Including L3,
The anti-idiotype antibody or antigen-binding fragment thereof according to any one of the present invention 1037 to 1045.
[Invention 1047]
The anti-idiotypic antibody of any one of the present invention 1037 to 1046 or its Antigen-binding fragment.
[Invention 1048]
CDR-H1 whose VH region contains the amino acid sequence shown in SEQ ID NO:52, CDR-H2 which contains the amino acid sequence shown in SEQ ID NO:53, and CDR whose VH region contains the amino acid sequence shown in SEQ ID NO:54. - Contains H3, and
CDR-L1 whose VL region contains the amino acid sequence shown in SEQ ID NO:59, CDR-L2 which contains the amino acid sequence shown in SEQ ID NO:60, and CDR whose VL region contains the amino acid sequence shown in SEQ ID NO:61. -Including L3,
The anti-idiotype antibody or antigen-binding fragment thereof according to any one of the present invention 1037 to 1045.
[Invention 1049]
The anti-idiotype antibody according to any one of the present invention 1037 to 1045 and 1048, wherein the VH region comprises the amino acid sequence shown in SEQ ID NO:62, and the VL region comprises the amino acid sequence shown in SEQ ID NO:63. or antigen-binding fragment thereof.
[Invention 1050]
The first anti-BCMA target antibody or antigen-binding fragment thereof binds to human BCMA, and the second anti-BCMA target antibody or antigen-binding fragment thereof binds to human BCMA. Anti-idiotype antibodies or antigen-binding fragments thereof.
[Present invention 1051]
The first anti-BCMA target antibody or antigen-binding fragment thereof comprises a VH region comprising the amino acid sequence shown in SEQ ID NO:11 and a VL region comprising the amino acid sequence shown in SEQ ID NO:12, and
The second anti-BCMA target antibody or antigen-binding fragment thereof comprises a VH region comprising the amino acid sequence shown in SEQ ID NO:25 and a VL region comprising the amino acid sequence shown in SEQ ID NO:26,
The anti-idiotype antibody or antigen-binding fragment thereof according to any one of the present invention 1037 to 1050.
[Invention 1052]
The present invention 1051, wherein the first anti-BCMA target antibody or antigen-binding fragment thereof is a first single-chain fragment, and the second anti-BCMA target antibody or antigen-binding fragment thereof is a second single-chain fragment. anti-idiotypic antibodies or antigen-binding fragments thereof.
[Present invention 1053]
the first single chain fragment comprises a flexible linker disposed between the VH region and the VL region, and the second single chain fragment comprises a flexible linker disposed between the VH region and the VL region , the anti-idiotypic antibody or antigen-binding fragment thereof of the present invention 1052.
[Invention 1054]
1053. The anti-idiotypic antibody or antigen-binding fragment thereof of the invention 1053, wherein the flexible linker of the first single-chain fragment and/or the second single-chain fragment comprises the amino acid sequence shown in SEQ ID NO:14.
[Present invention 1055]
The first single chain fragment of the first anti-BCMA target antibody or antigen-binding fragment thereof is a first scFv, and the second single chain fragment of the second anti-BCMA target antibody or antigen-binding fragment thereof is The anti-idiotypic antibody or antigen-binding fragment thereof of any of the invention 1052 to 1054, which is a second scFv.
[Invention 1056]
The anti-idiotypic antibody of the present invention 1055 or its antigen, wherein the first scFv comprises the amino acid sequence shown in SEQ ID NO:13, and the second scFv comprises the amino acid sequence shown in SEQ ID NO:27. Combined fragment.
[Present invention 1057]
an epitope within or comprising all or a portion of a CDR of a first anti-BCMA target antibody or antigen-binding fragment; and
an epitope within or comprising all or a portion of a CDR of a second anti-BCMA target antibody or antigen-binding fragment;
The anti-idiotypic antibody or antigen-binding fragment thereof of any of the invention 1037-1056, which binds to.
[Invention 1058]
the first anti-BCMA targeting antibody or antigen-binding fragment is within or included in the antigen-binding domain of the extracellular portion of the first CAR; and/or
the anti-idiotypic antibody or antigen-binding fragment is contained within or contained in the antigen-binding domain of the extracellular portion of the first CAR; Join,
The anti-idiotype antibody or antigen-binding fragment thereof according to any one of the present invention 1037 to 1057.
[Invention 1059]
a second anti-BCMA targeting antibody or antigen-binding fragment is present within or included in the antigen-binding domain of the extracellular portion of the second CAR; and/or
the anti-idiotypic antibody or antigen-binding fragment is contained within or contained in the antigen-binding domain of the extracellular portion of the second CAR; Join,
The anti-idiotype antibody or antigen-binding fragment thereof according to any one of the present invention 1037 to 1058.
[Invention 1060]
the first scFv is within or included in the extracellular portion of the first CAR; and/or
the anti-idiotypic antibody or antigen-binding fragment binds to a first scFv contained within or contained in the extracellular portion of the first CAR;
The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of the present invention 1037 to 1059.
[Present invention 1061]
the second scFv is present within or included in the extracellular portion of the second CAR; and/or
the anti-idiotypic antibody or antigen-binding fragment binds to a second scFv contained within or contained in the extracellular portion of the second CAR;
The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of the present invention 1037 to 1060.
[Invention 1062]
The first CAR further comprises a transmembrane domain linked to the antigen binding domain via a spacer, and/or the second CAR further comprises a transmembrane domain linked to the antigen binding domain via a spacer. , the anti-idiotypic antibody or antigen-binding fragment thereof according to any of the inventions 1058 to 1061.
[Invention 1063]
The spacer of the first CAR and/or the second CAR is an immunoglobulin spacer, and optionally the spacer comprises the amino acid sequence shown in SEQ ID NO:50, or the anti-idiotypic antibody of the invention 1062. Antigen-binding fragment.
[Present invention 1064]
The anti-idiotypic antibody or antigen-binding fragment thereof of the invention 1062 or the invention 1063, wherein the transmembrane domain of the first CAR and/or the second CAR comprises a transmembrane portion of CD28, optionally human CD28.
[Invention 1065]
An anti-idiotypic antibody or antigen-binding fragment thereof according to any of the invention 1062-1064, which does not bind to an epitope in the spacer domain of the first CAR and/or the second CAR.
[Invention 1066]
An anti-idiotypic antibody or antigen-binding fragment thereof of any of the invention 1037-1065 that does not bind or specifically bind to CD28, optionally human CD28, or a portion thereof.
[Invention 1067]
The anti-idiotypic antibody or antigen-binding fragment thereof of any of the invention 1037-1066, which does not bind to an epitope in the Fc domain, optionally in the human IgG1 Fc domain.
[Invention 1068]
The anti-idiotypic antibody or antigen-binding fragment thereof of any of the present invention 1037 to 1067, which specifically binds to the first anti-BCMA target antibody or antigen-binding fragment thereof and the second anti-BCMA target antibody or antigen-binding fragment thereof .
[Invention 1069]
Anti-idiotype antibodies are 100nM, 50nM, 40nM, 30nM, 25nM, 20nM, 19nM, 18nM, 17nM, 16nM, 15nM, 14nM, 13nM, 12nM, 11nM, 10nM, 9nM, 8nM, 7nM, 6nM, 5nM, 4nM, 3nM, 2nM or 1nM, or about 100nM, 50nM, 40nM, 30nM, 25nM, 20nM, 19nM, 18nM, 17nM, 16nM, 15nM, 14nM, 13nM, 12nM, 11nM, 10nM, 9nM, 8nM, 7nM, 6nM, 5nM, 4nM, 3nM, 2nM or 1nM, or 100nM, 50nM, 40nM, 30nM, 25nM, 20nM, 19nM, 18nM, 17nM, 16nM, 15nM, 14nM, 13nM, 12nM, 11nM, 10nM, 9nM, 8nM, 7nM, 6nM, 5nM , 4nM, 3nM, 2nM or less than 1nM, or about 100nM, 50nM, 40nM, 30nM, 25nM, 20nM, 19nM, 18nM, 17nM, 16nM, 15nM, 14nM, 13nM, 12nM, 11nM, 10nM, 9nM, 8nM, 7nM, The present invention has a dissociation constant for the first anti-BCMA target antibody or antigen-binding fragment thereof and/or the second anti-BCMA target antibody or antigen-binding fragment thereof that is less than 6 nM, 5 nM, 4 nM, 3 nM, 2 nM or 1 nM. Any anti-idiotypic antibody of 1037 to 1068 or an antigen-binding fragment thereof.
[Invention 1070]
an agonist of a first CAR comprising a first anti-BCMA targeting antibody or antigen-binding fragment thereof and/or an agonist of a second CAR comprising a second anti-BCMA targeting antibody or antigen-binding fragment thereof; The anti-idiotypic antibody or antigen-binding fragment thereof according to any of Inventions 1037 to 1069.
[Present invention 1071]
when in soluble form is an agonist of the first CAR and/or the second CAR, and/or
when immobilized on a support or stationary phase, is an agonist of the first CAR and/or the second CAR;
Optionally the support or stationary phase is a plate or a bead.
The anti-idiotypic antibody or antigen-binding fragment thereof of the present invention 1070.
[Invention 1072]
The binding of the anti-idiotypic antibody or antigen-binding fragment thereof to the first anti-BCMA target antibody or antigen-binding fragment thereof and/or the second anti-BCMA target antibody or antigen-binding fragment thereof may be caused by binding to human BCMA or human BCMA-Fc The anti-idiotype antibody or antigen-binding fragment thereof of any of the invention 1037-1071, which is not blocked by.
[Invention 1073]
The anti-idiotypic antibody or antigen-binding fragment thereof of any of the invention 1001-1072, comprising at least a portion of a heavy chain and/or light chain immunoglobulin constant region.
[Present invention 1074]
a heavy chain constant region comprising an Fc region, or a portion of said Fc comprising CH2 and CH3 domains, and/or
Light chain constant region, including CL domain
The anti-idiotype antibody or antigen-binding fragment thereof of the present invention 1073, comprising:
[Present invention 1075]
The anti-idiotypic antibody or antigen-binding fragment thereof of the invention 1073 or the invention 1074, wherein the immunoglobulin constant region is derived from IgG, optionally IgG1.
[Invention 1076]
The anti-idiotype antibody or antigen-binding fragment thereof of the invention 1074 or the invention 1075, wherein the light chain constant region is from a kappa light chain.
[Invention 1077]
The anti-idiotypic antibody or antigen-binding fragment thereof of any of the invention 1001-1076, which is an intact antibody or a full-length antibody.
[Invention 1078]
A heavy chain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:86 and at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:87. a light chain comprising an amino acid sequence having
A heavy chain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:116 and at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:117. A light chain comprising an amino acid sequence having
The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of the present invention 1001 to 1036 and 1073 to 1077, comprising:
[Invention 1079]
a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:86, and a light chain comprising the amino acid sequence set forth in SEQ ID NO:87, or
A heavy chain comprising the amino acid sequence shown in SEQ ID NO:116 and a light chain comprising the amino acid sequence shown in SEQ ID NO:117
The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of the present invention 1001 to 1036 and 1073 to 1078, comprising:
[Invention 1080]
Any of the present invention 1001-1009, 1012-1036, and 1073-1079, comprising a heavy chain comprising the amino acid sequence shown in SEQ ID NO:86 and a light chain comprising the amino acid sequence shown in SEQ ID NO:87. anti-idiotypic antibodies or antigen-binding fragments thereof.
[Present invention 1081]
Any of the present invention 1001-1007, 1010-1036, and 1073-1079, comprising a heavy chain comprising the amino acid sequence shown in SEQ ID NO:116 and a light chain comprising the amino acid sequence shown in SEQ ID NO:117. anti-idiotypic antibodies or antigen-binding fragments thereof.
[Present invention 1082]
a heavy chain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 40 or 64;
a light chain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:41 or 65;
The anti-idiotypic antibody or antigen-binding fragment thereof of any of the invention 1037-1077, comprising:
[Present invention 1083]
A heavy chain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:40 and at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:41. a light chain comprising an amino acid sequence having
A heavy chain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:64 and at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:65. A light chain comprising an amino acid sequence having
The anti-idiotypic antibody or antigen-binding fragment thereof of any of the invention 1037-1077 and 1082, comprising:
[Present invention 1084]
a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 40 or 64;
A light chain containing the amino acid sequence shown in SEQ ID NO:41 or 65
The anti-idiotypic antibody or antigen-binding fragment thereof of any of the invention 1037-1077, 1082, and 1083, comprising:
[Invention 1085]
a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:40, and a light chain comprising the amino acid sequence set forth in SEQ ID NO:41, or
A heavy chain comprising the amino acid sequence shown in SEQ ID NO:64 and a light chain comprising the amino acid sequence shown in SEQ ID NO:65
The anti-idiotypic antibody or antigen-binding fragment thereof of any of the present invention 1037-1077 and 1082-1084, comprising:
[Invention 1086]
Any of the present invention 1037-1047, 1050-1077, and 1082-1085, comprising a heavy chain comprising the amino acid sequence shown in SEQ ID NO:40 and a light chain comprising the amino acid sequence shown in SEQ ID NO:41. anti-idiotypic antibodies or antigen-binding fragments thereof.
[Present invention 1087]
Any of the present invention 1037-1045, 1048-1077, and 1082-1085, comprising a heavy chain comprising the amino acid sequence shown in SEQ ID NO:64 and a light chain comprising the amino acid sequence shown in SEQ ID NO:65. anti-idiotypic antibodies or antigen-binding fragments thereof.
[Present invention 1088]
The anti-idiotypic antibody or antigen-binding fragment thereof according to any of the inventions 1001 to 1076, which is an antigen-binding fragment.
[Invention 1089]
The anti-idiotypic antibody or antigen-binding fragment thereof of the invention 1088, wherein the antigen-binding fragment is selected from Fab fragments, F(ab') ₂ fragments, Fab' fragments, Fv fragments, scFv, and single domain antibodies.
[Invention 1090]
The anti-idiotypic antibody or antigen-binding fragment thereof according to any of the inventions 1001 to 1089, which is a humanized version.
[Present invention 1091]
The anti-idiotypic antibody or antigen-binding fragment thereof of any of the present inventions 1001 to 1090, which is a recombinant type.
[Invention 1092]
The anti-idiotypic antibody or antigen-binding fragment thereof according to any of the inventions 1001 to 1091, which is monoclonal.
[Present invention 1093]
A conjugate comprising an anti-idiotypic antibody or antigen-binding fragment thereof according to any of the inventions 1001 to 1092 and a heterologous molecule or a heterologous moiety.
[Present invention 1094]
A conjugate of the invention 1093, wherein the heterologous molecule or moiety is a label.
[Present invention 1095]
the label is selected from fluorescent dyes, fluorescent proteins, radioisotopes, chromophores, metal ions, gold particles, silver particles, magnetic particles, polypeptides, enzymes, streptavidin, biotin, luminescent compounds, and oligonucleotides; Conjugate of the invention 1094.
[Invention 1096]
A conjugate of the invention 1094, wherein the heterologous molecule or moiety is a protein, peptide, nucleic acid, or small molecule, optionally being or comprising a toxin or a Strep-Tag.
[Invention 1097]
A nucleic acid molecule encoding the heavy chain and/or light chain of the anti-idiotypic antibody or antigen-binding fragment thereof according to any of the inventions 1001 to 1092.
[Present invention 1098]
(i) a heavy chain variable region as set forth in SEQ ID NO:62; (ii) a heavy chain variable region having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:62; or (iii) a sequence of nucleotides encoding the degenerate sequence of (i) or (ii);
(iv) a light chain variable region as set forth in SEQ ID NO:63; (v) a light chain variable region having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:63; or (vi) a sequence of nucleotides encoding the degenerate sequence of (iv) or (v);
A nucleic acid molecule of the invention 1097 comprising.
[Invention 1099]
(i) a heavy chain variable region as set forth in SEQ ID NO:84; (ii) a heavy chain variable region having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:84; or (iii) a sequence of nucleotides encoding the degenerate sequence of (i) or (ii);
(iv) a light chain variable region as set forth in SEQ ID NO:85; (v) a light chain variable region having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:85; or (vi) a sequence of nucleotides encoding the degenerate sequence of (iv) or (v);
A nucleic acid molecule of the invention 1097 comprising.
[Invention 1100]
(i) a heavy chain variable region as set forth in SEQ ID NO:114; (ii) a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of the nucleotides set forth in SEQ ID NO:114; or ( iii) a sequence of nucleotides encoding the degenerate sequence of (i) or (ii);
(iv) a light chain variable region as set forth in SEQ ID NO:115; (v) a light chain variable region having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:115; or (vi) a sequence of nucleotides encoding the degenerate sequence of (iv) or (v);
A nucleic acid molecule of the invention 1097 comprising.
[Present invention 1101]
(i) a heavy chain variable region as set forth in SEQ ID NO:38; (ii) a heavy chain variable region having at least 90% sequence identity to the sequence of nucleotides set forth in SEQ ID NO:38; or (iii) ) a sequence of nucleotides encoding the degenerate sequence of (i) or (ii);
(iv) a light chain variable region as set forth in SEQ ID NO:39; (v) a light chain variable region having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:39; or (vi) a sequence of nucleotides encoding the degenerate sequence of (iv) or (v);
A nucleic acid molecule of the invention 1097 comprising.
[Present invention 1102]
(i) a heavy chain as set forth in SEQ ID NO:64, (ii) a heavy chain having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:64, or (iii) (i) or (ii) a sequence of nucleotides encoding a degenerate sequence of;
(iv) a light chain as set forth in SEQ ID NO:65, (v) a light chain having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:65, or (vi) (iv) or (v) a sequence of nucleotides encoding a degenerate sequence of
A nucleic acid molecule of the invention 1097 comprising.
[Present invention 1103]
(i) a heavy chain as set forth in SEQ ID NO:86, (ii) a heavy chain having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:86, or (iii) (i) or (ii) a sequence of nucleotides encoding a degenerate sequence of;
(iv) a light chain as set forth in SEQ ID NO:87; (v) a light chain having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:87; or (vi) (iv) or (v) a sequence of nucleotides encoding a degenerate sequence of
A nucleic acid molecule of the invention 1097 comprising.
[Present invention 1104]
(i) a heavy chain as set forth in SEQ ID NO:116, (ii) a heavy chain having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:116, or (iii) (i) or (ii) a sequence of nucleotides encoding a degenerate sequence of;
(iv) a light chain as set forth in SEQ ID NO:117, (v) a light chain having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:117, or (vi) (iv) or (v) a sequence of nucleotides encoding a degenerate sequence of
A nucleic acid molecule of the invention 1097 comprising.
[Present invention 1105]
(i) a heavy chain as set forth in SEQ ID NO:40, (ii) a heavy chain having at least 90% sequence identity to the sequence of nucleotides set forth in SEQ ID NO:40, or (iii) (i) or (ii) a sequence of nucleotides encoding the degenerate sequence of
(iv) a light chain as set forth in SEQ ID NO:41; (v) a light chain having at least 90% sequence identity to the sequence of nucleotides set forth in SEQ ID NO:41; or (vi) (iv) or (v) a sequence of nucleotides encoding a degenerate sequence of
A nucleic acid molecule of the invention 1097 comprising.
[Present invention 1106]
The nucleic acid molecule according to any of the inventions 1097-1105, wherein the nucleotide sequence encoding the heavy chain and/or light chain comprises a signal sequence.
[Present invention 1107]
Any of the invention 1097-1106, comprising one or more nucleotide sequences selected from the group consisting of SEQ ID NO: 42, 43, 48, 49, 66-69, 88-91, and 118-121. Nucleic acid molecule.
[Present invention 1108]
A vector comprising any of the nucleic acid molecules of the present invention 1097-1107.
[Present invention 1109]
A cell comprising an anti-idiotypic antibody or antigen-binding fragment thereof according to any of the inventions 1001 to 1092, a nucleic acid molecule according to any of the inventions 1097 to 1107, or a vector according to the invention 1108.
[Present invention 1110]
Expressing the heavy chain and/or light chain encoded by any of the nucleic acid molecules of the present invention 1097 to 1107 or the vector of the present invention 1108 in a suitable host cell, and recovering or isolating the antibody. A method of producing an anti-idiotypic antibody or antigen-binding fragment thereof, comprising:
[Present invention 1111]
A method for producing an anti-idiotypic antibody or an antigen-binding fragment thereof, comprising the steps of culturing the cells of the present invention 1109 under conditions in which the heavy chain and/or light chain are expressed, and collecting or isolating the antibody. .
[Present invention 1112]
An anti-idiotypic antibody or antigen-binding fragment thereof produced by the method of the invention 1110 or the invention 1111.
[Present invention 1113]
A composition comprising an anti-idiotypic antibody or antigen-binding fragment thereof of any of the inventions 1001-1092, a conjugate of any of the inventions 1093-1096, or a cell of the invention 1109.
[Present invention 1114]
The composition of the invention 1113 further comprising a pharmaceutically acceptable excipient.
[Present invention 1115]
One or more of the anti-idiotypic antibody or antigen-binding fragment thereof of any of the inventions 1001-1092, the conjugate of any of the inventions 1093-1096, and the nucleic acid molecule of any of the inventions 1097-1107. and, optionally, instructions for use.
[Present invention 1116]
The kit further comprises a reagent or support for immobilizing the anti-idiotypic antibody or antigen-binding fragment or conjugate thereof,
the reagent or support is a bead, column, microwell, stick, filter, strip, or soluble oligomeric streptavidin mutein reagent;
Kit of the present invention 1115.
[Present invention 1117]
(a) A composition comprising a target antibody or an antigen-binding fragment thereof, an anti-idiotypic antibody or an antigen-binding fragment thereof according to any one of the present inventions 1001 to 1092, or an antigen-binding fragment thereof that specifically binds to the target antibody or antigen-binding fragment thereof. contacting with the conjugate of any of Inventions 1093-1096; and
(b) Detecting the anti-idiotype antibody bound to the target antibody or antigen-binding fragment thereof
A method of detecting a target antibody or antigen-binding fragment thereof, comprising:
[Present invention 1118]
The target antibody or antigen-binding fragment thereof is bound to the cell or expressed on the surface of the cell, and the detecting step of (b) is a cell in which the anti-idiotypic antibody or antigen-binding fragment thereof binds. 1117. The method of the invention 1117, comprising detecting.
[Present invention 1119]
1118. The method of the invention 1118, wherein the cell expresses on its surface a CAR comprising a targeting antibody or antigen-binding fragment thereof.
[Invention 1120]
(a) The anti-idiotypic antibody or antigen-binding thereof according to any one of the present invention 1001 to 1092, which specifically binds a cell expressing a CAR containing the target antibody or antigen-binding fragment thereof to the target antibody or antigen-binding fragment thereof. contacting the fragment or the conjugate of any of the invention 1093-1096; and
(b) Detecting cells that bind to the anti-idiotype antibody or antigen-binding fragment thereof
A method for detecting a CAR comprising a target antibody or antigen-binding fragment thereof, comprising:
[Present invention 1121]
1120. The method of the invention 1120, wherein the anti-idiotypic antibody or antigen-binding fragment thereof is directly or indirectly labeled for detection.
[Invention 1122]
(a) The present invention which specifically binds to a target antibody or an antigen-binding fragment thereof, a cell population expressing a CAR containing the target antibody or an antigen-binding fragment thereof, or a cell that binds to the target antibody or an antigen-binding fragment thereof contacting with any of the anti-idiotypic antibodies or antigen-binding fragments thereof of 1001 to 1092 or the conjugates of any of the invention 1093 to 1096; and
(b) Selecting cells that bind to the anti-idiotypic antibody or antigen-binding fragment thereof
A method of selecting cells from a cell population, including:
[Invention 1123]
1122. The method of the invention 1122, wherein cells binding the anti-idiotypic antibody or antigen-binding fragment thereof are selected by affinity-based separation.
[Present invention 1124]
1123. The method of the invention 1123, wherein the affinity-based separation is an immunoaffinity-based separation.
[Invention 1125]
The method of invention 1123 or invention 1124, wherein the affinity-based separation is by flow cytometry.
[Present invention 1126]
The method of invention 1123 or invention 1124, wherein the affinity-based separation is by magnetically activated cell sorting.
[Present invention 1127]
The method of invention 1123 or invention 1124, wherein the affinity-based separation comprises affinity chromatography.
[Present invention 1128]
The method of invention 1126 or invention 1127, wherein the anti-idiotypic antibody or antigen-binding fragment thereof is reversibly bound or immobilized to a support or stationary phase.
[Present invention 1129]
An input composition containing cells expressing a CAR containing a target antibody or an antigen-binding fragment thereof is provided with an anti-idiotypic antibody or its anti-idiotypic antibody according to any one of the present inventions 1001 to 1092 that specifically binds to the target antibody or antigen-binding fragment thereof. incubating with an antigen-binding fragment or a conjugate of any of the invention 1093-1096, thereby producing an output composition comprising stimulated cells.
Methods of stimulating cells, including:
[Present invention 1130]
(a) introducing a nucleic acid molecule encoding a CAR into a cell, thereby creating an input composition; and
(b) The input composition is an anti-idiotypic antibody or an antigen-binding fragment thereof of any of the inventions 1001 to 1092 or a conjugate of any of the inventions 1093 to 1096 that specifically binds to the antigen receptor of the CAR. incubating with a gate, thereby producing a cell composition;
A method of producing a cell composition, comprising:
[Present invention 1131]
1130. The method of the invention 1130, wherein the step of introducing (a) comprises introducing the nucleic acid molecule into the cell by viral transduction, transfer, electroporation, or chemical transfection.
[Present invention 1132]
the step of introducing (a) comprises introducing the nucleic acid molecule into the cell by transduction with a viral vector comprising the nucleic acid molecule, optionally the viral vector being a retroviral vector or a lentiviral vector; The method of the invention 1130 or the invention 1131.
[Present invention 1133]
The method of the present invention 1130 or the present invention 1131, wherein the step of introducing (a) comprises introducing the nucleic acid molecule into the cell by transposition using a transposon containing the nucleic acid molecule.
[Present invention 1134]
The method of Invention 1130 or Invention 1131, wherein the step of introducing (a) comprises introducing the nucleic acid molecule into the cell by electroporation or transfection of a vector containing the nucleic acid molecule.
[Invention 1135]
The method of any of the inventions 1130-1134, further comprising the step of activating the cells before step (a).
[Present invention 1136]
1135. The method of the invention 1135, wherein activating the cell comprises contacting the cell with an agonist of CD3 and optionally an agonist of CD28.
[Present invention 1137]
1136. The method of the invention 1136, wherein activating the cells comprises contacting the cells with a reagent comprising anti-CD3 and anti-CD28 agonist antibodies.
[Present invention 1138]
The incubation is
Conditions under which the anti-idiotypic antibody or antigen-binding fragment thereof binds to the CAR, thereby inducing or modulating a signal in one or more cells in the input composition.
The method of any of the inventions 1130-1137, carried out in.
[Present invention 1139]
The method of any of the invention 1130-1138, wherein the cell comprises a T cell.
[Invention 1140]
1139 The method of the invention, wherein the T cells include CD4+ and/or CD8+ T cells.
[Present invention 1141]
The anti-idiotypic antibody or antigen-binding fragment thereof is immobilized on a solid support, and the solid support optionally has a plurality of bonds capable of reversibly binding the anti-idiotypic antibody or antigen-binding fragment thereof. 1140. The method of any of the inventions 1130-1140, comprising or conjugated to a reagent comprising a moiety.
[Invention 1142]
The anti-idiotypic antibody or antigen-binding fragment thereof is immobilized on a soluble reagent, optionally with a plurality of binding sites capable of reversibly binding the anti-idiotypic antibody or antigen-binding fragment thereof. The method of any of the inventions 1130-1140, comprising or comprising a binding site.
[Present invention 1143]
A method of invention 1141 or invention 1142, wherein the reagent comprises a streptavidin mutein.
[Present invention 1144]
the incubation is for at least 5 minutes, 10 minutes, 30 minutes, 60 minutes, 2 hours, 6 hours, 12 hours, 24 hours, 36 hours, 48 hours, 72 hours or 96 hours, or at least about 5 minutes, 10 minutes; The method of any of the inventions 1130 to 1143, wherein the duration is 30 minutes, 60 minutes, 2 hours, 6 hours, 12 hours, 24 hours, 36 hours, 48 hours, 72 hours or 96 hours.
[Invention 1145]
The input composition is less than or about 60%, less than or about 50%, less than or about 40%, less than or about 30% of the total cells in the composition. %, less than 20% or less than about 20%, or less than 10% or less than about 10% of the CAR-expressing cells.
[Present invention 1146]
The number of CAR-expressing cells in the output composition is 1.2 times, 1.5 times, 2.0 times, 3.0 times, 4.0 times, 5.0 times, 10 times or more compared to the number of CAR expressing cells in the input composition. significantly increase and/or
the percentage of CAR expression relative to all cells in the output composition is increased by more than 10%, 20%, 40%, 50%, 60%, 70%, 80% or more;
The method according to any one of the invention 1130 to 1145.
[Present invention 1147]
The method of any of the inventions 1130-1146, wherein the cells are neither selected nor enriched for CAR-expressing cells prior to the step of introducing and/or incubating.
[Present invention 1148]
(a) A composition comprising a target antibody or an antigen-binding fragment thereof, an anti-idiotypic antibody or an antigen-binding fragment thereof according to any one of the present inventions 1001 to 1092, or an antigen-binding fragment thereof that specifically binds to the target antibody or antigen-binding fragment thereof. contacting with the conjugate of any of Inventions 1093-1096; and
(b) isolating a complex containing the anti-idiotypic antibody or antigen-binding fragment thereof;
A method of purifying an anti-idiotypic antibody or antigen-binding fragment thereof, comprising:
[Present invention 1149]
1148. The method of the invention 1148, wherein the complex comprising the anti-idiotypic antibody or antigen-binding fragment thereof is isolated by affinity-based separation.
[Invention 1150]
1149. The method of the invention 1149, wherein the affinity-based separation is an immunoaffinity-based separation.
[Present invention 1151]
The method of the invention 1150, wherein the affinity-based separation is a magnetic-based separation.
[Present invention 1152]
The method of the invention 1150, wherein the affinity-based separation comprises affinity chromatography.
[Present invention 1153]
A composition comprising an anti-idiotype antibody or an antigen-binding fragment thereof according to any one of the present inventions 1001 to 1092 or a conjugate according to any one of the present inventions 1093 to 1096 that specifically binds to a target antibody or an antigen-binding fragment thereof, A method of depleting cells comprising administering to a subject a cell expressing a CAR comprising the target antibody or antigen-binding fragment thereof.
[Present invention 1154]
1153. The method of the invention 1153, wherein said depletion occurs by antibody-dependent cell-mediated cytotoxicity (ADCC).
[Present invention 1155]
The method of any of the inventions 1117-1154, wherein the target antibody or antigen-binding fragment thereof is a single chain fragment.
[Present invention 1156]
The method of the invention 1155, wherein the single chain fragment is an scFv.
[Present invention 1157]
Any of the present invention 1117 to 1156, wherein the target antibody or antigen-binding fragment thereof comprises a VH region comprising the amino acid sequence shown in SEQ ID NO:11 and a VL region comprising the amino acid sequence shown in SEQ ID NO:12. That method.
[Present invention 1158]
1157. The method of the present invention 1157, wherein the target antibody or antigen-binding fragment thereof is an scFv, and the VH region and VL region are connected by a flexible linker.
[Present invention 1159]
1158. The method of the invention 1158, wherein the flexible linker comprises the amino acid sequence set forth in SEQ ID NO:14 and the scFv comprises the amino acid sequence set forth in SEQ ID NO:13.
[Invention 1160]
Any of the present invention 1117 to 1156, wherein the target antibody or antigen-binding fragment thereof comprises a VH region comprising the amino acid sequence shown in SEQ ID NO:25 and a VL region comprising the amino acid sequence shown in SEQ ID NO:26. That method.
[Present invention 1161]
1160. The method of the invention 1160, wherein the target antibody or antigen-binding fragment thereof is an scFv, and the VH region and VL region are connected by a flexible linker.
[Invention 1162]
1161. The method of the invention 1161, wherein the flexible linker comprises the amino acid sequence set forth in SEQ ID NO:14 and the scFv comprises the amino acid sequence set forth in SEQ ID NO:27.
[Present invention 1163]
an anti-idiotypic antibody or antigen-binding fragment thereof according to any one of the present inventions 1001 to 1092 or a conjugate according to any one of the present inventions 1093 to 1096;
The anti-idiotypic antibody or antigen-binding fragment thereof is as follows:
detecting a target antibody or an antigen-binding fragment thereof, or detecting a CAR comprising a target antibody or an antigen-binding fragment thereof; and/or
selecting or enriching from a population of cells engineered cells that express a CAR comprising a target antibody or antigen-binding fragment thereof; and/or
stimulating an input composition comprising cells expressing a CAR comprising a target antibody or antigen-binding fragment thereof;
with instructions for use for
Articles of manufacture, including;
[Present invention 1164]
a binding reagent comprising an extracellular domain of a CAR comprising a target antibody or antigen-binding fragment thereof, wherein the extracellular domain or a portion thereof comprises the target antibody or antigen-binding fragment thereof;
with the anti-idiotypic antibody or antigen-binding fragment of any of the present inventions 1001 to 1092 or the conjugate of any of the present inventions 1093 to 1096.
Articles of manufacture, including;
[Present invention 1165]
1164. The article of manufacture of the invention 1164, wherein the binding reagent is a first binding reagent, and the article of manufacture further comprises a second binding reagent comprising an extracellular domain of a CAR or a portion thereof.
[Present invention 1166]
The article of manufacture of Invention 1164 or Invention 1165, wherein the extracellular domain of the CAR of the first binding reagent, or a portion thereof, and the extracellular domain of the CAR, or a portion thereof, of the second binding reagent are the same.
[Present invention 1167]
The article of manufacture is
Instructions for using a binding reagent, optionally a first binding reagent, and a second binding reagent, for assaying a sample for the presence or absence of molecules that bind to the binding reagent using an immunoassay.
further including;
Optionally, the immunoassay is a bridge immunoassay or a sandwich immunoassay, and optionally the sample is from a subject receiving cell therapy comprising cells engineered with a CAR comprising a targeting antibody or antigen-binding fragment thereof. ,
The article of manufacture of any of the inventions 1164-1166.
[Present invention 1168]
The article of manufacture of any of the inventions 1164-1167, wherein the binding reagent, optionally the first and/or second binding reagent, is detectably labeled or capable of producing a detectable signal.
[Present invention 1169]
one of the first binding reagent and the second binding reagent is bound to or capable of being bound to the solid support; The article of manufacture of any of the inventions 1165-1168, wherein the other of the binding reagents is a detectable label or is capable of producing a detectable signal.
[Invention 1170]
the article of manufacture further comprises a solid support;
Optionally, one of the first binding reagent and the second binding reagent is directly or indirectly linked to biotin, and the solid support comprises a streptavidin-coated surface.
Articles of manufacture according to the invention 1169.
[Present invention 1171]
The article of manufacture of any of the inventions 1163-1170, wherein the target antibody or antigen-binding fragment thereof is a single chain fragment.
[Invention 1172]
An article of manufacture according to the invention 1171, wherein the single chain fragment is an scFv.
[Present invention 1173]
Any of the present invention 1163 to 1172, wherein the target antibody or antigen-binding fragment thereof comprises a VH region comprising the amino acid sequence shown in SEQ ID NO:11 and a VL region comprising the amino acid sequence shown in SEQ ID NO:12. Articles of manufacture.
[Present invention 1174]
1173. The article of manufacture of the present invention 1173, wherein the target antibody or antigen-binding fragment thereof is an scFv, and the VH region and VL region are connected by a flexible linker.
[Present invention 1175]
1174. The article of manufacture of the invention 1174, wherein the flexible linker comprises the amino acid sequence set forth in SEQ ID NO:14 and the scFv comprises the amino acid sequence set forth in SEQ ID NO:13.
[Present invention 1176]
Any of the present invention 1163 to 1172, wherein the target antibody or antigen-binding fragment thereof comprises a VH region comprising the amino acid sequence shown in SEQ ID NO:25 and a VL region comprising the amino acid sequence shown in SEQ ID NO:26. Articles of manufacture.
[Invention 1177]
The article of manufacture of the present invention 1176, wherein the target antibody or antigen-binding fragment thereof is an scFv, and the VH region and VL region are connected by a flexible linker.
[Invention 1178]
1177. An article of manufacture of the invention 1177, wherein the flexible linker comprises the amino acid sequence set forth in SEQ ID NO:14 and the scFv comprises the amino acid sequence set forth in SEQ ID NO:27.

Claims (46)

An anti-idiotypic antibody or antigen-binding fragment thereof that binds to an anti-BCMA target antibody or antigen-binding fragment thereof,
(a) CDR-H1 comprising the amino acid sequence shown in SEQ ID NO:74 or 104, CDR-H2 comprising the amino acid sequence shown in SEQ ID NO:75 or 105, and CDR-H2 comprising the amino acid sequence shown in SEQ ID NO:76 or 106. a VH region comprising CDR-H3 comprising an amino acid sequence;
CDR-L1 containing the amino acid sequence shown in SEQ ID NO:81 or 111, CDR-L2 containing the amino acid sequence shown in SEQ ID NO:82 or 112, and amino acid sequence shown in SEQ ID NO:83 or 113. containing CDR-L3, containing VL region , or
(b) containing CDR-H1, CDR-H2, and CDR-H3, respectively, containing the amino acid sequences of CDR-H1, CDR-H2, and CDR-H3 contained within the amino acid sequence of SEQ ID NO: 84 or 114; VH area,
A VL region containing CDR-L1, CDR-L2, and CDR-L3 containing the amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 contained within the amino acid sequence of SEQ ID NO:85 or 115, respectively. or
(c) a VH region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:84 or 114;
a VL region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:85 or 115,
Anti-idiotype antibodies or antigen-binding fragments thereof. The VH region comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:84, and the VL region comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:85. contains an amino acid sequence with 90% sequence identity, or
The VH region comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:114, and the VL region comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:115. contains an amino acid sequence with 90% sequence identity , or
The VH region contains the amino acid sequence shown in SEQ ID NO:84, and the VL region contains the amino acid sequence shown in SEQ ID NO:85, or
The VH region contains the amino acid sequence shown in SEQ ID NO:114, and the VL region contains the amino acid sequence shown in SEQ ID NO:115,
The anti-idiotypic antibody or antigen-binding fragment thereof according to claim 1 . (a) CDR-H1 whose VH region includes the amino acid sequence shown in SEQ ID NO:74, CDR-H2 which includes the amino acid sequence shown in SEQ ID NO:75, and the amino acid sequence shown in SEQ ID NO:76 contains CDR-H3, and
CDR-L1 whose VL region contains the amino acid sequence shown in SEQ ID NO:81, CDR-L2 which contains the amino acid sequence shown in SEQ ID NO:82, and CDR whose VL region contains the amino acid sequence shown in SEQ ID NO:83. -contains L3, or
(b) CDR-H1 whose VH region contains the amino acid sequence shown in SEQ ID NO:104, CDR-H2 which contains the amino acid sequence shown in SEQ ID NO:105, and the amino acid sequence shown in SEQ ID NO:106. contains CDR-H3, and
CDR-L1 whose VL region contains the amino acid sequence shown in SEQ ID NO:111, CDR-L2 which contains the amino acid sequence shown in SEQ ID NO:112, and CDR whose VL region contains the amino acid sequence shown in SEQ ID NO:113. -Including L3,
The anti-idiotypic antibody or antigen-binding fragment thereof according to claim 1 or 2 . The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of claims 1 to 3 , wherein the anti-BCMA target antibody or antigen-binding fragment thereof binds human BCMA and/or is a single-chain fragment. Claims 1 to 4 , wherein the anti-BCMA target antibody or antigen-binding fragment thereof comprises a VH region comprising the amino acid sequence shown in SEQ ID NO:25 and a VL region comprising the amino acid sequence shown in SEQ ID NO:26. The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of . Claim 4 or 5 , wherein the single chain fragment comprises a flexible linker disposed between the VH region and the VL region, optionally said flexible linker comprising the amino acid sequence set forth in SEQ ID NO:14. anti-idiotypic antibodies or antigen-binding fragments thereof. Claims 4-6, wherein the single chain fragment of the anti-BCMA target antibody or antigen binding fragment thereof is a single chain variable region fragment (scFv), optionally said scFv comprising the amino acid sequence set forth in SEQ ID NO:27. The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of . the anti-BCMA targeting antibody or antigen-binding fragment is within or included in the antigen-binding domain of the extracellular portion of the CAR, and/or the anti-idiotypic antibody or antigen-binding fragment is binds to an anti-BCMA target antibody or antigen-binding fragment contained within or contained in the antigen-binding domain of the extracellular portion of the CAR;
The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of claims 1 to 7 . the scFv is present or contained within the extracellular portion of the CAR, and/or the anti-idiotypic antibody or antigen-binding fragment is contained within the extracellular portion of the CAR. or binds to the scFv contained in the extracellular portion,
8. The anti-idiotypic antibody or antigen-binding fragment thereof according to claim 7 . The CAR further comprises a transmembrane domain linked to the antigen binding domain via a spacer, the transmembrane domain comprising a transmembrane portion of CD28, optionally human CD28 , and optionally the spacer comprises a transmembrane portion of CD28, optionally human CD28. 10. The anti-idiotypic antibody or antigen-binding fragment thereof according to claim 8 or 9 , which is a globulin spacer, optionally comprising the amino acid sequence shown in SEQ ID NO:50. does not bind to an epitope in the spacer domain of the CAR and/or
does not bind or specifically bind to CD28, optionally human CD28, or a portion thereof ; and/or
does not bind to an epitope in the Fc domain, optionally in the human IgG1 Fc domain , and/or
does not cross-react with another anti-BCMA antibody and optionally with another anti-BCMA antibody contained in the extracellular antigen binding domain of another CAR;
The anti-idiotypic antibody or antigen-binding fragment thereof according to claim 10 . Said another anti-BCMA antibody,
(i) a VH region comprising an amino acid sequence having less than 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:25 and/or 90% to the amino acid sequence set forth in SEQ ID NO:26; and/or (ii) comprises an scFv that has less than 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:13, and/or or (iii) a CDR-H1 that has 1, 2, 3 or 4 or more amino acid residues that differ from the amino acid sequence of CDR-H1 shown in SEQ ID NO:15, and/or SEQ ID NO:16. CDR-H2 having 1, 2, 3 or 4 or more amino acid residues that differ from the amino acid sequence of CDR-H2 shown and/or differ from the amino acid sequence of CDR-H3 shown in SEQ ID NO:17 CDR-H3 having 1, 2, 3 or 4 or more amino acid residues and/or 1, 2, 3 or 4 or more amino acid residues different from the amino acid sequence of CDR-L1 shown in SEQ ID NO:22 or CDR-L2 having 1, 2, 3, or 4 or more amino acid residues different from the amino acid sequence of CDR-L2 shown in SEQ ID NO:23. , and/or comprises a CDR-L3 having 1, 2, 3 or 4 or more amino acid residues different from the amino acid sequence of CDR-L3 shown in SEQ ID NO:24, and/or (iv) an anti- binds to an epitope of human BCMA that is not the same epitope of human BCMA to which the BCMA targeting antibody or antigen-binding fragment thereof binds;
The anti-idiotypic antibody or antigen-binding fragment thereof according to claim 11 . an agonist of a CAR comprising an anti-BCMA targeting antibody or antigen-binding fragment thereof , optionally an agonist of said CAR when in soluble form and/or immobilized on a support or stationary phase; Item 13. The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of Items 1 to 12 . An anti -idiotypic antibody or antigen-binding fragment thereof that binds to an anti-BCMA target antibody or antigen-binding fragment thereof,
(a) CDR-H1 comprising the amino acid sequence shown in SEQ ID NO:28 or 52, CDR-H2 comprising the amino acid sequence shown in SEQ ID NO:29 or 53, and CDR-H2 comprising the amino acid sequence shown in SEQ ID NO:30 or 54. a VH region comprising CDR-H3 comprising an amino acid sequence;
CDR-L1 containing the amino acid sequence shown in SEQ ID NO:35 or 59, CDR-L2 containing the amino acid sequence shown in SEQ ID NO:36 or 60, and the amino acid sequence shown in SEQ ID NO:37 or 61. containing CDR-L3, containing VL region , or
(b) containing CDR-H1, CDR-H2, and CDR-H3, respectively, containing the amino acid sequences of CDR-H1, CDR-H2, and CDR-H3 contained within the amino acid sequence of SEQ ID NO: 38 or 62; VH area,
A VL region containing CDR-L1, CDR-L2, and CDR-L3 containing the amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 contained within the amino acid sequence of SEQ ID NO: 39 or 63, respectively. or
(c) a VH region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:38 or 62;
a VL region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:39 or 63,
Anti-idiotype antibodies or antigen-binding fragments thereof. The VH region comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:38, and the VL region comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:39. contains an amino acid sequence with 90% sequence identity, or
The VH region comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:62, and the VL region comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:63. contains an amino acid sequence with 90% sequence identity , or
The VH region contains the amino acid sequence shown in SEQ ID NO:38, and the VL region contains the amino acid sequence shown in SEQ ID NO:39, or
The VH region contains the amino acid sequence shown in SEQ ID NO:62, and the VL region contains the amino acid sequence shown in SEQ ID NO:63,
15. The anti-idiotypic antibody or antigen-binding fragment thereof according to claim 14 . (a) CDR-H1 whose VH region contains the amino acid sequence shown in SEQ ID NO:28, CDR-H2 which contains the amino acid sequence shown in SEQ ID NO:29, and the amino acid sequence shown in SEQ ID NO:30. contains CDR-H3, and
CDR-L1 whose VL region contains the amino acid sequence shown in SEQ ID NO:35, CDR-L2 which contains the amino acid sequence shown in SEQ ID NO:36, and CDR whose VL region contains the amino acid sequence shown in SEQ ID NO:37. -contains L3 , or
(b) CDR-H1 whose VH region contains the amino acid sequence shown in SEQ ID NO:52, CDR-H2 which contains the amino acid sequence shown in SEQ ID NO:53, and the amino acid sequence shown in SEQ ID NO:54. contains CDR-H3, and
CDR-L1 whose VL region contains the amino acid sequence shown in SEQ ID NO:59, CDR-L2 which contains the amino acid sequence shown in SEQ ID NO:60, and CDR whose VL region contains the amino acid sequence shown in SEQ ID NO:61. -Including L3,
The anti-idiotypic antibody or antigen-binding fragment thereof according to claim 14 or 15 . Any one of claims 14 to 16, wherein the anti-idiotypic antibody or antigen-binding fragment thereof binds to a first anti-BCMA target antibody or antigen-binding fragment thereof and a second anti-BCMA target antibody or antigen-binding fragment thereof. The anti-idiotypic antibody or antigen-binding fragment thereof as described in 2. the first anti-BCMA targeting antibody or antigen-binding fragment thereof binds human BCMA, and the second anti-BCMA targeting antibody or antigen-binding fragment thereof binds human BCMA, and/or
The binding of the anti-idiotypic antibody or antigen-binding fragment thereof to the first anti-BCMA target antibody or antigen-binding fragment thereof and/or the second anti-BCMA target antibody or antigen-binding fragment thereof may be caused by binding to human BCMA or human BCMA-Fc not blocked by,
18. The anti-idiotypic antibody or antigen-binding fragment thereof according to claim 17 . The first anti-BCMA target antibody or antigen-binding fragment thereof comprises a VH region comprising the amino acid sequence shown in SEQ ID NO:11 and a VL region comprising the amino acid sequence shown in SEQ ID NO:12, and The anti-BCMA target antibody or antigen-binding fragment thereof of No. 2 comprises a VH region comprising the amino acid sequence shown in SEQ ID NO:25 and a VL region comprising the amino acid sequence shown in SEQ ID NO:26,
The anti-idiotypic antibody or antigen-binding fragment thereof according to claim 17 or 18 . the first anti-BCMA targeting antibody or antigen-binding fragment thereof is a first single-chain fragment, and the second anti-BCMA targeting antibody or antigen-binding fragment thereof is a second single-chain fragment, optionally , the first single chain fragment comprises a flexible linker disposed between the VH region and the VL region, and the second single chain fragment comprises a flexible linker disposed between the VH region and the VL region. and optionally the flexible linker of the first single chain fragment and/or the second single chain fragment comprises the amino acid sequence shown in SEQ ID NO:14 or an anti-idiotypic antibody thereof according to claim 19 . Antigen-binding fragment. The first single chain fragment of the first anti-BCMA target antibody or antigen-binding fragment thereof is a first scFv, and the second single chain fragment of the second anti-BCMA target antibody or antigen-binding fragment thereof is 21. The anti-idiotype antibody or antigen-binding fragment thereof according to claim 20 , which is a second scFv. The anti-idiotypic antibody or its anti-idiotype antibody according to claim 21 , wherein the first scFv comprises the amino acid sequence shown in SEQ ID NO:13, and the second scFv comprises the amino acid sequence shown in SEQ ID NO:27. Antigen-binding fragment. an epitope within or comprising all or a portion of a CDR of a first anti-BCMA target antibody or antigen-binding fragment; and an epitope within or comprising all or a portion of a CDR of a second anti-BCMA target antibody or antigen-binding fragment; 23. The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of claims 17 to 22, which binds to an epitope comprising a portion thereof. the first anti-BCMA targeting antibody or antigen-binding fragment is within or included in the antigen-binding domain of the extracellular portion of the first CAR, and/or the anti-idiotypic antibody or the antigen-binding fragment binds to a first anti-BCMA targeting antibody or antigen-binding fragment contained within or contained in the antigen-binding domain of the extracellular portion of the first CAR , and /or
a second anti-BCMA targeting antibody or antigen-binding fragment is present within or included in the antigen-binding domain of the extracellular portion of the second CAR, and/or an anti-idiotypic antibody or the antigen-binding fragment binds to a second anti-BCMA targeting antibody or antigen-binding fragment contained within or contained in the antigen-binding domain of the extracellular portion of the second CAR;
The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of claims 17 to 23 . the first scFv is within or included in the extracellular portion of the first CAR, and/or the anti-idiotypic antibody or antigen-binding fragment is present in the extracellular portion of the first CAR; binds to a first scFv contained within or contained in the extracellular portion , and/or
the second scFv is within or included in the extracellular portion of the second CAR, and/or the anti-idiotypic antibody or antigen-binding fragment is present in or included in the extracellular portion of the second CAR; binds to a second scFv contained within or contained in the extracellular portion;
The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of claims 17 to 24 . The first CAR further comprises a transmembrane domain linked to the antigen binding domain via a spacer, and/or the second CAR further comprises a transmembrane domain linked to the antigen binding domain via a spacer. and optionally, the anti-idiotypic antibody or antigen-binding fragment thereof according to claim 24 or 25 , wherein the anti-idiotypic antibody or antigen-binding fragment thereof does not bind to an epitope in the spacer domain of the first CAR and/or the second CAR. Antigen-binding fragment. The spacer of the first CAR and/or the second CAR is an immunoglobulin spacer, optionally the spacer comprises the amino acid sequence set forth in SEQ ID NO:50 , and/or
the transmembrane domain of the first CAR and/or the second CAR comprises a transmembrane portion of CD28, optionally human CD28;
27. The anti-idiotypic antibody or antigen-binding fragment thereof according to claim 26 . does not bind or specifically bind to CD28, optionally human CD28, or a portion thereof ; and/or
does not bind or specifically bind to an epitope in the Fc domain, optionally in the human IgG1 Fc domain;
The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of claims 14 to 27 . is an agonist of a first CAR comprising a first anti-BCMA targeting antibody or antigen-binding fragment thereof, optionally in soluble form and/or immobilized on a support or stationary phase ; and/or
an agonist of a second CAR , optionally in soluble form and/or immobilized on a support or stationary phase, comprising a second anti-BCMA targeting antibody or antigen-binding fragment thereof;
The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of claims 17 to 28 . A heavy chain constant region comprising at least a portion of a heavy and/or light chain immunoglobulin constant region, optionally comprising an Fc region or a portion of said Fc comprising CH2 and CH3 domains, and/or a light chain comprising a CL domain. 30. The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of claims 1 to 29 , comprising a chain constant region, optionally wherein the light chain constant region is from a kappa light chain. A heavy chain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:86 and at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:87. a light chain comprising an amino acid sequence having
A heavy chain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:116 and at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:117. comprises a light chain comprising an amino acid sequence having
a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:86, and a light chain comprising the amino acid sequence set forth in SEQ ID NO:87, or
a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:116, and a light chain comprising the amino acid sequence set forth in SEQ ID NO:117;
The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of claims 1 to 13 and 30 . A heavy chain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:40 and at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:41. a light chain comprising an amino acid sequence having
A heavy chain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:64 and at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:65. a light chain comprising an amino acid sequence having
a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:40, and a light chain comprising the amino acid sequence set forth in SEQ ID NO:41, or
The anti-idiotype antibody according to any one of claims 14 to 30 , comprising a heavy chain comprising the amino acid sequence shown in SEQ ID NO:64 and a light chain comprising the amino acid sequence shown in SEQ ID NO:65, or its antigen-binding fragment. an antigen-binding fragment , and optionally the antigen-binding fragment is selected from Fab fragments, F(ab') ₂ fragments, Fab' fragments, Fv fragments, scFvs, and single domain antibodies. 30. The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of 30 . is humanized and/or
is recombinant and/or
is monoclonal and/or
is an intact or full-length antibody;
The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of claims 1 to 33 . A conjugate comprising an anti-idiotypic antibody or antigen-binding fragment thereof according to any one of claims 1 to 34 and a heterologous molecule or moiety. A nucleic acid molecule encoding the heavy chain and/or light chain of the anti-idiotypic antibody or antigen-binding fragment thereof according to any one of claims 1 to 34 . 37. A vector comprising the nucleic acid molecule of claim 36 . A cell comprising the anti-idiotypic antibody or antigen-binding fragment thereof according to any one of claims 1 to 34 , the nucleic acid molecule according to claim 36 , or the vector according to claim 37 . an anti-idiotypic antibody or antigen-binding fragment thereof according to any one of claims 1 to 34 , a conjugate according to claim 35 , or a cell according to claim 38 , optionally in a pharmaceutically acceptable form. A composition further comprising an excipient. The anti-idiotypic antibody or antigen-binding thereof according to any one of claims 1 to 34, wherein (a) a composition comprising a target antibody or antigen-binding fragment thereof specifically binds to the target antibody or antigen-binding fragment thereof. and ( b ) detecting anti-idiotypic antibodies bound to the target antibody or antigen-binding fragment thereof. How to detect. The anti-idiotypic antibody or anti-idiotypic antibody according to any one of claims 1 to 34, which (a) specifically binds to cells expressing a CAR containing the target antibody or antigen-binding fragment thereof. and ( b ) detecting cells that are bound to the anti-idiotypic antibody or antigen-binding fragment thereof. A method to detect CARs containing fragments. The anti-idiotype according to any one of claims 1 to 34 , wherein the input composition comprises cells expressing a CAR containing a target antibody or antigen-binding fragment thereof, which specifically binds to the target antibody or antigen-binding fragment thereof. 36. A method of stimulating cells, comprising incubating with an antibody or antigen-binding fragment thereof or a conjugate of claim 35 , thereby producing an output composition comprising stimulated cells. (a) introducing a nucleic acid molecule encoding a CAR into a cell, thereby creating an input composition; and (b) introducing the input composition specifically for the antigen receptor of the CAR. incubating with the anti-idiotypic antibody or antigen-binding fragment thereof according to any one of claims 1 to 34 or the conjugate according to claim 35 , thereby producing a cell composition. A method of producing a cell composition, comprising: The anti-idiotypic antibody or antigen-binding thereof according to any one of claims 1 to 34, wherein (a) a composition comprising a target antibody or antigen-binding fragment thereof specifically binds to the target antibody or antigen-binding fragment thereof. and ( b ) isolating a complex comprising the anti-idiotypic antibody or antigen-binding fragment thereof. How to refine. The anti-idiotype antibody or antigen-binding fragment thereof according to any one of claims 1 to 34 or the conjugate according to claim 35 , which specifically binds to a target antibody or antigen-binding fragment thereof , for depleting cells in a subject . A composition comprising a gate , wherein the subject has been administered cells expressing a CAR comprising the target antibody or antigen-binding fragment thereof. The anti-idiotypic antibody or antigen-binding fragment thereof according to any one of claims 1 to 34, which specifically binds to a target antibody or antigen-binding fragment thereof, for the manufacture of a medicament for depleting cells in a subject. 36. Use of a composition comprising a conjugate according to claim 35, wherein the subject has been administered cells expressing a CAR comprising the targeting antibody or antigen-binding fragment thereof.