# US 12,600,781 B2 · Tumor-targeted agonistic CD28 antigen binding molecules

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- Number: US 12,600,781 B2
- Document ID: US12600781B2
- Status: Granted
- Kind: B2
- Application number: 18/056,655
- Filed: 2022-11-17
- Published: 2023-12-28
- Granted: 2026-04-14
- Earliest claimed: 2018-12-21
- Assignees: Hoffmann-La Roche Inc.
- Applicants: Hoffmann-La Roche Inc.
- Inventors: Guy Georges; Thomas Hofer; Ralf Hosse; Christian Klein; Ekkehard Moessner; Johannes Sam; Pablo Umaña; Jenny Thom; Stephan Gasser; Jean-Baptiste Vallier; Tanja Fauti
- Web page: https://patentgrep.com/patent/US12600781B2

## CPC

- C07K 16/2818: {against CD28 or CD152}
- A61P 35/00: Antineoplastic agents
- C07K 16/2803: {against the immunoglobulin superfamily}
- C07K 16/2809: {against the T-cell receptor (TcR)-CD3 complex}
- C07K 16/2827: {against B7 molecules, e.g. CD80, CD86}
- C07K 16/286: {against neuromediator receptors, e.g. serotonin receptor, dopamine receptor}
- C07K 16/2878: {against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95}
- C07K 16/2896: {against molecules with a "CD"-designation, not provided for elsewhere}
- C07K 16/30: from tumour cells
- C07K 16/3007: {Carcino-embryonic Antigens}
- C07K 16/32: against translation products of oncogenes
- C07K 16/40: against enzymes
- A61K 2039/505: {comprising antibodies}
- A61K 2039/507: {Comprising a combination of two or more separate antibodies}
- C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
- C07K 2317/31: multispecific
- C07K 2317/35: Valency
- C07K 2317/52: Constant or Fc region; Isotype
- C07K 2317/526: CH3 domain
- C07K 2317/55: Fab or Fab'
- C07K 2317/565: Complementarity determining region [CDR]
- C07K 2317/73: Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
- C07K 2317/75: Agonist effect on antigen
- C07K 2317/92: Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
- C12N 5/10: Cells modified by introduction of foreign genetic material
- C12N 15/00: Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor (mutants or genetically engineered microorganisms, per se C12N1/00, C12N5/00, C12N7/00; new plants per se A01H; plant reproduction by tissue culture techniques A01H4/00; new animals per se A01K67/00; use of medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases, gene therapy A61K48/00)
- C12N 15/63: Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present invention relates to tumor targeted bispecific agonistic antigen binding molecules characterized by monovalent binding to CD28, methods for their production, pharmaceutical compositions containing these antibodies, and methods of using the same.

## Claims

### Claim 1

A polynucleotide or set of polynucleotides encoding a bispecific agonistic CD28 antigen binding molecule characterized by monovalent binding to CD28, wherein said bispecific agonistic CD28 antigen binding molecule comprises
(a) one antigen binding domain capable of specific binding to CD28, wherein the antigen binding domain capable of specific binding to CD28 comprises a heavy chain variable region (VHCD28) comprising a CDR-H1 of SEQ ID NO: 489, a CDR-H2 of SEQ ID NO: 490, and a CDR-H3 of SEQ ID NO:38 (SHYGX5DX6NFDV) wherein X5 is L and X6 is F, and a light chain variable region (VLCD28) comprising a CDR-L1 of SEQ ID NO: 492, a CDR-L2 of SEQ ID NO: 493 and a CDR-L3 of SEQ ID NO: 494;
(b) at least one antigen binding domain capable of specific binding to a tumor-associated antigen, and
(c) an Fc domain composed of a first and a second subunit capable of stable association comprising one or more amino acid substitution that reduces the binding affinity of the antigen binding molecule to an Fc receptor and/or effector function compared to a native IgG1 Fc domain.

### Claim 2 (depends on 1)

The polynucleotide or set of polynucleotides of claim 1, wherein the Fc domain is of human IgG1 subclass and comprises the amino acid mutations L234A, L235A and P329G, as numbered according to Kabat EU index.

### Claim 3 (depends on 1)

The polynucleotide or set of polynucleotides of claim 1, wherein the antigen binding domain capable of specific binding to CD28 comprises a heavy chain variable region (V H CD28) comprising the amino acid sequence of SEQ ID NO:47 and a light chain variable region (V L CD28) comprising the amino acid sequence of SEQ ID NO:54.

### Claim 4 (depends on 1)

The polynucleotide or set of polynucleotides of claim 1, wherein the tumor-associated antigen is selected from the group consisting of Fibroblast Activation Protein (FAP), Carcinoembryonic Antigen (CEA), Folate receptor alpha (FolR1), Melanoma-associated Chondroitin Sulfate Proteoglycan (MCSP), Epidermal Growth Factor Receptor (EGFR), human epidermal growth factor receptor 2 (HER2), p95HER2, epithelial cell adhesion molecule (EpCAM), HER3, CD30, TPBG (5T4), CD19, CD79b, CD20, CD22, CD37, CD38, BCMA and GPRC5D.

### Claim 5 (depends on 1)

The polynucleotide or set of polynucleotides of claim 1, wherein the at least one antigen binding domain capable of specific binding to a tumor-associated antigen is an antigen binding domain capable of specific binding to Carcinoembryonic Antigen (CEA).

### Claim 6 (depends on 5)

The polynucleotide or set of polynucleotides of claim 5, wherein the antigen binding domain capable of specific binding to CEA comprises
(i) a heavy chain variable region (V H CEA) comprising a CDR-H1 comprising the amino acid sequence of SEQ ID NO:180, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 181, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO:182, and a light chain variable region (V L CEA) comprising a CDR-L1 comprising the amino acid sequence of SEQ ID NO:183, a CDR-L2 comprising the amino acid sequence of SEQ ID NO:184, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO:185; or
(ii) a heavy chain variable region (V H CEA) comprising a CDR-H1 comprising the amino acid sequence of SEQ ID NO:127, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 128, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO:129, and a light chain variable region (V L CEA) comprising a CDR-L1 comprising the amino acid sequence of SEQ ID NO:130, a CDR-L2 comprising the amino acid sequence of SEQ ID NO:131, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 132; or
(iii) a heavy chain variable region (V H CEA) comprising a CDR-H1 comprising the amino acid sequence of SEQ ID NO:507, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:508, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO:509, and a light chain variable region (V L CEA) comprising a CDR-L1 comprising the amino acid sequence of SEQ ID NO:510, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 511, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO:512.

### Claim 7 (depends on 5)

The polynucleotide or set of polynucleotides of claim 5, wherein the antigen binding domain capable of specific binding to CEA comprises a heavy chain variable region (V H CEA) comprising the amino acid sequence of SEQ ID NO:186, and a light chain variable region (V L CEA) comprising the amino acid sequence of SEQ ID NO:187.

### Claim 8 (depends on 5)

The polynucleotide or set of polynucleotides of claim 5, wherein the antigen binding domain capable of specific binding to CEA comprises
(a) a heavy chain variable region (V H CEA) comprising the amino acid sequence of SEQ ID NO: 194 and a light chain variable region (V L CEA) comprising the amino acid sequence of SEQ ID NO:195, or
(b) a heavy chain variable region (V H CEA) comprising the amino acid sequence of SEQ ID NO: 196 and a light chain variable region (V L CEA) comprising the amino acid sequence of SEQ ID NO:197, or
(c) a heavy chain variable region (V H CEA) comprising the amino acid sequence of SEQ ID NO: 198 and a light chain variable region (V L CEA) comprising the amino acid sequence of SEQ ID NO:199, or
(d) a heavy chain variable region (V H CEA) comprising the amino acid sequence of SEQ ID NO: 200 and a light chain variable region (V L CEA) comprising the amino acid sequence of SEQ ID NO:201, or
(e) a heavy chain variable region (V H CEA) comprising the amino acid sequence of SEQ ID NO: 202 and a light chain variable region (V L CEA) comprising the amino acid sequence of SEQ ID NO:203, or
(f) a heavy chain variable region (V H CEA) comprising the amino acid sequence of SEQ ID NO: 204 and a light chain variable region (V L CEA) comprising the amino acid sequence of SEQ ID NO:205, or
(g) a heavy chain variable region (V H CEA) comprising the amino acid sequence of SEQ ID NO: 206 and a light chain variable region (V L CEA) comprising the amino acid sequence of SEQ ID NO:207, or
(h) a heavy chain variable region (V H CEA) comprising the amino acid sequence of SEQ ID NO: 208 and a light chain variable region (V L CEA) comprising the amino acid sequence of SEQ ID NO:209, or
(i) a heavy chain variable region (V H CEA) comprising the amino acid sequence of SEQ ID NO: 210 and a light chain variable region (V L CEA) comprising the amino acid sequence of SEQ ID NO:211, or
(j) a heavy chain variable region (V H CEA) comprising the amino acid sequence of SEQ ID NO: 212 and a light chain variable region (V L CEA) comprising the amino acid sequence of SEQ ID NO:213.

### Claim 9 (depends on 5)

The polynucleotide or set of polynucleotides of claim 5, wherein the antigen binding domain capable of specific binding to CEA comprises a heavy chain variable region (V H CEA) comprising the amino acid sequence of SEQ ID NO:200 and a light chain variable region (V L CEA) comprising the amino acid sequence of SEQ ID NO:201.

### Claim 10 (depends on 1)

The polynucleotide or set of polynucleotides of claim 1, wherein the at least one antigen binding domain capable of specific binding to a tumor-associated antigen is an antigen binding domain capable of specific binding to Fibroblast Activation Protein (FAP).

### Claim 11 (depends on 10)

The polynucleotide or set of polynucleotides of claim 10, wherein the antigen binding domain capable of specific binding to FAP comprises:
(a) a heavy chain variable region (V H FAP) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:12, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO: 13, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:14, and a light chain variable region (V L FAP) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO:15, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO:16, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:17, or
(b) a heavy chain variable region (V H FAP) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:4, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO: 5, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:6, and a light chain variable region (V L FAP) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO:7, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO:8, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:9.

### Claim 12 (depends on 10)

The polynucleotide or set of polynucleotides of claim 10, wherein the antigen binding domain capable of specific binding to FAP comprises a heavy chain variable region (V H FAP) comprising the amino acid sequence of SEQ ID NO:18 and a light chain variable region (V L FAP) comprising the amino acid sequence of SEQ ID NO:19.

### Claim 13 (depends on 1)

The polynucleotide or set of polynucleotides of claim 1, wherein the at least one antigen binding domain capable of specific binding to a tumor-associated antigen is an antigen binding domain capable of specific binding to epithelial cell adhesion molecule (EpCAM).

### Claim 14 (depends on 13)

The polynucleotide or set of polynucleotides of claim 13, wherein the antigen binding domain capable of specific binding to EpCAM comprises a heavy chain variable region (V H EpCAM) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:515, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:516, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:517, and a light chain variable region (V L EpCAM) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 518, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO:519, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:520.

### Claim 15 (depends on 13)

The polynucleotide or set of polynucleotides of claim 13, wherein the antigen binding domain capable of specific binding to EpCAM comprises a heavy chain variable region (V H EpCAM) comprising the amino acid sequence of SEQ ID NO:521 and a light chain variable region (V L EpCAM) comprising the amino acid sequence of SEQ ID NO: 522.

### Claim 16 (depends on 1)

The polynucleotide or set of polynucleotides of claim 1, wherein the at least one antigen binding domain capable of specific binding to a tumor-associated antigen is an antigen binding domain capable of specific binding to HER3.

### Claim 17 (depends on 16)

The polynucleotide or set of polynucleotides of claim 16, wherein the antigen binding domain capable of specific binding to HER3 comprises a heavy chain variable region (V H HER3) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:523, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:524, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:525, and a light chain variable region (V L HER3) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 526, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO:527, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:528.

### Claim 18 (depends on 16)

The polynucleotide or set of polynucleotides of claim 16, wherein the antigen binding domain capable of specific binding to HER3 comprises a heavy chain variable region (V H HER3) comprising the amino acid sequence of SEQ ID NO:529 and a light chain variable region (V L HER3) comprising the amino acid sequence of SEQ ID NO:530.

### Claim 19 (depends on 1)

The polynucleotide or set of polynucleotides of claim 1, wherein the at least one antigen binding domain capable of specific binding to a tumor-associated antigen is an antigen binding domain capable of specific binding to CD30.

### Claim 20 (depends on 19)

The polynucleotide or set of polynucleotides of claim 19, wherein the antigen binding domain capable of specific binding to CD30 comprises a heavy chain variable region (V H CD30) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:531, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:532, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:533, and a light chain variable region (V L CD30) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 534, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO:535, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:536.

### Claim 21 (depends on 19)

The polynucleotide or set of polynucleotides of claim 19, wherein the antigen binding domain capable of specific binding to CD30 comprises a heavy chain variable region (V H CD30) comprising the amino acid sequence of SEQ ID NO:537 and a light chain variable region (V L CD30) comprising the amino acid sequence of SEQ ID NO:538.

### Claim 22 (depends on 1)

The polynucleotide or set of polynucleotides of claim 1, wherein the at least one antigen binding domain capable of specific binding to a tumor-associated antigen is an antigen binding domain capable of specific binding to TBPG.

### Claim 23 (depends on 22)

The polynucleotide or set of polynucleotides of claim 22, wherein the antigen binding domain capable of specific binding to TBPG comprises a heavy chain variable region (V H TBPG) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:539, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:540, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:541, and a light chain variable region (V L TBPG) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 542, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO:543, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:544.

### Claim 24 (depends on 22)

The polynucleotide or set of polynucleotides of claim 22, wherein the antigen binding domain capable of specific binding to TBPG comprises a heavy chain variable region (V H TBPG) comprising the amino acid sequence of SEQ ID NO:545 and a light chain variable region (V L TBPG) comprising the amino acid sequence of SEQ ID NO:546.

### Claim 25 (depends on 1)

The polynucleotide or set of polynucleotides of claim 1, wherein the tumor-associated antigen is a Multiple Myeloma (MM) cell surface antigen selected from the group consisting of CD38, BCMA and GPRC5D.

### Claim 26 (depends on 25)

The polynucleotide or set of polynucleotides of claim 25, wherein the at least one antigen binding domain capable of specific binding to a tumor-associated antigen is an antigen binding domain capable of specific binding to GPRC5D.

### Claim 27 (depends on 26)

The polynucleotide or set of polynucleotides of claim 26, wherein the antigen binding domain capable of specific binding to GPRC5D comprises
(a) a heavy chain variable region (V H GPRC5D) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:563, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:564, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:565, and a light chain variable region (V L GPRC5D) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO:566, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 567, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:568, or
(b) a heavy chain variable region (V H GPRC5D) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:579, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:580, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:581, and a light chain variable region (V L GPRC5D) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO:582, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 583, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:584.

### Claim 28 (depends on 26)

The polynucleotide or set of polynucleotides of claim 26, wherein the antigen binding domain capable of specific binding to GPRC5D comprises a heavy chain variable region (V H GPRC5D) comprising the amino acid sequence of SEQ ID NO:569 and a light chain variable region (V L GPRC5D) comprising the amino acid sequence of SEQ ID NO: 570.

### Claim 29 (depends on 1)

The polynucleotide or set of polynucleotides of claim 1, wherein the at least one antigen binding domain capable of specific binding to a tumor-associated antigen is an antigen binding domain capable of specific binding to CD38.

### Claim 30 (depends on 29)

The polynucleotide or set of polynucleotides of claim 29, wherein the antigen binding domain capable of specific binding to CD38 comprises a heavy chain variable region (V H CD38) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:547, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:548, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:549, and a light chain variable region (V L CD38) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 550, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO:551, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:552.

### Claim 31 (depends on 29)

The polynucleotide or set of polynucleotides of claim 29, wherein the antigen binding domain capable of specific binding to CD38 comprises a heavy chain variable region (V H CD38) comprising the amino acid sequence of SEQ ID NO:553 and a light chain variable region (V L CD38) comprising the amino acid sequence of SEQ ID NO:554.

### Claim 32 (depends on 1)

The polynucleotide or set of polynucleotides of claim 1, wherein the at least one antigen binding domain capable of specific binding to a tumor-associated antigen is an antigen binding domain capable of specific binding to BCMA.

### Claim 33 (depends on 32)

The polynucleotide or set of polynucleotides of claim 32, wherein the antigen binding domain capable of specific binding to BCMA comprises a heavy chain variable region (V H BCMA) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:555, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:556, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:557, and a light chain variable region (V L BCMA) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 558, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO:559, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:560.

### Claim 34 (depends on 32)

The polynucleotide or set of polynucleotides of claim 32, wherein the antigen binding domain capable of specific binding to BCMA comprises a heavy chain variable region (V H BCMA) comprising the amino acid sequence of SEQ ID NO:561 and a light chain variable region (V L BCMA) comprising the amino acid sequence of SEQ ID NO: 562.

### Claim 35 (depends on 1)

The polynucleotide or set of polynucleotides of claim 1, wherein the tumor-associated antigen is a B cell surface antigen selected from the group consisting of CD19, CD79b, CD20, CD22 and CD37.

### Claim 36 (depends on 1)

The polynucleotide or set of polynucleotides of claim 1, wherein the at least one antigen binding domain capable of specific binding to a tumor-associated antigen is an antigen binding domain capable of specific binding to CD19.

### Claim 37 (depends on 36)

The polynucleotide or set of polynucleotides of claim 36, wherein the antigen binding domain capable of specific binding to CD19 comprises:
(a) a heavy chain variable region (V H CD19) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:406, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:407, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:408, and a light chain variable region (V L CD19) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO:409, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 410, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:411, or
(b) a heavy chain variable region (V H CD19) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:414, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:415, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:416, and a light chain variable region (V L CD19) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO:417, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 418, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:419.

### Claim 38 (depends on 36)

The polynucleotide or set of polynucleotides of claim 36, wherein the antigen binding domain capable of specific binding to CD19 comprises (a) a heavy chain variable region (V H CD19) comprising the amino acid sequence of SEQ ID NO:412, and a light chain variable region (V L CD19) comprising the amino acid sequence of SEQ ID NO: 413, or (b) a heavy chain variable region (V H CD19) comprising the amino acid sequence of SEQ ID NO:420, and a light chain variable region (V L CD19) comprising the amino acid sequence of SEQ ID NO:421.

### Claim 39 (depends on 36)

The polynucleotide or set of polynucleotides of claim 36, wherein the antigen binding domain capable of specific binding to CD19 comprises a heavy chain variable region (V H CD19) comprising the amino acid sequence of SEQ ID NO:412 and a light chain variable region (V L CD19) comprising the amino acid sequence of SEQ ID NO:413.

### Claim 40 (depends on 1)

The polynucleotide or set of polynucleotides of claim 1, wherein the at least one antigen binding domain capable of specific binding to a tumor-associated antigen is an antigen binding domain capable of specific binding to CD79b.

### Claim 41 (depends on 40)

The polynucleotide or set of polynucleotides of claim 40, wherein the antigen binding domain capable of specific binding to CD79b comprises a heavy chain variable region (V H CD79b) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:422, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:423, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:424, and a light chain variable region (V L CD79b) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 425, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO:426, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:427.

### Claim 42 (depends on 40)

The polynucleotide or set of polynucleotides of claim 40, wherein the antigen binding domain capable of binding to CD79b comprises a heavy chain variable region (V H CD79b) comprising the amino acid sequence of SEQ ID NO:428, and a light chain variable region (V L CD79b) comprising the amino acid sequence of SEQ ID NO: 429.

### Claim 43 (depends on 1)

The polynucleotide or set of polynucleotides of claim 1, wherein the bispecific agonistic CD28 antigen binding molecule comprises:
(a) one Fab fragment capable of specific binding to CD28,
(b) one crossFab fragment capable of specific binding to a tumor-associated antigen, and
(c) a Fc domain composed of a first and a second subunit capable of stable association comprising one or more amino acid substitution that reduces the binding affinity of the antigen binding molecule to an Fc receptor and/or effector function compared to a native IgG1 Fc domain.

### Claim 44 (depends on 1)

The polynucleotide or set of polynucleotides of claim 1, wherein the bispecific agonistic CD28 antigen binding molecule comprises:
(a) a first Fab fragment capable of specific binding to CD28,
(b) a second Fab fragment capable of specific binding to a tumor-associated antigen, and
(c) a Fc domain composed of a first and a second subunit capable of stable association comprising one or more amino acid substitution that reduces the binding affinity of the antigen binding molecule to an Fc receptor and/or effector function compared to a native IgG1 Fc domain,
wherein the first Fab fragment capable of specific binding to CD28 is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the second Fab fragment capable of specific binding to a tumor-associated antigen, which is in turn fused at its C-terminus to the N-terminus of one of the Fc domain subunits.

### Claim 45 (depends on 1)

The polynucleotide or set of polynucleotides of claim 1, wherein the bispecific agonistic CD28 antigen binding molecule comprises:
(a) a first Fab fragment capable of specific binding to CD28,
(b) a second and a third Fab fragment capable of specific binding to a tumor-associated antigen, and
(c) a Fc domain composed of a first and a second subunit capable of stable association comprising one or more amino acid substitution that reduces the binding affinity of the antigen binding molecule to an Fc receptor and/or effector function compared to a native IgG1 Fc domain,
wherein the first Fab fragment capable of specific binding to CD28 is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the second Fab fragment capable of specific binding to a tumor-associated antigen, which is in turn fused at its C-terminus to the N-terminus of the first Fc domain subunit, and the third Fab fragment capable of specific binding to a tumor-associated antigen is fused at the C-terminus of the Fab heavy chain to the N-terminus of the second Fc domain subunit.

### Claim 46 (depends on 1)

The polynucleotide or set of polynucleotides of claim 1, wherein the bispecific agonistic CD28 antigen binding molecule comprises:
(a) one antigen binding domain capable of specific binding to CD28, wherein the antigen binding domain capable of specific binding to CD28 comprises a heavy chain variable region (V H CD28) comprising the amino acid sequence of SEQ ID NO:47 and a light chain variable region (V L CD28) comprising the amino acid sequence of SEQ ID NO:54,
(b) at least one antigen binding domain capable of specific binding to CD19, and
(c) a Fc domain composed of a first and a second subunit capable of stable association, wherein the Fc domain is of human IgG1 subclass and comprises the amino acid mutations L234A, L235A and P329G, numbering according to Kabat EU index.

### Claim 47 (depends on 46)

The polynucleotide or set of polynucleotides of claim 46, wherein the at least one antigen binding domain capable of specific binding to CD19 comprises a heavy chain variable region (V H CD19) comprising the amino acid sequence of SEQ ID NO: 412 and a light chain variable region (V L CD19) comprising the amino acid sequence of SEQ ID NO:413.

### Claim 48

A polynucleotide or set of polynucleotides encoding a bispecific agonistic CD28 antigen binding molecule comprising a first light chain comprising the amino acid sequence of SEQ ID NO:122, a first heavy chain comprising the amino acid sequence of SEQ ID NO:114, a second heavy chain comprising the amino acid sequence of SEQ ID NO: 430 and a second light chain comprising the amino acid sequence of SEQ ID NO:431.

### Claim 49 (depends on 1)

A vector comprising the polynucleotide or set of polynucleotides of claim 1.

### Claim 50 (depends on 49)

A cell comprising the vector of claim 49.

### Claim 51 (depends on 50)

A method of producing a bispecific agonistic CD28 antigen binding molecule comprising culturing the host cell of claim 50 under conditions suitable for the expression of the bispecific agonistic CD28 antigen binding molecule, and recovering the bispecific agonistic CD28 antigen binding molecule.