Receptor-mediated endocytosis for targeted internalization and degradation of g protein-coupled receptors

Abstract

Disclosed are fusion proteins (homodimers and heterodimers thereof). Fusion proteins that can bind to a protein of interest that is a G Protein-Coupled Receptor (GPCR) and to an internalizing receptor on a cell surface (transferrin receptor). Once bound, the protein of interest can be internalized and / or degraded inside a cell.

Description

DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 RECEPTOR-MEDIATED ENDOCYTOSIS FOR TARGETED INTERNALIZATION AND DEGRADATION OF G PROTEIN-COUPLED RECEPTORS

[0001] This application is an International Application, which claims the benefit of priority from U.S. provisional patent application no.63 / 566,056, filed on March 15, 2024, U.S. provisional patent application no.63 / 566,069, filed on March 15, 2024, and U.S. provisional patent application no.63 / 566,076, filed on March 15, 2024, the entire contents of each which are incorporated herein by reference in their entireties.

[0002] All patents, patent applications and publications cited herein are hereby incorporated by reference in their entirety. The disclosures of these publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art as known to those skilled therein as of the date of the invention described and claimed herein.

[0003] This patent disclosure contains material that is subject to copyright protection. The copyright owner has no objection to the facsimile reproduction by anyone of the patent document or the patent disclosure as it appears in the U.S. Patent and Trademark Office patent file or records but otherwise reserves any and all copyright rights. FIELD

[0004] Aspects of the invention are drawn to compositions and methods for modulating molecules on the cell surface of cells, including G Protein-Coupled Receptors in cells to reversibly control cells activity to treat diseases related to G Protein-Coupled Receptors. SEQUENCE LISTING

[0005] The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on [ ], is named [ ] and is [ ] bytes in size. BACKGROUND

[0006] Traditional small molecule inhibitor-based therapies have certain disadvantages, such as toxicity and limited efficacy. Examples of inhibitor features that contribute to theirDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 disadvantages include a binding affinity that is the determinant of inhibitor potency; a therapeutic response that depends on sustained target binding; a limited functionality on the target protein; and subverted efficacy through target protein overexpression, native ligand competition, and development of target protein mutations that limit binding or facilitate resistance. Thus, a desirable therapeutic modality exhibits low toxicity and high efficacy. For example, a desirable therapeutic modality includes a variety of features that contribute to overall potency; a therapeutic response that does not entirely or mostly depend on sustained target binding; the ability to completely or substantially limit functionality of the target protein; and limited or no ability of a target cell to subvert efficacy, such as through target protein overexpression, native ligand competition, and / or development of target protein mutations that limit binding or facilitate resistance.

[0007] Membrane proteins are central to a myriad of cellular functions and serve as targets for over half of all drugs. Therefore, developing strategies to degrade membrane proteins is of exceptional interest for both basic research and therapeutic intervention purposes.

[0008] Targeted protein degradation (TPD) is a rapidly growing field in drug discovery and pharmacology. Complementing traditional drug modalities, TPD molecules offer a novel therapeutic mechanism to tackle challenging targets or increase the therapeutic potential of currently used drugs.

[0009] Eukaryotic cells use proteosomes to degrade proteins via the ubiquitin-proteosome pathway (UPP), and lysosomes degrade protein aggregates and whole organelles through endocytosis / phagocytosis. They also use lysosomes to degrade proteins and organelles through endocytosis, phagocytosis, and autophagy. A further component of this pathway is the E3 enzyme which recognizes the targeted protein. This interacts with a 26S proteosome which degrades the protein. TPD has a major role in immunology as it protects important mechanisms when pathogens, cancers, and other diseases target the mechanisms that protect and regulate key functions.

[0010] Proof-of-concept strategies for membrane receptor degradation have been described. These strategies use heterobifunctional biologics that recruit a specific “effector” protein such as a membrane E3 ligase (Cotton, A. D., Nguyen, D. P., Gramespacher, J. A., Seiple, I. B. & Wells, J. A. Development of Antibody-Based PROTACs for the Degradation of the Cell-Surface Immune Checkpoint Protein PD-L1. J Am Chem Soc 143, 593-598 (2021)) or a lysosomeDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 shuttling receptor (Ahn, G. et al. LYTACs that engage the asialoglycoprotein receptor for targeted protein degradation. Nat Chem Biol 17, 937-946 (2021)) to the protein of interest (POI) to induce lysosome-mediated protein degradation. However, the effectiveness of these biological effectors is often limited by their tissue-specific expression patterns. GalNAc-LYTAC, for instance, targets the hepatocyte-specific receptor asialoglycoprotein receptor (ASGPR), making it only suitable for treating liver disease or clearing circulating targets (Ahn, G. et al. Nat Chem Biol 17, 937-946 (2021); Zhou, Y., Teng, P., Montgomery, N. T., Li, X. & Tang, W. Development of Triantennary N-Acetylgalactosamine Conjugates as Degraders for Extracellular Proteins. ACS Cent Sci 7, 499-506 (2021)), while RNF43- or ZNRF3-based methods are more effective for treating Wnt-signaling upregulated disorders where RNF43 and ZNRF3 are expressed at high levels.

[0011] Current technologies are therefore not able to cover the full spectrum of diseases, and developing alternative effectors overexpressed in different diseases and tissues would greatly expand the range of cell surface targets that can be regulated and also increase the targeting specificity.

[0012] G protein-coupled receptors (GPCRs) are a large family of membrane proteins. The primary function of GPCRs is to transduce a wide and diverse array of extracellular stimuli such as biogenic amines, peptides, hormones, neurotransmitters, ions, odorants, and photons into intracellular signals that regulate a myriad of physiological processes including cell metabolism, cell differentiation, growth, neurotransmission, and sensory perception. GPCRs have also been implicated in a large number of diseases, such as type 2 diabetes mellitus (T2DM), Alzheimer’s disease, obesity, depression, and many others. In particular, the use of new molecules for treating of G-protein Couple Receptors (GPCR) related diseases is an area that is yet needed to be explored and developed. Accordingly, there is a need to improve targeted protein degradation, particularly of GPCRs to treat an array of diseases. SUMMARY

[0013] Disclosed here are new reagents (bispecific modulators, also called fusion proteins) and methods for regulating molecules on the surface of cells (e.g., such as proteins of interest). In some embodiments, the reagents and methods are used to control the levels of GPCR molecules on a cell surface. In some embodiments, the reagents and methods are used toDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 reversibly activate or inactivate a GPCR expressing cell (GPCR cell). In some embodiments, GPCR cell activation is modulated in vivo. In some embodiments, the reagents and methods are used to control the levels of receptors and / or coreceptors on the surface of GPCR cells (for examples of GPCRs, see G Protein-Coupled Receptor List | IUPHAR / BPS Guide to PHARMACOLOGY available at https: / / www.guidetopharmacology.org / GRAC / ReceptorFamiliesForward?type=GPCR that is incorporated by reference herein) to inhibit and / or modulate cell responses and cell signaling pathways affected by any of the GPCRs described herein. In some embodiments, disclosed are bispecific modulators as described herein (also called fusion proteins, fusion protein homodimers, fusion protein heterodimers), that target GPCRs. In some embodiments, a protein complex (e.g., an antigen) to which a cell-surface molecule (e.g., any of the GPCR receptors described herein; e.g., APJ, AT1R, RXFP1, A2AR, CCR6, GPR20, CCR4, PAR2, FZD1, CCR2, CXCR1 / 1, C5aR1, and the like) can bind or an antibody or antibody fragment that can bind to the cell-surface molecule is fused to a ligand for an internalizing receptor or an antibody or antibody fragment that can bind to the internalizing receptor or membrane protein. In embodiments, after binding, the bispecific modulator can cause the cell-surface molecule (e.g., proteins of interest, such as GPCR) to be internalized by the GPCR-expressing cell and, in some embodiments, the internalized cell-surface molecule can be degraded. In various embodiments, the bispecific modulators can target single-pass or multi-pass membrane proteins. In some embodiment, the bispecific modulators can target seven –(pass) membrane proteins.

[0014] In some embodiments, multimers of the bispecific modulators are described herein. The multimers can be homomultimers. The multimers can be heteromultimers. In some embodiments, the multimers can be dimers. In some embodiments, the dimers can be homodimers. In some embodiments, the dimers can be heterodimers. In some embodiments, the multimers (e.g., dimers) can have a means for multimerizing the bispecific modulators. In some embodiments, the means for multimerizing can be one or more disulfide bonds between cysteine amino acid residues of separate bispecific modulator molecules. In some embodiments, the multimers can include 3 monomers. In some embodiments, the multimers can include 4 multimers.

[0015] Disclosed are fusion proteins of Formula I, R1-R2-R3 (I), wherein R1 is a protein of interest (POI) binder (POIB) or POI binding means (also known as a means for binding POI) forDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 binding Apelin Receptor (APJ), Angiotensin II Receptor Type (AT1R), Relaxin Family Peptide Receptor 1 (RXFP1), Adenosine A2A Receptor (A2AR), C-C Chemokine Receptor Type 6 (CCR6), G Protein-Coupled Receptor 20 (GPR20), C-C Motif Chemokine Receptor 4 (CCR4), Protease-Activated Receptor 2 (PAR2), Frizzled Class Receptor 1 (FZD10), C-C Motif Chemokine Receptor 2 (CCR2), C-X-C Motif Chemokine Receptor 1 (CXCR1 / 1) or Complement Component 5a Receptor 1 (C5aR1). R2 is a linker of the formula R4-R5 or R5-R4, wherein, R4 is IgG Fc region, and R5 is an optional protease-sensitive linking means (also known as a means for linking), and R3 is a TR binding means (also known as a means for binding TR), wherein the TR binding means (also known as a means for binding TR) is non- competitive with endogenous transferrin for binding to the TR, and optionally, comprising a glycine-rich linker between R2 and R3.

[0016] The disclosed fusion proteins (or homodimers and heterodimers thereof) can have a TR binding means (also known as a means for binding TR) that is VHHA-12 (SEQ ID NO: 154), VHHA (SEQ ID NO: 146), VHHA-5 (SEQ ID NO: 155), VHHA-7 (SEQ ID NO: 156), or VHHB (SEQ ID NO: 148). The disclosed fusion proteins can have a protease-sensitive linking means (also known as a means for linking) that is a cathepsin-cleavable peptide. The cathepsin- cleavable peptide linker can be FK, VA, VK, SEQ ID NO: 7, 8, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144 and 145.

[0017] The disclosed fusion proteins can be homodimers. The TR binding means (also known as a means for binding TR) within the homodimer can have a binding affinity to TB (Kd) of about 0.001-50 nM.

[0018] The disclosed fusion proteins (e.g., R1-R2-R3 (I)) can be heterodimers linked to a fusion protein of Formula II, R4’-R3’ (II), wherein R4’ is IgG Fc region, and R3’ is a TR binding means (also known as a means for binding TR), and optionally, wherein a linkage between R3’ and R4’ is a protease-sensitive linking means (also known as a means for linking). The TR binding means (also known as a means for binding TR) within the heterodimer can have a binding affinity to TB (Kd) of about 0.1-200 nM.

[0019] The disclosed fusion proteins can have a TR binding means (also known as a means for binding TR) that is an antibody or polypeptide. The TR binding means (also known as a means for binding TR) can be an antibody fragment having a heavy chain variable region. TheDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 TR binding means (also known as a means for binding TR) can have: (a) a VH CDR1 having the amino acid sequence of GTDFSINF (SEQ ID NO: 175), a VH CDR2 having the amino acid sequence of ITATGNT (SEQ ID NO: 176), and a VH CDR3 having the amino acid sequence of YMLDK (SEQ ID NO: 177); (b) a VH CDR1 having the amino acid sequence of GTDFSINF (SEQ ID NO: 175), a VH CDR2 having the amino acid sequence of FTATGNT (SEQ ID NO: 179), and a VH CDR3 having the amino acid sequence of AMLDK (SEQ ID NO: 180); or (c) a VH CDR1 having the amino acid sequence of GTDFSINF (SEQ ID NO: 175), a VH CDR2 having the amino acid sequence of FTATGNT (SEQ ID NO: 179), and a VH CDR3 having the amino acid sequence of YMADK (SEQ ID NO: 183).

[0020] The disclosed fusion proteins can have a POIB or POI binding means (also known as a means for binding POI) that is an antibody. The antibody can bind to an extracellular domain of a transmembrane protein. The POIB or POI binding means (also known as a means for binding POI) can bind to an extracellular domain of APJ, AT1R, RXFP1, A2AR, CCR6, GPR20, CCR4, PAR2, FZD1, CCR2, CXCR1 / 1 or C5aR1.

[0021] The POIB or POI binding means (also known as a means for binding POI) of the disclosed fusion proteins can bind AT1R and can have a VH CDR1 that has the amino acid sequence of GYIYSRY (SEQ ID NO: 187), a VH CDR2 that has the amino acid sequence of SGGSS (SEQ ID NO: 188), and a VH CDR3 that has the amino acid sequence of YKIDSNPRVY (SEQ ID NO: 189). The POIB or POI binding means (also known as a means for binding POI) can bind APJ and can have a VH CDR1 comprising the amino acid sequence of GSTYSSH (SEQ ID NO: 190), a VH CDR2 that has the amino acid sequence of TRSRG (SEQ ID NO: 191), and a VH CDR3 that has the amino acid sequence of VPRAGIESGAYCKWNMKDSGS (SEQ ID NO: 192). The POIB or POI binding means (also known as a means for binding POI) can bind RXFP1 and can have a VH CDR1 that has the amino acid sequence of GNISRRV (SEQ ID NO: 193), a VH CDR2 that has the amino acid sequence of DLGGN (SEQ ID NO: 194), and a VH CDR3 that has the amino acid sequence of TYIDSDGYDYPHIY (SEQ ID NO: 195).

[0022] The disclosed fusion proteins can have a glycine-rich linker that is SEQ ID NO: 9, 10, 11, 12, 13, 14, 15 or 16.

[0023] Disclosed are homodimers of a fusion protein of Formula I, R1-R2-R3 (I),DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 Wherein R1 is a of interest (POI) binder (POIB) or POI binding means (also known as a means for binding POI) for binding APJ, AT1R, RXFP1, A2AR, CCR6, GPR20, CCR4, PAR2, FZD1, CCR2, CXCR1 / 1 or C5aR1, R2 is a linker of the formula R4-R5 or R5-R4, wherein R4 is IgG Fc region, and R5 is an optional protease-sensitive linking means (also known as a means for linking), and R3 is a TR binding means (also known as a means for binding TR), wherein the TR binding means (also known as a means for binding TR) is non-competitive with endogenous transferrin for binding to the TR, and optionally, the homodimer of the fusion protein has a glycine-rich linker between R2 and R3 is a glycine-rich linker. In the homodimer of a fusion protein TR binder or TR binding means can be VHHA-5 (SEQ ID NO: 155), VHHA (SEQ ID NO: 146), VHHA-7 (SEQ ID NO: 156), VHHA-12 (SEQ ID NO: 154) or VHHB (SEQ ID NO: 148). The homodimer of a fusion protein can have a disulfide bond between cysteine amino acids in R4 of separate fusion proteins.

[0024] Disclosed are homodimers of a fusion protein of Formula III, R1-R6-R3 (III), wherein R1 is a protein of interest (POI) binder (POIB) or POI binding means (also known as a means for binding POI) for binding APJ, AT1R, RXFP1, A2AR, CCR6, GPR20, CCR4, PAR2, FZD1, CCR2, CXCR1 / 1 or C5aR1, R6 is a dimerization means (also known as a means for dimerizing), and R3 is a TR binder or TR binding means (also known as a means for binding TR), wherein the TR binder or TR binding means (also known as a means for binding TR) is non-competitive with endogenous transferrin for binding to the TR and wherein the homodimer optionally has a protease-sensitive linking means (also known as a means for linking) between R1 and R6.

[0025] Disclosed are heterodimer of fusion proteins of Formula IV and V (R1-R2 (IV); R3- R2’ (V)) wherein R1 is at least one protein of interest (POI) binder (POIB) or POI binding means (also known as a means for binding POI) for binding APJ, AT1R, RXFP1, A2AR, CCR6, GPR20, CCR4, PAR2, FZD1, CCR2, CXCR1 / 1 or C5aR1, R2 and R2’ are linkers of the formula R4 or optionally R4-R5, wherein R4 is an IgG Fc region, R5 is an optional protease-sensitive linking means (also known as a means for linking), and R3 is at least one TR binder or TR binding means (also known as a means for binding TR, wherein the TR binding means (also known as a means for binding TR) is non-competitive with endogenous transferrin for binding to the TR. The R4 of separate fusion proteins in the heterodimer can have complementary knob and hole structures.DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17

[0026] Disclosed are heterodimers of fusion proteins of Formula I and VI (R1-R2-R3 (I); R2’ (VI)), wherein R1 is a protein of interest (POI) binder (POIB) or POI binding means (also known as a means for binding POI) for binding APJ, AT1R, RXFP1, A2AR, CCR6, GPR20, CCR4, PAR2, FZD1, CCR2, CXCR1 / 1 or C5aR1, R2 is a linker of the formula R4-R5 or R5-R4, wherein R4 is an IgG Fc region, R5 is an optional protease-sensitive linking means (also known as a means for linking), R3 is at least one TR binder or TR binding means (also known as a means for binding TR), wherein the TR binder or TR binding means (also known as a means for binding TR) is non-competitive with endogenous transferrin for binding to the TR, and R2’ is a linker of the formula R4 or optionally R4-R5.

[0027] Disclosed are nucleic acid sequences encoding the fusion proteins and homodimers and heterodimers thereof disclosed herein.

[0028] Disclosed are methods for treating a subject that has a cancer, cardiovascular disease, kidney disease, neurological disease, respiratory disease or autoimmune disease associated with a GPCR, the method being administering the disclosed fusion proteins / homodimers / heterodimers to the subject. The subject can have type 2 diabetes mellitus, Alzheimer’s disease, obesity or depression. The disclosed fusion proteins can be for use in treating a cancer, cardiovascular disease, kidney disease, neurological disease, respiratory disease or autoimmune disease associated with a GPCR in a patient. Disclosed are uses of the disclosed fusion proteins for treating a subject having a cancer, cardiovascular disease, kidney disease, neurological disease, respiratory disease or autoimmune disease associated with a GPCR.

[0029] Disclosed are pharmaceutical compositions of any of the fusion proteins, and homodimers or heterodimers thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Certain illustrations, charts, or flow charts are provided to allow for a better understanding for the present invention. It is to be noted, however, that the drawings illustrate only selected embodiments of the inventions and are therefore not to be considered limiting of scope. Additional and equally effective embodiments and applications of the present invention exist.

[0031] FIGS.1A, 1B-C, 1D illustrates PD-L1 degradation using TransTACs (fusion proteins, fusion protein homodimers, fusion protein heterodimers) variants with various geometries. FIG.DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 1A illustrates PD-L1 TransTAC variants. FIG.1B shows results of an SDS-PAGE showing protein expression of the TransTAC variants. FIG.1C illustrates results of Western Blot showing PD-L1 degradation for each of the TransTAC variants. FIG.1D shows PD-L1 dose curve degradation for variants v1.4 (top) and V1.2 (bottom).

[0032] FIG.2 illustrates PD-L1 degradation. FIG.2A illustrates Tf-TfR1 binding to TfR1 and shows PD-L1 degradation using VHHA (top) or VHHB (bottom) as TfR1 binders (transferrin receptor binders or TRBs). FIG.2B shows Tf-Tfr1 binding interface analysis for TransTACs including second moiety including H7, VHHA and VHHB binders.

[0033] FIGS.3A, 3B shows amino acid sequences corresponding to TransTAC variants v1.2 (FIG.3A, SEQ ID NO: 150-151) and v1.4 (FIG.3B, SEQ ID NO: 152-153). In the sequences, yellow is POIB (R1), blue is R4 (IgG Fc region), green is TRB (R3). Unshaded regions are either leader sequences or GS linkages.

[0034] FIGS.4A, 4B. FIG.4A shows IC50 values for PD-L1 degradation by TransTAC variants that include H7, VHHA or VHHB as second moiety binders (e.g., TRBs). FIG.4B shows IC50 for VHHA TransTAC variant.

[0035] FIGS.5A-D illustrates pharmacokinetics of TransTACs. FIG.5A shows a schematic of the experimental design used to assess the in vivo half-life and safety of EGFR-TransTACs. FIG.5B shows weight over time of mice treated as described in FIG.5A. N=3 per treatment group. FIG.5C shows mice erythrocyte cell counts and other relevant properties measured at day 5 of the experimental designed described in FIG.5A. N=3 per treatment group. RBC: Red blood cell counts; Hb: Hemoglobin levels; HCT: Hematocrit counts; MCV: Mean corpuscular volume, which refers to the average size and volume of a red blood cell; MCH: Mean corpuscular hemoglobin, which indicates the amount of hemoglobin per red blood cell; CHC: Cell hemoglobin concentration; RDW: Red cell distribution width. Results in b and c indicated no observable toxic effects from the injected proteins. The error bars represent the standard deviation. FIG.5D shows Western blot and quantification of plasma levels of the proteins injected in mice according to the different treatments described in FIG.5A. N = 3 per treatment group. The error bars represent standard deviation. i.p.: intraperitoneal injection

[0036] FIG.6 illustrates an example approach for controlling T cell activation using targeted CD19 binding receptor internalization and / or degradation (fusion proteins, including TransTACDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 molecules) (TransTAC is Transferrin receptor-mediated TArgeting Chimera), as disclosed herein.

[0037] FIG.7 illustrates an example schematic of TransTAC technology. When applied to cells of the system is called TransTAC (TransTAC is Transferrin receptor-mediated TArgeting Chimera, e.g., a type of fusion protein). Extracellular proteins (e.g., target membrane proteins with extracellular domains present in the surface of cells) can be selectively internalized and degraded by tethering, for example, an antibody to (or a ligand of) the target membrane protein to a transferrin receptor with a fusion protein, shown as a membrane protein-specific antibody- transferrin fusion protein.

[0038] FIGS.8A-B illustrates the characterization of EGFR-TransTAC cleavage in HEK cells expressing EGFR only, TfR1 only, or both EGFR and TfR1. Western blot (FIG.8A) and quantification of FIG.8A (FIG.8B). Recombinant TransTAC - / + cathepsin B was used as a control. Cells were treated with TransTACs for 24 hours. N = 2-3 biologically independent experiments.

[0039] FIGS.9A-B, 9C, 9D, 9E show TransTACs degraders for various proteins of interest (POIs). FIG.9A shows a schematic of exemplary membrane proteins targeted by TransTACs. These targets are either synthetic, or native, single- or multi-pass proteins expressed on cells surfaces. FIG.9B shows PD-L1 degradation by TransTACs in MDA-MB-231 cells analyzed by Western blot. A scFv format of atezolizumab is used as the PD-L1 binding moiety. FIG.9C shows EGFR degradation by TransTAC in A549 cells. An affibody is used as the EGFR binding moiety. FIG.9D shows CD20 degradation by TransTAC. A Fab format of rituximab is used as the CD20 binding moiety. FIG.9E shows CD19 binding receptor degradation by an TransTAC. An engineered CD19 ectodomain is used as the CAR binding moiety. All cells were incubated with TransTACs for 12-18 hours. Data are representative of 3 independent experiments. Controls without the TfR1 binding domain or the cathepsin cleavable linker showed no or little cleavage. Error bars represent standard deviations. P values were determined by unpaired two- tailed t tests. ns, not significant; * P ≤0.05; **P ≤0.01; ***P ≤0.001.

[0040] FIG.10 illustrates results showing expression of an anti-EGFR affibody-Fc-Tr TransTAC molecule (Left) and the effect on EGFR levels of incubating the TransTAC molecule with a MCF10A EGFR expressing cell line (Right).DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17

[0041] FIGS.11A-B illustrates results showing the effect on EGFR levels of incubating a TransTAC molecule with A549 cells (A) and killing of MCF10A EGFR cells (B).

[0042] FIG.12 illustrates results showing that TransTAC targeting effectively internalized the receptor.

[0043] FIG.13 illustrates example results showing expression of various TransTAC proteins in cultured cells.

[0044] FIG.14 illustrates results showing TransTAC targeting of T cells expressing a CD19 binding receptor (CD19 binding T cells) effectively decreased levels of the CD19 binding receptor (e.g., internalized the CD19 binding receptor).

[0045] FIG.15 illustrates example results showing that TransTAC targeting of a CD19 binding T cell effectively internalized the CD19 binding receptor.

[0046] FIG.16 illustrates example results showing that TransTAC targeting of a CD19 binding receptor on Jurkat cells, in the presence of K562 cells, inhibited activation of the Jurkat cells.

[0047] FIG.17 illustrates example results showing TransTAC targeting of a CD19 binding receptor internalized the CD19 binding receptor.

[0048] FIG.18 illustrates results showing that a TransTAC molecule specific for CD19 blocks the activation of CD19 binding T cells in presence of K562 cells and also shows that the TransTAC molecule had minimal effect on the Jurkat cells in absence of the K562 cells.

[0049] FIGS.19A-B, 19C illustrates a schematic diagram of an ImmuneTransTAC molecule that can bind a CD19 binding receptor and an internalizing receptor, and a dimer of a TransTAC molecule that can bind a T cell receptor and an internalizing receptor (A), and results of using the molecules on cells expressing the CD19 binding receptor on the levels of the CD19 binding receptor (B). FIG.19C illustrates fluorescence microscopy of the targeted CD19 binding receptors on the cells in FIG.19B.

[0050] FIGS.20A-B, 20C, 20D-E, 20F-G, 20H and 20I illustrate an embodiment where an internalized CD19 binding receptor is not degraded, but linker engineering (here, incorporating a cathepsin-sensitive linker) resulted in degradation of the CD19 binding receptor. (A) shows schematic diagrams of various molecules used in this study. GFLG indicates a Gly-Phe-Leu-Gly peptide linker which is sensitive to lysosomal cathepsin proteases. (B) shows Western blots of the targeted CD19 binding receptor (Anti-CD3z) and actin control (β-Actin) when various of theDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 molecules in (A) were used. (C) shows a graph of the data from (B), where CD19 binding receptor levels are normalized to β-Actin levels. (D) shows Western blots as above, using other of the molecules in (A). (E) shows a graph of the normalized data from (D). (F) shows a schematic of a dimer of a TransTAC molecule that can bind a CD19 binding receptor and an internalizing receptor, which also contains GFLG linkers. (G) shows Western blots using the molecule shown in (F). (H) shows a graph of the normalized data from (F). (I) shows results from screening additional cathepsin-sensitive TransTAC molecules that have improved inhibition potency.

[0051] FIGS.21A-B shows results demonstrating TransTAC inhibition of T cell activity is more potent than inhibition by ImmuneTrap (domain to which CD19 binding receptor can bind fused to an Fc region) in Jurkat cells (A) and in primary T cells (B).

[0052] FIG.22A and FIG.22B show results illustrating that CD19 binding receptor TransTAC that binds to CD19 receptor turns off A375 cell killing of the T cells expressing a CD19 binding receptor, and that A375 cell killing of the -T cells resumes when the TransTAC molecule is removed.

[0053] FIGS.23A-B shows schematic diagrams of molecules used in the study, including affibody-based EGFR TransTAC molecules (A). (B) shows results of removing EGFR from the surface of A549 cells using the molecules shown in (A). The data show that use of the affibody- based TransTAC molecule produces good results (approximately 10-50-fold improvement in IC50).

[0054] FIGS.23C-D shows results of inhibition of cell proliferation by the molecules shown in (A), as measured using the MTT cellular proliferation assay. The data show that use of the affibody-based TransTAC molecule produces good results (approximately 10-50-fold improvement in IC50).

[0055] FIGS.24A-B shows a schematic of molecules used in this study (B) and western blot results measuring internalization and degradation of the molecules (C).

[0056] FIGS.25A-B illustrates example data demonstrating protein internalization by TransTAC and reversibility of the internalization.

[0057] FIGS.26 shows example data demonstrating TransTAC can interfere with IFNγ production.DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17

[0058] FIGS.27A and 27B show example data of transferrin receptor expression on various cells.

[0059] FIGS.28 A-B illustrates CD20 degradation kinetics. FIG.28A shows time-course measurement of whole-cell CD20 levels in Raji B cells treated with 25 nM CD20-TransTAC. FIG.28B shows reversibility of degradation. Cells were treated with TransTAC for 12-16 hours before washout.

[0060] FIGS.29A and 29B show example data demonstrating that TransTACs can degrade EGFR in cells and the underlying cellular machinery that mediates the degradation.

[0061] FIGS.30A and 30B-C show EGFR internalization using with TransTACs.

[0062] FIG.31 shows example data demonstrating that TransTACs with the linker variants can degrade receptors in Jurkat cells.

[0063] FIGS.32A and 32B shows example data demonstrating that TransTACs can degrade PD-L1.

[0064] FIG.33 shows example data demonstrating that TransTAC can degrade CD20.

[0065] FIGS.34A-D show example TransTAC molecules that include protease-sensitive linkers and example data obtained with the molecules.

[0066] FIG.35 shows example data obtained with TransTAC molecules containing various protease-sensitive linkers.

[0067] FIGS.36A-C show example TransTAC molecules that include an antibody fragment specific for transferrin binding and example data obtained with the molecules.

[0068] FIGS.37A-B show examples of TransTAC molecules and example data obtained with the molecules.

[0069] FIGS.38A-E and 38F-H show an example overview of the TransTAC technology and TfR expression analysis. (A) Schematic of the example TransTAC technology. TransTAC induces close proximity of TfR and POI at the cell surface, leading to co-internalization of the complex to early endosomes (EE), where a cathepsin enzyme cleaves TransTAC and separates the POI from the TfR. The POI then traffics to late endosomes (LE) / lysosomes for degradation, while TfR is recycled back to the cell surface. (B) Illustration of an example TransTAC protein. Some example designs to make TransTACs efficient degraders include: (1) containing two anti- TfR binders for binding and priming a TfR dimer for endocytosis, (2) having a cathepsin B- sensitive linker between the anti-POI binder and the Fc for endosomal cleavage to separate theDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 POI from the recycling TfRs, and (3) using an antibody binder instead of a native TF ligand to reduce trafficking to the recycling endosomes (REs). (C) Relative cell surface TfR expression levels across various cell lines characterized by flow cytometry. Data are representative of 3 independent experiments. (D) Relative TFRC RNA expression levels in diverse tissues based on the MERAV database. (E) Relative TFRC RNA expression levels in native T cells. TfR is upregulated by approximately 6-fold in activated CD4 and CD8 T cells compared to inactivated T cells with statistical significance (p=1.25e-68 for CD4 T cells and 4.81e-68 for CD8 T cells, FIG.38H).

[0070] FIGS.39A-H, 39I-L shows example TransTAC degrader engineering. (A) Schematic of example CD19 binding receptor-TransTACs and control. TransTACv0.1 has a single CD19NT.1 domain, a single TF, and a knob-in-hole (KIH) Fc, v0.2 has two CD19NT.1s, two TFs, and a homodimeric Fc that connects the binders, v0.4 contains a cathepsin-sensitive linker between CD19NT.1 and Fc, v0.5 contains a H7 scFv for TfR binding, v1.0 contains both the H7 and the cathepsin sensitive linker. (B) Schematic of a myc-tagged anti-CD19 receptor. (C) Flow cytometry measurements of cell-surface CD19 binding receptor expression levels in -Jurkat cells treated with TransTACv0.1, v0.2, and control. TransTACv0.2 results in higher CD19 binding receptor clearance from cell surface than v0.1 and no hook effect. Data are representative of 2 independent experiments. (D) Characterization of whole-cell CD19 binding receptor levels by Western blot in Jurkat cells expressing the CD19 binding receptor and treated with TransTACs. TransTACv1.0 degrades approximately 80% of CD19 binding receptor; v0.2 didn’t result in significant CD19 binding receptor degradation. (E-H) Schematics showing different TransTACs alter intracellular trafficking of the POI. Cleavage of the cathepsin sensitive-linker in v0.4 and v1.0 leads to separation of POI from TfR, hence enhanced LE / lysosomal trafficking of the POI and degradation; the H7 scFv in v0.5 and v1.0 reduces trafficking of the complex to the REs, hence increases the proportion of POI in EEs and subsequent proteolytic processing, when a cleavage linker is present. (I, J) Representative fluorescence images of Hela cells co-expressing CD19 binding receptor-GFP (green) and endosomal / lysosomal markers-mCherry (red) treated with various TransTAC molecules. Cell nucleus is stained with Hochest (blue). Untreated (UT) or control-treated cells had CD19 binding receptor-GFP localized at the cell membrane. v0.5 and v1.0 led to efficient degradation of CD19 binding receptor-GFP, manifested by the significantly lower GFP signals. v0.2-treated cells predominantly trafficked CD19 binding receptor to theDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 REs, showing co-localization of CD19 binding receptor -GFP with mCherry-Rab11 (white arrows). v0.5-treated cells trafficked CD19 binding receptor to the EEs, showing co-localization of CD19 binding receptor -GFP with mCherry-Rab5 (white arrows). (K) Pearson correlation analysis of CD19 binding receptor -GFP colocalization with the Rab5 (EE), EEA1 (EE), and Rab11 (RE) markers. T-tests show Rab5, EEA1, Rab11 colocalization with CD19 binding receptor are statistically different for cells treated with v0.2 vs. v0.5. (L) Pearson correlation analysis of CD19 binding receptor -GFP colocalization with the Rab7 (LE) and Lamp1 (lysosome) markers. T-tests show Rab7 and Lamp1 colocalization with CD19 binding receptor are statistically significant for v0.2 vs. v0.4, and v0.5 vs. v1.0. For k and l, number of cells used for each analysis are as follows: For v0.2, N=12, N=12, and N=13 for the EEA1, Rab5 and Rab11 markers, respectively. For v0.5, N=10, N=22, and N=15 for the EEA1, Rab5 and Rab11 markers respectively. For v0.2, v0.4, v0.5, and v1.0 with the Lamp1 marker N=16, N=21, N=13, and N=13, respectively.

[0071] FIGS.40A-D shows examples of developing TransTACs degraders for various membrane targets. (A) Schematic of membrane proteins targeted by TransTACs in the present study. These targets are either synthetic, or native, single- or multi-pass proteins expressed on cell surface. (B) PD-L1 degradation by TransTACs in MDA-MB-231 cells analyzed by Western blot. A scFv or Fab format of atezolizumab is used as the PDL1 binding moiety. (C) EGFR degradation by TransTAC in A549 cells. An affibody is used as the EGFR binding moiety. (D) CD20 degradation by TransTAC. A Fab format of rituximab is used as the CD20 binding moiety.

[0072] FIGS.41A-H and 41I show example structure-activity relationship (SAR) studies of TransTACs, mechanisms, and in vivo characterizations. (A) Time-course measurement of cell surface CD19 binding receptor levels in CD19 binding receptor -Jurkats treated with TransTACs, revealing the fast kinetics of TransTAC-mediated CD19 binding receptor internalization. (B) Schematics of TransTAC variants consisting of one or two copies of anti-POI and anti-TfR binders in different protein geometries. (C) Cell surface CD19 binding receptor level measurements in CAR-Jurkats treated with TransTAC variants outlined in (B). The results highlight the impacts of having two vs. one TfR binders (v0.5 vs v0.7) and geometry (v0.8 vs. v0.9) in modulating protein internalization. Data are representative of 3 independent measurements. (D) Competition assay with a H7-Fc fusion protein. Concentration-dependentDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 reduction of receptor internalization is observed with H7-Fc, proving internalization is mediated through TfR. Data are representative of 3 independent measurements. (E) Study of underlying degradation pathways with TransTACs. Intact lysosomal function is critical for degradation, as degradation is fully inhibited by bafilomycin in A549 cells treated with EGFR-TransTACs. (F) Whole-cell TfR level measurement with TransTAC treatment. TfR level stays consistent while PD-L1 is degraded in MDA-MB-231 cells treated with PDL1 TransTAC. (G) Schematic of mouse experiments to assess TransTAC safety and serum half-life via IP injection. (H) Weight monitoring of mice over time after TransTAC or control IgG injection. Results reveal no observable effects on mouse weight over time, showing molecules are well tolerated. N=2 per treatment group. (I) Western blot quantification of plasma levels of CD20-TransTAC and IgG control over time. N=2 per treatment group.

[0073] FIGS.42A-C illustrates dependency of TransTACs on TfR1 for target degradation. FIG.42A shows cell-surface levels of TfR1 in siTfR-1 and siTfR-2 siRNAs treated cells. FIG. 42B shows EGFR degradation in siTfR-1 or 2 transfected PC9 Del19 cells. Cells were treated with TransTACs for 18-24 hours. FIG.42C shows EGFR degradation in A549 cells treated with bafilomycin and chloroquine (CQ) with EGFR-TransTACs, but not with MG132.10 µM MG132 conditions were not quantified due to observed cytotoxicity effects in the cells. Cells were treated with TransTACs for 12-16 hours and inhibitors were added 1 hour before TransTAC treatment.

[0074] FIGS.43A-B shows example characterization of Fc fusions of wildtype (WT) CD19 ectodomain and the variants. CD19ecto-WT-Fc shows aggregations in SDS-PAGE gel while the variants derived from yeast display do not. Among the four variants, CD19NT.1 is selected for TransTAC engineering given its high expression level.

[0075] FIGS.44A-E shows example different CAR degradation efficiencies mediated by TransTAC variants. (A) Schematics of different generations of CAR-TransTACs and the CD19NT.1-Fc control. (B) Western blots showing neither the control nor v0.2 leads to CAR degradation. (C) Western blots showing v0.4-GFLG, which contains a cathepsin sensitive GFLG linker between the CD19NT.1 and Fc domains, leads to approximately 40-50% of CAR degradation. v0.3-GFLG, which contains the cleavable linker between the Fc and the TF domains, doesn’t lead to significant receptor degradation, possibly due to Fc mediated receptor recycling. (D) Western blots showing different linker variants of v0.4 led to varying receptorDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 degradation efficiencies. (E) Western blots showing different linker variants of v1.0 led to varying receptor degradation efficiencies. Among all variants, linker GFLG-VR and VR show highest degradation.

[0076] FIGS.45A-B, 45C-D and 45E-F shows example colocalization analysis of internalized receptors with various endosomal / lysosomal markers. (A-C) Representative fluorescence images of Hela cells co-expressing Receptor-GFP (green) and endosomal / lysosomal markers-mCherry (red), treated with various TransTACs or controls. EEA1: EE marker, Rab7: LE marker, Lamp1: lysosomal marker. Cell nucleus is stained with Hochest (blue). (D) Pearson correlation analysis of Receptor-GFP colocalization with the five endosomal / lysosomal markers. T-tests show Rab5, EEA1, Rab11 colocalization with CAR are statistically different for cells treated with v0.2 vs. v0.5, and Rab7 and Lamp1 colocalization with CAR are statistically significant for v0.2 vs. v0.4, and v0.5 vs. v1.0. Cell numbers used for the analysis are as follows: For the control, UT, v0.2, v0.4, v0.5, and v1.0, N=5, N=4, N=12, N=15, N=10, and N=11, respectively for the EEA1 marker. For the control, UT, v0.2, v0.4, v0.5, and v1.0, N=12, N=11, N=12, N=15, N=22, and N=17, respectively for the Rab5 marker. For the control, UT, v0.2, v0.4, v0.5, and v1.0, N=9, N=7, N=8, N=12, N=26, and N=11, respectively for the Rab7 marker. For the control, UT, v0.2, v0.4, v0.5, and v1.0, N=9, N=6, N=13, N=13, N=15, and N=22, respectively for the Rab11 marker. For the control, UT, v0.2, v0.4, v0.5, and v1.0, N=17, N=12, N=16, N=21, N=13, and N=13, respectively for the Lamp1marker. (E-F) Incorporating a cathepsin sensitive linker into TransTAC enhances LE / lysosomal trafficking. Representative fluorescence images of Hela cells co-expressing Receptor-GFP (green) and mCherry-Rab7 or Lamp1-mCherry (red), treated with TransTACv0.2 vs v0.4. Cell nucleus is stained with Hochest (blue). The v0.4 GFP images were collected with 1000x more exposure than the v0.2 images to get sufficient GFP signals for the Pearson coefficient analysis in Fig.2i. v0.4-treated cells show colocalization of the internalized Receptor-GFP with mCherry-Rab7 and Lamp1 (white arrows).

[0077] FIGS.46A-C shows example characterizing TransTAC degraders for various membrane proteins. Different linkers and geometry designs lead to varied degradation efficiencies. (A) Western blots showing controls or v0.2 and v0.4 PDL1 TransTAC variants do not lead to much target degradation in MDA-MB-231 cells. (B) Western blots showing EGFR TransTACv0.2 does not cause much target degradation in A549 cells, whereas v0.4 or v1.0 withDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 different linkers cause varying degrees of degradation, with v1.0-EVR and GFLG-VR giving the highest degradation efficiency. (C) Western blots showing a rituximab-scFv-Fc control does not lead to significant CD20 degradation in Raji cells.

[0078] FIGS.47A-E shows examples of TransTAC regulating primary T cell activities. (A) Schematic of using -TransTAC to reversibly control T cells. TransTAC-mediated removal of CD19 binding receptor from cell surface prevents T cells from engaging with CD19+ cells, hence inhibits cytokine release and cytotoxicity. (B) Schematic of the setup of a primary T cell co-culture assay. Secreted IFN-^^ levels are measured to determine T cell activation levels in the presence if CD19+ A375 cells and TransTACs; live cell fluorescence microscopy is used to cell killing. (C) Measurement of human primary CD19 binding receptor T cell IFN-^^ release in the co-culture assay described in (b) with an IFN^^ split-luciferase assay (Promega). IFN-^^ secretion is inhibited by TransTACv0.4 in a dose-dependent manner. TransTAC shows an IC500f approximately 0.4 nM. Data are representative of 2 independent experiments. (D) Fluorescence microscopy of mCherry-labeled A375 cells showing T cell-mediated A375 killing reversibly controlled with CD19 binding receptor-TransTACv4. (E) Overlay of bright field and mCherry channel images showing T cell-mediated A375 killing activity was resumed after TransTAC washout over time.

[0079] FIGS.48A-B. FIG.48A illustrates PD-L1 degradation in MDA-MB-231 cells treated with bafilomycin and chloroquine (CQ), or MG132, and with PD-L1-TransTACs. Cells were treated with TransTACs for 12-16 hours and inhibitors were added 1 hour before TransTAC treatment. FIG.48B shows CD20 degradation in bafilomycin treated Raji B cells. Cells were treated with TransTACs for 12-16 hours and inhibitors were added 1 hour before TransTAC treatment.

[0080] FIGS.49A, 49B shows (A) Cleavage of the indicated linkers on yeast at pH 4.4 by recombinant cathepsin B as compared to cleavage of GFLGGVR (SEQ ID NO: 144). (B) Cleavage of the indicated linkers on yeast at pH 6.4 by recombinant cathepsin B as compared to GFLGGVR (SEQ ID NO: 144).

[0081] FIG.50 shows amino acid and nucleotide sequences for VHHA and VHHB.

[0082] FIG.51 illustrates mechanism of action of GPCR TransTACs.

[0083] FIG.52 illustrates a Doxorubicin inducible system for expression of FLAG-tagged GPCRs APJ, GPR20, CCR4 and PAR2.DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17

[0084] FIGS.53A-B illustrates degradation of AT1R or APJ by GPCR TransTACs. FIG.53A shows FLAG-AT1R degradation by GPCR TransTACs including a VHH that binds AT1R (AT1R VHH AT118-L), human IgG1 Fc (H435A), and second moiety binder that binds to TfR (TfR scFv H7) KR54 and SEQ ID NO: 155; FIG.57). A GPCR TransTAC without the TfR scFv H7 binder was used as a control (KR53 and SEQ ID NO: 154; FIG.57). Dox = doxorubicin. FLAG = tag. FIG.53B shows FLAG-APJ degradation by GPCR TransTACs including a VHH that binds APJ (APJ VHH JN241), human IgG1 Fc (F372L), and second moiety binder that binds to TfR (TfR scFv H7) (DP238 and SEQ ID NO: 157; FIG.58). A GPCR TransTAC without the TfR scFv H7 binder was used as a control (JD22 and SEQ ID NO: 156; FIG.58). Dox = doxorubicin. FLAG = tag.

[0085] FIG.54 illustrates validation of a titratable FLAG-GPCR expression system for measuring TransTAC binding and efficacy in HEK cells. Induction of FLAG-AT1R using doxorubicin (upper left panel). On-cell binding of AT1R-targeted TransTACs (upper right panel), AT1R depletion for various TransTAC doses (lower left panel), and AT1R depletion over time (lower right panel) using GPCR TransTACs including a VHH that binds AT1R (AT1R VHH AT118-L), human IgG1 Fc (H435A), and second moiety binder that binds to TfR (TfR scFv H7) (KR54 and SEQ ID NO: 155; FIG.57). A GPCR TransTAC without the TfR scFv H7 binder was used as a control (KR53 and SEQ ID NO: 154; FIG.57). Dox = doxorubicin. FLAG = tag.

[0086] FIGS.55A-B illustrates efficient TransTAC-mediated depletion of APJ (FIG.55A), RXFP1 (FIG.55B left panel), and FZD1 (FIG.55B right panel). FIG.55A shows FLAG-APJ depletion for various TransTAC doses (left panel), and FLAG-APJ depletion over time (right panel) using GPCR TransTACs including a VHH that binds APJ (APJ VHH JN241), human IgG1 Fc (F372L), and second moiety binder that binds to TfR (TfR scFv H7) (DP238 and SEQ ID NO: 157; FIG.58). A GPCR TransTAC without the TfR scFv H7 binder was used as a control (JD22 and SEQ ID NO: 156; FIG.58). Dox = doxorubicin. FLAG = tag. FIG.55B shows efficient FLAG-RXFP1 (left panel) depletion by GPCR TransTACs including a VHH that binds RXFP1 (RXFP1 VHH RX002), human IgG1 Fc (G236R, N297G, L328R, D356E, L358M), and second moiety binder that binds to TfR (TfR scFv H7) (KR98 and SEQ ID NO: 159; FIG.59). A GPCR TransTAC without the TfR scFv H7 binder was used as a control (KR98 and SEQ ID NO: 159; FIG.59). FIG.55B (right panel) shows efficient FLAG-HA-FZD1 depletion by GPCRDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 TransTACs including a FLAG HC M2, human IgG1 Fc (G236R, N297G, L328R, D356E, L358M), and second moiety binder that binds to TfR (TfR scFv H7) (KR62.1 and SEQ ID NO: 162; FIG.60); or by GPCR TransTACs including a FLAG HC M2, human IgG1 Fc (G236R, N297G, L328R, D356E, L358M) (KR61.1 and SEQ ID NO: 161; FIG.60)

[0087] FIG.56 illustrates efficient TransTAC-mediated depletion of FZD10. FIG.56 shows dose-response (left panel) and time course (right panel) for FLAG-HA-FZD1 depletion by GPCR TransTACs including a FLAG HC M2, human IgG1 Fc (G236R, N297G, L328R, D356E, L358M), and second moiety binder that binds to TfR (TfR scFv H7) (KR62.1 and SEQ ID NO: 162; FIG.60); or by GPCR TransTACs including a FLAG HC M2, human IgG1 Fc (G236R, N297G, L328R, D356E, L358M) (KR61.1 and SEQ ID NO: 161; FIG.60).

[0088] FIG.57 shows amino acid sequences for AT1R GPCR TransTACs KR53 (SEQ ID NO: 154) and KR54 (SEQ ID NO: 155). As shown in the sequences, yellow is POIB for AT1R (R1), violet is cleavable linker, green is IgG Fc region (R4), blue is TRB (R3), gray is linker.

[0089] FIG.58 shows amino acid sequences for APJ GPCR TransTACs JD22 (SEQ ID NO: 156) and DP238 (SEQ ID NO: 157). As shown in the sequences, yellow is POIB for RXFP1 (R1), violet is cleavable linker, green is IgG Fc region (R4), blue is TRB (R3), gray is linker.

[0090] FIG.59 shows amino acid sequences for RXFP1 GPCR TransTACs KR97 (SEQ ID NO: 158) and KR98 (SEQ ID NO: 159). As shown in the sequences, yellow is POIB for APJ (R1), violet is cleavable linker, green is IgG Fc region (R4), blue is TRB (R3), gray is linker.

[0091] FIG.60 shows amino acid sequences for FLAG TransTACs KR60 (SEQ ID NO: 160), KR61.1 (SEQ ID NO: 161) and KR62.1 (SEQ ID NO: 162). As shown in the sequences, yellow is POIB for FLAG (R1), violet is cleavable linker, green is IgG Fc region (R4), blue is TRB (R3), gray is linker.

[0092] FIG.61A shows PD-L1 degradation by binder and linker GPCR TransTACs variants.

[0001] FIG.61B provides more information on the homodimeric molecules in FIG.6A, which have different TfR binders.

[0093] FIG.62 shows AT1R internalization by GPCR TransTAC variants.

[0094] FIG.63 shows AngII / AT1R and AT1R signaling by GPCR TransTAC variants.

[0095] FIG.64 provides a schematic of AT1R TransTAC variants KR53 and KR54.DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17

[0096] FIG.65 shows amino acid sequences for AT1R TransTAC variants KR53 and KR54. As shown in the sequences, yellow is POIB for At1R (R1), violet is cleavable linker, green is IgG Fc region (R4), blue is TRB (R3), gray is linker.

[0097] FIG.66 shows A2AR internalization by GPCR TransTAC variants.

[0098] FIG.67 provides a schematic of A2AR TransTAC variant KR225.

[0099] FIG.68 provides data showing A2AR TransTAC on-cell binding (A) and cell surface levels (B).

[0100] FIG.69 shows amino acid sequences for A2AR TransTAC KR225. Amino acid sequences for JD27 and PIKA5 are also provided. As shown in the sequences, yellow is POIB for A2AR (R1), green is IgG Fc region (R4), blue is TRB (R3), gray is linker, violet and red are tags.

[0101] FIG.70 provides a western blot assay showing CCR6 is degraded by DJ1 heterodimeric fusion proteins (shown in inset) in doxycycline inducible CCR6-expressing Jurkat cells.

[0102] FIG.71 provides a western blot assay showing degradation of CCR6 by a CCR6 homodimeric fusion protein LS28 (VHH-A homodimer; shown in inset) in doxycycline inducible CCR6-expressing Jurkat cells.

[0103] FIG.72 provides a western blot assay showing no degradation of CCR6 by DJ2 control fusion protein (shown in inset) in doxycycline inducible CCR6-expressing Jurkat cells.

[0104] FIG.73 is a western blot assay showing degradation of CCR6 and prevention of TfR1 co-degradation by CCR6 heterodimeric fusion protein LS13 (VHHA-12 monomer; shown in inset) in doxycycline inducible CCR6-expressing Jurkat cells.

[0105] FIG.74 provides example protein of interest binder (POIB) sequences for CCR6. (A) Amino acid sequence for pLS15 Anti-CCR6_B368-G29A_HC-Knob_Fc-His is shown. (B) Amino acid sequence for pLS16-Anti-CCR6_B3G8-G29A_LC is shown. In the sequences, yellow is POIB for CCR6 (R1) and CDRs are underlined and bolded. Blue is IgG Fc region (R4) DETAILED DESCRIPTION

[0106] Targeted protein degradation (TPD) is a rapidly growing field in drug discovery and pharmacology. Complementing traditional drug modalities, TPD molecules offer a novel therapeutic mechanism to tackle challenging targets, increase the therapeutic potential ofDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 currently used drugs. While many efforts in this field have focused on small molecules for intracellular targets, inducing targeted degradation of extracellular proteins is a new opportunity. Developing strategies to degrade extracellular proteins is of exceptional interest for both basic research and therapeutic intervention purposes.

[0107] Iron is an essential element for cells, and its transportation is facilitated by transferrin receptor (TfR). TfR undergoes rapid endocytosis as a recycling receptor, with an average internalization rate of 500 molecules per cell per second, making it one of the fastest internalizing receptors known. Furthermore, TfR is upregulated in cells that have a high demand for iron. This includes rapidly dividing cells and activated T cells. TfR expression in these cells are higher than in non- or slowly- dividing normal tissues. TfR can be expressed on non-cancer cells at sufficient levels where the reagents and methods described herein can be used.

[0108] Herein, these features of TfR were leveraged and employed protein engineering strategies to develop a new technology for degrading membrane proteins. Herein, this technology is called receptor-mediated Targeting Chimeras (TransTACs; also, herein called fusion proteins, homodimers or heterodimers of fusion proteins). In some embodiments, TransTACs are heterobispecific antibodies that bring the protein of interest (POI) and TfR in close proximity at the cell surface and induce endocytosis of the POI / TfR complex and subsequent lysosomal- mediated POI degradation. In some embodiments, the POI is a G Protein-Coupled Receptor (GPCR). In some embodiments, the TransTAC targets, binds and / or interacts with a GPCR (GPCR Trans TAC) GPCR TransTACs are effective in degrading various types of membrane proteins, including multi-pass, native, and synthetic receptors, showing a degradation efficiency of over 80% for all targets in various cellular systems. A notable characteristic of TransTAC is its fast kinetics of targeted internalization, occurring on a timescale of minutes, making it a valuable molecular tool for rapidly knocking down cell-surface expression, offering temporal specificity for cell signaling studies that is not possible with genetic approaches. Moreover, TransTAC molecules are fully recombinant, modular and GPCR-specific. These properties make GPCR TransTACs a versatile technology for manipulating cell surface targets in disease-specific manners.

[0109] TransTAC can have broad applicability in both basic research and translational applications. Herein is demonstrated non-limiting applications of TransTACs in related disorders. TransTACs represent a new molecular archetype to control cell surface proteins.DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17

[0110] TransTAC is the first fusion protein technology that repurposes a recycling ligand / receptor interaction for targeted protein internalization and degradation, which can significantly expand the scope of effectors at the cell surface amenable for such purposes.

[0111] Herein are disclosed new, modular, and reversible strategies for modulating GPCR activity on cells that express GPCR (i.e., GPCR cells). In some embodiments, the methods require genetic engineering of GPCR cells. In some embodiments, these strategies are based on reversible internalization of cell surface molecules (e.g., GPCR) present in GPCR cells.

[0112] In some embodiments, a bispecific modulator (fusion protein), including for example a transferrin receptor-mediated targeting chimera or GPCR TransTAC molecule, can colocalize a GPCR receptor to an internalizing cell surface protein. In embodiments, fusion proteins can downregulate cell surface levels of GPCR receptors. In some embodiments, the fusion proteins can inhibit activation and / or function of GPCR cells. In some embodiments, the fusion proteins can activate functions of GPCR cells. In some embodiments, the fusion proteins can inhibit functions of GPCR cells. In some embodiments, the fusion proteins can inhibit signaling through intracellular pathways. In some embodiments, the fusion proteins can activate signaling through cellular pathways.

[0113] In embodiments, the fusion proteins can have a first portion or moiety (protein of interest binder or POIB or POI binding means (also known as a means for binding POI), (R1) that is an antibody or an antibody fragment that specifically binds to a target protein on a cell (GPCR), and a second portion or moiety (e.g., transferrin or an antibody or antibody fragment, transferrin receptor binder or TR binder or R3) that can bind to an internalizing molecule (transferrin receptor or TfR) on the cell surface (e.g., transferrin receptor or TR). In embodiments, the fusion proteins can have a first portion (R1) that is an antigen, or ligand, for a target protein on a cell (e.g., GPCR receptor). In embodiments, the fusion proteins can have a first portion (R1) that is an antibody or antibody fragment that can specifically bind a target protein (GPCR) on a cell. The fusion proteins can have a second portion (R3) that binds to an internalizing protein on the cell surface (e.g., transferrin receptor or TR). Binding of a fusion protein to the target protein and to the internalizing protein results in internalization of the target molecule.

[0114] In some embodiments, the fusion proteins can be applied to diseases related therapies that are already approved or in clinical development.DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17

[0115] In other embodiments, the fusion proteins can be reversible. Reversibility of fusion protein activity can provide for fine tuning of cell activities, for example for autoimmune related activities management and / or to activate / inactivate the cells for treatment. Reversibility can provide for fine tuning of GPCR cell activities, for example for toxicity management and / or to activate / inactivate the GPCR cells for continued treatment.

[0116] In some embodiments, fusion proteins can be tailored to GPCR cells that target different intracellular pathways, for example, by replacing the GPCR binding components used in the designs so the fusion protein can recognize any of the GPCR described herein.

[0117] Also disclosed are approaches for enhancing GPCR cells efficacy for controlling and / or treating GPCR related diseases. In some embodiments, by alternating GPCR cells status between an “active” and a “resting” state, the disclosed reversible GPCR modulators can control / treat GPCR related diseases.

[0118] Disclosed are development of GPCR TransTAC degraders to target GPCR related diseases. Herein is shown that (1) GPCR TransTAC can effectively degrade GPCR related proteins and hence treat GPCR disorders, (2) GPCR TransTAC can specifically target cells expressing a specific GPCR (e.g., any of the GPCR described herein) without affecting cells that do not express said GPCR, and (3) that the geometry of TransTAC molecules can influence TransTAC target degrading activities.

[0119] Detailed descriptions of one or more embodiments are provided herein. It is to be understood, however, that the present invention may be embodied in various forms. Therefore, specific details disclosed herein are not to be interpreted as limiting, but rather as a basis for the claims and as a representative basis for teaching one skilled in the art to employ the present invention in any appropriate manner.

[0120] The singular forms “a”, “an” and “the” include plural reference unless the context clearly dictates otherwise. The use of the word “a” or “an” when used in conjunction with the term “comprising” in the claims and / or the specification may mean “one,” but it is also consistent with the meaning of “one or more,” “at least one,” and “one or more than one.”

[0121] Wherever any of the phrases “for example,” “such as,” “including” and the like are used herein, the phrase “and without limitation” is understood to follow unless explicitly stated otherwise. Similarly, “an example,” “exemplary” and the like are understood to be nonlimiting.DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17

[0122] The term “substantially” allows for deviations from the descriptor that do not negatively impact the intended purpose. Descriptive terms are understood to be modified by the term “substantially” even if the word “substantially” is not explicitly recited.

[0123] The terms “comprising” and “including” and “having” and “involving” (and similarly “comprises”, “includes,” “has,” and “involves”) and the like are used interchangeably and have the same meaning. Specifically, each of the terms is defined consistent with the common United States patent law definition of “comprising” and is therefore interpreted to be an open term meaning “at least the following,” and is also interpreted not to exclude additional features, limitations, aspects, etc. Thus, for example, “a process involving steps a, b, and c” means that the process includes at least steps a, b and c. Wherever the terms “a” or “an” are used, “one or more” is understood, unless such interpretation is nonsensical in context.

[0124] As used herein, the term “about” can refer to approximately, roughly, around, or in the region of. When the term “about” is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the numerical values set forth. The term “about” is used herein to modify a numerical value above and below the stated value by a variance of 20 percent up or down (higher or lower). G Protein-Coupled Receptors (GPCR)

[0125] G-protein-coupled receptors (GPCRs), also known as seven-(pass)-transmembrane domain receptors, 7TM receptors, heptahelical receptors, serpentine receptors, and G protein- linked receptor (GPLR) are the largest and most diverse group of membrane receptors in eukaryotes. GPCRs are transmembrane cell surface receptors that can transduce extracellular signals into key physiological effects. GPCR’s endogenous ligands include, but are not limited to odors, neurotransmitters, hormones, chemokines, lipids, amines, carbohydrates, peptides and proteins. In some embodiments, the strategies and / or fusion proteins described herein can be used for regulating molecules present on the cell surface of any of the GPCR cells described herein (e.g., GPCRs including AT1R, APJ, CCR6, GPR20, CCR4, PAR2) and / or any regulatory activity of such molecules.

[0126] G Protein-Coupled Receptors (GPCRs) are a family of cell surface receptors involved in signal transduction and various physiological processes. They are activated by a wide range of ligands, including hormones, neurotransmitters, and other signaling molecules. GPCRs areDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 characterized by their seven-transmembrane domain structure and their ability to activate intracellular G proteins, which then trigger various downstream signaling pathways.

[0127] Upon activation by external signals, through coupling to different G proteins or arrestins, GPCRs elicit cyclic adenosine 3,5-monophosphate (cAMP) responses, phosphorylation of extracellular regulated protein kinases 1 / 2 (pERK1 / 2), or calcium mobilization. The two principal signaling transduction pathways involving the G protein-coupled receptors are the cAMP signal pathway and the phosphatidylinositol signaling pathway. In some embodiments, the strategies and / or fusion proteins described herein can be used for regulating cAMP signaling pathways. In some embodiments, the strategies and / or fusion proteins described herein can be used for regulating phosphatidylinositol signaling pathways.

[0128] The activation by GPCRs of diversified downstream signaling pathways makes GPCRs attractive for drug development (Yang et al., Signal Transduction and Targeted Therapy, volume 6, Article number: 7, 2021). In some embodiments, the strategies and / or fusion proteins described herein can be used for the treatment of diseases involving and / or related to GPCRs.

[0129] Upon ligand binding to the GPCR, there is a conformational change in the GPCR that can allow GPCRs to act as guanine nucleotide exchange factors (GEF). Then, the GPCR can activate an associated G protein by exchanging the GDP bound to the G protein for a GTP. The G protein's α subunit, together with the bound GTP, can then dissociate from the β and γ subunits to further affect intracellular signaling proteins or target functional proteins directly depending on the α subunit type. In some embodiments, the strategies and / or fusion proteins described herein can activate a GPCR associated G protein. In some embodiments, the strategies and / or fusion proteins described herein can induce GDP to GTP exchange and affect intracellular signaling.

[0130] GPCRs play a role in many functions in the human body, and increased understanding of these receptors has greatly affected modern medicine. All GPCRs have a common structure and mechanism of signal transduction; however, they can be classified and grouped into six classes based on sequence homology and functional similarity. One classification system groups GPCRs into groups A-F. Human GPCRs include groups A, B, C, and F. In some embodiments, the fusion proteins disclosed herein can target GPCRs that are present in humans.

[0131] In some embodiments, the strategies and / or fusion proteins described herein can include fusion proteins that can bind and / or interact with, and / or internalize, any of the class ADOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 GPCRs described herein. In some embodiments, the strategies and / or fusion proteins described herein can include fusion proteins that can bind and / or interact with, and / or internalize, any of the class B GPCRs described herein. In some embodiments, the strategies and / or fusion proteins described herein can include fusion proteins that can bind and / or interact with, and / or internalize, any of the class C GPCRs described herein. In some embodiments, the strategies and / or fusion proteins described herein can include fusion proteins that can bind and / or interact with any of the class F GPCRs described herein.

[0132] Class A GPCRs, also called “rhodopsin-like family” can be divided into several subgroups including aminergic, peptide, protein, lipid, melatonin, nucleotide, steroid, alicarboxylic acid, sensory, and orphan receptors. Class A GPCRs can be involved in a variety of medical conditions and disease including, but not limited to, allergies, cardiovascular diseases, hypertension, pulmonary diseases, multiple sclerosis, depression, migraine, narcolepsy-related daytime sleepiness, insomnia, glaucoma, Parkinson’s disease to schizophrenia, and fatigue.

[0133] Class B GPCRs can be divided into the secretin (B1) and the adhesion (B2) families. Members of the secretin subfamily can bind to a diversity of molecules including, but not limited to, growth hormone-releasing hormone (GHRH), vasoactive intestinal peptide (VIP), pituitary adenylate cyclase-activating peptide (PACAP), corticotropin-releasing factor (CRF, calcitonin gene-related peptide (CGRP), glucagon, and glucagon-like peptides (GLPs), and parathyroid peptide hormone (PTH).

[0134] In addition, there are nine subgroups in the adhesion subfamily, groups that present unique N-terminal domains including, but not limited to, epidermal growth factor, cadherin, and immunoglobulin domains. Members of the adhesion subfamily are distinguished from other GPCRs because of their roles in cell adhesion and migration. Other unique features of the B2 subfamily are the proteolysis site and the GPCR autoproteolysis-inducing domain that are responsible for signaling activation through a Stachel sequence (a tethered agonist) and producing N-terminal fragment (NTF) and C-terminal fragment.

[0135] Class B GPCRs include but are not limited to, receptors of glucagon family peptides, parathyroid hormone receptors (PTH), GHRH, CRF, VIP, GLP-1R, glucagon receptor (GCGR), glucose-dependent insulinotropic peptide receptor (GIPR), calcitonin gene-related peptide receptors (CGRP), ADGRG1 and ADGRF1, and PACAP. Class B GPCRs are involved in medical conditions and diseases including, but not limited to, obesity, T2DM, osteoporosis,DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 migraine, depression, anxiety, and diseases and conditions of the cardiovascular, respiratory, nervous, musculoskeletal, reproductive, renal, integumentary, sensory, endocrine, neurological and gastrointestinal systems.

[0136] Class C GPCRs are glutamate receptors and include, but are not limited to, calcium- sensing receptors (CaSR), gamma-aminobutyric acid (GABA) type B receptors (GABAB1 and GABAB2), taste 1 receptors (TS1R1–3), metabotropic glutamate receptors (mGluR1–8), and orphan GPCRs. This class is characterized by large ECD and obligated constitutive dimer for receptor activation. They can also include conserved venus fly trap (VFT) and cysteine-rich domain (CRD) on the ligand-binding site. Class F GPCRs can include conserved disulfide bonds between VFT domains that can stabilize the homodimers or heterodimers of class F GPCRs. Class C and F GPCRs can be involved in medical conditions and diseases including, but not limited to, schizophrenia, depression, and movement disorders fibrosis, and neurodegeneration.

[0137] Group F GPCRs include frizzled receptors.

[0138] In some embodiments, the strategies and / or GPCR TransTACs described herein can be used for regulating, preventing, and / or treating any of the GPCR related diseases described herein.

[0139] GPCRs are widely expressed in the cardiovascular system and play crucial roles in regulating cardiovascular function and morphology. Angiotensin II type 1 receptors (AT1Rs) are important GPCRs in cardiovascular function. In addition, there are many other GPCRs, such as apelin receptor (APJ), lysophosphatidic acid receptor (LPARs) and endothelin receptors (ETAR and ETBR), that play important roles in CVDs. Chronic activation by their endogenous ligands increases the workload of the heart, leading to harmful effects such as heart failure (HF). The angiotensin II receptors, (ATR1) and (ATR2), are a class of G protein-coupled receptors that bind angiotensin II (the ligand) (Li et al., Front. Cardiovasc. Med., 05 June 2023; Sec. Cardiovascular Pharmacology and Drug Discovery; Volume 10 – 2023). They are also important in the renin–angiotensin system where they are responsible for the signal transduction of the vasoconstricting stimulus of the main effector hormone, angiotensin II.

[0140] In some embodiments, the strategies and / or fusion proteins described herein can include fusion proteins that can bind and / or interact with AT1R. In some embodiments, the strategies and / or fusion proteins described herein can include fusion proteins that can bind and / or interact with APJ.DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17

[0141] Orphan G protein-coupled receptors (oGPCRs) are pathologically related to many human diseases including, but not limited to, hyperactivity, schizophrenia, type 2 diabetes, cognitive impairment, brain malformation, and Alzheimer’s disease. GPR20 is an orphan receptor for which the endogenous ligand remains unknown. GPR20 shows high level of constitutive activity in the absence of ligand, leading to continuous activation of its coupled Gi proteins.

[0142] Chemokine receptors include seven-transmembrane G-protein coupled receptors that can bind a diversity of chemokines including, but not limited to, CCL1-28, CXCL1-16, XCL1-2, and CX3CL1. The about 20 chemokine receptors described up to date can be divided into four main families: CC, CXC, XC, and CX3. Chemokine receptors can play roles in several diseases, including allergies, atherosclerosis, viruses, various infections, and inflammation.

[0143] CCC chemokine receptor 4 (CCR4) is important in the pathogenesis of many diseases including, but not limited to diabetes, multiple sclerosis, asthma, dermatitis, and pain from neuropathy. CCR4 ligands include, but are not limited to, CCL17, CCL22 and CCL2. CCR4 can be present in neurons, microglia, astroglia, dorsal root ganglia, spinal cord and brain. In some embodiments, the strategies and / or fusion proteins described herein can include fusion proteins that can bind and / or interact with CCR4.

[0144] Protease-activated receptors (PARs) are a family of four G protein-coupled receptors (GPCRs) that can be irreversibly activated through proteolytic cleavage of their N termini by serine proteases (e.g., thrombin, trypsin, plasmin). N termini cleavage can create new extracellular N termini, which serve as tethered ligands that intramolecularly activate the receptors and initiate complex intracellular signaling events. PAR1, one of the PARs receptors, binds thrombin and it is known for its role in the cardiovascular. In addition, PAR1 and three other PARs (PAR2–4) have been implicated in strikingly diverse pathophysiological functions including, but not limited to, stroke, inflammation, and reactive gliosis. In some embodiments, the strategies and / or fusion proteins described herein can include fusion proteins that can bind and / or interact with PAR1.

[0145] In some embodiments of the invention disclosed herein, fusion proteins (e.g., GPCR TransTAC molecules) are used to regulate GPCR expressing cells. In some embodiments, these fusion proteins can reversibly modulate GPCR expressing cells.DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17

[0146] In some embodiments, the GPCRs that can be targeted using bispecific modulators (i.e., the disclosed fusion proteins, and homodimers and heterodimers thereof) include Apelin Receptor (APJ), Angiotensin II Receptor Type (AT1R), Relaxin Family Peptide Receptor 1 (RXFP1), C-C Chemokine Receptor Type 6 (CCR6), Adenosine A2A Receptor (A2AR), G Protein-Coupled Receptor 20 (GPR20), C-C Motif Chemokine Receptor 4 (CCR4), Protease- Activated Receptor 2 (PAR2), Frizzled Class Receptor 1 (FZD10), C-C Motif Chemokine Receptor 2 (CCR2), C-X-C Motif Chemokine Receptor 1 (CXCR1 / 1) or Complement Component 5a Receptor 1 (C5aR1).

[0147] In some embodiments, the strategies and / or fusion proteins described herein can include fusion proteins that can bind and / or interact with a GPCR including a flag motif. In some embodiments, the flag motif can be a FLAG tag. In some embodiments, the flag motif can be an HA tag.

[0148] In some embodiments, the strategies and / or fusion proteins described herein can include fusion proteins that can bind and / or interact with FLAG-APJ. In some embodiments, the strategies and / or fusion proteins described herein can include fusion proteins that can bind and / or interact with FLAG-GPR20. In some embodiments, the strategies and / or fusion proteins described herein can include fusion proteins that can bind and / or interact with FLAG-CCR4. In some embodiments, the strategies and / or fusion proteins described herein can include fusion proteins that can bind and / or interact with FLAG-AT1R. In some embodiments, the strategies and / or fusion proteins described herein can include fusion proteins that can bind and / or interact with FLAG-PAR2. In some embodiments, the strategies and / or fusion proteins described herein can include fusion proteins that can bind and / or interact with FLAG-RXFP1.

[0149] In some embodiments, the strategies and / or fusion proteins described herein can include fusion proteins that can bind and / or interact with HA-FZD1.

[0150] In some embodiments, expression of any of the FLAG-GPCRs described herein can be inducible. In some embodiments, expression of any of the FLAG-GPCRs described herein can be induced with doxorubicin. In some embodiments, expression of any of the FLAG-GPCRs described herein can be induced with between about 1 to about 3 ng / ml of doxorubicin, between about 2 to about 4 ng / ml of doxorubicin, between about 3 to about 6 ng / ml of doxorubicin, between about 5 to about 8 ng / ml of doxorubicin. between about 7 to about 10 ng / ml of doxorubicin, between about 8 to about 15 ng / ml of doxorubicin, between about 13 to about 18DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 ng / ml of doxorubicin, between about 15 to about 20 ng / ml of doxorubicin, between about 18 to about 30 ng / ml of doxorubicin, between about 25 to about 50 ng / ml of doxorubicin, between about 50 to about 80 ng / ml of doxorubicin, between about 50 to about 80 ng / ml of doxorubicin, between about 70 to about 150 ng / ml of doxorubicin, between about 130 to about 300 ng / ml of doxorubicin, between about 250 to about 500 ng / ml of doxorubicin, between about 400 to about 800 ng / ml of doxorubicin, between about 600 to about 1000 ng / ml of doxorubicin, between about 900 to about 2000 ng / ml of doxorubicin. Fusion Proteins

[0151] In some embodiments, strategies disclosed herein for regulating a molecule on a cell surface (e.g., such as proteins of interest, GPCR) and / or regulatory activity of such proteins can use a fusion protein approach. In some embodiments, a fusion protein molecule can have at least two moieties. A first moiety (R1 or POI binder) can be a ligand that the cell-surface molecule(s) can bind or an antibody or antibody fragment that can bind to the cell-surface molecule(s) (e.g., GPCR). R1 can be a protein of interest (POI) binder (POIB) or POI binding means (also known as a means for binding POI). A second moiety (R3 or TR binder) can be a molecule that can bind to an internalizing receptor or membrane protein on a cell (e.g., transferrin receptor or TR). In some embodiments, R3 can bind transferrin receptor (TR) and can be called a transferrin receptor binder (TR binder). R3 can be a TR binding means (also known as a means for binding TR)). In embodiments, the second moiety (R3 or TR binder or TR binding means) can be an antibody or antibody fragment that binds to transferrin receptor on a cell. In embodiments, the second moiety can be a nanobody that binds to an internalizing receptor or membrane protein on a cell.

[0152] In some embodiments, R3 can be a means for binding a transferrin receptor (TR), wherein the TR binder or TR binding means (also known as a means for binding TR) is non- competitive with endogenous transferrin for binding to the TR. In some embodiments, the nanobody can include a VHH from a camelid. In embodiments, the fusion protein (or homodimer or heterodimer thereof) can be a bispecific antibody.

[0153] In some embodiments, the VHH includes VHHA (SEQ ID NO: 146-147). In some embodiments, the VHH includes VHHB (SEQ ID NO: 148-149). In some embodiments, the VHHA or VHHB can include amino acid substitutions. In some embodiments, the amino acidDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 substitutions can affect binding affinity of TR binder or TR binding means (also known as a means for binding TR) to transferrin receptor.

[0154] In embodiments, a fusion protein having these two moieties can bind, or be bound by, a cell-surface or other molecule (e.g., GPCRs) and can bind to an internalizing receptor which is TfR. After such binding, the TfR can cause the GPCR to be internalized into the cell (e.g., endocytosis). In embodiments, the internalized GPCR can be degraded. In embodiments, this internalization and / or degradation decreases the amount of the GPCR on the surface of a cell. In embodiments, the internalized GPCRs are not functional (e.g., they do not function as when they are present on a cell surface).

[0155] In some embodiments, administering the fusion proteins to a subject can be used for targeted internalization of GPCR. In some embodiments, administering the fusion proteins to a subject can be used for targeted degradation of membrane or other proteins.

[0156] In some embodiments, adding the fusion proteins to cells or administering to a patient can cause targeted internalization and / or degradation of proteins on the surface of a cell or outside of a cell. In some embodiments, this internalization / degradation is reversible. For example, when a cell is no longer exposed to fusion proteins, the membrane proteins to which the fusion proteins are specific are no longer internalized / degraded. Generally, the membrane proteins are still synthesized and trafficked to the cell membrane. Therefore, when the fusion proteins are removed or are no longer administered to a subject, there is not a stimulus to internalize / degrade the proteins. In some embodiments, a cellular membrane protein that can be internalized by a fusion protein, but not degraded, can be both internalized and degraded using a fusion protein that also contains an optional protease-sensitive linker or protease-sensitive linking means (also known as a means for linking). As discussed elsewhere, placement of an optional protease-sensitive linker or protease-sensitive linking means (also known as a means for linking) within a fusion protein can provide release of a targeted cellular protein of interest from the fusion protein inside of a cell.

[0157] In some embodiments, internalization and degradation of the GPCR can kill the cell. In some embodiments, internalization and degradation of the GPCR does not kill the cell.

[0158] In some embodiments, internalization of fusion proteins or parts thereof can involve receptor-mediated endocytosis, also called clathrin-mediated endocytosis. In someDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 embodiments, internalization of fusion proteins can involve clathrin-independent endocytosis. In some embodiments, internalization of fusion proteins can involve phagocytosis.

[0159] In some embodiments, the GPCR can be on any cell (e.g., a cell from the cardiac, neurological, gastrointestinal, urinary systems). In some embodiments, a strategy for regulating GPCR cell’s activities include internalizing GPCRs with a fusion protein. In some embodiments, the GPCR targeted by the first moiety can be any of the GPCR described herein, including, but not limited to, APJ, GPR20, CCR6, CCR2, CCR4, AT1R, CXCR1 / 1, PAR2, RXFP1, A2AR, FZD1, or any of the GPCR described herein.

[0160] The fusion proteins disclosed herein can be used for the treatment of diseases. In some embodiments, the GPCRs that are targeted in treatment of disease using the molecules disclosed herein can include Apelin Receptor (APJ), Angiotensin II Receptor Type (AT1R), Relaxin Family Peptide Receptor 1 (RXFP1), C-C Chemokine Receptor Type 6 (CCR6), Adenosine A2A Receptor (A2AR), G Protein-Coupled Receptor 20 (GPR20), C-C Motif Chemokine Receptor 4 (CCR4), Protease-Activated Receptor 2 (PAR2), Frizzled Class Receptor 1 (FZD1), C-C Motif Chemokine Receptor 2 (CCR2), C-X-C Motif Chemokine Receptor 1 (CXCR1 / 1) or Complement Component 5a Receptor 1 (C5aR1).

[0161] For example, the APJ receptor has been implicated in neurological diseases, respiratory diseases, and cardiovascular conditions. AT1R is associated with hypertension, heart failure, and kidney diseases. Autoantibodies against AT1R have been linked to various diseases, including cardiovascular diseases. RXFP1 is associated with fibrotic diseases. A2AR is involved in immune regulation and has been implicated in cancer, autoimmune diseases, and neurodegenerative disorders. GPR20 has been linked to various cancers. CCR4 has been associated with diseases like diabetes, cancer, asthma, and multiple sclerosis. PAR2 is involved in inflammatory responses and has been linked to autoimmune diseases. FZD1 is associated with certain cancers. CCR2 has been linked to certain inflammatory diseases.

[0162] In some embodiments, subjects having diseases including type 2 diabetes mellitus, Alzheimer’s disease, obesity and depression can be treated with the molecules disclosed herein.

[0163] In some embodiments, the molecule that is targeted by the first moiety of a fusion protein (i.e., protein of interest) can be a different molecule than the molecule targeted by the second moiety (e.g., transferrin receptor) of a fusion protein.DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17

[0164] In some embodiments, the fusion protein is a single molecule. In some embodiments, the fusion protein can be a single polypeptide. In some embodiments, a single polypeptide can contain both the first moiety and the second moiety (TR binder or TR binding means (also known as a means for binding TR)) of a fusion protein.

[0165] In some embodiments, the first moiety (R1) can be an antibody or antibody fragment (e.g., nanobody, peptide) that binds to any of the GPCR described herein present in a cell. In embodiments, the first moiety (R1) can be an antibody or antibody fragment (e.g., nanobody, peptide) that can bind to any one of a class A, B, C and F GPCR. In embodiments, the first moiety can be an antibody or antibody fragment (e.g., nanobody, peptide) that can bind to AT1R. In some embodiments, the first moiety (R1) can be an antibody or antibody fragment (e.g., nanobody, peptide) that can bind to APJ. In some embodiments, the first moiety (R1) can be an antibody or antibody fragment (e.g., nanobody, peptide) that can bind to CCR6. In some embodiments, the first moiety (R1) can be an antibody or antibody fragment (e.g., nanobody, peptide) that can bind to A2AR. In some embodiments, the first moiety (R1) can be an antibody or antibody fragment (e.g., nanobody, peptide) that can bind to GPR20. In some embodiments, the first moiety (R1) can be an antibody or antibody fragment (e.g., nanobody, peptide) that can bind to CCR4. In some embodiments, the first moiety (R1) can be an antibody or antibody fragment (e.g., nanobody, peptide) that can bind to PAR2. In some embodiments, the first moiety (R1) can be an antibody or antibody fragment (e.g., nanobody, peptide) that can bind to RXFP1. In some embodiments, the first moiety (R1) can be an antibody or antibody fragment (e.g., nanobody, peptide) that can bind to FZD1.

[0166] In some embodiments, a non-limiting list of membrane proteins that can be used in the fusion proteins includes any of the G-protein coupled receptors (GPCR) described herein and / or any of the GPCR described in G Protein-Coupled Receptor List | IUPHAR / BPS Guide to PHARMACOLOGY, Yang et al., Signal Transduction and Targeted Therapy volume 6, Article number: 7 (2021), Lin et al. Cell Discovery ( 2023) 9:23; McCoy et al., Mol Pharmacol 77:1005– 1015, 2010; Bogacka et al., Int. J. Mol. Sci.2022, 23, 1563 that are incorporated herein by reference. In some embodiments, GPCRs can include class A orphan receptors, opsin receptors, Taste 1 and Taste 2 receptors, 5-Hydroxytryptamine receptors, Acetylcholine receptors (muscarinic), Adenosine receptors, Adhesion Class GPCRs, angiotensin receptors, apelin receptors, bile acid receptors, bombesin receptors, Bradykinin receptors, calcitonin receptors,DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 Calcium-sensing receptors, cannabinoid receptors, Chemerin receptors, Cholecystokinin receptors, Class Frizzled GPCRs, Complement peptide receptors, Corticotropin-releasing factor receptors, Dopamine receptors, Endothelin receptors, G protein-coupled estrogen receptor, Formylpeptide receptors, Free fatty acid receptors, GABAB receptors, Galanin receptors, Ghrelin receptor, Glucagon receptor family, Glycoprotein hormone receptors, Gonadotrophin-releasing hormone receptors, GPR18, GPR55 and GPR119, Histamine receptors, Hydroxycarboxylic acid receptors, Kisspeptin receptor, Leukotriene receptors, Lysophospholipid (LPA) receptors, Melanin-concentrating hormone receptors, Melanocortin receptors, Metabotropic glutamate receptors, Motilin receptor, Neuromedin U receptors, Neuropeptide FF / neuropeptide AF receptors, Neuropeptide Y receptors, Neurotensin receptors, Opioid receptors, Orexin receptors, P2Y receptors , Oxoglutarate receptor, Parathyroid hormone receptors, Platelet-activating factor receptor, Prokineticin receptors, Prolactin-releasing peptide receptor, Prostanoid receptors, Proteinase-activated receptors, Relaxin family peptide receptors, Somatostatin receptors, Succinate receptor, Tachykinin receptors, Trace amine receptor, Urotensin receptor, Vasopressin and oxytocin receptors, and VIP, PACAP receptors, adrenoceptors, chemokine receptors, and coagulation receptors.

[0167] In some embodiments, the antibody that is the first moiety (POIB or POI binding means (also known as a means for binding POI)) can be an scFv, Fab, single-domain antibody, nanobody, monobody, DARPin or affibody. Antibody fragments and other molecules that can be used are described in the section titled “Antibodies” in this application.

[0168] In some embodiments, the first moiety (R1 or POI binding means (also known as a means for binding POI)) can be a nanobody that binds a GPCR (e.g., any of the GPCR described herein). In some embodiments, the first moiety includes a nanobody (VHH) that binds to AT1R. (VHH AT118-L; FIG.57). In some embodiments, the first moiety includes a nanobody (VHH) that binds to APJ. (VHH JN241; FIG.58). In some embodiments, the first moiety includes a nanobody (VHH) that binds to RXFP1 (VHH RX002; FIG.59). In some embodiments, the first moiety includes a peptide that binds to FLAG HC M2 (FLAG HC M2; FIG.60).

[0169] In some embodiments, the GPCR TransTAC can bind / deplete / degrade AT1R. In some embodiments, the AT1R GPCR TransTAC can include a VHH that binds AT1R (AT1R VHH AT118-L), a human IgG1 Fc (H435A), and a second moiety binder that can bind to TfR (TfRDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 scFv H7) KR54 and SEQ ID NO: 155; FIG.57). In some embodiments, AT1R GPCR TransTAC can bind and efficiently deplete AT1R from the cell surface.

[0170] In some embodiments, the GPCR TransTAC can bind / deplete / degrade APJ. In some embodiments, the APJ TransTAC can include a VHH that binds APJ (APJ VHH JN241), a human IgG1 Fc (F372L), and a second moiety binder that can bind to TfR (TfR scFv H7) (DP238 and SEQ ID NO: 157; FIG.58). In some embodiments, APJ GPCR TransTAC can bind and efficiently deplete APJ from the cell surface.

[0171] In some embodiments, the GPCR TransTAC can bind / deplete / degrade RXFP1. In some embodiments, the RXFP1 TransTAC can include a VHH that binds RXFP1 (RXFP1 VHH RX002), human IgG1 Fc (G236R, N297G, L328R, D356E, L358M), and second moiety binder that binds to TfR (TfR scFv H7) (KR98 and SEQ ID NO: 159; FIG.59). In some embodiments, RXFP1 GPCR TransTAC can bind and efficiently deplete RXFP1 from the cell surface.

[0172] In some embodiments, the GPCR TransTAC can bind / deplete / degrade FZD1. In some embodiments, the FZD1 TransTAC can include a FLAG HC M2 peptide, human IgG1 Fc (G236R, N297G, L328R, D356E, L358M), and second moiety binder that binds to TfR (TfR scFv H7) (KR62.1 and SEQ ID NO: 162; FIG.60). In some embodiments, FZD1 GPCR TransTAC can bind and efficiently deplete FZD1 from the cell surface.

[0173] In some embodiments, GPR20 GPCR TransTAC can bind and efficiently deplete GPR20 from the cell surface. In some embodiments, CCR4 GPCR TransTAC can bind and efficiently deplete CCR4 from the cell surface. In some embodiments, PAR2 GPCR TransTAC can bind and efficiently deplete PAR2 from the cell surface.

[0174] In some embodiments, any of the GPCR TransTAC described herein can deplete / degrade a GPCR from the cell surface in about 1 to 5 minutes, 4 to 6 minutes, 5 to 8 minutes, 7 to 10 minutes, 9 to 15 minutes, 12 to 17 minutes, 15 to 20 minutes, 18 to 25 minutes, 20 to 30 minutes, 25 to 35 minutes, 30 to 40 minutes, 35 to 50 minutes, 45 to 60 minutes, 50 minutes to 1.5 hours, 1.2 hours to 2 hours, 1.8 hours to 3 hours, 2 hours to 4 hours, 3 hours to 5 hours, 4 hours to 7 hours after adding the TransTAC.

[0175] In some embodiments, the GPCR TransTAC can effectively deplete / degrade a GPCR when the TransTAC at doses of about 1 to 5 nM, 4 to 6 nM, 5 to 8 nM, 7 to 10 nM, 9 to 15 nM, 12 to 17 nM, 15 to 20 nM, 18 to 25 nM, 20 to 30 nM, 25 to 35 nM, 30 to 40 nM, 35 to 50 nM, 45DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 to 60 nM, 50 nM to 100 nM, 90 to 120 nM, 100 to 140 nM, 130 to 160 nM, 150 to 200 nM, 170 to 300 nM, 250 to 500 nM, 400 to 800 nM, 500 to 1,000 nM.

[0176] In some embodiments, the second moiety (TR binder or TR binding means (also known as a means for binding TR)) binds to a transferrin receptor (R3 or transferrin receptor binder or TR binder or TR binding means (also known as a means for binding TR)) on a cell. In some embodiments, the receptor or membrane protein bound by the second moiety (i.e., TR binder or TR binding means (also known as a means for binding TR)) is an internalizing receptor or membrane protein. In some embodiments, the TR binder or TR binding means (also known as a means for binding TR) can mediate endocytosis. In embodiments, the second moiety (TR binder or TR binding means (also known as a means for binding TR)) can also be an antibody or antibody fragment that binds the transferrin receptor.

[0177] In some embodiments, the internalizing is a transferrin receptor. In embodiments, a ligand (e.g., second moiety (TR binder or TR binding means (also known as a means for binding TR))) that a transferrin receptor can bind can be transferrin or a fragment of transferrin. In some embodiments, the second moiety (TR binder or TR binding means (also known as a means for binding TR)) can be an antibody or antibody fragment that can bind to transferrin receptor. In some embodiments, the internalizing receptor or membrane protein can be a transferrin receptor (TfR). In some embodiments, the transferrin receptor can be transferrin receptor 1 or transferrin receptor 2.

[0178] In some embodiments, the transferrin receptor can have a high endocytosis rate of around 500 molecules per cell per second, making it good for inducing protein endocytosis. In some embodiments, transferrin receptor can be expressed in healthy tissues. In some embodiments, transferrin receptor expression can be expressed in non-healthy (e.g., diseases) tissues. In some embodiments, transferrin receptor expression can efficiently endocytose and degrade target proteins (POI) when the transferrin receptor is expressed. Transferrin receptor can be expressed in tissues from organs including, but not limited to brain, liver, breast, lung, colon, pancreas, skin, spleen, lymph nodes, bone marrow, thymus, kidneys, and heart.

[0179] In some embodiments, the second moiety (TR binder or TR binding means (also known as a means for binding TR)) can be an antibody, antibody fragment or other molecule that can bind the internalizing receptor or membrane protein. In some embodiments, the secondDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 moiety (TR binder or TR binding means (also known as a means for binding TR)) can be a scFv, Fab, single-domain antibody, nanobody, monobody, DARPin or affibody.

[0180] In some embodiments, H7 is competitive with transferrin for binding to TfR. M16 is competitive with transferrin for binding to TfR.

[0181] In some embodiments, the second moiety (R3) can bind to a transferrin receptor (TfR) and is a non-competitive transferrin receptor binder that does not compete or competes insignificantly with endogenous (wild-type, full-length transferrin) transferrin for binding to transferrin receptor. In some embodiments, the TR binder or TR binding means (also known as a means for binding TR) that does not compete or competes insignificantly with endogenous Tf for binding to TfR can be more efficient in binding, internalizing and / or degrading cell surface molecular targets (e.g., protein of interest or POI) than TransTAC molecules having a TR binder or TR binding means (also known as a means for binding TR) that is competitive with endogenous Tf for binding to TfR.

[0182] In some embodiments, a competitive inhibitor of Tf binding to TfR can prevent or reduce Tf binding to TfR. In some embodiments, a competitive inhibitor of Tf binding to TfR does not bind to a TfR simultaneously with the non-competitive inhibitor (they both cannot bind to the same receptor). In some embodiments, a non-competitive inhibitor of Tf binding to TfR can bind to a TfR simultaneously with Tf binding to TfR (both molecules can simultaneously bind to a TfR). In some embodiments, a non-competitive inhibitor can insignificantly compete with Tf for binding to TfR. In some embodiments, insignificantly compete means that that less than 50, 40, 30, 20, 10, 5 or 1% of non-competitive inhibitor molecules that bind to TfR inhibit a Tf from binding to TfR. In some embodiments, the non-competitive transferrin receptor binder binds to a different region of the transferrin receptor than does Tf. In some embodiments, the transferrin receptor binders used in the molecules disclosed herein are VHHA, VHHA-12, VHHA-5, VHHA-7 or VHHB. Other non-competitive transferrin receptor binders can be used.

[0183] In some embodiments, when a non-competitive inhibitor is added to TfR in the presence of Tf, binding of Tf to TfR is not affected. In some embodiments, when increasing amounts of a non-competitive inhibitor are added to TfR in the presence of Tf, binding of Tf to TfR is affected less than if increasing amounts of a competitive inhibitor, instead of the non- competitive inhibitor, were used.DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17

[0184] In some embodiments, the second moiety (R3) can be an antibody or antibody fragment that can bind transferrin receptor. In some embodiments, the R3 does not compete or competes insignificantly with endogenous Tf for binding to TfR. In some embodiments, the antibody or antibody fragment thereof (e.g., nanobody) that does not compete or competes insignificantly with endogenous Tf for binding to TfR can be VHHA or VHHB. VHHA or VHHB that have certain amino acid substitutions (i.e., VHHA-12, VHHA-5, and VHHA-7) can also be non-competitive with endogenous transferrin for binding to transferrin receptor. In some embodiments, VHHA can be more efficient in binding, internalizing and / or degrading POI targets than TransTAC molecules having a TR binder or TR binding means (also known as a means for binding TR) that is competitive with endogenous Tf for binding to TfR. In some embodiments, VHHB can be more efficient in binding, internalizing and / or degrading POI targets than TransTAC molecules having a TR binder or TR binding means (also known as a means for binding TR) that is competitive with endogenous Tf for binding to TfR.

[0185] Variable Domain of Camelid Heavy Chain-only (VHH) antibodies that bind to transferrin receptor are disclosed in U.S. Patent Publication Number 2023 / 0414780 A1, published on December 28, 2023, and filed on June 22, 2023, and which is incorporated herein by reference.

[0186] In one embodiment, an amino acid sequence of a competitive TRB, H7, can include the molecules below, or can include molecules at least 60, 65, 70, 75, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequences below:

[0187] H7 scFV-LC (SEQ ID NO: 1):

[0188] SELTQDPAVSVALGQTVRITCQGDSLRSYYASWYQQKPGQAPVLVMYGRNE RPSGVPDRFSGSKSGTSASLAISGLQPEDEANYYCAGWDDSLTGPVFGGGTKLTVLG*

[0189] H7 scFV-HC (SEQ ID NO: 2):

[0190] QVQLQESGGGVVQPGRSLRLSCAASRFTFSSYAMHWVRQAPGKGLEWVAVI SYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDLSGYGDYPD YWGQGTLVTVSS

[0191] H7 -scFv (SEQ ID NO: 3)

[0192] SELTQDPAVSVALGQTVRITCQGDSLRSYYASWYQQKPGQAPVLVMYGRNE RPSGVPDRFSGSKSGTSASLAISGLQPEDEANYYCAGWDDSLTGPVFGGGTKLTVLGGG GGSGGGGSGGGGSQVQLQESGGGVVQPGRSLRLSCAASRFTFSSYAMHWVRQAPGKG LEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDLSG YGDYPDYWGQGTLVTVSSDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17

[0193] M16 (SEQ ID NO: 4):

[0194] SELTQDPAVSVALGQTVRITCQGDSLRSYYASWYQQKPGQAPVLVMYGRNE RPSGVPDRFSGSKSGTSASLAISGLQPEDEANYYCAGWDDSLTGPVFGGGTKLTVLGGG GGSGGGGSGGGGSQVQLQESGGGVVQPGRSLRLSCAASRYPFHHHDHHWVRQAPGKG LEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDLSG YGDYPDYWGQGTLVTVSS

[0195] TransTAC designs can employ binders that compete with natural transferrin (Tf) for binding to TfR1 (Tf-competitive binder), which may affect iron import and cause toxicities to certain cells and tissues. Binders recognizing a non-competitive epitope on TfR1 (Tf non- competitive) can be effective at degrading a POI while reducing toxicity effects. Nanobodies VHHA and VHHB can bind to TfR1 in an epitope that is non-competitive with Tf. VHHA an VHHB can drive PD-L1 degradation. VHHA and VHHB can induce POI degradation potently and effectively.

[0196] TfR1 includes epitopes that are recognized and bound by the natural TFR1 ligand Transferrin (Tf) (Tf-competitive epitopes) and epitopes that are not recognized / bound by Tf (Tf non-competitive epitopes). TransTAC designs including second moieties (R3) with binders that compete with natural transferrin (Tf) for binding to Tf competitive epitopes (called Tf competitive TransTACs, e.g., v1.0 including H7 as a binder) can affect iron import and cause toxicities to certain cells and tissues. TransTAC designs including second moieties (R3) that do not compete with endogenous transferrin for binding to non-competitive epitopes (non- competitive TransTACs) can be effective at degrading a POI while reducing toxicity. These binders can be non-competitive with transferrin in binding to transferrin receptor. In some embodiments, non-competitive TR binders or TR binding means (also known as a means for binding TR) can limit toxicity that can occur when transferrin binding to transferrin receptor is affected (as when R3 is competitive with transferrin for binding to TfR).

[0197] In some embodiments, an antibody or antibody fragment that can bind a transferrin receptor can be a nanobody. This nanobody can be called “VHHA.” In some embodiments, VHHA can bind to an epitope in TfR1 that does not compete for binding with natural / endogenous Tf. In some embodiments, a TransTAC including a first moiety including a molecule that can bind a GPCR and a second moiety (TR binders or TR binding means (also known as a means for binding TR)) including VHHA can bind to an epitope of TfR and can drive GPCR degradation independently of the presence of natural transferrin (Tf). In someDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 embodiments binding of the TransTAC including VHHA to TfR1 is non-competitive with natural Tf. In some embodiments, non-competitive TR binders or TR binding means (also known as a means for binding TR) can limit toxicity that can occur if transferrin binding to TfR is affected by using an R3 that competes with transferrin for binding to TfR.

[0198] In some embodiments, an antibody or antibody fragment that can bind a transferrin receptor can be a nanobody. This nanobody can be called “VHHB.” In some embodiments, VHHB can bind to a non-competitive TfR1 epitope.

[0199] Nanobodies VHHA and VHHB can bind to non-competitive TfR1 epitopes. TransTACs including VHHA and / or VHHB are non-competitive TransTACs that can drive POI degradation independently of the presence of Tf. VHHA and VHHB TransTACs can induce POI degradation potently and effectively.

[0200] In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind any of the POI described herein, and a second moiety (R3) including a molecule (e.g., a peptide, a nanobody, VHHA, VHHB) that can bind to a non-competitive epitope in an internalizing molecule (e.g., a non-competitive epitope in TfR1) can drive the internalization and degradation of the POI.

[0201] In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind any of the POI described herein, and a second moiety (R3) including VHHA can bind to TfR and drive POI internalization / degradation. In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind any of the POI described herein, and a second moiety (R3) including VHHA can bind to a non-competitive epitope in TfR and drive POI degradation. In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind any of the POI described herein, and a second moiety (R3) including H7 can bind to a competitive epitope in TfR and drive POI degradation. In some embodiments, the TransTAC including a first moiety (R1) including a molecule that can bind any of the POI described herein, and a second moiety (R3) including VHHA can bind to TfR and drive POI degradation more efficiently and potently than a TransTAC including the same POI and a second moiety (R3) including a moiety that can bind a competitive TfR1 epitope (e.g., H7).

[0202] In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind any of the POI described herein, and a second moiety (R3) including VHHB can bind to TfR and drive POI internalization / degradation. In some embodiments, a TransTACDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 including a first moiety (R1) including a molecule that can bind any of the POI described herein, and a second moiety (R3) including VHHB can bind to a non-competitive epitope in TfR and drive POI degradation. In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind any of the POI described herein, and a second moiety (R3) including a TRB that is non-competitive with transferrin in binding a transferrin receptor can bind to an epitope in TfR and drive POI degradation. In some embodiments, the TransTAC including a first moiety (R1) including a molecule that can bind any of the POI described herein, and a second moiety (R3) including VHHB can bind to TfR and drive POI degradation more efficiently and potently than a TransTAC including the same POI and a second moiety (R3) including a moiety that can bind a competitive TfR1 epitope (e.g., H7).

[0203] In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHA can bind to a non-competitive epitope in TfR and drive GPCR degradation. In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHA can bind to TfR and drive POI degradation. In some embodiments, the TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHA can bind to TfR and drive GPCR degradation more efficiently and potently than a TransTAC including a GPCR and a second moiety (R3) that can bind a competitive TfR1 epitope (e.g., H7). In some embodiments, a TR binder or TR binding means (also known as a means for binding TR) that binds a transferrin non-competitive epitope may not bind to transferrin receptor better or substantially better than a TR binder that binds a transferrin competitive epitope. In some embodiments, a TR binder or TR binding means (also known as a means for binding TR) that binds a transferrin non-competitive epitope may drive internalization and / or degradation of a POI better than a TR binder that binds a transferrin competitive epitope.

[0204] In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHA can bind to a non-competitive epitope in TfR and drive GPCR degradation. In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHA can bind to TfR and drive GPCR degradation. In some embodiments, the TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and aDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 second moiety (R3) including VHHA can bind to TfR and drive GPCR degradation more efficiently and potently than a TransTAC including a GPCR and a second moiety (R3) that can bind a competitive TfR1 epitope (e.g., H7). In some embodiments, a TR binder or TR binding means (also known as a means for binding TR) that binds a transferrin non-competitive epitope may not bind to transferrin receptor better or substantially better than a TR binder that binds a transferrin competitive epitope. In some embodiments, a TR binder or TR binding means (also known as a means for binding TR) that binds a transferrin non-competitive epitope may drive internalization and / or degradation of a POI better than a TR binder that binds a transferrin competitive epitope. In some embodiments, non-competitive TR binders or TR binding means (also known as a means for binding TR) can limit toxicity that can occur when transferrin binding to TfR is affected by using an R3 that competes with transferrin for binding to TfR.

[0205] In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a CAR, and a second moiety (R3) including VHHA can bind to a non-competitive epitope in TfR and drive CAR degradation. In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind CAR, and a second moiety (R3) including VHHA can bind to TfR and drive CAR degradation. In some embodiments, the TransTAC including a first moiety (R1) including a molecule that can bind CAR, and a second moiety (R3) including VHHA can bind to TfR and drive CAR degradation more efficiently and potently than a TransTAC including CAR and a second moiety (R3) that can bind a competitive TfR1 epitope (e.g., H7). In some embodiments, a TR binder or TR binding means (also known as a means for binding TR) that binds a transferrin non-competitive epitope may not bind to transferrin receptor better or substantially better than a TR binder that binds a transferrin competitive epitope. In some embodiments, a TR binder or TR binding means (also known as a means for binding TR) that binds a transferrin non-competitive epitope may drive internalization and / or degradation of a POI better than a TR binder that binds a transferrin competitive epitope.

[0206] In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHB can bind to a non-competitive epitope in TfR and drive GPCR degradation. In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHB can bind to TfR and drive POI degradation. In some embodiments, the TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and aDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 second moiety (R3) including VHHB can bind to TfR and drive GPCR degradation more efficiently and potently than a TransTAC including a GPCR and a second moiety (R3) that can bind a competitive TfR1 epitope (e.g., H7). In some embodiments, a TR binder or TR binding means (also known as a means for binding TR) that binds a transferrin non-competitive epitope may not bind to transferrin receptor better or substantially better than a TR binder that binds a transferrin competitive epitope. In some embodiments, a TR binder or TR binding means (also known as a means for binding TR) that binds a transferrin non-competitive epitope may drive internalization and / or degradation of a POI better than a TR binder that binds a transferrin competitive epitope.

[0207] In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a CAR, and a second moiety (R3) including VHHB can bind to a non-competitive epitope in TfR and drive CAR degradation. In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind CAR, and a second moiety (R3) including VHHB can bind to TfR and drive CAR degradation. In some embodiments, the TransTAC including a first moiety (R1) including a molecule that can bind CAR, and a second moiety (R3) including VHHB can bind to TfR and drive CAR degradation more efficiently and potently than a TransTAC including CAR and a second moiety (R3) that can bind a competitive TfR1 epitope (e.g., H7). In some embodiments, a TR binder or TR binding means (also known as a means for binding TR) that binds a transferrin non-competitive epitope may not bind to transferrin receptor better or substantially better than a TR binder that binds a transferrin competitive epitope. In some embodiments, a TR binder or TR binding means (also known as a means for binding TR) that binds a transferrin non-competitive epitope may drive internalization and / or degradation of a POI better than a TR binder that binds a transferrin competitive epitope.

[0208] In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHA can bind to TfR1 with a Kd at least about 2, 4, 5, 10, 30, 50, 100, 300, 400, 600, 1000, 1500, 2000 times the Kd of a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including a binder that can bind to H7. In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHA can bind to TfR1 with a Kd between about 10 and about 100 times the Kd of a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and aDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 second moiety (R3) including a binder that can bind to H7. In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHA can bind to an internalizing molecule (e.g., TfR1) with a Kd at least about 100 times the Kd of a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including a binder (e.g., H7) that can bind to a competitive epitope in an internalizing molecule (e.g. TfR1).

[0209] In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHA can bind to an internalizing molecule (e.g., TfR1) with a Kd at least about 100 times the Kd of a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including a binder that can bind to a competitive epitope in an internalizing molecule (e.g. TfR1). In some embodiments, the TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHA can bind to TfR1 with a Kd of about 1.7 nM (nanomolar) and the TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including a binder that can bind to a competitive epitope (H7) in an internalizing molecule (e.g. TfR1) can be about 160 pM (picomolar). In some embodiments, a TR binder or TR binding means (also known as a means for binding TR) that binds a transferrin non-competitive epitope (e.g., VHHA) may not bind to transferrin receptor better or substantially better than a TR binder that binds a transferrin competitive epitope. In some embodiments, a binder or TR binding means (also known as a means for binding TR) that binds a transferrin non-competitive epitope may drive internalization and / or degradation of a POI better than a TR binder that binds a transferrin competitive epitope.

[0210] In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHB can bind to TfR1 with a Kd at least about 2, 4, 5, 10, 30, 50, 100, 300, 400, 600, 1000, 1500, 2000 times the Kd of a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including a binder that can bind to H7. In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHB can bind to TfR1 with a Kd between about 10 and about 100 times the Kd of a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including a binder that can bind to H7. In some embodiments, a TransTAC including a firstDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHB can bind to an internalizing molecule (e.g., TfR1) with a Kd at least about 100 times the Kd of a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including a binder (e.g., H7) that can bind to a competitive epitope in an internalizing molecule (e.g. TfR1).

[0211] In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHB can bind to an internalizing molecule (e.g., TfR1) with a Kd at least about 100 times the Kd of a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including a binder that can bind to a competitive epitope in an internalizing molecule (e.g. TfR1). In some embodiments, the TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHB can bind to TfR1 with a Kd of about 2.7 nM (nanomolar) and the TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including a binder that can bind to a competitive epitope (H7) in an internalizing molecule (e.g. TfR1) can be about 160 pM (picomolar). In some embodiments, a TR binder or TR binding means (also known as a means for binding TR) that binds a transferrin non-competitive epitope (e.g., VHHB) may not bind to transferrin receptor better or substantially better than a TR binder that binds a transferrin competitive epitope. In some embodiments, a TR binder or TR binding means (also known as a means for binding TR) that binds a transferrin non-competitive epitope may drive internalization and / or degradation of a POI better than a TR binder that binds a transferrin competitive epitope.

[0212] In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHA can bind to TfR1 with an affinity at least about 1, 5, 10, 50, 100, 2000, 500, 1000 higher that a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including a binder that can bind to H7. In some embodiments, a TR binder or TR binding means (also known as a means for binding TR) that binds a transferrin non-competitive epitope may not bind to transferrin receptor better or substantially better than a TR binder that binds a transferrin competitive epitope. In some embodiments, a TR binder or TR binding means (also known as a means for binding TR) that binds a transferrin non-competitive epitope may drive internalization and / or degradation of a POI better than a TR binder that binds a transferrin competitive epitope.DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17

[0213] In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHB can bind to TfR1 with an affinity at least about 1, 5, 10, 50, 100, 2000, 500, 1000 higher that a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including a binder that can bind to H7. In some embodiments, a TR binder or TR binding means (also known as a means for binding TR) that binds a transferrin non-competitive epitope may not bind to transferrin receptor better or substantially better than a TR binder that binds a transferrin competitive epitope. In some embodiments, a TR binder or TR binding means (also known as a means for binding TR) that binds a transferrin non-competitive epitope may drive internalization and / or degradation of a POI better than a TR binder that binds a transferrin competitive epitope.

[0214] In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR and a second moiety (R3) including VHHA can bind to TfR and drive GPCR degradation independently of the presence of natural transferrin (Tf). In some embodiments binding of the TransTAC including VHHA to TfR1 is non-competitive with natural Tf.

[0215] In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR and a second moiety (R3) including VHHB can bind to TfR and drive GPCR degradation independently of the presence of natural transferrin (Tf). In some embodiments binding of the TransTAC including VHHB to TfR1 is non-competitive with natural Tf.

[0216] In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHA can internalize and / or degrade GPCR (e.g., TfR1) with an IC50 at least about 1, 2, 4, 5, 10, 30 times smaller than the IC50 of a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including a binder (e.g., H7) that binds to a competitive epitope in an internalizing molecule (e.g. TfR1). In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHA can internalize and / or degrade the GPCR with an IC50 between about 0.05 to about 0.5 nM. In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including H7 can internalize and / or degrade the GPCR with an IC50 of about 1.1 mM. In some embodiments, a TransTAC including a first moiety (R1)DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 including a molecule that can bind a GPCR, and a second moiety (R3) including H7 can internalize and / or degrade the GPCR with an IC50 of about 1.2 mM. In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including H7 can internalize and / or degrade the GPCR with an IC50 of about 1.7 mM.

[0217] In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHB can internalize and / or degrade the GPCR with an IC50 at least about 1, 2, 4, 5, 10, 30 times smaller than the IC50 of a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including a binder (e.g., H7) that binds to a competitive epitope in an internalizing molecule (e.g. TfR1). In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHB can internalize and / or degrade the GPCR with an IC50 between about 0.05 to about 0.5 nM. In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including H7 can internalize and / or degrade the GPCR with an IC50 of about 0.26 mM. In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including H7 can internalize and / or degrade the GPCR with an IC50 of about 0.1 mM.

[0218] In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHA can bind to, internalize and / or degrade the POI using an internalizing molecule (e.g. TfR1) with at least about 1, 5, 10, 50, 100, 200, 500, or 1000 times higher than a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including a binder (e.g., H7) that can bind to a competitive epitope in an internalizing molecule (e.g. TfR1).

[0219] In some embodiments, a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including VHHB can bind to an internalizing molecule (e.g. TfR1) can bind to, internalize and / or degrade the POI with an affinity at least about 1, 5, 10, 50, 100, 200, 500, or 1000 times higher than the affinity of a TransTAC including a first moiety (R1) including a molecule that can bind a GPCR, and a second moiety (R3) including a binder (e.g., H7) that can bind to a competitive epitope in an internalizing molecule (e.g. TfR1).DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17

[0220] In some embodiments, a TransTAC including VHHA can degrade a POI with a potency of about between 0.01 and 0.03 nM EC50, about between 0.02 and 0.05 nM EC50, about between 0.04 and 0.1 nM EC50, about between 0.08 and 0.2 nM EC50, about between 0.15 and 0.3 nM EC50, about between 0.25 and 0.5 nM EC50, about between 0.4 and 1 nM EC50. In some embodiments, a TransTAC including VHHA (SEQ ID NO: 144-145) can degrade a POI with a potency of 0.2 nM EC50. In some embodiments, a TransTAC including VHHA (SEQ ID NO: 144-145) and a first moiety that can bind PD-LI can degrade PD-L1 with a potency of 0.2 nM EC50.

[0221] In some embodiments, a TransTAC including VHHA can degrade a POI with an efficiency of between about 50% and about 60%, between about 55% and about 65%, between about 65% to about 75%, between about 70% and about 80%, between about 80% to 90%, or between about 85% and about 95%, or between about 90% and about 100%. In some embodiments, a TransTAC including VHHA can degrade a POI with an efficiency of about 80%.

[0222] In some embodiments, a TransTAC including VHHB can degrade a POI with a potency of about between 0.01 and 0.03 nM IC50, about between 0.02 and 0.05 nM IC50, about between 0.04 and 0.1 nM IC50, about between 0.08 and 0.2 nM IC50, about between 0.15 and 0.3 nM IC50, about between 0.25 and 0.5 nM IC50, about between 0.4 and 1 nM IC50. In some embodiments, a TransTAC including VHHB (SEQ ID NO: 148-149) can degrade a POI with a potency of 0.1 nM IC50. In some embodiments, a TransTAC including VHHB (SEQ ID NO: 148-149) and a first moiety (R1) that can bind a GPCR and can degrade the GPCR with a potency of about 0.1 nM IC50. In some embodiments, a TransTAC including VHHB (SEQ ID NO: 148-149) and a first moiety (R1) that can bind a GPCR and can degrade the GPCR with a potency of about 0.26 nM IC50.

[0223] In some embodiments, a TransTAC including VHHB can degrade a POI with an efficiency of between about 50% and about 60%, between about 55% and about 65%, between about 65% to about 75%, between about 70% and about 80%, between about 80% to 90%, or between about 85% and about 95%, or between about 90% and about 100%. In some embodiments, a TransTAC including VHHB can degrade a POI with an efficiency of about 100%. In some embodiments, a TransTAC including VHHB can degrade a POI with an efficiency of about 99%. In some embodiments, a TransTAC including VHHB can degrade a POI with an efficiency of about 98%. In some embodiments, a TransTAC including VHHB canDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 degrade a POI with an efficiency of about 97%. In some embodiments, a TransTAC including VHHB can degrade a POI with an efficiency of about 96%.

[0224] In some embodiments, the TransTAC includes a first moiety that binds to any of the first moiety targets described herein, and a second moiety (e.g., VHHA) that binds to an internalizing receptor (e.g., TfR). In some embodiments, the TransTAC includes a first moiety that binds to any of the first moiety targets described herein, and a second moiety (e.g., VHHA) that binds transferrin with an EC50 of about 0.15. In some embodiments, the TransTAC includes a first moiety that binds to a GPCR, and a second moiety that binds to TfR with an EC50 of about 0.15.

[0225] In some embodiments, an antibody or antibody fragment that can bind transferrin receptor can be a nanobody. This nanobody can be called “VHHB.” In some embodiments, VHHB can bind to an epitope in TfR1 that does not compete for binding with natural / endogenous Tf. In some embodiments, a TransTAC including a first moiety including a molecule that can bind a GPCR and a second moiety (TR binder or TR binding means (also known as a means for binding TR)) including VHHB can bind to an epitope of TfR and can drive GPCR degradation independently of the presence of natural transferrin (Tf). In some embodiments binding of the TransTAC including VHHB to TfR1 is non-competitive with natural Tf.

[0226] In some embodiments, a TransTAC including VHHB can degrade a POI with a potency of about between 0.01 and 0.03 nM EC50, about between 0.02 and 0.05 nM EC50, about between 0.04 and 0.1 nM EC50, about between 0.08 and 0.2 nM EC50, about between 0.15 and 0.3 nM EC50, about between 0.25 and 0.5 nM EC50, about between 0.4 and 1 nM EC50. In some embodiments, a TransTAC including VHHB (SEQ ID NO: 146-147) can degrade a POI with a potency of 0.1 nM EC50. In some embodiments, a TransTAC including VHHB (SEQ ID NO: 146-147) and a first moiety that can bind GPCR can degrade GPCR with a potency of 0.1 nM EC50.

[0227] In some embodiments, a TransTAC including VHHB can degrade a POI with an efficiency of between about 50% and about 60%, between about 55% and about 65%, between about 65% to about 75%, between about 70% and about 80%, between about 80% to 90%, or between about 85% and about 95%, or between about 90% and about 100%. In some embodiments, a TransTAC including VHHB can degrade a POI with an efficiency of aboutDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 100%. In some embodiments, a TransTAC including VHHB can degrade a POI with an efficiency of about 99%. In some embodiments, a TransTAC including VHHB can degrade a POI with an efficiency of about 98%. In some embodiments, a TransTAC including VHHB can degrade a POI with an efficiency of about 97%. In some embodiments, a TransTAC including VHHB can degrade a POI with an efficiency of about 96%.

[0228] In some embodiments, a TransTAC including VHHA can bind to TfR1 with a KD of about 2.7 nM. In some embodiments, a TransTAC including VHHB can bind to TfR1 with a KD of about 1.7 nM. In some embodiments, a TransTAC including H7 can bind to TfR1 with a KD of 160 pM.

[0229] In some embodiments, a TransTAC including VHHA and a first motif that binds to a POI can degrade a POI more potently than a TransTAC including H7 and a first motif that binds to the POI. In some embodiments, a TransTAC including VHHB and a first motif that binds to a POI can degrade a POI more potently than a TransTAC including H7 and a first motif that binds to the POI. In some embodiments, a TransTAC including VHHA and a first motif that binds to a POI can degrade a POI more efficiently than a TransTAC including H7 and a first motif that binds to the POI. In some embodiments, a TransTAC including VHHB and a first motif that binds to a POI can degrade a POI more efficiently than a TransTAC including H7 and a first motif that binds to the POI.

[0230] In some embodiments, a TransTAC including VHHA (SEQ ID NO: 146-147) can bind to TfR1 more efficiently than a TransTAC including H7. In some embodiments, a TransTAC including VHHB (SEQ ID NO: 148-149) can bind to TfR1 more efficiently than a TransTAC including H7.

[0231] In some embodiments, a TransTAC including VHHA can degrade a POI more potently than a TransTAC including H7. In some embodiments, a TransTAC including VHHB can degrade a POI more potently than a TransTAC including H7. In some embodiments, a TransTAC including VHHA can degrade a POI more efficiently than a TransTAC including H7. In some embodiments, a TransTAC including VHHB can degrade a POI more efficiently than a TransTAC including H7.

[0232] In one embodiment, an amino acid sequence of VHHA molecules can include the molecules below, or can include molecules at least 60, 65, 70, 75, 80, 81, 82, 83, 84, 85, 86, 87,DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequences below:

[0233] EVQLVESGGGVVQPGGSLKLSCVASGTDFSINFIRWYRQAPGKQREFVAGFTA TGNTNYADSMKGRFTISRDNTKNAVYLQIDSLKPEDTAVYYCYMLDKWGQGTQVTVS S***(SEQ ID NO: 146)

[0234] In one embodiment, a nucleotide sequence of VHHA molecules can include the molecules below, or can include molecules at least 60, 65, 70, 75, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the nucleotide sequences below:

[0235] GAGGTGcAGCTGGTGGAGTCTGGGGGAGGCGTGGTGCAGCCTGGGGGGTC TCTAAAACTCTCCTGCGTAGCCTCGGGAACGGACTTCAGTATCAATTTTATACGCTG GTACCGCCAGGCTCCAGGGAAGCAGCGCGAGTTCGTCGCAGGATTTACTGCGACTG GTAACACAAACTATGCAGACTCCATGAAGGGGCGATTCACCATCTCCAGAGACAAC ACCAAGAACGCGGTGTATCTGCAAATAGACAGCCTGAAACCTGAGGACACGGCCGT GTATTACTGCTATATGTTGGACAAGTGGGGCCAGGGGACCCAGGTCACAGTATCCTC C*** (SEQ ID NO: 147 and FIG.60).

[0236] In embodiments, the VHHA TR binder or TR binding means (also known as a means for binding TR) can be modified as follows: EVQLVESGGGVVQPGGSLKLSCVASGTDFSINFIRWYRQAPGKQREFVAGFTATGNTN YADSMKGRFTISRDNTKNAVYLQIDSLKPEDTAVYYCYMLDKWGQGTQVTVSS*** (111 amino acids; SEQ ID NO: 146). The italicized, bolded and underlined amino acid sections correspond to CDR1, CDR2, and CDR3, respectively. In embodiments, the shaded amino acids in CDR2 and CDR3 can be substituted for a neutral amino acid(s). In embodiments, the modifications can be amino acid substitutions in CDR2. In embodiments, the modifications can be amino acid substitutions in CDR3.

[0237] Table 1. Amino Acid Sequences of VHHA Molecules Disclosed Herein. VHHA (SEQ ID NO: 146) EVQLVESGGGVVQPGGSLKLSCVASGTDFSINFIRWYRQAPGKQREFVAGFTATGNT NYADSMKGRFTISRDNTKNAVYLQIDSLKPEDTAVYYCYMLDKWGQGTQVTVSSDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 T T T

[0238] Table 2. Amino Acid Sequences of Complementarity Determining Regions (CDRs) of VHHA Molecules Disclosed Herein. VHHA Molecule CDR1 CDR2 CDR3, molecules below, or can include molecules at least 60, 65, 70, 75, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequences below:

[0240] EVQLVESGGGVVQPGGSLRLSCAASGEIFSINFMRWYRQAPGKQREWVAGFT RDGSTNYPDSAKGRFTISRDNAKNTVYLQIDSLKPEDTAVYYCYMLDTWGQGTQVTVS S*** (SEQ ID NO: 148 and FIG.60).

[0241] In one embodiment, a nucleotide sequence of VHHB molecules can include the molecules below, or can include molecules at least 60, 65, 70, 75, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the nucleotide sequences below:DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17

[0242] GAgGTGCAGCTGGTGGAGTCTGGGGGagGCGTGGTGCAGCCTGGGGGGTCT CTGAGACTCTCCTGTGCAGCCTCTGGAGAGATCTTCAGTATCAATTTTATGCGCTGGT ACCGCCAGGCTCCAGGGAAGCAGCGCGAGTGGGTCGCAGGTTTTACTAGGGATGGA AGCACAAACTATCCAGACTCCGCGAAGGGCCGATTCACCATCTCTAGAGACAACGC CAAGAACACGGTGTATCTGCAAATAGACAGCCTGAAACCTGAGGACACGGCCGTCT ATTATTGTTATATGTTGGACACCTGGGGCCAGGGGACCCAGGTCACAGTATCCTCC** * (SEQ ID NO: 149 and FIG.60)

[0243] Herein, we disclose novel mechanisms for modulating proteins at the cell membrane of GPCR cells. Endocytosis is a common machinery of regulating membrane protein recycling and degradation. Among the various transmembrane proteins regulated by endocytosis, transferrin receptor (TfR) is a well-characterized recycling receptor with a rapid internalization rate (500 molecules / cell / s). TfR imports iron by binding to a plasma protein transferrin (Tf) in complex with iron. TfR is expressed in various cell types and tissues.

[0244] The method is a novel and generic archetype to degrade proteins with fully recombinant biological molecules. A universal approach for degrading membrane / extracellular proteins would open up unlimited possibilities to manipulate cell behaviors, thus serving as important research tools as well as expanding the PROTAC field’s attempts to target challenging extracellular targets. The fully recombinant nature of TransTAC allows for simple generalization to broad range of targets and optimization of binding properties.

[0245] In some embodiments, the fusion proteins disclosed herein have an antigen to which a receptor can bind and a ligand for an internalizing receptor or membrane protein (e.g., transferrin receptor). In some embodiments, the fusion proteins disclosed herein have an antibody that can bind to a GPCR receptor and a ligand for an internalizing receptor or membrane protein (e.g., a nanobody including VHHA and VHHB).

[0246] In some embodiments, the fusion proteins can be fusion proteins of the formula R1- R2-R3. In some embodiments, the fusion proteins can be fusion proteins of the formula R3-R2- R1. For example, R1 or R3 can be located at the C-terminus or N-terminus of fusion proteins disclosed herein. In some embodiments, the fusion proteins can be dimers of R1-R2-R3 or R3- R2-R1 (homodimers).

[0247] In some embodiments, R1 can be a protein of interest binder (POIB), or a POI binding means (also known as a means for binding POI). In some embodiments, the POIB or POIDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 binding means (also known as a means for binding POI) can be an antibody that binds to the POI. The POIB or POI binding means (also known as a means for binding POI) can bind to a GPCR on a cell surface. In some embodiments, the POIB or POI binding means (also known as a means for binding POI) can bind to an extracellular domain of the GPCR. In some embodiments the POIB POI binding means (also known as a means for binding POI) can be an antibody (e.g., scFv) that can bind to an extracellular domain of the GPCR. Antibodies that bind to an extracellular domain of many different GPCRs are known in the art. In some embodiments, the POIB or POI binding means (also known as a means for binding POI) can bind to an extracellular domain of a GPCR including, but not limited to, APJ, GPR20, CCR6, CCR4, AT1R, PAR2, RXFP1, A2AR, FZD1, CCR2, CXCR1 / 1 or C5aR1. In some embodiments, the POIB or POI binding means (also known as a means for binding POI) can bind to an extracellular domain of any of the GPCR described herein. The POI binding means (also known as a means for binding POI) (R1), as part of a fusion protein (R1-R2-R3), can bind to a GPCR on the surface of a cell and can result in the GPCR being internalized and, optionally degraded, as described below. Amino acid sequences of example POIBs or POI binding means (also known as a means for binding POI) are shown in Table 3. Amino acid sequences of CDRs from these POIBs or POI binding means (also known as a means for binding POI) are shown in Table 4.

[0248] Table 3. Amino Acid Sequences of POIBs or POI binding means (also known as a means for binding POI) Disclosed Herein. AT1R (SEQ ID NO: 184) EVQLVESGGGLVQPGGSLRLSCAASGYIYSRYRMGWYRQAPGKGREFVAAISGGSST Q M N GDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 CDRs are underlined, bolded and

[0249] Table 4. Amino Acid Sequences of Complementarity Determining Regions (CDRs) of POIBs or POI binding means (also known as a means for binding POI) Disclosed Herein. POIB CDR1 CDR2 CDR3 S

[0250] In some embodiments, the POI binding means (also known as a means for binding POI) can bind to an extracellular domain of any of the GPCRs included in any of the GPCR classes, families, and types described herein. In some embodiments, R3 can be a transferrin receptor (TR) binding means (also known as a means for binding TR). The TR binding means (also known as a means for binding TR) binds to a transferrin receptor on the surface of cells.

[0251] In some embodiments, the TR binder or TR binding means (also known as a means for binding TR) (R3) can be a molecule that binds to a transferrin receptor (TfR). In some embodiments, the TR binder or TR binding means (also known as a means for binding TR) can be transferrin or a part of transferrin that can bind to a TfR. In some embodiments, a TR binder or TR binding means (also known as a means for binding TR) can be non-competitive for binding to a TfR (e.g., VHHA or VHHB). The TR binder or TR binding means (also known as a means for binding TR) (R3), as part of a fusion protein molecule (e.g., R1-R2-R3), can bind to transferrin receptor (TfR) on the surface of a cell and be internalized into the cell. When the protein of interest binder (POIB; R1) or POI binding means (also known as a means for binding POI) in the fusion protein binds a GPCR on a cell surface, internalization of the TR binder or TR binding means (also known as a means for binding TR) can also cause internalization of the GPCR. Internalized POIs can be degraded once internalized by a cell. A TR binder or TR binding means binds to TfR such that the TR binder or TR binding means (also known as a means for binding TR) can be internalized, and optionally degraded. Within the context of aDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 fusion protein, the TR binder or TR binding means binds to TfR, is internalized, and also results in internalization of a GPCR bound by the POIB or POI binding means (also known as a means for binding POI) portion (R1) of the fusion protein. The internalized GPCR can be degraded.

[0252] In some embodiments, the TR binder or TR binding means (also known as a means for binding TR) can be a nanobody that binds to the transferrin receptor. In some embodiments, the TR binder or TR binding means (also known as a means for binding TR) can be a nanobody that binds to the transferrin receptor without competing with endogenous Tf for binding to the TfR1. In some embodiments, the TR binder or TR binding means (also known as a means for binding TR) can be VHHA (SEQ IDs NO: 146-147). In some embodiments, the TR binder or TR binding means (also known as a means for binding TR) can be VHHB (SEQ IDs NO: 148-149). See FIG. 50.

[0253] In some embodiments, a TR binder or TR binding means (also known as a means for binding TR can have CDRs. In some embodiments, the CDRs can be a VH CDR1 comprising the amino acid sequence of GTDFSINF (SEQ ID NO: 175), a VH CDR2 comprising the amino acid sequence of ITATGNT (SEQ ID NO: 176), and a VH CDR3 comprising the amino acid sequence of YMLDK (SEQ ID NO: 177), a VH CDR1 comprising the amino acid sequence of GTDFSINF (SEQ ID NO: 175), a VH CDR2 comprising the amino acid sequence of FTATGNT (SEQ ID NO: 179), and a VH CDR3 comprising the amino acid sequence of AMLDK (SEQ ID NO: 180), or a VH CDR1 comprising the amino acid sequence of GTDFSINF (SEQ ID NO: 175), a VH CDR2 comprising the amino acid sequence of FTATGNT (SEQ ID NO: 179), and a VH CDR3 comprising the amino acid sequence of YMADK (SEQ ID NO: 183).

[0254] In some embodiments, R2 can be a linker of the formula R4-R5 or R5-R4. In some embodiments, R4 can be an Fc region from an antibody. In some embodiments, R4 can be an Fc region from IgG. In some embodiments, R5 is an optional protein-sensitive linking means (also known as a means for linking). In some embodiments, an Fc region can be (SEQ ID NO: 6):

[0255] DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPA PIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 6).DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17

[0256] In some embodiments, the Fc regions can dimerize, forming homodimers or heterodimer structures. In some embodiments, the Fc regions can have, or can be modified to have, cysteine amino acids that are capable of forming disulfide bonds. In some embodiments, dimers of the R1-R2-R3 fusion proteins can form through disulfide bonds (1 or more, such as 2 disulfide bonds) between cysteine residues in R2 regions of separate fusion protein molecules. In some embodiments, the disulfide bonds form between R4 in separate fusion molecules (e.g., Fc with disulfides can be a type of dimerization domain). In some embodiments, disulfide bonds form between fusion proteins of a homodimer.

[0257] In some embodiments (e.g., heterodimers), one monomer of the heterodimer can have a “knob”, and the second monomer of the heterodimer can have a “hole” structure. In some embodiments, the knob and hole structures of two monomers can be in the Fc regions (R4) of the monomers. Knob-into-hole technologies are well known in the art (Xu et al., MAbs.2015 Jan- Feb; 7(1): 231–242).

[0258] In certain embodiments, the Fc region can be a variant comprising an amino acid substitution which alters antigen-independent effector functions, like the circulating half-life of a molecule to which it is linked. Molecules linked to these Fc regions can exhibit either increased or decreased binding to FcRn compared to Fc regions lacking these substitutions and can have an increased or decreased half-life in serum, respectively. Fc variants with improved affinity for FcRn are anticipated to have longer serum half-lives, and such molecules have useful applications in methods long half-life of the linked molecule is desired. In contrast, Fc variants with decreased FcRn binding affinity are expected to have shorter half-lives, and such molecules are also useful, for example, where a shortened circulation time can be advantageous. Fc variants with decreased FcRn binding affinity are also less likely to cross the placenta. In addition, other applications in which reduced FcRn binding affinity can be desired include those applications in which localization to the brain, kidney, and / or liver is desired. In one embodiment, the Fc variant-linked molecules can exhibit reduced transport across the epithelium of kidney glomeruli from the vasculature.

[0259] In another embodiment, the Fc variant-linked molecules can exhibit reduced transport across the blood brain barrier (BBB) from the brain, into the vascular space. In one embodiment, an Fc region with altered FcRn binding comprises an Fc domain having one or more amino acid substitutions within the "FcRn binding loop" of an Fc domain. The FcRn binding loop isDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 comprised of amino acid residues 280-299 (according to EU numbering). Exemplary amino acid substitutions with altered FcRn binding activity are disclosed in PCT Publication No. WO05 / 047327 which is incorporated by reference herein. In certain exemplary embodiments, the fusion proteins disclosed herein comprise an Fc domain having one or more of the following substitutions: V284E, H285E, N286D, K290E and S304D (EU numbering).

[0260] In some embodiments, a molecule disclosed herein can be linked to an Fc variant comprising an amino acid substitution which alters glycosylation. For example, the Fc variant can have reduced glycosylation (e.g., N- or O-linked glycosylation). In some embodiments, the Fc variant comprises reduced glycosylation of the N-linked glycan normally found at amino acid position 297 (EU numbering). In another embodiment, the molecules can have an amino acid substitution near or within a glycosylation motif, for example, an N-linked glycosylation motif that contains the amino acid sequence NXT or NXS. In a particular embodiment, the Fc variant can have amino acid substitution at amino acid position 228 or 299 (EU numbering). Exemplary amino acid substitutions which confer reduced or altered glycosylation are described in PCT Publication No, WO05 / 018572, which is incorporated by reference herein in its entirety.

[0261] In some embodiments, the molecules disclosed herein can be modified to eliminate glycosylation and can be referred to as "agly" molecules. Exemplary agly molecules, can have an aglycosylated Fc region of an IgG4 antibody which is devoid of Fc-effector function thereby eliminating the potential for Fc mediated toxicity to the normal vital tissues and cells. In yet other embodiments, the molecules disclosed herein can have an altered glycan. For example, there can be a reduced number of fucose residues on an N-glycan at Asn297 of the Fc region, i.e., is afucosylated. In some embodiments, there can be an altered number of sialic acid residues on the N-glycan at Asn297 of the Fc region.

[0262] In some embodiments, the CH2 or CH3 region of the Fc antibody domain can be truncated or modified to adjust the half-life of the molecule. In some embodiments, an Fc truncation includes CH3 or CH2 (e.g., Gehlsen, Kurt R., et al. "Pharmacokinetics of engineered human monomeric and dimeric CH2 domains." MAbs. Vol.4. No.4. Taylor & Francis, 2012; Ying, Tianlei, et al. "Engineered soluble monomeric IgG1 CH3 domain: generation, mechanisms of function, and implications for design of biological therapeutics." Journal of Biological Chemistry 288.35 (2013): 25154-25164).DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17

[0263] In some embodiments, R4 can be a dimerization domain. The dimerization domain can be any region that can associate with another dimerization domain, through covalent or non- covalent bonds, to form a dimer (e.g., a fusion protein that is a homodimer or heterodimer).

[0264] There are many protein dimerization domains known in the art (e.g., see Dang, Dung Thanh. "Molecular Approaches to Protein Dimerization: Opportunities for Supramolecular Chemistry." Frontiers in Chemistry 10 (2022): 829312). An example dimerization domain can include zipper motifs, like a leucine zipper.

[0265] In some embodiments, the dimerization can form between regions of the fusion proteins that are not R4 regions.

[0266] In some embodiments, R5 can be a protease-sensitive linking means (also known as a means for linking). In some embodiments, R5 is an optional feature. In some embodiments, the protease-sensitive linking means (also known as a means for linking) is an amino acid sequence that can be cleaved by a protease. In some embodiments, the protease can be a protease in an endosome or lysosome. In some embodiments, the protease can be a cathepsin (e.g., cathepsin B) and the protease-sensitive linking means (also known as a means for linking) can be a cathepsin-cleavable peptide. The protease-sensitive linking means (also known as a means for linking) can be an amino acid sequence that can be cleaved by a protease. In some embodiments, the protease-sensitive linker has an amino acid sequence that can be recognized by a cathepsin protease. In some embodiments, cleavage of R5 within the context of a fusion protein can result in the internalized GPCR being degraded. In some embodiments, recycling of the internalized TfR can also be improved.

[0267] Some example protease-sensitive linking means (also known as a means for linking) are shown in FIG.35. In some embodiments, the protease-sensitive linking means (also known as a means for linking) can be GGFLGGVRGVDG (SEQ ID NO: 7) or GSGSGGEVRGVDG (SEQ ID NO: 8). Sequences of other protease-sensitive linking means (also known as a means for linking) are disclosed herein (e.g., FK, VA, VK, SEQ ID NO: 7, 8, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145). In some embodiments, sequences that are at least 80, 84, 88, 92 or 96 percent identical to these sequences can be used. Functional alternative amino acid sequences that can be cleaved by a protease (e.g., cathepsin) are known in the art. Use of other protease- sensitive linkers are contemplated.DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17

[0268] In some embodiments, a linkage noted as a “-“ (e.g., not R2) can be located between various sections of the R1-R2-R3 fusion protein. In some embodiments, a linkage can be located between R2 and R3. In some embodiments, the fusion proteins described herein comprise a linker between R2 and R3. In some embodiments, this linkage can be located between R1 and R2. In some embodiments, the linkage can be a glycine-rich linker (“GS linker). In some embodiments, a “GS” linker can be a combination of glycine and serine amino acids. In some embodiments, the GS linker can be GSSGGSGGSGGS (SEQ ID NO: 9). Other sequences are possible. In some embodiments, the GS linker can be SGGGG (SEQ ID NO: 10), SGGGSGGG (SEQ ID NO: 11), GSSGGSGGSGGS (SEQ ID NO: 12), GSGS (SEQ ID NO: 13), GSGGS (SEQ ID NO: 14), GSSGSS (SEQ ID NO: 15), GSSSSSS (SEQ ID NO: 16) and the like. In some embodiments, a GS linker can have at least 4 amino acids that are glycine and / or serine. In some embodiments, other amino acids can be part of a GS linker, as long as glycine and serine are in the majority. In some embodiments, optionally, a linker (e.g., glycine-rich) can be comprised between R2 and R3. In some embodiments, optionally, a linker can be comprised between R1 and R2.

[0269] In some embodiments, the GS linkers are one to 50 amino acids in length (e.g., 1, 2, 3, 4, 5, 10, 12, 1-10, 1-12, or 1-20, or more amino acids) containing various types of amino acids (e.g., US Patent No.11,041,023, incorporated by reference herein in its entirety on March 14, 2024). In some embodiments, the GS linker is 12 amino acids in length. In some embodiments, the GS linker can be 20, 25, 30, 35, 40, 45 or 50 amino acids in length. In embodiments, the amino acids include a combination of one or more glycine(s) and / or serine(s). In embodiments, the peptide linker comprises the amino acid sequence comprising GGGGS (SEQ ID NO: 200). In embodiments, the peptide linker comprises or consists of 2, 3, 4, 5, 6, or 7 consecutive repeats of the amino acid sequence GGGGS (SEQ ID NO: 200). Particularly in embodiments, the peptide linker comprises or consists of the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO: 201).

[0270] In some embodiments, fusion proteins that can bind to a dimeric TfR (e.g., that can bind to two TfRs) can be more efficient at internalizing and / or degrading a POI than a fusion protein that can bind a monomeric TfR (e.g., that can bind to one TfR). In some embodiments, fusion proteins can include one copy of R3. In some embodiments, fusion proteins can includeDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 two copies of R3. In some embodiments, fusion proteins can include three or more copies of R3. In some embodiments, different R3s can be combined in a fusion protein.

[0271] In some embodiments, a fusion protein with two R3 can be more efficient at internalizing and / or degrading a POI than a fusion protein including two R1 (e.g., v0.6 vs. v0.7, FIG.37A). In some embodiments, a fusion protein with two R3 can be at least between about 10 to about 20%, about 15 to about 25%, about 20% to about 30%, about 25 to about 35%, about 30 and about 40%, about 35 and about 45%, about 40% and about 50%, about 45 and about 55%, about 50 to about 60%, about 55 to about 65%, about 60 to about 70%, about 65 to about 75%, about 70% to about 80%, about 75% to about 85%, about 80 to about 90%, about 85% to about 95%, about 90 to about 100% more efficient at internalizing and / or degrading a POI than a fusion protein including two R1.

[0272] In some embodiments, fusion proteins including one monomer including from N terminus to C terminus one copy of R1, a linker (not R2), an Fc (knob), a linker (not R2), and one copy of R3, respectively, and another monomer including an Fc (hole), a linker (not R2), and one copy of R3, wherein both monomers are bound by knob / hole interactions (e.g., v0.6 in FIG. 37A) can be more efficient at internalizing and / or degrading a POI (e.g., APJ, GPR20, CCR6, CCR2, CCR4, AT1R, CXCR1 / 1, PAR2, RXFP1, A2AR, FZD1, C5aR1 or any of the GPCR described herein) than fusion proteins including one monomer including from N terminus to C terminus one copy of R1, a linker (not R2), and an Fc (hole), and a second monomer including from N terminus to C terminus one copy of R1, a linker (not R2), an Fc (knob), a linker (not R2), and one copy of R3, respectively, wherein both monomers are bound by knob / hole interactions (e.g., v0.7 in FIG.37A).

[0273] In some embodiments, fusion proteins including one monomer including from N terminus to C terminus one copy of R1, a linker (not R2), an Fc (knob), a linker (not R2), and one copy of R3, respectively; and another monomer including an Fc (hole), a linker (not R2), and one copy of R3, respectively, wherein both monomers are bound by knob / hole interactions (e.g., v0.6) can be at least between about 10 to about 20%, about 15 to about 25%, about 20% to about 30%, about 25 to about 35%, about 30 and about 40%, about 35 and about 45%, about 40% and about 50%, about 45 and about 55%, about 50 to about 60%, about 55 to about 65%, about 60 to about 70%, about 65 to about 75%, about 70% to about 80%, about 75% to about 85%, about 80 to about 90%, about 85% to about 95%, about 90 to about 100% more efficient at internalizingDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 and / or degrading a POI (e.g., APJ, GPR20, CCR6, CCR2, CCR4, AT1R, CXCR1 / 1, PAR2, RXFP1, A2AR, FZD1, C5aR1, or any of the GPCR described herein) than fusion proteins including one monomer including from N terminus to C terminus one copy of R1, a linker (not R2), and an Fc (hole), and a second monomer including from N terminus to C terminus one copy of R1, a linker (not R2), an Fc (knob), a linker (not R2), and one copy of R3, respectively, wherein both monomers are bound by knob / hole interactions (e.g., v0.7).

[0274] In some embodiments, fusion proteins including one monomer including from N terminus to C terminus one copy of CD19NT.1, a linker (not R2), an Fc (knob), a linker (not R2), and one copy of R3, respectively; and another monomer including an Fc (hole) a linker (not R2) and one copy of R3, respectively, wherein both monomers are bound by knob / hole interactions (e.g., v0.6) can be more efficient at internalizing and / or degrading a POI than heterodimeric fusion proteins including one monomer including from N terminus to C terminus one copy of CD19NT.1, a linker (not R2), and an Fc (hole), and a second monomer including from N terminus to C terminus one copy of CD19NT.1, a linker (not R2), an Fc (knob), a linker (not R2) and one copy of R3, respectively, wherein both monomers are bound by knob / hole interactions (e.g., v0.7).

[0275] In some embodiments, fusion proteins including one monomer including from N terminus to C terminus one copy of CD19NT.1, a linker (not R2), an Fc (knob), a linker (not R2), and one copy of R3, respectively, and another monomer including an Fc (hole) a linker (not R2) and one copy of R3, respectively, wherein both monomers are bound by knob / hole interactions (e.g., v0.6) can be can be at least between about 10 to about 20%, about 15 to about 25%, about 20% to about 30%, about 25 to about 35%, about 30 and about 40%, about 35 and about 45%, about 40% and about 50%, about 45 and about 55%, about 50 to about 60%, about 55 to about 65%, about 60 to about 70%, about 65 to about 75%, about 70% to about 80%, about 75% to about 85%, about 80 to about 90%, about 85% to about 95%, about 90 to about 100% more efficient at internalizing and / or degrading a POI (e.g., APJ, GPR20, CCR6, CCR2, CCR4, AT1R, CXCR1 / 1, PAR2, RXFP1, A2AR, FZD1, C5aR1, or any of the GPCR described herein) than heterodimeric fusion proteins including one monomer including from N terminus to C terminus one copy of CD19NT.1, a linker (not R2), and an Fc (hole), and a second monomer including from N terminus to C terminus one copy of CD19NT.1, a linker (not R2), an FcDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 (knob), a linker (not R2) and one copy of R3, respectively, wherein both monomers are bound by knob / hole interactions (e.g., v0.7).

[0276] In some embodiments, a fusion protein with two R3 can be more efficient at internalizing and / or degrading a POI than a fusion protein including one R3 (e.g., FIG.37A). In some embodiments, a fusion protein with two R3 can be at least between about 10 to about 20%, about 15 to about 25%, about 20% to about 30%, about 25 to about 35%, about 30 and about 40%, about 35 and about 45%, about 40% and about 50%, about 45 and about 55%, about 50 to about 60%, about 55 to about 65%, about 60 to about 70%, about 65 to about 75%, about 70% to about 80%, about 75% to about 85%, about 80 to about 90%, about 85% to about 95%, about 90 to about 100% more efficient at internalizing and / or degrading a POI than a fusion protein including one R3.

[0277] In some embodiments, fusion proteins including one monomer including from N terminus to C terminus one copy of R1, a linker (not R2), an Fc, a linker (not R2), and one copy of R3, respectively, and another monomer including one copy of R1, a linker (not R2), an Fc, a linker (not R2), and one copy of R3, respectively, wherein both monomers are bound by the Fc (e.g., v0.5 FIG.37A) can be more efficient at internalizing and / or degrading a POI (e.g., APJ, GPR20, CCR6, CCR2, CCR4, AT1R, CXCR1 / 1, PAR2, RXFP1, A2AR, FZD1, C5aR1, or any of the GPCR described herein) than fusion proteins including one monomer including from N terminus to C terminus one copy of R1, a linker (not R2), and an Fc (hole), and a second monomer including from N terminus to C terminus one copy of R1, a linker (not R2), an Fc (knob), a linker (not R2), and one copy of R3, respectively, wherein both monomers are bound by knob / hole interactions (e.g., v0.7 in FIG.37A).

[0278] In some embodiments, fusion proteins including one monomer including from N terminus to C terminus one copy of R1, a linker (not R2), an Fc, a linker (not R2), and one copy of R3, respectively, and another monomer including one copy of R1, a linker (not R2), an Fc, a linker (not R2), and one copy of R3, respectively, wherein both monomers are bound by the Fc (e.g., v0.5, FIG.37A) can be at least between about 10 to about 20%, about 15 to about 25%, about 20% to about 30%, about 25 to about 35%, about 30 and about 40%, about 35 and about 45%, about 40% and about 50%, about 45 and about 55%, about 50 to about 60%, about 55 to about 65%, about 60 to about 70%, about 65 to about 75%, about 70% to about 80%, about 75% to about 85%, about 80 to about 90%, about 85% to about 95%, about 90 to about 100% moreDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 efficient at internalizing and / or degrading a POI (e.g., APJ, GPR20, CCR6, CCR2, CCR4, AT1R, CXCR1 / 1, PAR2, RXFP1, A2AR, FZD1, C5aR1, or any of the GPCR described herein) than fusion proteins including one monomer including from N terminus to C terminus one copy of R1, a linker (not R2), and an Fc, and a second monomer including from N terminus to C terminus one copy of R1, a linker (not R2), an Fc, a linker (not R2), and one copy of R3, respectively, wherein both monomers are bound by knob / hole interactions (e.g., v0.7 in FIG. 37A).

[0279] In some embodiments, fusion proteins including one monomer including from N terminus to C terminus one copy of a CD19NT.1 binder, a linker (not R2), an Fc, a linker (not R2), and one copy of R3, respectively, and another monomer including one copy of a CD19NT.1 binder, a linker (not R2), an Fc, a linker (not R2), and one copy of R3, respectively, wherein both monomers are bound by the Fc (e.g., v0.5 FIG.37A) can be more efficient at internalizing and / or degrading a POI (e.g., APJ, GPR20, CCR6, CCR2, CCR4, AT1R, CXCR1 / 1, PAR2, RXFP1, A2AR, FZD1, C5aR1, or any of the GPCR described herein) than fusion proteins including one monomer including from N terminus to C terminus one copy of a CD19NT.1 binder, a linker (not R2), and an Fc, and a second monomer including from N terminus to C terminus one copy of a CD19NT.1 binder, a linker (not R2), an Fc, a linker (not R2), and one copy of R3, respectively, wherein both monomers are bound by knob / hole interactions (e.g., v0.7 in FIG. 37A).

[0280] In some embodiments, fusion proteins including one monomer including from N terminus to C terminus one copy of a CD19NT.1 binder, a linker (not R2), an Fc, a linker (not R2), and one copy of R3, respectively, and another monomer including one copy of a CD19NT.1 binder, a linker (not R2), an Fc, a linker (not R2), and one copy of R3, respectively, wherein both monomers are bound by the Fc (e.g., v0.5 FIG.37A) can be at least between about 10 to about 20%, about 15 to about 25%, about 20% to about 30%, about 25 to about 35%, about 30 and about 40%, about 35 and about 45%, about 40% and about 50%, about 45 and about 55%, about 50 to about 60%, about 55 to about 65%, about 60 to about 70%, about 65 to about 75%, about 70% to about 80%, about 75% to about 85%, about 80 to about 90%, about 85% to about 95%, about 90 to about 100% more efficient more efficient at internalizing and / or degrading a POI (e.g., APJ, GPR20, CCR6, CCR2, CCR4, AT1R, CXCR1 / 1, PAR2, RXFP1, A2AR, FZD1, C5aR1, or any of the GPCR described herein) than fusion proteins including one monomerDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 including from N terminus to C terminus one copy of a CD19NT.1 binder, a linker (not R2), and an Fc, respectively, and a second monomer including from N terminus to C terminus one copy of a CD19NT.1 binder, a linker (not R2), an Fc, a linker (not R2), and one copy of R3, respectively, wherein both monomers are bound by knob / hole interactions (e.g., v0.7 in FIG.37A).

[0281] In some embodiments, fusion proteins including one monomer including from N terminus to C terminus one copy of R1, a linker (not R2), and an Fc (hole), respectively, and a second monomer including from N terminus to C terminus one copy of R3, a linker (not R2) and an Fc (knob), respectively, wherein both monomers are bound by knob / hole interactions (e.g., v0.8; FIG.37A), can be more efficient at internalizing and / or degrading a POI than fusion proteins including one monomer including from N terminus to C terminus one copy of R1, a linker (not R2), an Fc (hole), a linker (not R2), and one copy of R3, respectively, and another monomer including an Fc (knob), respectively, wherein both monomers are bound by knob / hole interactions (e.g., v0.9; FIG.37A).

[0282] In some embodiments, fusion proteins including one monomer including from N terminus to C terminus one copy of R1, a linker (not R2), and an Fc (hole), and a second monomer including from N terminus to C terminus one copy of R3, a linker (not R2), and an Fc (knob), respectively, wherein both monomers are bound by knob / hole interactions (e.g., v0.8), can be at least between about 10 to about 20%, about 15 to about 25%, about 20% to about 30%, about 25 to about 35%, about 30 and about 40%, about 35 and about 45%, about 40% and about 50%, about 45 and about 55%, about 50 to about 60%, about 55 to about 65%, about 60 to about 70%, about 65 to about 75%, about 70% to about 80%, about 75% to about 85%, about 80 to about 90%, about 85% to about 95%, about 90 to about 100% more efficient at internalizing and / or degrading a POI (e.g., APJ, GPR20, CCR6, CCR2, CCR4, AT1R, CXCR1 / 1, PAR2, RXFP1, A2AR, FZD1, C5aR1, or any of the GPCR described herein) than fusion proteins including one monomer including from N terminus to C terminus one copy of R1, a linker (not R2), an Fc (hole), a linker (not R2), and one copy of R3, respectively, and another monomer including an Fc (knob), wherein both monomers are bound by knob / hole interactions (e.g., v0.9).

[0283] In some embodiments, fusion proteins including one monomer including from N terminus to C terminus one copy of CD19NT.1, a linker (not R2), and an Fc (hole), and a second monomer including from N terminus to C terminus one copy of R3, a linker (not R2) and an Fc (knob), respectively, wherein both monomers are bound by knob / hole interactions (e.g., v0.8),DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 can be more efficient at internalizing and / or degrading a POI than heterodimeric fusion proteins including one monomer including from N terminus to C terminus one copy of CD19NT.1, a linker (not R2), an Fc (hole), a linker (not R2), and one copy of R3, respectively, and another monomer including an Fc (knob), wherein both monomers are bound by knob / hole interactions (e.g., v0.9).

[0284] In some embodiments, heterodimeric fusion proteins including one monomer including from N terminus to C terminus one copy of CD19NT.1, a linker (not R2), and an Fc (hole), respectively, and a second monomer including from N terminus to C terminus one copy of R3, a linker (not R2), and an Fc (knob), respectively, wherein both monomers are bound by knob / hole interactions (e.g., v0.8), can be at least between about 10 to about 20%, about 15 to about 25%, about 20% to about 30%, about 25 to about 35%, about 30 and about 40%, about 35 and about 45%, about 40% and about 50%, about 45 and about 55%, about 50 to about 60%, about 55 to about 65%, about 60 to about 70%, about 65 to about 75%, about 70% to about 80%, about 75% to about 85%, about 80 to about 90%, about 85% to about 95%, about 90 to about 100% more efficient at internalizing and / or degrading a POI (e.g., APJ, GPR20, CCR6, CCR2, CCR4, AT1R, CXCR1 / 1, PAR2, RXFP1, A2AR, FZD1, C5aR1, or any of the GPCR described herein) than fusion proteins including one monomer including from N terminus to C terminus one copy of CD19NT.1, a linker (not R2), an Fc (hole), a linker (not R2), and one copy of R3, respectively, and another monomer including an Fc (knob), wherein both monomers are bound by knob / hole interactions (e.g., v0.9).

[0285] In some embodiments, a fusion protein can include two or more copies of an R3s located in tandem (e.g., two or more R3 contiguous to each other; e.g., v1.2 in FIG.1A). In some embodiments, a fusion protein can include three in tandem copies of R3. In some embodiments, a fusion protein can include four in tandem copies of R3. In some embodiments, a fusion protein can include five in tandem copies of R3. In some embodiments, a fusion protein can include six in tandem copies of R3. In some embodiments, a fusion protein can include a plurality of in tandem copies of R3.

[0286] In some embodiments, a fusion protein with two or more R3s located in tandem (e.g., two or more R3 contiguous to each other (e.g., v1.2 in FIG.1A and SEQ ID NO: 150-151, FIG. 3A) can be more efficient at internalizing and / or degrading a POI than a fusion protein including two R3 that are not in tandem (e.g., v1.4 in FIG.1A; SEQ IDs NO: 152-153; FIG.3B; and v1.0DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 in FIG.1A). In some embodiments, a fusion protein with two R3s located in tandem (e.g., two R3 contiguous to each other (e.g., v1.2 in FIG.1A and SEQ ID NO: 150-151, FIG.3A) can be at least between about 10 to about 20%, about 15 to about 25%, about 20% to about 30%, about 25 to about 35%, about 30 and about 40%, about 35 and about 45%, about 40% and about 50%, about 45 and about 55%, about 50 to about 60%, about 55 to about 65%, about 60 to about 70%, about 65 to about 75%, about 70% to about 80%, about 75% to about 85%, about 80 to about 90%, about 85% to about 95%, about 90 to about 100% more efficient at internalizing and / or degrading a POI than a fusion protein including two R3 that are not in tandem (e.g., v1.4 in FIG. 1A; SEQ IDs NO: 152-153; FIG.3B; and v1.0 in FIG.1A).

[0287] In some embodiments, fusion proteins including one monomer including from N terminus to C terminus one copy of R1, a linker (not R2), an Fc (knob), respectively, and another monomer including from N terminus to C terminus two in tandem copies of R3, and an Fc (knob), respectively, wherein both monomers are bound by knob / hole interactions (e.g., v1.2; FIG.1A) can be more efficient at internalizing and / or degrading a POI than fusion proteins including one monomer including from N terminus to C terminus one copy of R1, a linker (not R2), an Fc (knob), and one copy of R3, respectively, and a second monomer including from N terminus to C terminus an Fc (hole) and one copy of R3, respectively, wherein both monomers are bound by knob / hole interactions (e.g., v1.4; FIG.1 and FIG.3).

[0288] In some embodiments, fusion proteins including one monomer including from N terminus to C terminus one copy of R1, a linker (not R2), an Fc (knob), respectively, and another monomer including from N terminus to C terminus two in tandem copies of R3, and an Fc (knob), respectively, wherein both monomers are bound by knob / hole interactions (e.g., v1.2), can be at least between about 10 to about 20%, about 15 to about 25%, about 20% to about 30%, about 25 to about 35%, about 30 and about 40%, about 35 and about 45%, about 40% and about 50%, about 45 and about 55%, about 50 to about 60%, about 55 to about 65%, about 60 to about 70%, about 65 to about 75%, about 70% to about 80%, about 75% to about 85%, about 80 to about 90%, about 85% to about 95%, about 90 to about 100% more efficient at internalizing and / or degrading a POI than fusion proteins including one monomer including from N terminus to C terminus one copy of R1, a linker (not R2), an Fc (knob), and one copy of R3, respectively, and a second monomer including from N terminus to C terminus an Fc (hole) and one copy ofDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 R3, respectively, wherein both monomers are bound by knob / hole interactions (e.g., v1.4; FIG.1 and FIG.3).

[0289] In some embodiments, fusion proteins including one monomer including from N terminus to C terminus one copy of a GPCR binder, a linker (not R2), an Fc (knob), respectively, and another monomer including from N terminus to C terminus two in tandem copies of R3, and an Fc (knob), respectively, wherein both monomers are bound by knob / hole interactions (e.g., v1.2), can be more efficient at internalizing and / or degrading a POI than heterodimeric fusion proteins including one monomer including from N terminus to C terminus one copy of an GPCR binder, a linker (not R2), an Fc (knob), and one copy of R3, respectively, and a second monomer including from N terminus to C terminus an Fc (hole) and one copy of R3, respectively, wherein both monomers are bound by knob / hole interactions (e.g., v1.4; FIG.1 and FIG.3).

[0290] In some embodiments, fusion proteins including one monomer including from N terminus to C terminus one copy of a GPCR binder, a linker (not R2), an Fc (knob), respectively, and another monomer including from N terminus to C terminus two in tandem copies of R3, and an FC (knob), respectively, wherein both monomers are bound by knob / hole interactions (e.g., v1.2), can be at least between about 10 to about 20%, about 15 to about 25%, about 20% to about 30%, about 25 to about 35%, about 30 and about 40%, about 35 and about 45%, about 40% and about 50%, about 45 and about 55%, about 50 to about 60%, about 55 to about 65%, about 60 to about 70%, about 65 to about 75%, about 70% to about 80%, about 75% to about 85%, about 80 to about 90%, about 85% to about 95%, about 90 to about 100% more efficient at internalizing and / or degrading a POI than heterodimeric fusion proteins including one monomer including from N terminus to C terminus one copy of an GPCR binder, a linker (not R2), an Fc (knob), and one copy of R3, respectively, and a second monomer including from N terminus to C terminus an Fc (hole) and one copy of R3, respectively, wherein both monomers are bound by knob / hole interactions (e.g., v1.4; FIG.1 and FIG.3).

[0291] In some embodiments, the location of the cathepsin sensitive linker within a fusion protein can modulate the efficiency of the fusion protein at internalizing and / or degrading a POI. In some embodiment, a fusion protein can include a cathepsin B-sensitive linker between the Fc domain and the R3 (e.g., v0.3, FIG.34). In some embodiments, a fusion protein can include a cathepsin B-sensitive linker between R1 and the Fc domain (e.g., v0.4, FIG.34). In some embodiments, a fusion protein including a cathepsin B-sensitive linker between R1 and the FcDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 domain (e.g., v0.4) can be more efficient at internalizing and / or degrading a POI than a fusion protein including a cathepsin B-sensitive linker between the Fc domain and the R3 (e.g., v0.3; FIG.34D).

[0292] In some embodiments, a fusion protein including a cathepsin B-sensitive linker between R1 and the Fc domain (e.g., v0.4) can be at least between about 10 to about 20%, about 15 to about 25%, about 20% to about 30%, about 25 to about 35%, about 30 and about 40%, about 35 and about 45%, about 40% and about 50%, about 45 and about 55%, about 50 to about 60%, about 55 to about 65%, about 60 to about 70%, about 65 to about 75%, about 70% to about 80%, about 75% to about 85%, about 80 to about 90%, about 85% to about 95%, about 90 to about 100% more efficient at internalizing and / or degrading a POI than a fusion protein including a cathepsin B-sensitive linker between the Fc domain and the R3 (e.g., v0.3; FIG. 34B).

[0293] In some embodiments, a fusion protein including two monomers bound by Fc interactions, each monomer including from N terminus to C terminus and R1, a cathepsin linker, and Fc, and an R3 (e.g., v0.4) can be more efficient at internalizing and / or degrading a POI than a fusion protein including two monomers bound by Fc interactions, each monomer including from N terminus to C terminus and R1, and Fc, a cathepsin linker, and an R3 (e.g., v0.3; FIG. 34).

[0294] In some embodiments, a fusion protein including two monomers bound by Fc interactions, each monomer including from N terminus to C terminus and R1, a cathepsin linker, and Fc, and an R3 (e.g., v0.4) can be at least between about 10 to about 20%, about 15 to about 25%, about 20% to about 30%, about 25 to about 35%, about 30 and about 40%, about 35 and about 45%, about 40% and about 50%, about 45 and about 55%, about 50 to about 60%, about 55 to about 65%, about 60 to about 70%, about 65 to about 75%, about 70% to about 80%, about 75% to about 85%, about 80 to about 90%, about 85% to about 95%, about 90 to about 100% more efficient at internalizing and / or degrading a POI (e.g., APJ, GPR20, CCR6, CCR2, CCR4, AT1R, CXCR1 / 1, PAR2, RXFP1, A2AR, FZD1, C5aR1, or any of the GPCR described herein) than a fusion protein including two monomers bound by Fc interactions, each monomer including from N terminus to C terminus and R1, and Fc, a cathepsin linker, and an R3 (e.g., v0.3; FIG.34B).DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17

[0295] In some embodiments, R3 is a transferrin receptor (TR) binding means (also known as a means for binding TR). The TR binder or TR binding means (also known as a means for binding TR) binds to a transferrin receptor on the surface of cells. In some embodiments, the TR binder or TR binding means (also known as a means for binding TR) is a molecule that binds to a transferrin receptor (TfR). In some embodiments, the TR binder or TR binding means (also known as a means for binding TR) is transferrin or a part of transferrin that can bind to a TfR. In some embodiments, the TR binder or TR binding means (also known as a means for binding TR) is an antibody that binds to the transferrin receptor. In some embodiments, a TR binder or TR binding means (also known as a means for binding TR) is competitive with transferrin for binding to a TfR (e.g., H7 or M16). In some embodiments, a TR binder or TR binding means (also known as a means for binding TR) is non-competitive for binding to a TfR (e.g., VHHA or VHHB). The TR binder or TR binding means (also known as a means for binding TR) (R3), as part of a fusion protein molecule (e.g., R1-R2-R3), binds to transferrin (Tf) on the surface of a cell and is internalized into the cell. When the protein of interest binder (POIB or POI binding means (also known as a means for binding POI); R1) in the fusion protein binds a protein of interest (POI) on a cell surface, internalization of the TR binder or TR binding means (also known as a means for binding TR) can also cause internalization of the POI. Internalized POIs can be degraded once internalized by a cell.

[0296] The TR binder or TR binding means (also known as a means for binding TR) (R3) binds to a transferrin receptor on the surface of cells. In some embodiments, the TR binder or TR binding means (also known as a means for binding TR) can be an antibody that binds to the transferrin receptor (e.g., H7 or M16). In some embodiments, the TR binder or TR binding means (also known as a means for binding TR) can be a nanobody that binds to the transferrin receptor. In some embodiments, the TR binder or TR binding means (also known as a means for binding TR) can be a nanobody that binds to the transferrin receptor without competing with endogenous Tf for binding to the TfR1. In some embodiments, the TR binder or TR binding means (also known as a means for binding TR) can be VHHA (SEQ IDs NO: 146-147). In some embodiments, the TR binder or TR binding means (also known as a means for binding TR) can be VHHB (SEQ IDs NO: 148-149).

[0297] In some embodiments, the fusion proteins disclosed herein can include the following nucleotide and amino acid sequences, and molecules at least 60, 65, 70, 75, 80, 81, 82, 83, 84,DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the nucleotide and amino acid sequences below. Specifically, the amino acid sequences of the fusion proteins can be labeled as follows:

[0298] Times New Roman font underlined is signal peptide;

[0299] Times New Roman font bolded is anti-protein of interest Fab-heavy chain, scFv, or affibody;

[0300] Times New Roman italicized font is linker encoded by restriction enzyme site creation;

[0301] Times New Roman underlined and bolded font is GS linker;

[0302] Times New Roman underlined, italicized, and bolded font is cleavable linker;

[0303] Courier New font is H7-scFv;

[0304] Courier New underlined font is Fc domain;

[0305] Courier New bolded font is TEV site;

[0306] Courier New italicized font is fragment from Transferrin;

[0307] Courier New underlined and bolded font is His-Tag;

[0308] Courier New bolded and italicized is light chain, and

[0309] Courier New underlined, italicized, and bolded font is Avi-Tag.

[0310] pDP14-CD19 ETD_Hole Fc Nucleic Acid Sequence (SEQ ID NO: 17)

[0311] atgcgAatgcagctgctgctgctgattgcgctgagcctggcgctggtgaccaacagcactagtcccgaggaacctctagtg gtgaaggtggaagagggagataacgctgtgctgcagtgcctcaaggggacctcagatggccccactcagcagctgacctggtctcggga gtccccgcttaaacccttcttaaaactcagcctggggctgccaggcctgggaatccacatgaggcccctggccatctggcttttcatcttcaa cgtctctcaacagatggggggcttctacctgtgccagccggggcccccctctgagaaggcctggcagcctggctggacagtcaatgtgga gggcagcggggagctgttccggtggaatgtttcggacctaggtggcctgggctgtggcctgaagaacaggtcctcagagggccccagct ccccttccgggaagctcatgagccccaagctgtatgtgtgggccaaagaccgccctgagatctgggagggagagcctccgtgtctcccac cgagggacagcctgaaccagagcctcagccaggacctcaccatggcccctggctccacactctggctgtcctgtggggtaccccctgact ctgtgtccaggggccccctctcctggacccatgtgcaccccaaggggcctaagtcattgctgagcctagagctgaaggacgatcgcccgg ccagagatatgtgggtaatggagacgggtctgttgttgccccgggccacagctcaagacgctggaaagtattattgtcaccgtggcaacctg accatgtcattccacctggagatcactgctcggccagtactatggcactggctgctgaggactggtggctggaagactagtTCTGGTG GTGGTGGTGAGAATCTGTACTTTCAGAGCTCGGGCGGAGGATCgggtggaggcgagcccaaatc ttgtgacaaaactcacacatgcCCCCCCTGCCCAGCGCCAGAATTGCTGGGCGGACCCAGCGTGT TCCTGTTCCCCCCCAAACCTAAAGACACCCTGATGATCAGCCGAACCCCTGAGGTGA CCTGCGTGGTGGTGGACGTGAGCCACGAGGACCCCGAGGTGAAGTTCAACTGGTAT GTGGACGGCGTGGAGGTCCACAATGCCAAAACGAAGCCCAGGGAGGAGCAGTACA ACAGCACCTACAGGGTAGTGAGCGTCTTGACCGTGCTGCACCAGGACTGGCTGAAC GGCAAGGAATACAAATGCAAGGTCAGCAATAAGGCTCTGCCGGCTCCTATCGAGAA GACAATCAGCAAGGCAAAGGGCCAGCCACGCGAACCGCAGGTGTATACTCTGCCCCDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 CCAGCCGGGACGAGCTGACCAAGAACCAGGTGTCCCTGAGCTGTGCCGTGAAAGGC TTCTACCCCAGCGACATCGCTGTGGAGTGGGAGAGTAACGGGCAGCCCGAGAACAA CTACAAGACCACGCCTCCTGTGCTGGACAGCGACGGCAGCTTCTTCCTGGTGAGCAA GCTCACCGTGGACAAGAGCAGGTGGCAACAGGGCAACGTGTTCAGCTGCTCTGTGA TGCACGAGGCCCTGCACAACCATTACACCCAGAAGAGTCTCAGTCTGAGCCCGGGA AAGGGTGGAGGCGGATCCGGCCTGAACGACATCTTCGAGGCTCAGAAAATCGAATG GCACGAAGGCtaa

[0312] pDP14-CD19 ETD_Hole Fc Amino Acid Sequence (SEQ ID NO: 18)

[0313] MRMQLLLLIALSLALVTNSTSPEEPLVVKVEEGDNAVLQCLKGTSDGPTQQ LTWSRESPLKPFLKLSLGLPGLGIHMRPLAIWLFIFNVSQQMGGFYLCQPGPPSEK AWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGKLMSPKLYVW AKDRPEIWEGEPPCLPPRDSLNQSLSQDLTMAPGSTLWLSCGVPPDSVSRGPLSWT HVHPKGPKSLLSLELKDDRPARDMWVMETGLLLPRATAQDAGKYYCHRGNLTMS FHLEITARPVLWHWLLRTGGWKTSSGGGGENLYFQSSGGGSGGGEPKSCDKTHTCPP CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPR EEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRD ELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSLSLSPGKGGGGSGLNDIFEAQKIEWHEG*

[0314] pDP16-EGFR-Affibody-FC-Tf Nucleic Acid Sequence (SEQ ID NO: 19)

[0315] ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTC ACGAATTCGCTGCAGGTAGATAACAAATTCAACAAAGAAATGTGGGCGGCGTGGGA AGAAATTCGCAACCTGCCGAACCTGAACGGCTGGCAGATGACCGCGTTTATTGCGA GCCTGGTGGATGACCCAAGCCAAAGCGCTAACTTGCTAGCAGAAGCTAAAAAGCTA AATGATGCTCAGGCGCCGAAAGTAGACGGCAGCGGCAGCGACAAAACTCACACATG CCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCC AAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGG TGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTG GAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACC GTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTAC AAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAA AGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATG AGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGC GACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCA CGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGG ACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCT CTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAAggatcctctggggg aagtggaggtagcggtggttctgtgcccgataagacagtgcggtggtgcgccgtgtctgagcacgaggccaccaagtgccagagcttcc gggaccacatgaagtccgtgatccccagcgacggccctagcgtggcctgtgtgaagaaggccagctacctggactgcatccgggccatt gccgccaatgaggccgacgccgtgacactggatgccggcctggtgtacgatgcctacctggcccccaacaacctgaagcccgtggtggc cgagttctacggcagcaaagaggacccccagaccttctactacgccgtggccgtggtcaagaaggacagcggcttccagatgaaccagc tgcggggcaagaagtcctgtcacaccggcctgggcagaagcgccggctggaacatccccatcggcctgctgtactgcgatctgcccgag ccccggaagcctctggaaaaggccgtggccaacttcttcagcggcagctgcgccccttgtgctgacggaaccgacttcccccagctgtgtc agctgtgccccggctgtggctgcagcaccctgaaccagtacttcggctacagcggcgccttcaagtgcctgaaggacggcgctggcgac gtggccttcgtgaagcacagcaccatcttcgagaacctggccaacaaggccgaccgggaccagtacgagctgctgtgcctggacaacacDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 cagaaagcccgtggacgagtacaaggactgccacctcgcccaggtgccatctcacacagtggtggcccggtccatgggcggcaaagag gatctgatctgggagctgctgaaccaggcccaggaacacttcggcaaggacaagagcaaagagttccagctgttcagcagcccccacgg caaggatctgctgttcaaggacagcgcccacggctttctgaaggtgccccccagaatggacgccaagatgtacctgggctacgagtacgt gaccgccatccggaacctgagagagggcacctgtcccgaggcccccaccgatgagtgcaagcccgtgaagtggtgcgccctgagccac cacgagcggctgaagtgcgacgagtggagcgtgaacagcgtgggcaagatcgagtgcgtgagcgccgagacaaccgaggactgtatc gccaagatcatgaacggcgaggccgatgccatgagcctggacggcggcttcgtgtacattgccggcaagtgcggcctggtgcctgtgctg gccgagaactacaacaagagcgacaactgcgaggatacccccgaggccggctactttgccatcgcagtcgtgaagaagtccgccagcg acctgacctgggacaatctgaagggcaagaaaagctgccacaccgccgtgggaaggaccgccgggtggaatattcctatggggctgctg tacaacaagatcaaccactgcagattcgacgagttcttcagcgagggctgcgctcccggcagcaagaaagacagcagcctgtgcaagctg tgcatgggcagcggcctgaacctgtgcgagcccaacaacaaagagggctactacggctacacaggggccttccggtgtctggtggagaa gggggacgtggcttttgtgaaacaccagaccgtgccccagaacaccggcggcaagaaccccgacccctgggccaagaacctgaacga gaaggactacgaactgctgtgtctcgacggcacccggaagccagtggaggaatacgccaactgtcacctggccagagcccccaatcacg ccgtggtcacccggaaggacaaagaggcctgcgtccacaagatcctgcggcagcagcagcacctgttcggcagcaacgtgaccgactg cagcggcaacttctgcctgttcagaagcgagacaaaggacctcctgttccgggacgacaccgtgtgtctggccaagctgcacgaccggaa cacctacgagaagtacctgggcgaggaatatgtgaaggccgtgggcaatctgcggaagtgcagcacctctagcctgctggaagcctgca cctttcgcagacctTAA

[0316] pDP16-EGFR-Affibody-FC-Tf Amino Acid Sequence (SEQ ID NO: 20)

[0317] MYRMQLLSCIALSLALVTNSLQVDNKFNKEMWAAWEEIRNLPNLNGWQM TAFIASLVDDPSQSANLLAEAKKLNDAQAPKVDGSGSDKTHTCPPCPAPELLGGPSVFLF PPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGKGSSGGSGGSGGSVPDKTVRWCAVSEHEATKCQSFRDHMKSVIPSDGPSVAC VKKASYLDCIRAIAANEADAVTLDAGLVYDAYLAPNNLKPVVAEFYGSKEDPQTFYYAVAVVKK DSGFQMNQLRGKKSCHTGLGRSAGWNIPIGLLYCDLPEPRKPLEKAVANFFSGSCAPCADGTDF PQLCQLCPGCGCSTLNQYFGYSGAFKCLKDGAGDVAFVKHSTIFENLANKADRDQYELLCLDNT RKPVDEYKDCHLAQVPSHTVVARSMGGKEDLIWELLNQAQEHFGKDKSKEFQLFSSPHGKDLLF KDSAHGFLKVPPRMDAKMYLGYEYVTAIRNLREGTCPEAPTDECKPVKWCALSHHERLKCDEWS VNSVGKIECVSAETTEDCIAKIMNGEADAMSLDGGFVYIAGKCGLVPVLAENYNKSDNCEDTPE AGYFAIAVVKKSASDLTWDNLKGKKSCHTAVGRTAGWNIPMGLLYNKINHCRFDEFFSEGCAPG SKKDSSLCKLCMGSGLNLCEPNNKEGYYGYTGAFRCLVEKGDVAFVKHQTVPQNTGGKNPDPWA KNLNEKDYELLCLDGTRKPVEEYANCHLARAPNHAVVTRKDKEACVHKILRQQQHLFGSNVTDC SGNFCLFRSETKDLLFRDDTVCLAKLHDRNTYEKYLGEEYVKAVGNLRKCSTSSLLEACTFRRP*

[0318] pDP18-EGFR-affibody Nucleic Acid Sequence (SEQ ID NO: 21)

[0319] ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTC ACGAATTCGCTGCAGGTAGATAACAAATTCAACAAAGAAATGTGGGCGGCGTGGGA AGAAATTCGCAACCTGCCGAACCTGAACGGCTGGCAGATGACCGCGTTTATTGCGA GCCTGGTGGATGACCCAAGCCAAAGCGCTAACTTGCTAGCAGAAGCTAAAAAGCTA AATGATGCTCAGGCGCCGAAAGTAGACGGTGAGAATCTGTACTTTCAGAGCTCGGG CGGAGGATCGGGTGGAGGCCACCACCATCATCACCACCATCACGGATCCGGCCTGA ACGACATCTTCGAGGCTCAGAAAATCGAATGGCACGAAGGCggatcctctgggggaagtggaggt agcggtggttctgtgcccgataagacagtgcggtggtgcgccgtgtctgagcacgaggccaccaagtgccagagcttccgggaccacat gaagtccgtgatccccagcgacggccctagcgtggcctgtgtgaagaaggccagctacctggactgcatccgggccattgccgccaatg aggccgacgccgtgacactggatgccggcctggtgtacgatgcctacctggcccccaacaacctgaagcccgtggtggccgagttctacDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 ggcagcaaagaggacccccagaccttctactacgccgtggccgtggtcaagaaggacagcggcttccagatgaaccagctgcggggca agaagtcctgtcacaccggcctgggcagaagcgccggctggaacatccccatcggcctgctgtactgcgatctgcccgagccccggaag cctctggaaaaggccgtggccaacttcttcagcggcagctgcgccccttgtgctgacggaaccgacttcccccagctgtgtcagctgtgcc ccggctgtggctgcagcaccctgaaccagtacttcggctacagcggcgccttcaagtgcctgaaggacggcgctggcgacgtggccttcg tgaagcacagcaccatcttcgagaacctggccaacaaggccgaccgggaccagtacgagctgctgtgcctggacaacaccagaaagcc cgtggacgagtacaaggactgccacctcgcccaggtgccatctcacacagtggtggcccggtccatgggcggcaaagaggatctgatct gggagctgctgaaccaggcccaggaacacttcggcaaggacaagagcaaagagttccagctgttcagcagcccccacggcaaggatct gctgttcaaggacagcgcccacggctttctgaaggtgccccccagaatggacgccaagatgtacctgggctacgagtacgtgaccgccat ccggaacctgagagagggcacctgtcccgaggcccccaccgatgagtgcaagcccgtgaagtggtgcgccctgagccaccacgagcg gctgaagtgcgacgagtggagcgtgaacagcgtgggcaagatcgagtgcgtgagcgccgagacaaccgaggactgtatcgccaagatc atgaacggcgaggccgatgccatgagcctggacggcggcttcgtgtacattgccggcaagtgcggcctggtgcctgtgctggccgagaa ctacaacaagagcgacaactgcgaggatacccccgaggccggctactttgccatcgcagtcgtgaagaagtccgccagcgacctgacct gggacaatctgaagggcaagaaaagctgccacaccgccgtgggaaggaccgccgggtggaatattcctatggggctgctgtacaacaa gatcaaccactgcagattcgacgagttcttcagcgagggctgcgctcccggcagcaagaaagacagcagcctgtgcaagctgtgcatgg gcagcggcctgaacctgtgcgagcccaacaacaaagagggctactacggctacacaggggccttccggtgtctggtggagaaggggga cgtggcttttgtgaaacaccagaccgtgccccagaacaccggcggcaagaaccccgacccctgggccaagaacctgaacgagaaggac tacgaactgctgtgtctcgacggcacccggaagccagtggaggaatacgccaactgtcacctggccagagcccccaatcacgccgtggt cacccggaaggacaaagaggcctgcgtccacaagatcctgcggcagcagcagcacctgttcggcagcaacgtgaccgactgcagcgg caacttctgcctgttcagaagcgagacaaaggacctcctgttccgggacgacaccgtgtgtctggccaagctgcacgaccggaacaccta cgagaagtacctgggcgaggaatatgtgaaggccgtgggcaatctgcggaagtgcagcacctctagcctgctggaagcctgcacctttcg cagacctTAA

[0320] pDP18-EGFR-affibody Amino Acid Sequence (SEQ ID NO: 22)

[0321] MYRMQLLSCIALSLALVTNSLQVDNKFNKEMWAAWEEIRNLPNLNGWQM TAFIASLVDDPSQSANLLAEAKKLNDAQAPKVDGENLYFQSSGGGSGGGHHHHHHHHG SGLNDIFEAQKIEWHEGGSSGGSGGSGGSVPDKTVRWCAVSEHEATKCQSFRDHMKSVIPSDG PSVACVKKASYLDCIRAIAANEADAVTLDAGLVYDAYLAPNNLKPVVAEFYGSKEDPQTFYYAV AVVKKDSGFQMNQLRGKKSCHTGLGRSAGWNIPIGLLYCDLPEPRKPLEKAVANFFSGSCAPCA DGTDFPQLCQLCPGCGCSTLNQYFGYSGAFKCLKDGAGDVAFVKHSTIFENLANKADRDQYELL CLDNTRKPVDEYKDCHLAQVPSHTVVARSMGGKEDLIWELLNQAQEHFGKDKSKEFQLFSSPHG KDLLFKDSAHGFLKVPPRMDAKMYLGYEYVTAIRNLREGTCPEAPTDECKPVKWCALSHHERLK CDEWSVNSVGKIECVSAETTEDCIAKIMNGEADAMSLDGGFVYIAGKCGLVPVLAENYNKSDNC EDTPEAGYFAIAVVKKSASDLTWDNLKGKKSCHTAVGRTAGWNIPMGLLYNKINHCRFDEFFSE GCAPGSKKDSSLCKLCMGSGLNLCEPNNKEGYYGYTGAFRCLVEKGDVAFVKHQTVPQNTGGKN PDPWAKNLNEKDYELLCLDGTRKPVEEYANCHLARAPNHAVVTRKDKEACVHKILRQQQHLFGS NVTDCSGNFCLFRSETKDLLFRDDTVCLAKLHDRNTYEKYLGEEYVKAVGNLRKCSTSSLLEAC TFRRP*

[0322] pDP20-EGFR-Affibody-FC Nucleic Acid Sequence (SEQ ID NO: 23)

[0323] ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTC ACGAATTCGCTGCAGGTAGATAACAAATTCAACAAAGAAATGTGGGCGGCGTGGGA AGAAATTCGCAACCTGCCGAACCTGAACGGCTGGCAGATGACCGCGTTTATTGCGA GCCTGGTGGATGACCCAAGCCAAAGCGCTAACTTGCTAGCAGAAGCTAAAAAGCTA AATGATGCTCAGGCGCCGAAAGTAGACGGCAGCGGCAGCGACAAAACTCACACATG CCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCC AAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 TGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTG GAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACC GTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTAC AAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAA AGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATG AGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGC GACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCA CGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGG ACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCT CTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATAA

[0324] pDP20-EGFR-Affibody-FC Amino Acid Sequence (SEQ ID NO: 24)

[0325] MYRMQLLSCIALSLALVTNSLQVDNKFNKEMWAAWEEIRNLPNLNGWQM TAFIASLVDDPSQSANLLAEAKKLNDAQAPKVDGSGSDKTHTCPPCPAPELLGGPSVFLF PPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK*

[0326] pDP22-EGFR-affibody Nucleic Acid Sequence (SEQ ID NO: 25)

[0327] ATGCGAATGCAGCTGCTGCTGCTGATTGCGCTGAGCCTGGCGCTGGTGACC AACAGCACTAGTCTGCAGGTAGATAACAAATTCAACAAAGAAATGTGGGCGGCGTG GGAAGAAATTCGCAACCTGCCGAACCTGAACGGCTGGCAGATGACCGCGTTTATTG CGAGCCTGGTGGATGACCCAAGCCAAAGCGCTAACTTGCTAGCAGAAGCTAAAAAG CTAAATGATGCTCAGGCGCCGAAAGTAGACGGCAGCGGCAGCACTAGTTCTGGTGG TGGTGGTGAGAATCTGTACTTTCAGAGCTCGGGCGGAGGATCGGGTGGAGGCCACC ACCATCATCACCACCATCACGGATCCGGCCTGAACGACATCTTCGAGGCTCAGAAA ATCGAATGGCACGAAGGCTAA

[0328] pDP22-EGFR-affibody Amino Acid Sequence (SEQ ID NO: 26)

[0329] MRMQLLLLIALSLALVTNSTSLQVDNKFNKEMWAAWEEIRNLPNLNGWQ MTAFIASLVDDPSQSANLLAEAKKLNDAQAPKVDGSGSTSSGGGGENLYFQSSGGGS GGGHHHHHHHHGSGLNDIFEAQKIEWHEG*

[0330] pDP24-CD19-FC-Tf Nucleic Acid Sequence (SEQ ID NO: 27)

[0331] ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTC ACGAATTCGcccgaggaacctctagtggtgaaggtggaagagggagataacgctgtgctgcagtgcctcaaggggacctcagat ggccccactcagcagctgacctggtctcgggagtccccgcttaaacccttcttaaaactcagcctggggctgccaggcctgggaatccaca tgaggcccctggccatctggcttttcatcttcaacgtctctcaacagatggggggcttctacctgtgccagccggggcccccctctgagaag gcctggcagcctggctggacagtcaatgtggagggcagcggggagctgttccggtggaatgtttcggacctaggtggcctgggctgtggc ctgaagaacaggtcctcagagggccccagctccccttccgggaagctcatgagccccaagctgtatgtgtgggccaaagaccgccctga gatctgggagggagagcctccgtgtctcccaccgagggacagcctgaaccagagcctcagccaggacctcaccatggcccctggctcca cactctggctgtcctgtggggtaccccctgactctgtgtccaggggccccctctcctggacccatgtgcaccccaaggggcctaagtcattg ctgagcctagagctgaaggacgatcgcccggccagagatatgtgggtaatggagacgggtctgttgttgccccgggccacagctcaaga cgctggaaagtattattgtcaccgtggcaacctgaccatgtcattccacctggagatcactgctcggccagtactatggcactggctgctgag gactggtggctggaagGTAGACGGCAGCGGCAGCGACAAAACTCACACATGCCCACCGTGCCDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 CAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGG ACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCC ACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAAT GCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGT CCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCT CCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAG CCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAA CCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGA GTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGG ACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGC AGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACA CGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAAggatcctctgggggaagtggaggtagcggtggttctgtgc ccgataagacagtgcggtggtgcgccgtgtctgagcacgaggccaccaagtgccagagcttccgggaccacatgaagtccgtgatcccc agcgacggccctagcgtggcctgtgtgaagaaggccagctacctggactgcatccgggccattgccgccaatgaggccgacgccgtga cactggatgccggcctggtgtacgatgcctacctggcccccaacaacctgaagcccgtggtggccgagttctacggcagcaaagaggac ccccagaccttctactacgccgtggccgtggtcaagaaggacagcggcttccagatgaaccagctgcggggcaagaagtcctgtcacac cggcctgggcagaagcgccggctggaacatccccatcggcctgctgtactgcgatctgcccgagccccggaagcctctggaaaaggcc gtggccaacttcttcagcggcagctgcgccccttgtgctgacggaaccgacttcccccagctgtgtcagctgtgccccggctgtggctgca gcaccctgaaccagtacttcggctacagcggcgccttcaagtgcctgaaggacggcgctggcgacgtggccttcgtgaagcacagcacc atcttcgagaacctggccaacaaggccgaccgggaccagtacgagctgctgtgcctggacaacaccagaaagcccgtggacgagtaca aggactgccacctcgcccaggtgccatctcacacagtggtggcccggtccatgggcggcaaagaggatctgatctgggagctgctgaac caggcccaggaacacttcggcaaggacaagagcaaagagttccagctgttcagcagcccccacggcaaggatctgctgttcaaggacag cgcccacggctttctgaaggtgccccccagaatggacgccaagatgtacctgggctacgagtacgtgaccgccatccggaacctgagag agggcacctgtcccgaggcccccaccgatgagtgcaagcccgtgaagtggtgcgccctgagccaccacgagcggctgaagtgcgacg agtggagcgtgaacagcgtgggcaagatcgagtgcgtgagcgccgagacaaccgaggactgtatcgccaagatcatgaacggcgagg ccgatgccatgagcctggacggcggcttcgtgtacattgccggcaagtgcggcctggtgcctgtgctggccgagaactacaacaagagc gacaactgcgaggatacccccgaggccggctactttgccatcgcagtcgtgaagaagtccgccagcgacctgacctgggacaatctgaa gggcaagaaaagctgccacaccgccgtgggaaggaccgccgggtggaatattcctatggggctgctgtacaacaagatcaaccactgca gattcgacgagttcttcagcgagggctgcgctcccggcagcaagaaagacagcagcctgtgcaagctgtgcatgggcagcggcctgaac ctgtgcgagcccaacaacaaagagggctactacggctacacaggggccttccggtgtctggtggagaagggggacgtggcttttgtgaaa caccagaccgtgccccagaacaccggcggcaagaaccccgacccctgggccaagaacctgaacgagaaggactacgaactgctgtgt ctcgacggcacccggaagccagtggaggaatacgccaactgtcacctggccagagcccccaatcacgccgtggtcacccggaaggaca aagaggcctgcgtccacaagatcctgcggcagcagcagcacctgttcggcagcaacgtgaccgactgcagcggcaacttctgcctgttca gaagcgagacaaaggacctcctgttccgggacgacaccgtgtgtctggccaagctgcacgaccggaacacctacgagaagtacctggg cgaggaatatgtgaaggccgtgggcaatctgcggaagtgcagcacctctagcctgctggaagcctgcacctttcgcagacctTAA

[0332] pDP24-CD19-FC-Tf Amino Acid Sequence (SEQ ID NO: 28)

[0333] MYRMQLLSCIALSLALVTNSPEEPLVVKVEEGDNAVLQCLKGTSDGPTQQ LTWSRESPLKPFLKLSLGLPGLGIHMRPLAIWLFIFNVSQQMGGFYLCQPGPPSEK AWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGKLMSPKLYVW AKDRPEIWEGEPPCLPPRDSLNQSLSQDLTMAPGSTLWLSCGVPPDSVSRGPLSWT HVHPKGPKSLLSLELKDDRPARDMWVMETGLLLPRATAQDAGKYYCHRGNLTMS FHLEITARPVLWHWLLRTGGWKVDGSGSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTL MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAV EWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 SPGKGSSGGSGGSGGSVPDKTVRWCAVSEHEATKCQSFRDHMKSVIPSDGPSVACVKKASYLD CIRAIAANEADAVTLDAGLVYDAYLAPNNLKPVVAEFYGSKEDPQTFYYAVAVVKKDSGFQMNQ LRGKKSCHTGLGRSAGWNIPIGLLYCDLPEPRKPLEKAVANFFSGSCAPCADGTDFPQLCQLCP GCGCSTLNQYFGYSGAFKCLKDGAGDVAFVKHSTIFENLANKADRDQYELLCLDNTRKPVDEYK DCHLAQVPSHTVVARSMGGKEDLIWELLNQAQEHFGKDKSKEFQLFSSPHGKDLLFKDSAHGFL KVPPRMDAKMYLGYEYVTAIRNLREGTCPEAPTDECKPVKWCALSHHERLKCDEWSVNSVGKIE CVSAETTEDCIAKIMNGEADAMSLDGGFVYIAGKCGLVPVLAENYNKSDNCEDTPEAGYFAIAV VKKSASDLTWDNLKGKKSCHTAVGRTAGWNIPMGLLYNKINHCRFDEFFSEGCAPGSKKDSSLC KLCMGSGLNLCEPNNKEGYYGYTGAFRCLVEKGDVAFVKHQTVPQNTGGKNPDPWAKNLNEKDY ELLCLDGTRKPVEEYANCHLARAPNHAVVTRKDKEACVHKILRQQQHLFGSNVTDCSGNFCLFR SETKDLLFRDDTVCLAKLHDRNTYEKYLGEEYVKAVGNLRKCSTSSLLEACTFRRP*

[0334] pDP25-pFUSE-Tf-knob-Fc Nucleic Acid Sequence (SEQ ID NO: 29)

[0335] ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTC ACGAATTCGgtgcccgataagacagtgcggtggtgcgccgtgtctgagcacgaggccaccaagtgccagagcttccgggacca catgaagtccgtgatccccagcgacggccctagcgtggcctgtgtgaagaaggccagctacctggactgcatccgggccattgccgccaa tgaggccgacgccgtgacactggatgccggcctggtgtacgatgcctacctggcccccaacaacctgaagcccgtggtggccgagttcta cggcagcaaagaggacccccagaccttctactacgccgtggccgtggtcaagaaggacagcggcttccagatgaaccagctgcggggc aagaagtcctgtcacaccggcctgggcagaagcgccggctggaacatccccatcggcctgctgtactgcgatctgcccgagccccggaa gcctctggaaaaggccgtggccaacttcttcagcggcagctgcgccccttgtgctgacggaaccgacttcccccagctgtgtcagctgtgc cccggctgtggctgcagcaccctgaaccagtacttcggctacagcggcgccttcaagtgcctgaaggacggcgctggcgacgtggccttc gtgaagcacagcaccatcttcgagaacctggccaacaaggccgaccgggaccagtacgagctgctgtgcctggacaacaccagaaagc ccgtggacgagtacaaggactgccacctcgcccaggtgccatctcacacagtggtggcccggtccatgggcggcaaagaggatctgatc tgggagctgctgaaccaggcccaggaacacttcggcaaggacaagagcaaagagttccagctgttcagcagcccccacggcaaggatct gctgttcaaggacagcgcccacggctttctgaaggtgccccccagaatggacgccaagatgtacctgggctacgagtacgtgaccgccat ccggaacctgagagagggcacctgtcccgaggcccccaccgatgagtgcaagcccgtgaagtggtgcgccctgagccaccacgagcg gctgaagtgcgacgagtggagcgtgaacagcgtgggcaagatcgagtgcgtgagcgccgagacaaccgaggactgtatcgccaagatc atgaacggcgaggccgatgccatgagcctggacggcggcttcgtgtacattgccggcaagtgcggcctggtgcctgtgctggccgagaa ctacaacaagagcgacaactgcgaggatacccccgaggccggctactttgccatcgcagtcgtgaagaagtccgccagcgacctgacct gggacaatctgaagggcaagaaaagctgccacaccgccgtgggaaggaccgccgggtggaatattcctatggggctgctgtacaacaa gatcaaccactgcagattcgacgagttcttcagcgagggctgcgctcccggcagcaagaaagacagcagcctgtgcaagctgtgcatgg gcagcggcctgaacctgtgcgagcccaacaacaaagagggctactacggctacacaggggccttccggtgtctggtggagaaggggga cgtggcttttgtgaaacaccagaccgtgccccagaacaccggcggcaagaaccccgacccctgggccaagaacctgaacgagaaggac tacgaactgctgtgtctcgacggcacccggaagccagtggaggaatacgccaactgtcacctggccagagcccccaatcacgccgtggt cacccggaaggacaaagaggcctgcgtccacaagatcctgcggcagcagcagcacctgttcggcagcaacgtgaccgactgcagcgg caacttctgcctgttcagaagcgagacaaaggacctcctgttccgggacgacaccgtgtgtctggccaagctgcacgaccggaacaccta cgagaagtacctgggcgaggaatatgtgaaggccgtgggcaatctgcggaagtgcagcacctctagcctgctggaagcctgcacctttcg cagacctCCATGGgagcccaaatcttgtgacaaaactcacacatgcCCCCCCTGCCCAGCGCCAGAATTGCT GGGCGGACCCAGCGTGTTCCTGTTCCCCCCCAAACCTAAAGACACCCTGATGATCAG CCGAACCCCTGAGGTGACCTGCGTGGTGGTGGACGTGAGCCACGAGGACCCCGAGG TGAAGTTCAACTGGTATGTGGACGGCGTGGAGGTCCACAATGCCAAAACGAAGCCC AGGGAGGAGCAGTACGGAAGCACCTACAGGGTAGTGAGCGTCTTGACCGTGCTGCA CCAGGACTGGCTGAACGGCAAGGAATACAAATGCAAGGTCAGCAATAAGGCTCTGC CGGCTCCTATCGAGAAGACAATCAGCAAGGCAAAGGGCCAGCCACGCGAACCGCA GGTGTATACTCTGCCCCCCAGCCGGGACGAGCTGACCAAGAACCAGGTGTCCCTGT GGTGTCTGGTGAAAGGCTTCTACCCCAGCGACATCGCTGTGGAGTGGGAGAGTAACDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 GGGCAGCCCGAGAACAACTACAAGACCACGCCTCCTGTGCTGGACAGCGACGGCAG CTTCTTCCTGTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAACAGGGCAACG TGTTCAGCTGCTCTGTGATGCACGAGGCCCTGCACAACCATTACACCCAGAAGAGTC TCAGTCTGAGCCCGGGAAAGggtggctctcatcatcaccatcaccactga

[0336] pDP25-pFUSE-Tf-knob-Fc Amino Acid Sequence (SEQ ID NO: 30)

[0337] MYRMQLLSCIALSLALVTNSVPDKTVRWCAVSEHEATKCQSFRDHMKSVIPSDGPS VACVKKASYLDCIRAIAANEADAVTLDAGLVYDAYLAPNNLKPVVAEFYGSKEDPQTFYYAVAV VKKDSGFQMNQLRGKKSCHTGLGRSAGWNIPIGLLYCDLPEPRKPLEKAVANFFSGSCAPCADG TDFPQLCQLCPGCGCSTLNQYFGYSGAFKCLKDGAGDVAFVKHSTIFENLANKADRDQYELLCL DNTRKPVDEYKDCHLAQVPSHTVVARSMGGKEDLIWELLNQAQEHFGKDKSKEFQLFSSPHGKD LLFKDSAHGFLKVPPRMDAKMYLGYEYVTAIRNLREGTCPEAPTDECKPVKWCALSHHERLKCD EWSVNSVGKIECVSAETTEDCIAKIMNGEADAMSLDGGFVYIAGKCGLVPVLAENYNKSDNCED TPEAGYFAIAVVKKSASDLTWDNLKGKKSCHTAVGRTAGWNIPMGLLYNKINHCRFDEFFSEGC APGSKKDSSLCKLCMGSGLNLCEPNNKEGYYGYTGAFRCLVEKGDVAFVKHQTVPQNTGGKNPD PWAKNLNEKDYELLCLDGTRKPVEEYANCHLARAPNHAVVTRKDKEACVHKILRQQQHLFGSNV TDCSGNFCLFRSETKDLLFRDDTVCLAKLHDRNTYEKYLGEEYVKAVGNLRKCSTSSLLEACTF RRPPWEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK FNWYVDGVEVHNAKTKPREEQYGSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDG SFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGGSHHHHHH*

[0338] pDP32-CD19-FC-CD19 Nucleic Acid Sequence (SEQ ID NO: 31)

[0339] atgcgAatgcagctgctgctgctgattgcgctgagcctggcgctggtgaccaacagcactagtcccgaggaacctctagtg gtgaaggtggaagagggagataacgctgtgctgcagtgcctcaaggggacctcagatggccccactcagcagctgacctggtctcggga gtccccgcttaaacccttcttaaaactcagcctggggctgccaggcctgggaatccacatgaggcccctggccatctggcttttcatcttcaa cgtctctcaacagatggggggcttctacctgtgccagccggggcccccctctgagaaggcctggcagcctggctggacagtcaatgtgga gggcagcggggagctgttccggtggaatgtttcggacctaggtggcctgggctgtggcctgaagaacaggtcctcagagggccccagct ccccttccgggaagctcatgagccccaagctgtatgtgtgggccaaagaccgccctgagatctgggagggagagcctccgtgtctcccac cgagggacagcctgaaccagagcctcagccaggacctcaccatggcccctggctccacactctggctgtcctgtggggtaccccctgact ctgtgtccaggggccccctctcctggacccatgtgcaccccaaggggcctaagtcattgctgagcctagagctgaaggacgatcgcccgg ccagagatatgtgggtaatggagacgggtctgttgttgccccgggccacagctcaagacgctggaaagtattattgtcaccgtggcaacctg accatgtcattccacctggagatcactgctcggccagtactatggcactggctgctgaggactggtggctggaagactagtTCTGGTG GTGGTGGTGAGAATCTGTACTTTCAGAGCTCGGGCGGAGGATCgggtggaggcgagcccaaatc ttgtgacaaaactcacacatgcCCCCCCTGCCCAGCGCCAGAATTGCTGGGCGGACCCAGCGTGT TCCTGTTCCCCCCCAAACCTAAAGACACCCTGATGATCAGCCGAACCCCTGAGGTGA CCTGCGTGGTGGTGGACGTGAGCCACGAGGACCCCGAGGTGAAGTTCAACTGGTAT GTGGACGGCGTGGAGGTCCACAATGCCAAAACGAAGCCCAGGGAGGAGCAGTACA ACAGCACCTACAGGGTAGTGAGCGTCTTGACCGTGCTGCACCAGGACTGGCTGAAC GGCAAGGAATACAAATGCAAGGTCAGCAATAAGGCTCTGCCGGCTCCTATCGAGAA GACAATCAGCAAGGCAAAGGGCCAGCCACGCGAACCGCAGGTGTATACTCTGCCCC CCAGCCGGGACGAGCTGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAAGGC TTCTACCCCAGCGACATCGCTGTGGAGTGGGAGAGTAACGGGCAGCCCGAGAACAA CTACAAGACCACGCCTCCTGTGCTGGACAGCGACGGCAGCTTCTTCCTGTATAGCAA GCTCACCGTGGACAAGAGCAGGTGGCAACAGGGCAACGTGTTCAGCTGCTCTGTGA TGCACGAGGCCCTGCACAACCATTACACCCAGAAGAGTCTCAGTCTGAGCCCGGGADOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 AAGGGTGGAGGCGGATCCggaggtagcggtggttctGGAcccgaggaacctctagtggtgaaggtggaagagggag ataacgctgtgctgcagtgcctcaaggggacctcagatggccccactcagcagctgacctggtctcgggagtccccgcttaaacccttctta aaactcagcctggggctgccaggcctgggaatccacatgaggcccctggccatctggcttttcatcttcaacgtctctcaacagatgggggg cttctacctgtgccagccggggcccccctctgagaaggcctggcagcctggctggacagtcaatgtggagggcagcggggagctgttcc ggtggaatgtttcggacctaggtggcctgggctgtggcctgaagaacaggtcctcagagggccccagctccccttccgggaagctcatga gccccaagctgtatgtgtgggccaaagaccgccctgagatctgggagggagagcctccgtgtctcccaccgagggacagcctgaaccag agcctcagccaggacctcaccatggcccctggctccacactctggctgtcctgtggggtaccccctgactctgtgtccaggggccccctctc ctggacccatgtgcaccccaaggggcctaagtcattgctgagcctagagctgaaggacgatcgcccggccagagatatgtgggtaatgga gacgggtctgttgttgccccgggccacagctcaagacgctggaaagtattattgtcaccgtggcaacctgaccatgtcattccacctggaga tcactgctcggccagtactatggcactggctgctgaggactggtggctggaagGGCCTGAACGACATCTTCGAGGCT CAGAAAATCGAATGGCACGAAGGCtaa

[0340] pDP32-CD19-FC-CD19 Amino Acid Sequence (SEQ ID NO: 32)

[0341] MRMQLLLLIALSLALVTNSTSPEEPLVVKVEEGDNAVLQCLKGTSDGPTQQ LTWSRESPLKPFLKLSLGLPGLGIHMRPLAIWLFIFNVSQQMGGFYLCQPGPPSEK AWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGKLMSPKLYVW AKDRPEIWEGEPPCLPPRDSLNQSLSQDLTMAPGSTLWLSCGVPPDSVSRGPLSWT HVHPKGPKSLLSLELKDDRPARDMWVMETGLLLPRATAQDAGKYYCHRGNLTMS FHLEITARPVLWHWLLRTGGWKTSSGGGGENLYFQSSGGGSGGGEPKSCDKTHTCPP CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPR EEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRD ELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSLSLSPGKGGGGSGGSGGSGPEEPLVVKVEEGDNAVLQCL KGTSDGPTQQLTWSRESPLKPFLKLSLGLPGLGIHMRPLAIWLFIFNVSQQMGGFY LCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGK LMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSQDLTMAPGSTLWLSCGVPPDS VSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMETGLLLPRATAQDAGKYY CHRGNLTMSFHLEITARPVLWHWLLRTGGWKGLNDIFEAQKIEWHEG*

[0342] pDP44-CD19 NT.1-FC-Tf Nucleic Acid Sequence (SEQ ID NO: 33)

[0343] ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTC ACGAATTCGcccgaggaacctctagtggtgaaggtggaagagggagataccgctgctctgtggtgcctcaaggggacctcagat ggccccactcagcagctgacctggtctcgggagtccccgcttaaacccttcttaaaatacagcctgggggtgccaggcctgggagtccac gtcaggcccgatgccatctctgtcgtcatcaggaacgtctctcaacagatggggggcttctacctgtgccagccggggcccccctctgaga aggcctggcagcctggctggacagtcaatgtggagggcagcggggagctgttccggtggaatgtttcggacctaggtggcctgggctgtg gcctgaagaacaggtcctcagagggccccagctccccttccgggaagctcatgagccccaagctgtatgtgtgggccaaagaccgccct gagatctgggagggagagcctccgtgtctcccaccgagggacagcctgaaccagagcctcagcagggacctcaccgtagcccctggct ccacactctggctgtcctgtggggtaccccctgactctgtgtccaggggccccctctcctggacccatgtgcaccccaaggggcctaagtc attgctgagcctagagctgaaggacgatcgcccggccagagatatgtgggtaatgggtacgtcactgatgttgccccgggccacagctca agacgctggaaagtggtattgtcaccgtggcaacgtaaccacctcattccacctggaggtaatcgctcggccagtaaaggctcactcagac ctgaggactggtggctggaagGTAGACGGCAGCGGCAGCGACAAAACTCACACATGCCCACCGT GCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCA AGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTG AGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAAggatcctctgggggaagtggaggtagcggtggt tctgtgcccgataagacagtgcggtggtgcgccgtgtctgagcacgaggccaccaagtgccagagcttccgggaccacatgaagtccgtg atccccagcgacggccctagcgtggcctgtgtgaagaaggccagctacctggactgcatccgggccattgccgccaatgaggccgacgc cgtgacactggatgccggcctggtgtacgatgcctacctggcccccaacaacctgaagcccgtggtggccgagttctacggcagcaaaga ggacccccagaccttctactacgccgtggccgtggtcaagaaggacagcggcttccagatgaaccagctgcggggcaagaagtcctgtc acaccggcctgggcagaagcgccggctggaacatccccatcggcctgctgtactgcgatctgcccgagccccggaagcctctggaaaa ggccgtggccaacttcttcagcggcagctgcgccccttgtgctgacggaaccgacttcccccagctgtgtcagctgtgccccggctgtggc tgcagcaccctgaaccagtacttcggctacagcggcgccttcaagtgcctgaaggacggcgctggcgacgtggccttcgtgaagcacag caccatcttcgagaacctggccaacaaggccgaccgggaccagtacgagctgctgtgcctggacaacaccagaaagcccgtggacgag tacaaggactgccacctcgcccaggtgccatctcacacagtggtggcccggtccatgggcggcaaagaggatctgatctgggagctgctg aaccaggcccaggaacacttcggcaaggacaagagcaaagagttccagctgttcagcagcccccacggcaaggatctgctgttcaagga cagcgcccacggctttctgaaggtgccccccagaatggacgccaagatgtacctgggctacgagtacgtgaccgccatccggaacctga gagagggcacctgtcccgaggcccccaccgatgagtgcaagcccgtgaagtggtgcgccctgagccaccacgagcggctgaagtgcg acgagtggagcgtgaacagcgtgggcaagatcgagtgcgtgagcgccgagacaaccgaggactgtatcgccaagatcatgaacggcg aggccgatgccatgagcctggacggcggcttcgtgtacattgccggcaagtgcggcctggtgcctgtgctggccgagaactacaacaag agcgacaactgcgaggatacccccgaggccggctactttgccatcgcagtcgtgaagaagtccgccagcgacctgacctgggacaatct gaagggcaagaaaagctgccacaccgccgtgggaaggaccgccgggtggaatattcctatggggctgctgtacaacaagatcaaccact gcagattcgacgagttcttcagcgagggctgcgctcccggcagcaagaaagacagcagcctgtgcaagctgtgcatgggcagcggcctg aacctgtgcgagcccaacaacaaagagggctactacggctacacaggggccttccggtgtctggtggagaagggggacgtggcttttgtg aaacaccagaccgtgccccagaacaccggcggcaagaaccccgacccctgggccaagaacctgaacgagaaggactacgaactgctg tgtctcgacggcacccggaagccagtggaggaatacgccaactgtcacctggccagagcccccaatcacgccgtggtcacccggaagg acaaagaggcctgcgtccacaagatcctgcggcagcagcagcacctgttcggcagcaacgtgaccgactgcagcggcaacttctgcctg ttcagaagcgagacaaaggacctcctgttccgggacgacaccgtgtgtctggccaagctgcacgaccggaacacctacgagaagtacct gggcgaggaatatgtgaaggccgtgggcaatctgcggaagtgcagcacctctagcctgctggaagcctgcacctttcgcagacctTAA

[0344] pDP44-CD19 NT.1-FC-Tf Amino Acid Sequence (SEQ ID NO: 34)

[0345] MYRMQLLSCIALSLALVTNSPEEPLVVKVEEGDTAALWCLKGTSDGPTQQ LTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFYLCQPGPPSEK AWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGKLMSPKLYVW AKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDSVSRGPLSWT HVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKWYCHRGNVTT SFHLEVIARPVKAHSDLRTGGWKVDGSGSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTL MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAV EWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSL SPGKGSSGGSGGSGGSVPDKTVRWCAVSEHEATKCQSFRDHMKSVIPSDGPSVACVKKASYLD CIRAIAANEADAVTLDAGLVYDAYLAPNNLKPVVAEFYGSKEDPQTFYYAVAVVKKDSGFQMNQ LRGKKSCHTGLGRSAGWNIPIGLLYCDLPEPRKPLEKAVANFFSGSCAPCADGTDFPQLCQLCP GCGCSTLNQYFGYSGAFKCLKDGAGDVAFVKHSTIFENLANKADRDQYELLCLDNTRKPVDEYK DCHLAQVPSHTVVARSMGGKEDLIWELLNQAQEHFGKDKSKEFQLFSSPHGKDLLFKDSAHGFLDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 KVPPRMDAKMYLGYEYVTAIRNLREGTCPEAPTDECKPVKWCALSHHERLKCDEWSVNSVGKIE CVSAETTEDCIAKIMNGEADAMSLDGGFVYIAGKCGLVPVLAENYNKSDNCEDTPEAGYFAIAV VKKSASDLTWDNLKGKKSCHTAVGRTAGWNIPMGLLYNKINHCRFDEFFSEGCAPGSKKDSSLC KLCMGSGLNLCEPNNKEGYYGYTGAFRCLVEKGDVAFVKHQTVPQNTGGKNPDPWAKNLNEKDY ELLCLDGTRKPVEEYANCHLARAPNHAVVTRKDKEACVHKILRQQQHLFGSNVTDCSGNFCLFR SETKDLLFRDDTVCLAKLHDRNTYEKYLGEEYVKAVGNLRKCSTSSLLEACTFRRP*

[0346] pDP49-CD19 NT.1-FC-GFLG-Tf Nucleic Acid Sequence (SEQ ID NO: 35)

[0347] ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTC ACGAATTCGcccgaggaacctctagtggtgaaggtggaagagggagataccgctgctctgtggtgcctcaaggggacctcagat ggccccactcagcagctgacctggtctcgggagtccccgcttaaacccttcttaaaatacagcctgggggtgccaggcctgggagtccac gtcaggcccgatgccatctctgtcgtcatcaggaacgtctctcaacagatggggggcttctacctgtgccagccggggcccccctctgaga aggcctggcagcctggctggacagtcaatgtggagggcagcggggagctgttccggtggaatgtttcggacctaggtggcctgggctgtg gcctgaagaacaggtcctcagagggccccagctccccttccgggaagctcatgagccccaagctgtatgtgtgggccaaagaccgccct gagatctgggagggagagcctccgtgtctcccaccgagggacagcctgaaccagagcctcagcagggacctcaccgtagcccctggct ccacactctggctgtcctgtggggtaccccctgactctgtgtccaggggccccctctcctggacccatgtgcaccccaaggggcctaagtc attgctgagcctagagctgaaggacgatcgcccggccagagatatgtgggtaatgggtacgtcactgatgttgccccgggccacagctca agacgctggaaagtggtattgtcaccgtggcaacgtaaccacctcattccacctggaggtaatcgctcggccagtaaaggctcactcagac ctgaggactggtggctggaagGTAGACGGCAGCGGCAGCGACAAAACTCACACATGCCCACCGT GCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCA AGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTG AGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAAggatcctctggggggTTCCTGggaagcgg tggttctgtgcccgataagacagtgcggtggtgcgccgtgtctgagcacgaggccaccaagtgccagagcttccgggaccacatgaagtc cgtgatccccagcgacggccctagcgtggcctgtgtgaagaaggccagctacctggactgcatccgggccattgccgccaatgaggccg acgccgtgacactggatgccggcctggtgtacgatgcctacctggcccccaacaacctgaagcccgtggtggccgagttctacggcagca aagaggacccccagaccttctactacgccgtggccgtggtcaagaaggacagcggcttccagatgaaccagctgcggggcaagaagtc ctgtcacaccggcctgggcagaagcgccggctggaacatccccatcggcctgctgtactgcgatctgcccgagccccggaagcctctgg aaaaggccgtggccaacttcttcagcggcagctgcgccccttgtgctgacggaaccgacttcccccagctgtgtcagctgtgccccggctg tggctgcagcaccctgaaccagtacttcggctacagcggcgccttcaagtgcctgaaggacggcgctggcgacgtggccttcgtgaagca cagcaccatcttcgagaacctggccaacaaggccgaccgggaccagtacgagctgctgtgcctggacaacaccagaaagcccgtggac gagtacaaggactgccacctcgcccaggtgccatctcacacagtggtggcccggtccatgggcggcaaagaggatctgatctgggagct gctgaaccaggcccaggaacacttcggcaaggacaagagcaaagagttccagctgttcagcagcccccacggcaaggatctgctgttca aggacagcgcccacggctttctgaaggtgccccccagaatggacgccaagatgtacctgggctacgagtacgtgaccgccatccggaac ctgagagagggcacctgtcccgaggcccccaccgatgagtgcaagcccgtgaagtggtgcgccctgagccaccacgagcggctgaagt gcgacgagtggagcgtgaacagcgtgggcaagatcgagtgcgtgagcgccgagacaaccgaggactgtatcgccaagatcatgaacg gcgaggccgatgccatgagcctggacggcggcttcgtgtacattgccggcaagtgcggcctggtgcctgtgctggccgagaactacaac aagagcgacaactgcgaggatacccccgaggccggctactttgccatcgcagtcgtgaagaagtccgccagcgacctgacctgggacaaDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 tctgaagggcaagaaaagctgccacaccgccgtgggaaggaccgccgggtggaatattcctatggggctgctgtacaacaagatcaacc actgcagattcgacgagttcttcagcgagggctgcgctcccggcagcaagaaagacagcagcctgtgcaagctgtgcatgggcagcggc ctgaacctgtgcgagcccaacaacaaagagggctactacggctacacaggggccttccggtgtctggtggagaagggggacgtggctttt gtgaaacaccagaccgtgccccagaacaccggcggcaagaaccccgacccctgggccaagaacctgaacgagaaggactacgaactg ctgtgtctcgacggcacccggaagccagtggaggaatacgccaactgtcacctggccagagcccccaatcacgccgtggtcacccggaa ggacaaagaggcctgcgtccacaagatcctgcggcagcagcagcacctgttcggcagcaacgtgaccgactgcagcggcaacttctgcc tgttcagaagcgagacaaaggacctcctgttccgggacgacaccgtgtgtctggccaagctgcacgaccggaacacctacgagaagtac ctgggcgaggaatatgtgaaggccgtgggcaatctgcggaagtgcagcacctctagcctgctggaagcctgcacctttcgcagacctacc ggtCACCACCATCATCACCACCATCACTAA

[0348] pDP49-CD19 NT.1-FC-GFLG-Tf Amino Acid Sequence (SEQ ID NO: 36)

[0349] MYRMQLLSCIALSLALVTNSPEEPLVVKVEEGDTAALWCLKGTSDGPTQQ LTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFYLCQPGPPSEK AWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGKLMSPKLYVW AKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDSVSRGPLSWT HVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKWYCHRGNVTT SFHLEVIARPVKAHSDLRTGGWKVDGSGSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTL MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAV EWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSL SPGKGSSGGFLGSGGSVPDKTVRWCAVSEHEATKCQSFRDHMKSVIPSDGPSVACVKKASYLD CIRAIAANEADAVTLDAGLVYDAYLAPNNLKPVVAEFYGSKEDPQTFYYAVAVVKKDSGFQMNQ LRGKKSCHTGLGRSAGWNIPIGLLYCDLPEPRKPLEKAVANFFSGSCAPCADGTDFPQLCQLCP GCGCSTLNQYFGYSGAFKCLKDGAGDVAFVKHSTIFENLANKADRDQYELLCLDNTRKPVDEYK DCHLAQVPSHTVVARSMGGKEDLIWELLNQAQEHFGKDKSKEFQLFSSPHGKDLLFKDSAHGFL KVPPRMDAKMYLGYEYVTAIRNLREGTCPEAPTDECKPVKWCALSHHERLKCDEWSVNSVGKIE CVSAETTEDCIAKIMNGEADAMSLDGGFVYIAGKCGLVPVLAENYNKSDNCEDTPEAGYFAIAV VKKSASDLTWDNLKGKKSCHTAVGRTAGWNIPMGLLYNKINHCRFDEFFSEGCAPGSKKDSSLC KLCMGSGLNLCEPNNKEGYYGYTGAFRCLVEKGDVAFVKHQTVPQNTGGKNPDPWAKNLNEKDY ELLCLDGTRKPVEEYANCHLARAPNHAVVTRKDKEACVHKILRQQQHLFGSNVTDCSGNFCLFR SETKDLLFRDDTVCLAKLHDRNTYEKYLGEEYVKAVGNLRKCSTSSLLEACTFRRPTGHHHHHH HH*ACGAATTCGGGCCACCACCATCATCACCACCATCACaccggtcccgaggaacctctagtggtgaaggtg gaagagggagataccgctgctctgtggtgcctcaaggggacctcagatggccccactcagcagctgacctggtctcgggagtccccgctt aaacccttcttaaaatacagcctgggggtgccaggcctgggagtccacgtcaggcccgatgccatctctgtcgtcatcaggaacgtctctca acagatggggggcttctacctgtgccagccggggcccccctctgagaaggcctggcagcctggctggacagtcaatgtggagggcagc ggggagctgttccggtggaatgtttcggacctaggtggcctgggctgtggcctgaagaacaggtcctcagagggccccagctccccttcc gggaagctcatgagccccaagctgtatgtgtgggccaaagaccgccctgagatctgggagggagagcctccgtgtctcccaccgaggga cagcctgaaccagagcctcagcagggacctcaccgtagcccctggctccacactctggctgtcctgtggggtaccccctgactctgtgtcc aggggccccctctcctggacccatgtgcaccccaaggggcctaagtcattgctgagcctagagctgaaggacgatcgcccggccagaga tatgtgggtaatgggtacgtcactgatgttgccccgggccacagctcaagacgctggaaagtggtattgtcaccgtggcaacgtaaccacct cattccacctggaggtaatcgctcggccagtaaaggctcactcagacctgaggactggtggctggaagggtgggTTCCTGggaggc GTAGACGGCGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 GGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCG GACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCA AGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGG GAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCA GGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAG CCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTG TACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTG CCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGC AGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCT TCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTC TCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCC CTGTCTCCGGGTAAAggatcctctgggggaagtggaggtagcggtggttctgtgcccgataagacagtgcggtggtgcgcc gtgtctgagcacgaggccaccaagtgccagagcttccgggaccacatgaagtccgtgatccccagcgacggccctagcgtggcctgtgt gaagaaggccagctacctggactgcatccgggccattgccgccaatgaggccgacgccgtgacactggatgccggcctggtgtacgatg cctacctggcccccaacaacctgaagcccgtggtggccgagttctacggcagcaaagaggacccccagaccttctactacgccgtggcc gtggtcaagaaggacagcggcttccagatgaaccagctgcggggcaagaagtcctgtcacaccggcctgggcagaagcgccggctgg aacatccccatcggcctgctgtactgcgatctgcccgagccccggaagcctctggaaaaggccgtggccaacttcttcagcggcagctgc gccccttgtgctgacggaaccgacttcccccagctgtgtcagctgtgccccggctgtggctgcagcaccctgaaccagtacttcggctaca gcggcgccttcaagtgcctgaaggacggcgctggcgacgtggccttcgtgaagcacagcaccatcttcgagaacctggccaacaaggcc gaccgggaccagtacgagctgctgtgcctggacaacaccagaaagcccgtggacgagtacaaggactgccacctcgcccaggtgccat ctcacacagtggtggcccggtccatgggcggcaaagaggatctgatctgggagctgctgaaccaggcccaggaacacttcggcaagga caagagcaaagagttccagctgttcagcagcccccacggcaaggatctgctgttcaaggacagcgcccacggctttctgaaggtgccccc cagaatggacgccaagatgtacctgggctacgagtacgtgaccgccatccggaacctgagagagggcacctgtcccgaggcccccacc gatgagtgcaagcccgtgaagtggtgcgccctgagccaccacgagcggctgaagtgcgacgagtggagcgtgaacagcgtgggcaag atcgagtgcgtgagcgccgagacaaccgaggactgtatcgccaagatcatgaacggcgaggccgatgccatgagcctggacggcggct tcgtgtacattgccggcaagtgcggcctggtgcctgtgctggccgagaactacaacaagagcgacaactgcgaggatacccccgaggcc ggctactttgccatcgcagtcgtgaagaagtccgccagcgacctgacctgggacaatctgaagggcaagaaaagctgccacaccgccgt gggaaggaccgccgggtggaatattcctatggggctgctgtacaacaagatcaaccactgcagattcgacgagttcttcagcgagggctg cgctcccggcagcaagaaagacagcagcctgtgcaagctgtgcatgggcagcggcctgaacctgtgcgagcccaacaacaaagaggg ctactacggctacacaggggccttccggtgtctggtggagaagggggacgtggcttttgtgaaacaccagaccgtgccccagaacaccgg cggcaagaaccccgacccctgggccaagaacctgaacgagaaggactacgaactgctgtgtctcgacggcacccggaagccagtgga ggaatacgccaactgtcacctggccagagcccccaatcacgccgtggtcacccggaaggacaaagaggcctgcgtccacaagatcctgc ggcagcagcagcacctgttcggcagcaacgtgaccgactgcagcggcaacttctgcctgttcagaagcgagacaaaggacctcctgttcc gggacgacaccgtgtgtctggccaagctgcacgaccggaacacctacgagaagtacctgggcgaggaatatgtgaaggccgtgggcaa tctgcggaagtgcagcacctctagcctgctggaagcctgcacctttcgcagacctTAA

[0352] pDP50-His8-CD19 NT.1-GFLG-FC-Tf Amino Acid Sequence (SEQ ID NO: 38)

[0353] MYRMQLLSCIALSLALVTNSGHHHHHHHHTGPEEPLVVKVEEGDTAALWCL KGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFY LCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGK LMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDS VSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKW YCHRGNVTTSFHLEVIARPVKAHSDLRTGGWKGGFLGGVDGDKTHTCPPCPAPELLG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVS LTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 HEALHNHYTQKSLSLSPGKGSSGGSGGSGGSVPDKTVRWCAVSEHEATKCQSFRDHMKSVIPS DGPSVACVKKASYLDCIRAIAANEADAVTLDAGLVYDAYLAPNNLKPVVAEFYGSKEDPQTFYY AVAVVKKDSGFQMNQLRGKKSCHTGLGRSAGWNIPIGLLYCDLPEPRKPLEKAVANFFSGSCAP CADGTDFPQLCQLCPGCGCSTLNQYFGYSGAFKCLKDGAGDVAFVKHSTIFENLANKADRDQYE LLCLDNTRKPVDEYKDCHLAQVPSHTVVARSMGGKEDLIWELLNQAQEHFGKDKSKEFQLFSSP HGKDLLFKDSAHGFLKVPPRMDAKMYLGYEYVTAIRNLREGTCPEAPTDECKPVKWCALSHHER LKCDEWSVNSVGKIECVSAETTEDCIAKIMNGEADAMSLDGGFVYIAGKCGLVPVLAENYNKSD NCEDTPEAGYFAIAVVKKSASDLTWDNLKGKKSCHTAVGRTAGWNIPMGLLYNKINHCRFDEFF SEGCAPGSKKDSSLCKLCMGSGLNLCEPNNKEGYYGYTGAFRCLVEKGDVAFVKHQTVPQNTGG KNPDPWAKNLNEKDYELLCLDGTRKPVEEYANCHLARAPNHAVVTRKDKEACVHKILRQQQHLF GSNVTDCSGNFCLFRSETKDLLFRDDTVCLAKLHDRNTYEKYLGEEYVKAVGNLRKCSTSSLLE ACTFRRP*

[0354] pDP85-cd20-scfv-GFLG-FC-Tf Nucleic Acid Sequence (SEQ ID NO: 39)

[0355] ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTC ACGAATTCGCAGATTGTTCTGAGCCAGTCCCCGGCAATCCTCTCTGCCAGCCCAGGC GAAAAGGTGACAATGACTTGCCGAGCGAGTTCCAGTGTCTCTTATATCCACTGGTTC CAGCAAAAGCCGGGAAGCAGCCCTAAACCATGGATATATGCAACGTCTAACCTGGC GAGCGGGGTCCCAGTGAGATTTTCCGGAAGCGGCAGCGGAACTAGTTACTCTTTGAC AATAAGCAGAGTGGAGGCTGAGGACGCTGCTACTTACTATTGCCAGCAATGGACGA GTAACCCGCCGACGTTTGGAGGTGGAACGAAGCTGGAGATTAAAGGTGGAGGTGGT TCTGGCGGAGGTGGTTCCGGTGGTGGTGGAAGTCAGGTGCAGCTCCAACAGCCTGG TGCCGAACTTGTCAAACCTGGGGCTAGTGTGAAGATGAGTTGCAAAGCTTCAGGGT ACACGTTTACGTCATACAACATGCATTGGGTAAAGCAAACACCAGGACGCGGCTTG GAATGGATCGGCGCGATATATCCAGGAAACGGTGACACTTCTTATAACCAGAAGTT CAAGGGGAAAGCTACTCTCACAGCGGACAAATCTTCTTCAACAGCGTATATGCAGTT GTCAAGCCTTACTAGCGAGGACAGTGCTGTTTATTACTGCGCCCGGTCCACCTATTA TGGGGGTGATTGGTACTTTAATGTTTGGGGCGCGGGTACTACCGTTACTGTGTCCGC GGGTGGCAGCGGCAGCggtgggTTCCTGggaGGCGTAGACGGCGACAAAACTCACACAT GCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCC CAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTG GTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGT GGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTAC CGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTA CAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCA AAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAT GAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAG CGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACC ACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTG GACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGC TCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAAggatcctctgggg gaagtggaggtagcggtggttctgtgcccgataagacagtgcggtggtgcgccgtgtctgagcacgaggccaccaagtgccagagcttc cgggaccacatgaagtccgtgatccccagcgacggccctagcgtggcctgtgtgaagaaggccagctacctggactgcatccgggccat tgccgccaatgaggccgacgccgtgacactggatgccggcctggtgtacgatgcctacctggcccccaacaacctgaagcccgtggtgg ccgagttctacggcagcaaagaggacccccagaccttctactacgccgtggccgtggtcaagaaggacagcggcttccagatgaaccag ctgcggggcaagaagtcctgtcacaccggcctgggcagaagcgccggctggaacatccccatcggcctgctgtactgcgatctgcccgaDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 gccccggaagcctctggaaaaggccgtggccaacttcttcagcggcagctgcgccccttgtgctgacggaaccgacttcccccagctgtg tcagctgtgccccggctgtggctgcagcaccctgaaccagtacttcggctacagcggcgccttcaagtgcctgaaggacggcgctggcga cgtggccttcgtgaagcacagcaccatcttcgagaacctggccaacaaggccgaccgggaccagtacgagctgctgtgcctggacaaca ccagaaagcccgtggacgagtacaaggactgccacctcgcccaggtgccatctcacacagtggtggcccggtccatgggcggcaaaga ggatctgatctgggagctgctgaaccaggcccaggaacacttcggcaaggacaagagcaaagagttccagctgttcagcagcccccacg gcaaggatctgctgttcaaggacagcgcccacggctttctgaaggtgccccccagaatggacgccaagatgtacctgggctacgagtacg tgaccgccatccggaacctgagagagggcacctgtcccgaggcccccaccgatgagtgcaagcccgtgaagtggtgcgccctgagcca ccacgagcggctgaagtgcgacgagtggagcgtgaacagcgtgggcaagatcgagtgcgtgagcgccgagacaaccgaggactgtat cgccaagatcatgaacggcgaggccgatgccatgagcctggacggcggcttcgtgtacattgccggcaagtgcggcctggtgcctgtgct ggccgagaactacaacaagagcgacaactgcgaggatacccccgaggccggctactttgccatcgcagtcgtgaagaagtccgccagc gacctgacctgggacaatctgaagggcaagaaaagctgccacaccgccgtgggaaggaccgccgggtggaatattcctatggggctgct gtacaacaagatcaaccactgcagattcgacgagttcttcagcgagggctgcgctcccggcagcaagaaagacagcagcctgtgcaagct gtgcatgggcagcggcctgaacctgtgcgagcccaacaacaaagagggctactacggctacacaggggccttccggtgtctggtggaga agggggacgtggcttttgtgaaacaccagaccgtgccccagaacaccggcggcaagaaccccgacccctgggccaagaacctgaacg agaaggactacgaactgctgtgtctcgacggcacccggaagccagtggaggaatacgccaactgtcacctggccagagcccccaatcac gccgtggtcacccggaaggacaaagaggcctgcgtccacaagatcctgcggcagcagcagcacctgttcggcagcaacgtgaccgact gcagcggcaacttctgcctgttcagaagcgagacaaaggacctcctgttccgggacgacaccgtgtgtctggccaagctgcacgaccgga acacctacgagaagtacctgggcgaggaatatgtgaaggccgtgggcaatctgcggaagtgcagcacctctagcctgctggaagcctgc acctttcgcagacctTAA

[0356] pDP85-cd20-scfv-GFLG-FC-Tf Amino Acid Sequence (SEQ ID NO: 40)

[0357] MYRMQLLSCIALSLALVTNSQIVLSQSPAILSASPGEKVTMTCRASSSVSYIH WFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATYYCQQ WTSNPPTFGGGTKLEIKGGGGSGGGGSGGGGSQVQLQQPGAELVKPGASVKMSC KASGYTFTSYNMHWVKQTPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSS TAYMQLSSLTSEDSAVYYCARSTYYGGDWYFNVWGAGTTVTVSAGGSGSGGFLGG VDGDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDG VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSK LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGSSGGSGGSGGSVPDKTVRWCAVSE HEATKCQSFRDHMKSVIPSDGPSVACVKKASYLDCIRAIAANEADAVTLDAGLVYDAYLAPNNL KPVVAEFYGSKEDPQTFYYAVAVVKKDSGFQMNQLRGKKSCHTGLGRSAGWNIPIGLLYCDLPE PRKPLEKAVANFFSGSCAPCADGTDFPQLCQLCPGCGCSTLNQYFGYSGAFKCLKDGAGDVAFV KHSTIFENLANKADRDQYELLCLDNTRKPVDEYKDCHLAQVPSHTVVARSMGGKEDLIWELLNQ AQEHFGKDKSKEFQLFSSPHGKDLLFKDSAHGFLKVPPRMDAKMYLGYEYVTAIRNLREGTCPE APTDECKPVKWCALSHHERLKCDEWSVNSVGKIECVSAETTEDCIAKIMNGEADAMSLDGGFVY IAGKCGLVPVLAENYNKSDNCEDTPEAGYFAIAVVKKSASDLTWDNLKGKKSCHTAVGRTAGWN IPMGLLYNKINHCRFDEFFSEGCAPGSKKDSSLCKLCMGSGLNLCEPNNKEGYYGYTGAFRCLV EKGDVAFVKHQTVPQNTGGKNPDPWAKNLNEKDYELLCLDGTRKPVEEYANCHLARAPNHAVVT RKDKEACVHKILRQQQHLFGSNVTDCSGNFCLFRSETKDLLFRDDTVCLAKLHDRNTYEKYLGE EYVKAVGNLRKCSTSSLLEACTFRRP*

[0358] pDP95-Herceptin_HC_Fc-N297G,S427C Nucleic Acid Sequence (SEQ ID NO: 41)

[0359] ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTC ACGAATTCGgaggttcagctggtggagtctggcggtggcctggtgcagccagggggctcactccgtttgtcctgtgcagcttctgg cttcaacATCAAGGACACCTATATCcactgggtgcgtcaggccccgggtaagggcctggaatgggttgcaCGCATCDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 TACCCGACGAATGGCTACACGCGCTatgccgatAGCgtcaagggccgtttcactataagcGCAGACACA TCCAAAAACACAGCCtacctacaaatgaacagcttaagagctgaggacactgccgtctattattgtTCACGCTGGGG GGGAGATGGGTTTTATGCAATGgactactggggtcaaggaaccctggtcaccgtctcctcggcctccaccaagggtc catcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaac cggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcag cagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtcgaca aAAAGGTCgagcccaaatcttgtgacaaaactcacacatgcCCCCCCTGCCCAGCGCCAGAATTGCTGG GCGGACCCAGCGTGTTCCTGTTCCCCCCCAAACCTAAAGACACCCTGATGATCAGCC GAACCCCTGAGGTGACCTGCGTGGTGGTGGACGTGAGCCACGAGGACCCCGAGGTG AAGTTCAACTGGTATGTGGACGGCGTGGAGGTCCACAATGCCAAAACGAAGCCCAG GGAGGAGCAGTACGGCAGCACCTACAGGGTAGTGAGCGTCTTGACCGTGCTGCACC AGGACTGGCTGAACGGCAAGGAATACAAATGCAAGGTCAGCAATAAGGCTCTGCCG GCTCCTATCGAGAAGACAATCAGCAAGGCAAAGGGCCAGCCACGCGAACCGCAGGT GTATACTCTGCCCCCCAGCCGGGACGAGCTGACCAAGAACCAGGTGTCCCTGACCT GTCTGGTGAAAGGCTTCTACCCCAGCGACATCGCTGTGGAGTGGGAGAGTAACGGG CAGCCCGAGAACAACTACAAGACCACGCCTCCTGTGCTGGACAGCGACGGCAGCTT CTTCCTGTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAACAGGGCAACGTGT TCTGCTGCTCTGTGATGCACGAGGCCCTGCACAACCATTACACCCAGAAGAGTCTCA GTCTGAGCCCGGGAAAGtaa

[0360] pDP95-Herceptin_HC_Fc-N297G,S427C Amino Acid Sequence (SEQ ID NO: 42)

[0361] MYRMQLLSCIALSLALVTNSEVQLVESGGGLVQPGGSLRLSCAASGFNIKD TYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSL RAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRT PEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYGSTYRVVSVLTVLHQDWLNGKEYK CKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFCCSVMHEALHNHYTQKSLSLSPGK*

[0362] pDP69-His8-CD19 NT.1-2XGFLG-FC-Tf Nucleic Acid Sequence (SEQ ID NO: 43)

[0363] ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTC ACGAATTCGGGCCACCACCATCATCACCACCATCACaccggtcccgaggaacctctagtggtgaaggtg gaagagggagataccgctgctctgtggtgcctcaaggggacctcagatggccccactcagcagctgacctggtctcgggagtccccgctt aaacccttcttaaaatacagcctgggggtgccaggcctgggagtccacgtcaggcccgatgccatctctgtcgtcatcaggaacgtctctca acagatggggggcttctacctgtgccagccggggcccccctctgagaaggcctggcagcctggctggacagtcaatgtggagggcagc ggggagctgttccggtggaatgtttcggacctaggtggcctgggctgtggcctgaagaacaggtcctcagagggccccagctccccttcc gggaagctcatgagccccaagctgtatgtgtgggccaaagaccgccctgagatctgggagggagagcctccgtgtctcccaccgaggga cagcctgaaccagagcctcagcagggacctcaccgtagcccctggctccacactctggctgtcctgtggggtaccccctgactctgtgtcc aggggccccctctcctggacccatgtgcaccccaaggggcctaagtcattgctgagcctagagctgaaggacgatcgcccggccagaga tatgtgggtaatgggtacgtcactgatgttgccccgggccacagctcaagacgctggaaagtggtattgtcaccgtggcaacgtaaccacct cattccacctggaggtaatcgctcggccagtaaaggctcactcagacctgaggactggtggctggaagggtggaTTtCTGggcggcg ggTTCCTGggaggcGTAGACGGCGACAAAACTCACACATGCCCACCGTGCCCAGCACCT GAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTC ATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGA CCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGADOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 CAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACC GTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAA AGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAG AACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTC AGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGA GAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCG ACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAG GGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAG AAGAGCCTCTCCCTGTCTCCGGGTAAAggatcctctgggggaagtggaggtagcggtggttctgtgcccgataag acagtgcggtggtgcgccgtgtctgagcacgaggccaccaagtgccagagcttccgggaccacatgaagtccgtgatccccagcgacg gccctagcgtggcctgtgtgaagaaggccagctacctggactgcatccgggccattgccgccaatgaggccgacgccgtgacactggat gccggcctggtgtacgatgcctacctggcccccaacaacctgaagcccgtggtggccgagttctacggcagcaaagaggacccccagac cttctactacgccgtggccgtggtcaagaaggacagcggcttccagatgaaccagctgcggggcaagaagtcctgtcacaccggcctgg gcagaagcgccggctggaacatccccatcggcctgctgtactgcgatctgcccgagccccggaagcctctggaaaaggccgtggccaa cttcttcagcggcagctgcgccccttgtgctgacggaaccgacttcccccagctgtgtcagctgtgccccggctgtggctgcagcaccctga accagtacttcggctacagcggcgccttcaagtgcctgaaggacggcgctggcgacgtggccttcgtgaagcacagcaccatcttcgaga acctggccaacaaggccgaccgggaccagtacgagctgctgtgcctggacaacaccagaaagcccgtggacgagtacaaggactgcc acctcgcccaggtgccatctcacacagtggtggcccggtccatgggcggcaaagaggatctgatctgggagctgctgaaccaggcccag gaacacttcggcaaggacaagagcaaagagttccagctgttcagcagcccccacggcaaggatctgctgttcaaggacagcgcccacgg ctttctgaaggtgccccccagaatggacgccaagatgtacctgggctacgagtacgtgaccgccatccggaacctgagagagggcacctg tcccgaggcccccaccgatgagtgcaagcccgtgaagtggtgcgccctgagccaccacgagcggctgaagtgcgacgagtggagcgt gaacagcgtgggcaagatcgagtgcgtgagcgccgagacaaccgaggactgtatcgccaagatcatgaacggcgaggccgatgccatg agcctggacggcggcttcgtgtacattgccggcaagtgcggcctggtgcctgtgctggccgagaactacaacaagagcgacaactgcga ggatacccccgaggccggctactttgccatcgcagtcgtgaagaagtccgccagcgacctgacctgggacaatctgaagggcaagaaaa gctgccacaccgccgtgggaaggaccgccgggtggaatattcctatggggctgctgtacaacaagatcaaccactgcagattcgacgagt tcttcagcgagggctgcgctcccggcagcaagaaagacagcagcctgtgcaagctgtgcatgggcagcggcctgaacctgtgcgagcc caacaacaaagagggctactacggctacacaggggccttccggtgtctggtggagaagggggacgtggcttttgtgaaacaccagaccgt gccccagaacaccggcggcaagaaccccgacccctgggccaagaacctgaacgagaaggactacgaactgctgtgtctcgacggcac ccggaagccagtggaggaatacgccaactgtcacctggccagagcccccaatcacgccgtggtcacccggaaggacaaagaggcctg cgtccacaagatcctgcggcagcagcagcacctgttcggcagcaacgtgaccgactgcagcggcaacttctgcctgttcagaagcgaga caaaggacctcctgttccgggacgacaccgtgtgtctggccaagctgcacgaccggaacacctacgagaagtacctgggcgaggaatat gtgaaggccgtgggcaatctgcggaagtgcagcacctctagcctgctggaagcctgcacctttcgcagacctTAA

[0364] pDP69-His8-CD19 NT.1-2XGFLG-FC-Tf Amino Acid Sequence (SEQ ID NO: 44)

[0365] MYRMQLLSCIALSLALVTNSGHHHHHHHHTGPEEPLVVKVEEGDTAALWCL KGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFY LCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGK LMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDS VSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKW YCHRGNVTTSFHLEVIARPVKAHSDLRTGGWKGGFLGGGFLGGVDGDKTHTCPPCPA PELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQ YNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGKGSSGGSGGSGGSVPDKTVRWCAVSEHEATKCQSFRDHMK SVIPSDGPSVACVKKASYLDCIRAIAANEADAVTLDAGLVYDAYLAPNNLKPVVAEFYGSKEDP QTFYYAVAVVKKDSGFQMNQLRGKKSCHTGLGRSAGWNIPIGLLYCDLPEPRKPLEKAVANFFSDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 GSCAPCADGTDFPQLCQLCPGCGCSTLNQYFGYSGAFKCLKDGAGDVAFVKHSTIFENLANKAD RDQYELLCLDNTRKPVDEYKDCHLAQVPSHTVVARSMGGKEDLIWELLNQAQEHFGKDKSKEFQ LFSSPHGKDLLFKDSAHGFLKVPPRMDAKMYLGYEYVTAIRNLREGTCPEAPTDECKPVKWCAL SHHERLKCDEWSVNSVGKIECVSAETTEDCIAKIMNGEADAMSLDGGFVYIAGKCGLVPVLAEN YNKSDNCEDTPEAGYFAIAVVKKSASDLTWDNLKGKKSCHTAVGRTAGWNIPMGLLYNKINHCR FDEFFSEGCAPGSKKDSSLCKLCMGSGLNLCEPNNKEGYYGYTGAFRCLVEKGDVAFVKHQTVP QNTGGKNPDPWAKNLNEKDYELLCLDGTRKPVEEYANCHLARAPNHAVVTRKDKEACVHKILRQ QQHLFGSNVTDCSGNFCLFRSETKDLLFRDDTVCLAKLHDRNTYEKYLGEEYVKAVGNLRKCST SSLLEACTFRRP*

[0366] pDP70-His8-CD19 NT.1-3XGFLG-FC-Tf Nucleic Acid Sequence (SEQ ID NO: 45)

[0367] ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTC ACGAATTCGGGCCACCACCATCATCACCACCATCACaccggtcccgaggaacctctagtggtgaaggtg gaagagggagataccgctgctctgtggtgcctcaaggggacctcagatggccccactcagcagctgacctggtctcgggagtccccgctt aaacccttcttaaaatacagcctgggggtgccaggcctgggagtccacgtcaggcccgatgccatctctgtcgtcatcaggaacgtctctca acagatggggggcttctacctgtgccagccggggcccccctctgagaaggcctggcagcctggctggacagtcaatgtggagggcagc ggggagctgttccggtggaatgtttcggacctaggtggcctgggctgtggcctgaagaacaggtcctcagagggccccagctccccttcc gggaagctcatgagccccaagctgtatgtgtgggccaaagaccgccctgagatctgggagggagagcctccgtgtctcccaccgaggga cagcctgaaccagagcctcagcagggacctcaccgtagcccctggctccacactctggctgtcctgtggggtaccccctgactctgtgtcc aggggccccctctcctggacccatgtgcaccccaaggggcctaagtcattgctgagcctagagctgaaggacgatcgcccggccagaga tatgtgggtaatgggtacgtcactgatgttgccccgggccacagctcaagacgctggaaagtggtattgtcaccgtggcaacgtaaccacct cattccacctggaggtaatcgctcggccagtaaaggctcactcagacctgaggactggtggctggaagggtggaTTCCTGggcggc gggTTtCTGggaggcggaTTtCTGggaggcGTAGACGGCGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGT GAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGC ATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTC AGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAA GGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAG GGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACC AAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCC GTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGT GCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAG GTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCA CTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAAggatcctctgggggaagtggaggtagcggtg gttctgtgcccgataagacagtgcggtggtgcgccgtgtctgagcacgaggccaccaagtgccagagcttccgggaccacatgaagtccg tgatccccagcgacggccctagcgtggcctgtgtgaagaaggccagctacctggactgcatccgggccattgccgccaatgaggccgac gccgtgacactggatgccggcctggtgtacgatgcctacctggcccccaacaacctgaagcccgtggtggccgagttctacggcagcaaa gaggacccccagaccttctactacgccgtggccgtggtcaagaaggacagcggcttccagatgaaccagctgcggggcaagaagtcctg tcacaccggcctgggcagaagcgccggctggaacatccccatcggcctgctgtactgcgatctgcccgagccccggaagcctctggaaa aggccgtggccaacttcttcagcggcagctgcgccccttgtgctgacggaaccgacttcccccagctgtgtcagctgtgccccggctgtgg ctgcagcaccctgaaccagtacttcggctacagcggcgccttcaagtgcctgaaggacggcgctggcgacgtggccttcgtgaagcaca gcaccatcttcgagaacctggccaacaaggccgaccgggaccagtacgagctgctgtgcctggacaacaccagaaagcccgtggacga gtacaaggactgccacctcgcccaggtgccatctcacacagtggtggcccggtccatgggcggcaaagaggatctgatctgggagctgct gaaccaggcccaggaacacttcggcaaggacaagagcaaagagttccagctgttcagcagcccccacggcaaggatctgctgttcaagg acagcgcccacggctttctgaaggtgccccccagaatggacgccaagatgtacctgggctacgagtacgtgaccgccatccggaacctg agagagggcacctgtcccgaggcccccaccgatgagtgcaagcccgtgaagtggtgcgccctgagccaccacgagcggctgaagtgcDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 gacgagtggagcgtgaacagcgtgggcaagatcgagtgcgtgagcgccgagacaaccgaggactgtatcgccaagatcatgaacggc gaggccgatgccatgagcctggacggcggcttcgtgtacattgccggcaagtgcggcctggtgcctgtgctggccgagaactacaacaa gagcgacaactgcgaggatacccccgaggccggctactttgccatcgcagtcgtgaagaagtccgccagcgacctgacctgggacaatc tgaagggcaagaaaagctgccacaccgccgtgggaaggaccgccgggtggaatattcctatggggctgctgtacaacaagatcaaccac tgcagattcgacgagttcttcagcgagggctgcgctcccggcagcaagaaagacagcagcctgtgcaagctgtgcatgggcagcggcct gaacctgtgcgagcccaacaacaaagagggctactacggctacacaggggccttccggtgtctggtggagaagggggacgtggcttttgt gaaacaccagaccgtgccccagaacaccggcggcaagaaccccgacccctgggccaagaacctgaacgagaaggactacgaactgct gtgtctcgacggcacccggaagccagtggaggaatacgccaactgtcacctggccagagcccccaatcacgccgtggtcacccggaag gacaaagaggcctgcgtccacaagatcctgcggcagcagcagcacctgttcggcagcaacgtgaccgactgcagcggcaacttctgcct gttcagaagcgagacaaaggacctcctgttccgggacgacaccgtgtgtctggccaagctgcacgaccggaacacctacgagaagtacc tgggcgaggaatatgtgaaggccgtgggcaatctgcggaagtgcagcacctctagcctgctggaagcctgcacctttcgcagacctTAA

[0368] pDP70-His8-CD19 NT.1-3XGFLG-FC-Tf Amino Acid Sequence (SEQ ID NO: 46)

[0369] MYRMQLLSCIALSLALVTNSGHHHHHHHHTGPEEPLVVKVEEGDTAALWCL KGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFY LCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGK LMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDS VSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKW YCHRGNVTTSFHLEVIARPVKAHSDLRTGGWKGGFLGGGFLGGGFLGGVDGDKTHT CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPP SRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPGKGSSGGSGGSGGSVPDKTVRWCAVSEHEATKCQS FRDHMKSVIPSDGPSVACVKKASYLDCIRAIAANEADAVTLDAGLVYDAYLAPNNLKPVVAEFY GSKEDPQTFYYAVAVVKKDSGFQMNQLRGKKSCHTGLGRSAGWNIPIGLLYCDLPEPRKPLEKA VANFFSGSCAPCADGTDFPQLCQLCPGCGCSTLNQYFGYSGAFKCLKDGAGDVAFVKHSTIFEN LANKADRDQYELLCLDNTRKPVDEYKDCHLAQVPSHTVVARSMGGKEDLIWELLNQAQEHFGKD KSKEFQLFSSPHGKDLLFKDSAHGFLKVPPRMDAKMYLGYEYVTAIRNLREGTCPEAPTDECKP VKWCALSHHERLKCDEWSVNSVGKIECVSAETTEDCIAKIMNGEADAMSLDGGFVYIAGKCGLV PVLAENYNKSDNCEDTPEAGYFAIAVVKKSASDLTWDNLKGKKSCHTAVGRTAGWNIPMGLLYN KINHCRFDEFFSEGCAPGSKKDSSLCKLCMGSGLNLCEPNNKEGYYGYTGAFRCLVEKGDVAFV KHQTVPQNTGGKNPDPWAKNLNEKDYELLCLDGTRKPVEEYANCHLARAPNHAVVTRKDKEACV HKILRQQQHLFGSNVTDCSGNFCLFRSETKDLLFRDDTVCLAKLHDRNTYEKYLGEEYVKAVGN LRKCSTSSLLEACTFRRP*

[0370] pDP71-His8-CD19 NT.1-GFLG-FK-FC-Tf Nucleic Acid Sequence (SEQ ID NO: 47)

[0371] ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTC ACGAATTCGGGCCACCACCATCATCACCACCATCACaccggtcccgaggaacctctagtggtgaaggtg gaagagggagataccgctgctctgtggtgcctcaaggggacctcagatggccccactcagcagctgacctggtctcgggagtccccgctt aaacccttcttaaaatacagcctgggggtgccaggcctgggagtccacgtcaggcccgatgccatctctgtcgtcatcaggaacgtctctca acagatggggggcttctacctgtgccagccggggcccccctctgagaaggcctggcagcctggctggacagtcaatgtggagggcagc ggggagctgttccggtggaatgtttcggacctaggtggcctgggctgtggcctgaagaacaggtcctcagagggccccagctccccttcc gggaagctcatgagccccaagctgtatgtgtgggccaaagaccgccctgagatctgggagggagagcctccgtgtctcccaccgaggga cagcctgaaccagagcctcagcagggacctcaccgtagcccctggctccacactctggctgtcctgtggggtaccccctgactctgtgtcc aggggccccctctcctggacccatgtgcaccccaaggggcctaagtcattgctgagcctagagctgaaggacgatcgcccggccagaga tatgtgggtaatgggtacgtcactgatgttgccccgggccacagctcaagacgctggaaagtggtattgtcaccgtggcaacgtaaccacctDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 cattccacctggaggtaatcgctcggccagtaaaggctcactcagacctgaggactggtggctggaagggtgggTTCCTGggaggc TTCAAAggcGTAGACGGCGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAA CTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATG ATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCC TGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAA AGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTC CTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGC CCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAAC CACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGC CTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAG CAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACG GCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGG AACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAG AGCCTCTCCCTGTCTCCGGGTAAAggatcctctgggggaagtggaggtagcggtggttctgtgcccgataagacagt gcggtggtgcgccgtgtctgagcacgaggccaccaagtgccagagcttccgggaccacatgaagtccgtgatccccagcgacggcccta gcgtggcctgtgtgaagaaggccagctacctggactgcatccgggccattgccgccaatgaggccgacgccgtgacactggatgccggc ctggtgtacgatgcctacctggcccccaacaacctgaagcccgtggtggccgagttctacggcagcaaagaggacccccagaccttctac tacgccgtggccgtggtcaagaaggacagcggcttccagatgaaccagctgcggggcaagaagtcctgtcacaccggcctgggcagaa gcgccggctggaacatccccatcggcctgctgtactgcgatctgcccgagccccggaagcctctggaaaaggccgtggccaacttcttca gcggcagctgcgccccttgtgctgacggaaccgacttcccccagctgtgtcagctgtgccccggctgtggctgcagcaccctgaaccagt acttcggctacagcggcgccttcaagtgcctgaaggacggcgctggcgacgtggccttcgtgaagcacagcaccatcttcgagaacctgg ccaacaaggccgaccgggaccagtacgagctgctgtgcctggacaacaccagaaagcccgtggacgagtacaaggactgccacctcgc ccaggtgccatctcacacagtggtggcccggtccatgggcggcaaagaggatctgatctgggagctgctgaaccaggcccaggaacact tcggcaaggacaagagcaaagagttccagctgttcagcagcccccacggcaaggatctgctgttcaaggacagcgcccacggctttctga aggtgccccccagaatggacgccaagatgtacctgggctacgagtacgtgaccgccatccggaacctgagagagggcacctgtcccga ggcccccaccgatgagtgcaagcccgtgaagtggtgcgccctgagccaccacgagcggctgaagtgcgacgagtggagcgtgaacag cgtgggcaagatcgagtgcgtgagcgccgagacaaccgaggactgtatcgccaagatcatgaacggcgaggccgatgccatgagcctg gacggcggcttcgtgtacattgccggcaagtgcggcctggtgcctgtgctggccgagaactacaacaagagcgacaactgcgaggatac ccccgaggccggctactttgccatcgcagtcgtgaagaagtccgccagcgacctgacctgggacaatctgaagggcaagaaaagctgcc acaccgccgtgggaaggaccgccgggtggaatattcctatggggctgctgtacaacaagatcaaccactgcagattcgacgagttcttcag cgagggctgcgctcccggcagcaagaaagacagcagcctgtgcaagctgtgcatgggcagcggcctgaacctgtgcgagcccaacaa caaagagggctactacggctacacaggggccttccggtgtctggtggagaagggggacgtggcttttgtgaaacaccagaccgtgcccc agaacaccggcggcaagaaccccgacccctgggccaagaacctgaacgagaaggactacgaactgctgtgtctcgacggcacccgga agccagtggaggaatacgccaactgtcacctggccagagcccccaatcacgccgtggtcacccggaaggacaaagaggcctgcgtcca caagatcctgcggcagcagcagcacctgttcggcagcaacgtgaccgactgcagcggcaacttctgcctgttcagaagcgagacaaagg acctcctgttccgggacgacaccgtgtgtctggccaagctgcacgaccggaacacctacgagaagtacctgggcgaggaatatgtgaag gccgtgggcaatctgcggaagtgcagcacctctagcctgctggaagcctgcacctttcgcagacctTAA

[0372] pDP71-His8-CD19 NT.1-GFLG-FK-FC-Tf Amino Acid Sequence (SEQ ID NO: 48)

[0373] MYRMQLLSCIALSLALVTNSGHHHHHHHHTGPEEPLVVKVEEGDTAALWCL KGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFY LCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGK LMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDS VSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKW YCHRGNVTTSFHLEVIARPVKAHSDLRTGGWKGGFLGGFKGVDGDKTHTCPPCPAPE LLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN QVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGKGSSGGSGGSGGSVPDKTVRWCAVSEHEATKCQSFRDHMKSV IPSDGPSVACVKKASYLDCIRAIAANEADAVTLDAGLVYDAYLAPNNLKPVVAEFYGSKEDPQT FYYAVAVVKKDSGFQMNQLRGKKSCHTGLGRSAGWNIPIGLLYCDLPEPRKPLEKAVANFFSGS CAPCADGTDFPQLCQLCPGCGCSTLNQYFGYSGAFKCLKDGAGDVAFVKHSTIFENLANKADRD QYELLCLDNTRKPVDEYKDCHLAQVPSHTVVARSMGGKEDLIWELLNQAQEHFGKDKSKEFQLF SSPHGKDLLFKDSAHGFLKVPPRMDAKMYLGYEYVTAIRNLREGTCPEAPTDECKPVKWCALSH HERLKCDEWSVNSVGKIECVSAETTEDCIAKIMNGEADAMSLDGGFVYIAGKCGLVPVLAENYN KSDNCEDTPEAGYFAIAVVKKSASDLTWDNLKGKKSCHTAVGRTAGWNIPMGLLYNKINHCRFD EFFSEGCAPGSKKDSSLCKLCMGSGLNLCEPNNKEGYYGYTGAFRCLVEKGDVAFVKHQTVPQN TGGKNPDPWAKNLNEKDYELLCLDGTRKPVEEYANCHLARAPNHAVVTRKDKEACVHKILRQQQ HLFGSNVTDCSGNFCLFRSETKDLLFRDDTVCLAKLHDRNTYEKYLGEEYVKAVGNLRKCSTSS LLEACTFRRP*

[0374] pDP72-His8-CD19 NT.1-GFLG-VA-FC-Tf Nucleic Acid Sequence (SEQ ID NO: 49)

[0375] ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTC ACGAATTCGGGCCACCACCATCATCACCACCATCACaccggtcccgaggaacctctagtggtgaaggtg gaagagggagataccgctgctctgtggtgcctcaaggggacctcagatggccccactcagcagctgacctggtctcgggagtccccgctt aaacccttcttaaaatacagcctgggggtgccaggcctgggagtccacgtcaggcccgatgccatctctgtcgtcatcaggaacgtctctca acagatggggggcttctacctgtgccagccggggcccccctctgagaaggcctggcagcctggctggacagtcaatgtggagggcagc ggggagctgttccggtggaatgtttcggacctaggtggcctgggctgtggcctgaagaacaggtcctcagagggccccagctccccttcc gggaagctcatgagccccaagctgtatgtgtgggccaaagaccgccctgagatctgggagggagagcctccgtgtctcccaccgaggga cagcctgaaccagagcctcagcagggacctcaccgtagcccctggctccacactctggctgtcctgtggggtaccccctgactctgtgtcc aggggccccctctcctggacccatgtgcaccccaaggggcctaagtcattgctgagcctagagctgaaggacgatcgcccggccagaga tatgtgggtaatgggtacgtcactgatgttgccccgggccacagctcaagacgctggaaagtggtattgtcaccgtggcaacgtaaccacct cattccacctggaggtaatcgctcggccagtaaaggctcactcagacctgaggactggtggctggaagggtgggTTCCTGggaggc GTAgctggcGTAGACGGCGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAC TCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGA TCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCT GAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAA GCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCC TGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCC CTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACC ACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCC TGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGC AATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGG CTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGA ACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGA GCCTCTCCCTGTCTCCGGGTAAAggatcctctgggggaagtggaggtagcggtggttctgtgcccgataagacagtg cggtggtgcgccgtgtctgagcacgaggccaccaagtgccagagcttccgggaccacatgaagtccgtgatccccagcgacggccctag cgtggcctgtgtgaagaaggccagctacctggactgcatccgggccattgccgccaatgaggccgacgccgtgacactggatgccggcc tggtgtacgatgcctacctggcccccaacaacctgaagcccgtggtggccgagttctacggcagcaaagaggacccccagaccttctact acgccgtggccgtggtcaagaaggacagcggcttccagatgaaccagctgcggggcaagaagtcctgtcacaccggcctgggcagaag cgccggctggaacatccccatcggcctgctgtactgcgatctgcccgagccccggaagcctctggaaaaggccgtggccaacttcttcag cggcagctgcgccccttgtgctgacggaaccgacttcccccagctgtgtcagctgtgccccggctgtggctgcagcaccctgaaccagtaDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 cttcggctacagcggcgccttcaagtgcctgaaggacggcgctggcgacgtggccttcgtgaagcacagcaccatcttcgagaacctggc caacaaggccgaccgggaccagtacgagctgctgtgcctggacaacaccagaaagcccgtggacgagtacaaggactgccacctcgcc caggtgccatctcacacagtggtggcccggtccatgggcggcaaagaggatctgatctgggagctgctgaaccaggcccaggaacactt cggcaaggacaagagcaaagagttccagctgttcagcagcccccacggcaaggatctgctgttcaaggacagcgcccacggctttctga aggtgccccccagaatggacgccaagatgtacctgggctacgagtacgtgaccgccatccggaacctgagagagggcacctgtcccga ggcccccaccgatgagtgcaagcccgtgaagtggtgcgccctgagccaccacgagcggctgaagtgcgacgagtggagcgtgaacag cgtgggcaagatcgagtgcgtgagcgccgagacaaccgaggactgtatcgccaagatcatgaacggcgaggccgatgccatgagcctg gacggcggcttcgtgtacattgccggcaagtgcggcctggtgcctgtgctggccgagaactacaacaagagcgacaactgcgaggatac ccccgaggccggctactttgccatcgcagtcgtgaagaagtccgccagcgacctgacctgggacaatctgaagggcaagaaaagctgcc acaccgccgtgggaaggaccgccgggtggaatattcctatggggctgctgtacaacaagatcaaccactgcagattcgacgagttcttcag cgagggctgcgctcccggcagcaagaaagacagcagcctgtgcaagctgtgcatgggcagcggcctgaacctgtgcgagcccaacaa caaagagggctactacggctacacaggggccttccggtgtctggtggagaagggggacgtggcttttgtgaaacaccagaccgtgcccc agaacaccggcggcaagaaccccgacccctgggccaagaacctgaacgagaaggactacgaactgctgtgtctcgacggcacccgga agccagtggaggaatacgccaactgtcacctggccagagcccccaatcacgccgtggtcacccggaaggacaaagaggcctgcgtcca caagatcctgcggcagcagcagcacctgttcggcagcaacgtgaccgactgcagcggcaacttctgcctgttcagaagcgagacaaagg acctcctgttccgggacgacaccgtgtgtctggccaagctgcacgaccggaacacctacgagaagtacctgggcgaggaatatgtgaag gccgtgggcaatctgcggaagtgcagcacctctagcctgctggaagcctgcacctttcgcagacctTAA

[0376] pDP72-His8-CD19 NT.1-GFLG-VA-FC-Tf Amino Acid Sequence (SEQ ID NO: 50)

[0377] MYRMQLLSCIALSLALVTNSGHHHHHHHHTGPEEPLVVKVEEGDTAALWCL KGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFY LCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGK LMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDS VSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKW YCHRGNVTTSFHLEVIARPVKAHSDLRTGGWKGGFLGGVAGVDGDKTHTCPPCPAPE LLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN QVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGKGSSGGSGGSGGSVPDKTVRWCAVSEHEATKCQSFRDHMKSV IPSDGPSVACVKKASYLDCIRAIAANEADAVTLDAGLVYDAYLAPNNLKPVVAEFYGSKEDPQT FYYAVAVVKKDSGFQMNQLRGKKSCHTGLGRSAGWNIPIGLLYCDLPEPRKPLEKAVANFFSGS CAPCADGTDFPQLCQLCPGCGCSTLNQYFGYSGAFKCLKDGAGDVAFVKHSTIFENLANKADRD QYELLCLDNTRKPVDEYKDCHLAQVPSHTVVARSMGGKEDLIWELLNQAQEHFGKDKSKEFQLF SSPHGKDLLFKDSAHGFLKVPPRMDAKMYLGYEYVTAIRNLREGTCPEAPTDECKPVKWCALSH HERLKCDEWSVNSVGKIECVSAETTEDCIAKIMNGEADAMSLDGGFVYIAGKCGLVPVLAENYN KSDNCEDTPEAGYFAIAVVKKSASDLTWDNLKGKKSCHTAVGRTAGWNIPMGLLYNKINHCRFD EFFSEGCAPGSKKDSSLCKLCMGSGLNLCEPNNKEGYYGYTGAFRCLVEKGDVAFVKHQTVPQN TGGKNPDPWAKNLNEKDYELLCLDGTRKPVEEYANCHLARAPNHAVVTRKDKEACVHKILRQQQ HLFGSNVTDCSGNFCLFRSETKDLLFRDDTVCLAKLHDRNTYEKYLGEEYVKAVGNLRKCSTSS LLEACTFRRP*

[0378] pDP73-His8-CD19 NT.1-GFLG-VK-FC-Tf Nucleic Acid Sequence (SEQ ID NO: 51)

[0379] ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTC ACGAATTCGGGCCACCACCATCATCACCACCATCACaccggtcccgaggaacctctagtggtgaaggtg gaagagggagataccgctgctctgtggtgcctcaaggggacctcagatggccccactcagcagctgacctggtctcgggagtccccgctt aaacccttcttaaaatacagcctgggggtgccaggcctgggagtccacgtcaggcccgatgccatctctgtcgtcatcaggaacgtctctcaDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 acagatggggggcttctacctgtgccagccggggcccccctctgagaaggcctggcagcctggctggacagtcaatgtggagggcagc ggggagctgttccggtggaatgtttcggacctaggtggcctgggctgtggcctgaagaacaggtcctcagagggccccagctccccttcc gggaagctcatgagccccaagctgtatgtgtgggccaaagaccgccctgagatctgggagggagagcctccgtgtctcccaccgaggga cagcctgaaccagagcctcagcagggacctcaccgtagcccctggctccacactctggctgtcctgtggggtaccccctgactctgtgtcc aggggccccctctcctggacccatgtgcaccccaaggggcctaagtcattgctgagcctagagctgaaggacgatcgcccggccagaga tatgtgggtaatgggtacgtcactgatgttgccccgggccacagctcaagacgctggaaagtggtattgtcaccgtggcaacgtaaccacct cattccacctggaggtaatcgctcggccagtaaaggctcactcagacctgaggactggtggctggaagggtgggTTCCTGggaggc GTAAAAggcGTAGACGGCGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAA CTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATG ATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCC TGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAA AGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTC CTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGC CCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAAC CACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGC CTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAG CAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACG GCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGG AACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAG AGCCTCTCCCTGTCTCCGGGTAAAggatcctctgggggaagtggaggtagcggtggttctgtgcccgataagacagt gcggtggtgcgccgtgtctgagcacgaggccaccaagtgccagagcttccgggaccacatgaagtccgtgatccccagcgacggcccta gcgtggcctgtgtgaagaaggccagctacctggactgcatccgggccattgccgccaatgaggccgacgccgtgacactggatgccggc ctggtgtacgatgcctacctggcccccaacaacctgaagcccgtggtggccgagttctacggcagcaaagaggacccccagaccttctac tacgccgtggccgtggtcaagaaggacagcggcttccagatgaaccagctgcggggcaagaagtcctgtcacaccggcctgggcagaa gcgccggctggaacatccccatcggcctgctgtactgcgatctgcccgagccccggaagcctctggaaaaggccgtggccaacttcttca gcggcagctgcgccccttgtgctgacggaaccgacttcccccagctgtgtcagctgtgccccggctgtggctgcagcaccctgaaccagt acttcggctacagcggcgccttcaagtgcctgaaggacggcgctggcgacgtggccttcgtgaagcacagcaccatcttcgagaacctgg ccaacaaggccgaccgggaccagtacgagctgctgtgcctggacaacaccagaaagcccgtggacgagtacaaggactgccacctcgc ccaggtgccatctcacacagtggtggcccggtccatgggcggcaaagaggatctgatctgggagctgctgaaccaggcccaggaacact tcggcaaggacaagagcaaagagttccagctgttcagcagcccccacggcaaggatctgctgttcaaggacagcgcccacggctttctga aggtgccccccagaatggacgccaagatgtacctgggctacgagtacgtgaccgccatccggaacctgagagagggcacctgtcccga ggcccccaccgatgagtgcaagcccgtgaagtggtgcgccctgagccaccacgagcggctgaagtgcgacgagtggagcgtgaacag cgtgggcaagatcgagtgcgtgagcgccgagacaaccgaggactgtatcgccaagatcatgaacggcgaggccgatgccatgagcctg gacggcggcttcgtgtacattgccggcaagtgcggcctggtgcctgtgctggccgagaactacaacaagagcgacaactgcgaggatac ccccgaggccggctactttgccatcgcagtcgtgaagaagtccgccagcgacctgacctgggacaatctgaagggcaagaaaagctgcc acaccgccgtgggaaggaccgccgggtggaatattcctatggggctgctgtacaacaagatcaaccactgcagattcgacgagttcttcag cgagggctgcgctcccggcagcaagaaagacagcagcctgtgcaagctgtgcatgggcagcggcctgaacctgtgcgagcccaacaa caaagagggctactacggctacacaggggccttccggtgtctggtggagaagggggacgtggcttttgtgaaacaccagaccgtgcccc agaacaccggcggcaagaaccccgacccctgggccaagaacctgaacgagaaggactacgaactgctgtgtctcgacggcacccgga agccagtggaggaatacgccaactgtcacctggccagagcccccaatcacgccgtggtcacccggaaggacaaagaggcctgcgtcca caagatcctgcggcagcagcagcacctgttcggcagcaacgtgaccgactgcagcggcaacttctgcctgttcagaagcgagacaaagg acctcctgttccgggacgacaccgtgtgtctggccaagctgcacgaccggaacacctacgagaagtacctgggcgaggaatatgtgaag gccgtgggcaatctgcggaagtgcagcacctctagcctgctggaagcctgcacctttcgcagacctTAA

[0380] pDP73-His8-CD19 NT.1-GFLG-VK-FC-Tf Amino Acid Sequence (SEQ ID NO: 52)DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17

[0381] MYRMQLLSCIALSLALVTNSGHHHHHHHHTGPEEPLVVKVEEGDTAALWCL KGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFY LCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGK LMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDS VSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKW YCHRGNVTTSFHLEVIARPVKAHSDLRTGGWKGGFLGGVKGVDGDKTHTCPPCPAPE LLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN QVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGKGSSGGSGGSGGSVPDKTVRWCAVSEHEATKCQSFRDHMKSV IPSDGPSVACVKKASYLDCIRAIAANEADAVTLDAGLVYDAYLAPNNLKPVVAEFYGSKEDPQT FYYAVAVVKKDSGFQMNQLRGKKSCHTGLGRSAGWNIPIGLLYCDLPEPRKPLEKAVANFFSGS CAPCADGTDFPQLCQLCPGCGCSTLNQYFGYSGAFKCLKDGAGDVAFVKHSTIFENLANKADRD QYELLCLDNTRKPVDEYKDCHLAQVPSHTVVARSMGGKEDLIWELLNQAQEHFGKDKSKEFQLF SSPHGKDLLFKDSAHGFLKVPPRMDAKMYLGYEYVTAIRNLREGTCPEAPTDECKPVKWCALSH HERLKCDEWSVNSVGKIECVSAETTEDCIAKIMNGEADAMSLDGGFVYIAGKCGLVPVLAENYN KSDNCEDTPEAGYFAIAVVKKSASDLTWDNLKGKKSCHTAVGRTAGWNIPMGLLYNKINHCRFD EFFSEGCAPGSKKDSSLCKLCMGSGLNLCEPNNKEGYYGYTGAFRCLVEKGDVAFVKHQTVPQN TGGKNPDPWAKNLNEKDYELLCLDGTRKPVEEYANCHLARAPNHAVVTRKDKEACVHKILRQQQ HLFGSNVTDCSGNFCLFRSETKDLLFRDDTVCLAKLHDRNTYEKYLGEEYVKAVGNLRKCSTSS LLEACTFRRP*

[0382] pDP74-His8-CD19 NT.1-GFLG-VR-FC-Tf Nucleic Acid Sequence (SEQ ID NO: 53)

[0383] ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTC ACGAATTCGGGCCACCACCATCATCACCACCATCACaccggtcccgaggaacctctagtggtgaaggtg gaagagggagataccgctgctctgtggtgcctcaaggggacctcagatggccccactcagcagctgacctggtctcgggagtccccgctt aaacccttcttaaaatacagcctgggggtgccaggcctgggagtccacgtcaggcccgatgccatctctgtcgtcatcaggaacgtctctca acagatggggggcttctacctgtgccagccggggcccccctctgagaaggcctggcagcctggctggacagtcaatgtggagggcagc ggggagctgttccggtggaatgtttcggacctaggtggcctgggctgtggcctgaagaacaggtcctcagagggccccagctccccttcc gggaagctcatgagccccaagctgtatgtgtgggccaaagaccgccctgagatctgggagggagagcctccgtgtctcccaccgaggga cagcctgaaccagagcctcagcagggacctcaccgtagcccctggctccacactctggctgtcctgtggggtaccccctgactctgtgtcc aggggccccctctcctggacccatgtgcaccccaaggggcctaagtcattgctgagcctagagctgaaggacgatcgcccggccagaga tatgtgggtaatgggtacgtcactgatgttgccccgggccacagctcaagacgctggaaagtggtattgtcaccgtggcaacgtaaccacct cattccacctggaggtaatcgctcggccagtaaaggctcactcagacctgaggactggtggctggaagggtgggTTCCTGggaggc GTACGGggcGTAGACGGCGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAA CTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATG ATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCC TGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAA AGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTC CTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGC CCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAAC CACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGC CTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAG CAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACG GCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 AACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAG AGCCTCTCCCTGTCTCCGGGTAAAggatcctctgggggaagtggaggtagcggtggttctgtgcccgataagacagt gcggtggtgcgccgtgtctgagcacgaggccaccaagtgccagagcttccgggaccacatgaagtccgtgatccccagcgacggcccta gcgtggcctgtgtgaagaaggccagctacctggactgcatccgggccattgccgccaatgaggccgacgccgtgacactggatgccggc ctggtgtacgatgcctacctggcccccaacaacctgaagcccgtggtggccgagttctacggcagcaaagaggacccccagaccttctac tacgccgtggccgtggtcaagaaggacagcggcttccagatgaaccagctgcggggcaagaagtcctgtcacaccggcctgggcagaa gcgccggctggaacatccccatcggcctgctgtactgcgatctgcccgagccccggaagcctctggaaaaggccgtggccaacttcttca gcggcagctgcgccccttgtgctgacggaaccgacttcccccagctgtgtcagctgtgccccggctgtggctgcagcaccctgaaccagt acttcggctacagcggcgccttcaagtgcctgaaggacggcgctggcgacgtggccttcgtgaagcacagcaccatcttcgagaacctgg ccaacaaggccgaccgggaccagtacgagctgctgtgcctggacaacaccagaaagcccgtggacgagtacaaggactgccacctcgc ccaggtgccatctcacacagtggtggcccggtccatgggcggcaaagaggatctgatctgggagctgctgaaccaggcccaggaacact tcggcaaggacaagagcaaagagttccagctgttcagcagcccccacggcaaggatctgctgttcaaggacagcgcccacggctttctga aggtgccccccagaatggacgccaagatgtacctgggctacgagtacgtgaccgccatccggaacctgagagagggcacctgtcccga ggcccccaccgatgagtgcaagcccgtgaagtggtgcgccctgagccaccacgagcggctgaagtgcgacgagtggagcgtgaacag cgtgggcaagatcgagtgcgtgagcgccgagacaaccgaggactgtatcgccaagatcatgaacggcgaggccgatgccatgagcctg gacggcggcttcgtgtacattgccggcaagtgcggcctggtgcctgtgctggccgagaactacaacaagagcgacaactgcgaggatac ccccgaggccggctactttgccatcgcagtcgtgaagaagtccgccagcgacctgacctgggacaatctgaagggcaagaaaagctgcc acaccgccgtgggaaggaccgccgggtggaatattcctatggggctgctgtacaacaagatcaaccactgcagattcgacgagttcttcag cgagggctgcgctcccggcagcaagaaagacagcagcctgtgcaagctgtgcatgggcagcggcctgaacctgtgcgagcccaacaa caaagagggctactacggctacacaggggccttccggtgtctggtggagaagggggacgtggcttttgtgaaacaccagaccgtgcccc agaacaccggcggcaagaaccccgacccctgggccaagaacctgaacgagaaggactacgaactgctgtgtctcgacggcacccgga agccagtggaggaatacgccaactgtcacctggccagagcccccaatcacgccgtggtcacccggaaggacaaagaggcctgcgtcca caagatcctgcggcagcagcagcacctgttcggcagcaacgtgaccgactgcagcggcaacttctgcctgttcagaagcgagacaaagg acctcctgttccgggacgacaccgtgtgtctggccaagctgcacgaccggaacacctacgagaagtacctgggcgaggaatatgtgaag gccgtgggcaatctgcggaagtgcagcacctctagcctgctggaagcctgcacctttcgcagacctTAA

[0384] pDP74-His8-CD19 NT.1-GFLG-VR-FC-Tf Amino Acid Sequence (SEQ ID NO: 54)

[0385] MYRMQLLSCIALSLALVTNSGHHHHHHHHTGPEEPLVVKVEEGDTAALWCL KGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFY LCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGK LMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDS VSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKW YCHRGNVTTSFHLEVIARPVKAHSDLRTGGWKGGFLGGVRGVDGDKTHTCPPCPAPE LLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN QVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGKGSSGGSGGSGGSVPDKTVRWCAVSEHEATKCQSFRDHMKSV IPSDGPSVACVKKASYLDCIRAIAANEADAVTLDAGLVYDAYLAPNNLKPVVAEFYGSKEDPQT FYYAVAVVKKDSGFQMNQLRGKKSCHTGLGRSAGWNIPIGLLYCDLPEPRKPLEKAVANFFSGS CAPCADGTDFPQLCQLCPGCGCSTLNQYFGYSGAFKCLKDGAGDVAFVKHSTIFENLANKADRD QYELLCLDNTRKPVDEYKDCHLAQVPSHTVVARSMGGKEDLIWELLNQAQEHFGKDKSKEFQLF SSPHGKDLLFKDSAHGFLKVPPRMDAKMYLGYEYVTAIRNLREGTCPEAPTDECKPVKWCALSH HERLKCDEWSVNSVGKIECVSAETTEDCIAKIMNGEADAMSLDGGFVYIAGKCGLVPVLAENYN KSDNCEDTPEAGYFAIAVVKKSASDLTWDNLKGKKSCHTAVGRTAGWNIPMGLLYNKINHCRFD EFFSEGCAPGSKKDSSLCKLCMGSGLNLCEPNNKEGYYGYTGAFRCLVEKGDVAFVKHQTVPQN TGGKNPDPWAKNLNEKDYELLCLDGTRKPVEEYANCHLARAPNHAVVTRKDKEACVHKILRQQQDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 HLFGSNVTDCSGNFCLFRSETKDLLFRDDTVCLAKLHDRNTYEKYLGEEYVKAVGNLRKCSTSS LLEACTFRRP*

[0386] pDP75-His8-CD19 NT.1-GFLG-GGFG-FC-Tf Nucleic Acid Sequence (SEQ ID NO: 55)

[0387] ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTC ACGAATTCGGGCCACCACCATCATCACCACCATCACaccggtcccgaggaacctctagtggtgaaggtg gaagagggagataccgctgctctgtggtgcctcaaggggacctcagatggccccactcagcagctgacctggtctcgggagtccccgctt aaacccttcttaaaatacagcctgggggtgccaggcctgggagtccacgtcaggcccgatgccatctctgtcgtcatcaggaacgtctctca acagatggggggcttctacctgtgccagccggggcccccctctgagaaggcctggcagcctggctggacagtcaatgtggagggcagc ggggagctgttccggtggaatgtttcggacctaggtggcctgggctgtggcctgaagaacaggtcctcagagggccccagctccccttcc gggaagctcatgagccccaagctgtatgtgtgggccaaagaccgccctgagatctgggagggagagcctccgtgtctcccaccgaggga cagcctgaaccagagcctcagcagggacctcaccgtagcccctggctccacactctggctgtcctgtggggtaccccctgactctgtgtcc aggggccccctctcctggacccatgtgcaccccaaggggcctaagtcattgctgagcctagagctgaaggacgatcgcccggccagaga tatgtgggtaatgggtacgtcactgatgttgccccgggccacagctcaagacgctggaaagtggtattgtcaccgtggcaacgtaaccacct cattccacctggaggtaatcgctcggccagtaaaggctcactcagacctgaggactggtggctggaagggtgggTTCCTGggaggc ggtgggTTCgggGTAGACGGCGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGA ACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCAT GATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACC CTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACA AAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGT CCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAG CCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAA CCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAG CCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGA GCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGAC GGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGG GAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAA GAGCCTCTCCCTGTCTCCGGGTAAAggatcctctgggggaagtggaggtagcggtggttctgtgcccgataagaca gtgcggtggtgcgccgtgtctgagcacgaggccaccaagtgccagagcttccgggaccacatgaagtccgtgatccccagcgacggcc ctagcgtggcctgtgtgaagaaggccagctacctggactgcatccgggccattgccgccaatgaggccgacgccgtgacactggatgcc ggcctggtgtacgatgcctacctggcccccaacaacctgaagcccgtggtggccgagttctacggcagcaaagaggacccccagacctt ctactacgccgtggccgtggtcaagaaggacagcggcttccagatgaaccagctgcggggcaagaagtcctgtcacaccggcctgggca gaagcgccggctggaacatccccatcggcctgctgtactgcgatctgcccgagccccggaagcctctggaaaaggccgtggccaacttct tcagcggcagctgcgccccttgtgctgacggaaccgacttcccccagctgtgtcagctgtgccccggctgtggctgcagcaccctgaacc agtacttcggctacagcggcgccttcaagtgcctgaaggacggcgctggcgacgtggccttcgtgaagcacagcaccatcttcgagaacc tggccaacaaggccgaccgggaccagtacgagctgctgtgcctggacaacaccagaaagcccgtggacgagtacaaggactgccacct cgcccaggtgccatctcacacagtggtggcccggtccatgggcggcaaagaggatctgatctgggagctgctgaaccaggcccaggaa cacttcggcaaggacaagagcaaagagttccagctgttcagcagcccccacggcaaggatctgctgttcaaggacagcgcccacggcttt ctgaaggtgccccccagaatggacgccaagatgtacctgggctacgagtacgtgaccgccatccggaacctgagagagggcacctgtcc cgaggcccccaccgatgagtgcaagcccgtgaagtggtgcgccctgagccaccacgagcggctgaagtgcgacgagtggagcgtgaa cagcgtgggcaagatcgagtgcgtgagcgccgagacaaccgaggactgtatcgccaagatcatgaacggcgaggccgatgccatgagc ctggacggcggcttcgtgtacattgccggcaagtgcggcctggtgcctgtgctggccgagaactacaacaagagcgacaactgcgagga tacccccgaggccggctactttgccatcgcagtcgtgaagaagtccgccagcgacctgacctgggacaatctgaagggcaagaaaagct gccacaccgccgtgggaaggaccgccgggtggaatattcctatggggctgctgtacaacaagatcaaccactgcagattcgacgagttctt cagcgagggctgcgctcccggcagcaagaaagacagcagcctgtgcaagctgtgcatgggcagcggcctgaacctgtgcgagcccaaDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 caacaaagagggctactacggctacacaggggccttccggtgtctggtggagaagggggacgtggcttttgtgaaacaccagaccgtgc cccagaacaccggcggcaagaaccccgacccctgggccaagaacctgaacgagaaggactacgaactgctgtgtctcgacggcaccc ggaagccagtggaggaatacgccaactgtcacctggccagagcccccaatcacgccgtggtcacccggaaggacaaagaggcctgcgt ccacaagatcctgcggcagcagcagcacctgttcggcagcaacgtgaccgactgcagcggcaacttctgcctgttcagaagcgagacaa aggacctcctgttccgggacgacaccgtgtgtctggccaagctgcacgaccggaacacctacgagaagtacctgggcgaggaatatgtg aaggccgtgggcaatctgcggaagtgcagcacctctagcctgctggaagcctgcacctttcgcagacctTAA

[0388] pDP75-His8-CD19 NT.1-GFLG-GGFG-FC-Tf Amino Acid Sequence (SEQ ID NO: 56)

[0389] MYRMQLLSCIALSLALVTNSGHHHHHHHHTGPEEPLVVKVEEGDTAALWCL KGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFY LCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGK LMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDS VSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKW YCHRGNVTTSFHLEVIARPVKAHSDLRTGGWKGGFLGGGGFGVDGDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS CSVMHEALHNHYTQKSLSLSPGKGSSGGSGGSGGSVPDKTVRWCAVSEHEATKCQSFRDHMKS VIPSDGPSVACVKKASYLDCIRAIAANEADAVTLDAGLVYDAYLAPNNLKPVVAEFYGSKEDPQ TFYYAVAVVKKDSGFQMNQLRGKKSCHTGLGRSAGWNIPIGLLYCDLPEPRKPLEKAVANFFSG SCAPCADGTDFPQLCQLCPGCGCSTLNQYFGYSGAFKCLKDGAGDVAFVKHSTIFENLANKADR DQYELLCLDNTRKPVDEYKDCHLAQVPSHTVVARSMGGKEDLIWELLNQAQEHFGKDKSKEFQL FSSPHGKDLLFKDSAHGFLKVPPRMDAKMYLGYEYVTAIRNLREGTCPEAPTDECKPVKWCALS HHERLKCDEWSVNSVGKIECVSAETTEDCIAKIMNGEADAMSLDGGFVYIAGKCGLVPVLAENY NKSDNCEDTPEAGYFAIAVVKKSASDLTWDNLKGKKSCHTAVGRTAGWNIPMGLLYNKINHCRF DEFFSEGCAPGSKKDSSLCKLCMGSGLNLCEPNNKEGYYGYTGAFRCLVEKGDVAFVKHQTVPQ NTGGKNPDPWAKNLNEKDYELLCLDGTRKPVEEYANCHLARAPNHAVVTRKDKEACVHKILRQQ QHLFGSNVTDCSGNFCLFRSETKDLLFRDDTVCLAKLHDRNTYEKYLGEEYVKAVGNLRKCSTS SLLEACTFRRP*

[0390] pDP76-His8-CD19 NT.1-FK-FC-Tf Nucleic Acid Sequence (SEQ ID NO: 57)

[0391] ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTC ACGAATTCGGGCCACCACCATCATCACCACCATCACaccggtcccgaggaacctctagtggtgaaggtg gaagagggagataccgctgctctgtggtgcctcaaggggacctcagatggccccactcagcagctgacctggtctcgggagtccccgctt aaacccttcttaaaatacagcctgggggtgccaggcctgggagtccacgtcaggcccgatgccatctctgtcgtcatcaggaacgtctctca acagatggggggcttctacctgtgccagccggggcccccctctgagaaggcctggcagcctggctggacagtcaatgtggagggcagc ggggagctgttccggtggaatgtttcggacctaggtggcctgggctgtggcctgaagaacaggtcctcagagggccccagctccccttcc gggaagctcatgagccccaagctgtatgtgtgggccaaagaccgccctgagatctgggagggagagcctccgtgtctcccaccgaggga cagcctgaaccagagcctcagcagggacctcaccgtagcccctggctccacactctggctgtcctgtggggtaccccctgactctgtgtcc aggggccccctctcctggacccatgtgcaccccaaggggcctaagtcattgctgagcctagagctgaaggacgatcgcccggccagaga tatgtgggtaatgggtacgtcactgatgttgccccgggccacagctcaagacgctggaaagtggtattgtcaccgtggcaacgtaaccacct cattccacctggaggtaatcgctcggccagtaaaggctcactcagacctgaggactggtggctggaagggcTTCAAAggcGTAG ACGGCGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGA CCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACC CCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 CAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGG AGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACT GGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCC ATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACAC CCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGT CAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGG AGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCT ACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGC TCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCT CCGGGTAAAggatcctctgggggaagtggaggtagcggtggttctgtgcccgataagacagtgcggtggtgcgccgtgtctgag cacgaggccaccaagtgccagagcttccgggaccacatgaagtccgtgatccccagcgacggccctagcgtggcctgtgtgaagaagg ccagctacctggactgcatccgggccattgccgccaatgaggccgacgccgtgacactggatgccggcctggtgtacgatgcctacctgg cccccaacaacctgaagcccgtggtggccgagttctacggcagcaaagaggacccccagaccttctactacgccgtggccgtggtcaag aaggacagcggcttccagatgaaccagctgcggggcaagaagtcctgtcacaccggcctgggcagaagcgccggctggaacatcccca tcggcctgctgtactgcgatctgcccgagccccggaagcctctggaaaaggccgtggccaacttcttcagcggcagctgcgccccttgtgc tgacggaaccgacttcccccagctgtgtcagctgtgccccggctgtggctgcagcaccctgaaccagtacttcggctacagcggcgccttc aagtgcctgaaggacggcgctggcgacgtggccttcgtgaagcacagcaccatcttcgagaacctggccaacaaggccgaccgggacc agtacgagctgctgtgcctggacaacaccagaaagcccgtggacgagtacaaggactgccacctcgcccaggtgccatctcacacagtg gtggcccggtccatgggcggcaaagaggatctgatctgggagctgctgaaccaggcccaggaacacttcggcaaggacaagagcaaag agttccagctgttcagcagcccccacggcaaggatctgctgttcaaggacagcgcccacggctttctgaaggtgccccccagaatggacg ccaagatgtacctgggctacgagtacgtgaccgccatccggaacctgagagagggcacctgtcccgaggcccccaccgatgagtgcaa gcccgtgaagtggtgcgccctgagccaccacgagcggctgaagtgcgacgagtggagcgtgaacagcgtgggcaagatcgagtgcgt gagcgccgagacaaccgaggactgtatcgccaagatcatgaacggcgaggccgatgccatgagcctggacggcggcttcgtgtacattg ccggcaagtgcggcctggtgcctgtgctggccgagaactacaacaagagcgacaactgcgaggatacccccgaggccggctactttgcc atcgcagtcgtgaagaagtccgccagcgacctgacctgggacaatctgaagggcaagaaaagctgccacaccgccgtgggaaggaccg ccgggtggaatattcctatggggctgctgtacaacaagatcaaccactgcagattcgacgagttcttcagcgagggctgcgctcccggcag caagaaagacagcagcctgtgcaagctgtgcatgggcagcggcctgaacctgtgcgagcccaacaacaaagagggctactacggctac acaggggccttccggtgtctggtggagaagggggacgtggcttttgtgaaacaccagaccgtgccccagaacaccggcggcaagaacc ccgacccctgggccaagaacctgaacgagaaggactacgaactgctgtgtctcgacggcacccggaagccagtggaggaatacgccaa ctgtcacctggccagagcccccaatcacgccgtggtcacccggaaggacaaagaggcctgcgtccacaagatcctgcggcagcagcag cacctgttcggcagcaacgtgaccgactgcagcggcaacttctgcctgttcagaagcgagacaaaggacctcctgttccgggacgacacc gtgtgtctggccaagctgcacgaccggaacacctacgagaagtacctgggcgaggaatatgtgaaggccgtgggcaatctgcggaagtg cagcacctctagcctgctggaagcctgcacctttcgcagacctTAA

[0392] pDP76-His8-CD19 NT.1-FK-FC-Tf Amino Acid Sequence (SEQ ID NO: 58)

[0393] MYRMQLLSCIALSLALVTNSGHHHHHHHHTGPEEPLVVKVEEGDTAALWCL KGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFY LCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGK LMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDS VSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKW YCHRGNVTTSFHLEVIARPVKAHSDLRTGGWKGFKGVDGDKTHTCPPCPAPELLGGPS VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTC LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEA LHNHYTQKSLSLSPGKGSSGGSGGSGGSVPDKTVRWCAVSEHEATKCQSFRDHMKSVIPSDGP SVACVKKASYLDCIRAIAANEADAVTLDAGLVYDAYLAPNNLKPVVAEFYGSKEDPQTFYYAVADOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 VVKKDSGFQMNQLRGKKSCHTGLGRSAGWNIPIGLLYCDLPEPRKPLEKAVANFFSGSCAPCAD GTDFPQLCQLCPGCGCSTLNQYFGYSGAFKCLKDGAGDVAFVKHSTIFENLANKADRDQYELLC LDNTRKPVDEYKDCHLAQVPSHTVVARSMGGKEDLIWELLNQAQEHFGKDKSKEFQLFSSPHGK DLLFKDSAHGFLKVPPRMDAKMYLGYEYVTAIRNLREGTCPEAPTDECKPVKWCALSHHERLKC DEWSVNSVGKIECVSAETTEDCIAKIMNGEADAMSLDGGFVYIAGKCGLVPVLAENYNKSDNCE DTPEAGYFAIAVVKKSASDLTWDNLKGKKSCHTAVGRTAGWNIPMGLLYNKINHCRFDEFFSEG CAPGSKKDSSLCKLCMGSGLNLCEPNNKEGYYGYTGAFRCLVEKGDVAFVKHQTVPQNTGGKNP DPWAKNLNEKDYELLCLDGTRKPVEEYANCHLARAPNHAVVTRKDKEACVHKILRQQQHLFGSN VTDCSGNFCLFRSETKDLLFRDDTVCLAKLHDRNTYEKYLGEEYVKAVGNLRKCSTSSLLEACT FRRP*

[0394] pDP77-His8-CD19 NT.1-VA-FC-Tf Nucleic Acid Sequence (SEQ ID NO: 59)

[0395] ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTC ACGAATTCGGGCCACCACCATCATCACCACCATCACaccggtcccgaggaacctctagtggtgaaggtg gaagagggagataccgctgctctgtggtgcctcaaggggacctcagatggccccactcagcagctgacctggtctcgggagtccccgctt aaacccttcttaaaatacagcctgggggtgccaggcctgggagtccacgtcaggcccgatgccatctctgtcgtcatcaggaacgtctctca acagatggggggcttctacctgtgccagccggggcccccctctgagaaggcctggcagcctggctggacagtcaatgtggagggcagc ggggagctgttccggtggaatgtttcggacctaggtggcctgggctgtggcctgaagaacaggtcctcagagggccccagctccccttcc gggaagctcatgagccccaagctgtatgtgtgggccaaagaccgccctgagatctgggagggagagcctccgtgtctcccaccgaggga cagcctgaaccagagcctcagcagggacctcaccgtagcccctggctccacactctggctgtcctgtggggtaccccctgactctgtgtcc aggggccccctctcctggacccatgtgcaccccaaggggcctaagtcattgctgagcctagagctgaaggacgatcgcccggccagaga tatgtgggtaatgggtacgtcactgatgttgccccgggccacagctcaagacgctggaaagtggtattgtcaccgtggcaacgtaaccacct cattccacctggaggtaatcgctcggccagtaaaggctcactcagacctgaggactggtggctggaagggcGTAgctggcGTAGA CGGCGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGAC CGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCC CTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTC AACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGG AGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACT GGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCC ATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACAC CCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGT CAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGG AGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCT ACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGC TCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCT CCGGGTAAAggatcctctgggggaagtggaggtagcggtggttctgtgcccgataagacagtgcggtggtgcgccgtgtctgag cacgaggccaccaagtgccagagcttccgggaccacatgaagtccgtgatccccagcgacggccctagcgtggcctgtgtgaagaagg ccagctacctggactgcatccgggccattgccgccaatgaggccgacgccgtgacactggatgccggcctggtgtacgatgcctacctgg cccccaacaacctgaagcccgtggtggccgagttctacggcagcaaagaggacccccagaccttctactacgccgtggccgtggtcaag aaggacagcggcttccagatgaaccagctgcggggcaagaagtcctgtcacaccggcctgggcagaagcgccggctggaacatcccca tcggcctgctgtactgcgatctgcccgagccccggaagcctctggaaaaggccgtggccaacttcttcagcggcagctgcgccccttgtgc tgacggaaccgacttcccccagctgtgtcagctgtgccccggctgtggctgcagcaccctgaaccagtacttcggctacagcggcgccttc aagtgcctgaaggacggcgctggcgacgtggccttcgtgaagcacagcaccatcttcgagaacctggccaacaaggccgaccgggacc agtacgagctgctgtgcctggacaacaccagaaagcccgtggacgagtacaaggactgccacctcgcccaggtgccatctcacacagtg gtggcccggtccatgggcggcaaagaggatctgatctgggagctgctgaaccaggcccaggaacacttcggcaaggacaagagcaaag agttccagctgttcagcagcccccacggcaaggatctgctgttcaaggacagcgcccacggctttctgaaggtgccccccagaatggacg ccaagatgtacctgggctacgagtacgtgaccgccatccggaacctgagagagggcacctgtcccgaggcccccaccgatgagtgcaaDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 gcccgtgaagtggtgcgccctgagccaccacgagcggctgaagtgcgacgagtggagcgtgaacagcgtgggcaagatcgagtgcgt gagcgccgagacaaccgaggactgtatcgccaagatcatgaacggcgaggccgatgccatgagcctggacggcggcttcgtgtacattg ccggcaagtgcggcctggtgcctgtgctggccgagaactacaacaagagcgacaactgcgaggatacccccgaggccggctactttgcc atcgcagtcgtgaagaagtccgccagcgacctgacctgggacaatctgaagggcaagaaaagctgccacaccgccgtgggaaggaccg ccgggtggaatattcctatggggctgctgtacaacaagatcaaccactgcagattcgacgagttcttcagcgagggctgcgctcccggcag caagaaagacagcagcctgtgcaagctgtgcatgggcagcggcctgaacctgtgcgagcccaacaacaaagagggctactacggctac acaggggccttccggtgtctggtggagaagggggacgtggcttttgtgaaacaccagaccgtgccccagaacaccggcggcaagaacc ccgacccctgggccaagaacctgaacgagaaggactacgaactgctgtgtctcgacggcacccggaagccagtggaggaatacgccaa ctgtcacctggccagagcccccaatcacgccgtggtcacccggaaggacaaagaggcctgcgtccacaagatcctgcggcagcagcag cacctgttcggcagcaacgtgaccgactgcagcggcaacttctgcctgttcagaagcgagacaaaggacctcctgttccgggacgacacc gtgtgtctggccaagctgcacgaccggaacacctacgagaagtacctgggcgaggaatatgtgaaggccgtgggcaatctgcggaagtg cagcacctctagcctgctggaagcctgcacctttcgcagacctTAA

[0396] pDP77-His8-CD19 NT.1-VA-FC-Tf Amino Acid Sequence (SEQ ID NO: 60)

[0397] MYRMQLLSCIALSLALVTNSGHHHHHHHHTGPEEPLVVKVEEGDTAALWCL KGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFY LCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGK LMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDS VSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKW YCHRGNVTTSFHLEVIARPVKAHSDLRTGGWKGVAGVDGDKTHTCPPCPAPELLGGPS VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTC LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEA LHNHYTQKSLSLSPGKGSSGGSGGSGGSVPDKTVRWCAVSEHEATKCQSFRDHMKSVIPSDGP SVACVKKASYLDCIRAIAANEADAVTLDAGLVYDAYLAPNNLKPVVAEFYGSKEDPQTFYYAVA VVKKDSGFQMNQLRGKKSCHTGLGRSAGWNIPIGLLYCDLPEPRKPLEKAVANFFSGSCAPCAD GTDFPQLCQLCPGCGCSTLNQYFGYSGAFKCLKDGAGDVAFVKHSTIFENLANKADRDQYELLC LDNTRKPVDEYKDCHLAQVPSHTVVARSMGGKEDLIWELLNQAQEHFGKDKSKEFQLFSSPHGK DLLFKDSAHGFLKVPPRMDAKMYLGYEYVTAIRNLREGTCPEAPTDECKPVKWCALSHHERLKC DEWSVNSVGKIECVSAETTEDCIAKIMNGEADAMSLDGGFVYIAGKCGLVPVLAENYNKSDNCE DTPEAGYFAIAVVKKSASDLTWDNLKGKKSCHTAVGRTAGWNIPMGLLYNKINHCRFDEFFSEG CAPGSKKDSSLCKLCMGSGLNLCEPNNKEGYYGYTGAFRCLVEKGDVAFVKHQTVPQNTGGKNP DPWAKNLNEKDYELLCLDGTRKPVEEYANCHLARAPNHAVVTRKDKEACVHKILRQQQHLFGSN VTDCSGNFCLFRSETKDLLFRDDTVCLAKLHDRNTYEKYLGEEYVKAVGNLRKCSTSSLLEACT FRRP*

[0398] pDP78-His8-CD19 NT.1-VK-FC-Tf Nucleic Acid Sequence (SEQ ID NO: 61)

[0399] ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTC ACGAATTCGGGCCACCACCATCATCACCACCATCACaccggtcccgaggaacctctagtggtgaaggtg gaagagggagataccgctgctctgtggtgcctcaaggggacctcagatggccccactcagcagctgacctggtctcgggagtccccgctt aaacccttcttaaaatacagcctgggggtgccaggcctgggagtccacgtcaggcccgatgccatctctgtcgtcatcaggaacgtctctca acagatggggggcttctacctgtgccagccggggcccccctctgagaaggcctggcagcctggctggacagtcaatgtggagggcagc ggggagctgttccggtggaatgtttcggacctaggtggcctgggctgtggcctgaagaacaggtcctcagagggccccagctccccttcc gggaagctcatgagccccaagctgtatgtgtgggccaaagaccgccctgagatctgggagggagagcctccgtgtctcccaccgaggga cagcctgaaccagagcctcagcagggacctcaccgtagcccctggctccacactctggctgtcctgtggggtaccccctgactctgtgtcc aggggccccctctcctggacccatgtgcaccccaaggggcctaagtcattgctgagcctagagctgaaggacgatcgcccggccagagaDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 tatgtgggtaatgggtacgtcactgatgttgccccgggccacagctcaagacgctggaaagtggtattgtcaccgtggcaacgtaaccacct cattccacctggaggtaatcgctcggccagtaaaggctcactcagacctgaggactggtggctggaagggcGTAAAAggcGTA GACGGCGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGG ACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGAC CCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGT TCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAG GAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGA CTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCC CCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTAC ACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCT GGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGC CGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCC TCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCA TGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTG TCTCCGGGTAAAggatcctctgggggaagtggaggtagcggtggttctgtgcccgataagacagtgcggtggtgcgccgtgtc tgagcacgaggccaccaagtgccagagcttccgggaccacatgaagtccgtgatccccagcgacggccctagcgtggcctgtgtgaaga aggccagctacctggactgcatccgggccattgccgccaatgaggccgacgccgtgacactggatgccggcctggtgtacgatgcctac ctggcccccaacaacctgaagcccgtggtggccgagttctacggcagcaaagaggacccccagaccttctactacgccgtggccgtggt caagaaggacagcggcttccagatgaaccagctgcggggcaagaagtcctgtcacaccggcctgggcagaagcgccggctggaacat ccccatcggcctgctgtactgcgatctgcccgagccccggaagcctctggaaaaggccgtggccaacttcttcagcggcagctgcgcccc ttgtgctgacggaaccgacttcccccagctgtgtcagctgtgccccggctgtggctgcagcaccctgaaccagtacttcggctacagcggc gccttcaagtgcctgaaggacggcgctggcgacgtggccttcgtgaagcacagcaccatcttcgagaacctggccaacaaggccgaccg ggaccagtacgagctgctgtgcctggacaacaccagaaagcccgtggacgagtacaaggactgccacctcgcccaggtgccatctcaca cagtggtggcccggtccatgggcggcaaagaggatctgatctgggagctgctgaaccaggcccaggaacacttcggcaaggacaagag caaagagttccagctgttcagcagcccccacggcaaggatctgctgttcaaggacagcgcccacggctttctgaaggtgccccccagaat ggacgccaagatgtacctgggctacgagtacgtgaccgccatccggaacctgagagagggcacctgtcccgaggcccccaccgatgag tgcaagcccgtgaagtggtgcgccctgagccaccacgagcggctgaagtgcgacgagtggagcgtgaacagcgtgggcaagatcgagt gcgtgagcgccgagacaaccgaggactgtatcgccaagatcatgaacggcgaggccgatgccatgagcctggacggcggcttcgtgta cattgccggcaagtgcggcctggtgcctgtgctggccgagaactacaacaagagcgacaactgcgaggatacccccgaggccggctact ttgccatcgcagtcgtgaagaagtccgccagcgacctgacctgggacaatctgaagggcaagaaaagctgccacaccgccgtgggaag gaccgccgggtggaatattcctatggggctgctgtacaacaagatcaaccactgcagattcgacgagttcttcagcgagggctgcgctccc ggcagcaagaaagacagcagcctgtgcaagctgtgcatgggcagcggcctgaacctgtgcgagcccaacaacaaagagggctactacg gctacacaggggccttccggtgtctggtggagaagggggacgtggcttttgtgaaacaccagaccgtgccccagaacaccggcggcaag aaccccgacccctgggccaagaacctgaacgagaaggactacgaactgctgtgtctcgacggcacccggaagccagtggaggaatacg ccaactgtcacctggccagagcccccaatcacgccgtggtcacccggaaggacaaagaggcctgcgtccacaagatcctgcggcagca gcagcacctgttcggcagcaacgtgaccgactgcagcggcaacttctgcctgttcagaagcgagacaaaggacctcctgttccgggacga caccgtgtgtctggccaagctgcacgaccggaacacctacgagaagtacctgggcgaggaatatgtgaaggccgtgggcaatctgcgga agtgcagcacctctagcctgctggaagcctgcacctttcgcagacctTAA

[0400] pDP78-His8-CD19 NT.1-VK-FC-Tf Amino Acid Sequence (SEQ ID NO: 62)

[0401] MYRMQLLSCIALSLALVTNSGHHHHHHHHTGPEEPLVVKVEEGDTAALWCL KGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFY LCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGK LMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDS VSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKW YCHRGNVTTSFHLEVIARPVKAHSDLRTGGWKGVKGVDGDKTHTCPPCPAPELLGGPSDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTC LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEA LHNHYTQKSLSLSPGKGSSGGSGGSGGSVPDKTVRWCAVSEHEATKCQSFRDHMKSVIPSDGP SVACVKKASYLDCIRAIAANEADAVTLDAGLVYDAYLAPNNLKPVVAEFYGSKEDPQTFYYAVA VVKKDSGFQMNQLRGKKSCHTGLGRSAGWNIPIGLLYCDLPEPRKPLEKAVANFFSGSCAPCAD GTDFPQLCQLCPGCGCSTLNQYFGYSGAFKCLKDGAGDVAFVKHSTIFENLANKADRDQYELLC LDNTRKPVDEYKDCHLAQVPSHTVVARSMGGKEDLIWELLNQAQEHFGKDKSKEFQLFSSPHGK DLLFKDSAHGFLKVPPRMDAKMYLGYEYVTAIRNLREGTCPEAPTDECKPVKWCALSHHERLKC DEWSVNSVGKIECVSAETTEDCIAKIMNGEADAMSLDGGFVYIAGKCGLVPVLAENYNKSDNCE DTPEAGYFAIAVVKKSASDLTWDNLKGKKSCHTAVGRTAGWNIPMGLLYNKINHCRFDEFFSEG CAPGSKKDSSLCKLCMGSGLNLCEPNNKEGYYGYTGAFRCLVEKGDVAFVKHQTVPQNTGGKNP DPWAKNLNEKDYELLCLDGTRKPVEEYANCHLARAPNHAVVTRKDKEACVHKILRQQQHLFGSN VTDCSGNFCLFRSETKDLLFRDDTVCLAKLHDRNTYEKYLGEEYVKAVGNLRKCSTSSLLEACT FRRP*

[0402] pDP79-His8-CD19 NT.1-VR-FC-Tf Nucleic Acid Sequence (SEQ ID NO: 63)

[0403] ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTC ACGAATTCGGGCCACCACCATCATCACCACCATCACaccggtcccgaggaacctctagtggtgaaggtg gaagagggagataccgctgctctgtggtgcctcaaggggacctcagatggccccactcagcagctgacctggtctcgggagtccccgctt aaacccttcttaaaatacagcctgggggtgccaggcctgggagtccacgtcaggcccgatgccatctctgtcgtcatcaggaacgtctctca acagatggggggcttctacctgtgccagccggggcccccctctgagaaggcctggcagcctggctggacagtcaatgtggagggcagc ggggagctgttccggtggaatgtttcggacctaggtggcctgggctgtggcctgaagaacaggtcctcagagggccccagctccccttcc gggaagctcatgagccccaagctgtatgtgtgggccaaagaccgccctgagatctgggagggagagcctccgtgtctcccaccgaggga cagcctgaaccagagcctcagcagggacctcaccgtagcccctggctccacactctggctgtcctgtggggtaccccctgactctgtgtcc aggggccccctctcctggacccatgtgcaccccaaggggcctaagtcattgctgagcctagagctgaaggacgatcgcccggccagaga tatgtgggtaatgggtacgtcactgatgttgccccgggccacagctcaagacgctggaaagtggtattgtcaccgtggcaacgtaaccacct cattccacctggaggtaatcgctcggccagtaaaggctcactcagacctgaggactggtggctggaagggcGTACGGggcGTA GACGGCGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGG ACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGAC CCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGT TCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAG GAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGA CTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCC CCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTAC ACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCT GGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGC CGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCC TCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCA TGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTG TCTCCGGGTAAAggatcctctgggggaagtggaggtagcggtggttctgtgcccgataagacagtgcggtggtgcgccgtgtc tgagcacgaggccaccaagtgccagagcttccgggaccacatgaagtccgtgatccccagcgacggccctagcgtggcctgtgtgaaga aggccagctacctggactgcatccgggccattgccgccaatgaggccgacgccgtgacactggatgccggcctggtgtacgatgcctac ctggcccccaacaacctgaagcccgtggtggccgagttctacggcagcaaagaggacccccagaccttctactacgccgtggccgtggt caagaaggacagcggcttccagatgaaccagctgcggggcaagaagtcctgtcacaccggcctgggcagaagcgccggctggaacat ccccatcggcctgctgtactgcgatctgcccgagccccggaagcctctggaaaaggccgtggccaacttcttcagcggcagctgcgcccc ttgtgctgacggaaccgacttcccccagctgtgtcagctgtgccccggctgtggctgcagcaccctgaaccagtacttcggctacagcggcDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 gccttcaagtgcctgaaggacggcgctggcgacgtggccttcgtgaagcacagcaccatcttcgagaacctggccaacaaggccgaccg ggaccagtacgagctgctgtgcctggacaacaccagaaagcccgtggacgagtacaaggactgccacctcgcccaggtgccatctcaca cagtggtggcccggtccatgggcggcaaagaggatctgatctgggagctgctgaaccaggcccaggaacacttcggcaaggacaagag caaagagttccagctgttcagcagcccccacggcaaggatctgctgttcaaggacagcgcccacggctttctgaaggtgccccccagaat ggacgccaagatgtacctgggctacgagtacgtgaccgccatccggaacctgagagagggcacctgtcccgaggcccccaccgatgag tgcaagcccgtgaagtggtgcgccctgagccaccacgagcggctgaagtgcgacgagtggagcgtgaacagcgtgggcaagatcgagt gcgtgagcgccgagacaaccgaggactgtatcgccaagatcatgaacggcgaggccgatgccatgagcctggacggcggcttcgtgta cattgccggcaagtgcggcctggtgcctgtgctggccgagaactacaacaagagcgacaactgcgaggatacccccgaggccggctact ttgccatcgcagtcgtgaagaagtccgccagcgacctgacctgggacaatctgaagggcaagaaaagctgccacaccgccgtgggaag gaccgccgggtggaatattcctatggggctgctgtacaacaagatcaaccactgcagattcgacgagttcttcagcgagggctgcgctccc ggcagcaagaaagacagcagcctgtgcaagctgtgcatgggcagcggcctgaacctgtgcgagcccaacaacaaagagggctactacg gctacacaggggccttccggtgtctggtggagaagggggacgtggcttttgtgaaacaccagaccgtgccccagaacaccggcggcaag aaccccgacccctgggccaagaacctgaacgagaaggactacgaactgctgtgtctcgacggcacccggaagccagtggaggaatacg ccaactgtcacctggccagagcccccaatcacgccgtggtcacccggaaggacaaagaggcctgcgtccacaagatcctgcggcagca gcagcacctgttcggcagcaacgtgaccgactgcagcggcaacttctgcctgttcagaagcgagacaaaggacctcctgttccgggacga caccgtgtgtctggccaagctgcacgaccggaacacctacgagaagtacctgggcgaggaatatgtgaaggccgtgggcaatctgcgga agtgcagcacctctagcctgctggaagcctgcacctttcgcagacctTAA

[0404] pDP79-His8-CD19 NT.1-VR-FC-Tf Amino Acid Sequence (SEQ ID NO: 64)

[0405] MYRMQLLSCIALSLALVTNSGHHHHHHHHTGPEEPLVVKVEEGDTAALWCL KGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFY LCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGK LMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDS VSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKW YCHRGNVTTSFHLEVIARPVKAHSDLRTGGWKGVRGVDGDKTHTCPPCPAPELLGGPS VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTC LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEA LHNHYTQKSLSLSPGKGSSGGSGGSGGSVPDKTVRWCAVSEHEATKCQSFRDHMKSVIPSDGP SVACVKKASYLDCIRAIAANEADAVTLDAGLVYDAYLAPNNLKPVVAEFYGSKEDPQTFYYAVA VVKKDSGFQMNQLRGKKSCHTGLGRSAGWNIPIGLLYCDLPEPRKPLEKAVANFFSGSCAPCAD GTDFPQLCQLCPGCGCSTLNQYFGYSGAFKCLKDGAGDVAFVKHSTIFENLANKADRDQYELLC LDNTRKPVDEYKDCHLAQVPSHTVVARSMGGKEDLIWELLNQAQEHFGKDKSKEFQLFSSPHGK DLLFKDSAHGFLKVPPRMDAKMYLGYEYVTAIRNLREGTCPEAPTDECKPVKWCALSHHERLKC DEWSVNSVGKIECVSAETTEDCIAKIMNGEADAMSLDGGFVYIAGKCGLVPVLAENYNKSDNCE DTPEAGYFAIAVVKKSASDLTWDNLKGKKSCHTAVGRTAGWNIPMGLLYNKINHCRFDEFFSEG CAPGSKKDSSLCKLCMGSGLNLCEPNNKEGYYGYTGAFRCLVEKGDVAFVKHQTVPQNTGGKNP DPWAKNLNEKDYELLCLDGTRKPVEEYANCHLARAPNHAVVTRKDKEACVHKILRQQQHLFGSN VTDCSGNFCLFRSETKDLLFRDDTVCLAKLHDRNTYEKYLGEEYVKAVGNLRKCSTSSLLEACT FRRP*

[0406] pDP80-His8-CD19 NT.1-GGFG-FC-Tf Nucleic Acid Sequence (SEQ ID NO: 65)

[0407] ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTC ACGAATTCGGGCCACCACCATCATCACCACCATCACaccggtcccgaggaacctctagtggtgaaggtg gaagagggagataccgctgctctgtggtgcctcaaggggacctcagatggccccactcagcagctgacctggtctcgggagtccccgctt aaacccttcttaaaatacagcctgggggtgccaggcctgggagtccacgtcaggcccgatgccatctctgtcgtcatcaggaacgtctctcaDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 acagatggggggcttctacctgtgccagccggggcccccctctgagaaggcctggcagcctggctggacagtcaatgtggagggcagc ggggagctgttccggtggaatgtttcggacctaggtggcctgggctgtggcctgaagaacaggtcctcagagggccccagctccccttcc gggaagctcatgagccccaagctgtatgtgtgggccaaagaccgccctgagatctgggagggagagcctccgtgtctcccaccgaggga cagcctgaaccagagcctcagcagggacctcaccgtagcccctggctccacactctggctgtcctgtggggtaccccctgactctgtgtcc aggggccccctctcctggacccatgtgcaccccaaggggcctaagtcattgctgagcctagagctgaaggacgatcgcccggccagaga tatgtgggtaatgggtacgtcactgatgttgccccgggccacagctcaagacgctggaaagtggtattgtcaccgtggcaacgtaaccacct cattccacctggaggtaatcgctcggccagtaaaggctcactcagacctgaggactggtggctggaagggtgggTTCggaggcGTA GACGGCGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGG ACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGAC CCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGT TCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAG GAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGA CTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCC CCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTAC ACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCT GGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGC CGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCC TCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCA TGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTG TCTCCGGGTAAAggatcctctgggggaagtggaggtagcggtggttctgtgcccgataagacagtgcggtggtgcgccgtgtc tgagcacgaggccaccaagtgccagagcttccgggaccacatgaagtccgtgatccccagcgacggccctagcgtggcctgtgtgaaga aggccagctacctggactgcatccgggccattgccgccaatgaggccgacgccgtgacactggatgccggcctggtgtacgatgcctac ctggcccccaacaacctgaagcccgtggtggccgagttctacggcagcaaagaggacccccagaccttctactacgccgtggccgtggt caagaaggacagcggcttccagatgaaccagctgcggggcaagaagtcctgtcacaccggcctgggcagaagcgccggctggaacat ccccatcggcctgctgtactgcgatctgcccgagccccggaagcctctggaaaaggccgtggccaacttcttcagcggcagctgcgcccc ttgtgctgacggaaccgacttcccccagctgtgtcagctgtgccccggctgtggctgcagcaccctgaaccagtacttcggctacagcggc gccttcaagtgcctgaaggacggcgctggcgacgtggccttcgtgaagcacagcaccatcttcgagaacctggccaacaaggccgaccg ggaccagtacgagctgctgtgcctggacaacaccagaaagcccgtggacgagtacaaggactgccacctcgcccaggtgccatctcaca cagtggtggcccggtccatgggcggcaaagaggatctgatctgggagctgctgaaccaggcccaggaacacttcggcaaggacaagag caaagagttccagctgttcagcagcccccacggcaaggatctgctgttcaaggacagcgcccacggctttctgaaggtgccccccagaat ggacgccaagatgtacctgggctacgagtacgtgaccgccatccggaacctgagagagggcacctgtcccgaggcccccaccgatgag tgcaagcccgtgaagtggtgcgccctgagccaccacgagcggctgaagtgcgacgagtggagcgtgaacagcgtgggcaagatcgagt gcgtgagcgccgagacaaccgaggactgtatcgccaagatcatgaacggcgaggccgatgccatgagcctggacggcggcttcgtgta cattgccggcaagtgcggcctggtgcctgtgctggccgagaactacaacaagagcgacaactgcgaggatacccccgaggccggctact ttgccatcgcagtcgtgaagaagtccgccagcgacctgacctgggacaatctgaagggcaagaaaagctgccacaccgccgtgggaag gaccgccgggtggaatattcctatggggctgctgtacaacaagatcaaccactgcagattcgacgagttcttcagcgagggctgcgctccc ggcagcaagaaagacagcagcctgtgcaagctgtgcatgggcagcggcctgaacctgtgcgagcccaacaacaaagagggctactacg gctacacaggggccttccggtgtctggtggagaagggggacgtggcttttgtgaaacaccagaccgtgccccagaacaccggcggcaag aaccccgacccctgggccaagaacctgaacgagaaggactacgaactgctgtgtctcgacggcacccggaagccagtggaggaatacg ccaactgtcacctggccagagcccccaatcacgccgtggtcacccggaaggacaaagaggcctgcgtccacaagatcctgcggcagca gcagcacctgttcggcagcaacgtgaccgactgcagcggcaacttctgcctgttcagaagcgagacaaaggacctcctgttccgggacga caccgtgtgtctggccaagctgcacgaccggaacacctacgagaagtacctgggcgaggaatatgtgaaggccgtgggcaatctgcgga agtgcagcacctctagcctgctggaagcctgcacctttcgcagacctTAA

[0408] pDP80-His8-CD19 NT.1-GGFG-FC-Tf Amino Acid Sequence (SEQ ID NO: 66)DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17

[0409] MYRMQLLSCIALSLALVTNSGHHHHHHHHTGPEEPLVVKVEEGDTAALWCL KGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFY LCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGK LMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDS VSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKW YCHRGNVTTSFHLEVIARPVKAHSDLRTGGWKGGFGGVDGDKTHTCPPCPAPELLGG PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSL TCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSPGKGSSGGSGGSGGSVPDKTVRWCAVSEHEATKCQSFRDHMKSVIPSD GPSVACVKKASYLDCIRAIAANEADAVTLDAGLVYDAYLAPNNLKPVVAEFYGSKEDPQTFYYA VAVVKKDSGFQMNQLRGKKSCHTGLGRSAGWNIPIGLLYCDLPEPRKPLEKAVANFFSGSCAPC ADGTDFPQLCQLCPGCGCSTLNQYFGYSGAFKCLKDGAGDVAFVKHSTIFENLANKADRDQYEL LCLDNTRKPVDEYKDCHLAQVPSHTVVARSMGGKEDLIWELLNQAQEHFGKDKSKEFQLFSSPH GKDLLFKDSAHGFLKVPPRMDAKMYLGYEYVTAIRNLREGTCPEAPTDECKPVKWCALSHHERL KCDEWSVNSVGKIECVSAETTEDCIAKIMNGEADAMSLDGGFVYIAGKCGLVPVLAENYNKSDN CEDTPEAGYFAIAVVKKSASDLTWDNLKGKKSCHTAVGRTAGWNIPMGLLYNKINHCRFDEFFS EGCAPGSKKDSSLCKLCMGSGLNLCEPNNKEGYYGYTGAFRCLVEKGDVAFVKHQTVPQNTGGK NPDPWAKNLNEKDYELLCLDGTRKPVEEYANCHLARAPNHAVVTRKDKEACVHKILRQQQHLFG SNVTDCSGNFCLFRSETKDLLFRDDTVCLAKLHDRNTYEKYLGEEYVKAVGNLRKCSTSSLLEA CTFRRP*

[0410] pDP86-PDL1-scfv-GFLG-fc-Tf Nucleic Acid Sequence (SEQ ID NO: 67)

[0411] ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTC ACGAATTCGGATATACAGATGACCCAATCCCCATCCAGCTTGTCCGCTAGCGTAGGC GATAGAGTAACTATTACATGCCGCGCTAGTCAAGACGTGTCAACTGCAGTCGCGTGG TACCAACAAAAGCCTGGCAAAGCTCCGAAACTGCTGATTTACAGCGCGTCTTTCCTT TACTCTGGGGTACCTAGCCGATTTTCTGGGTCTGGTAGCGGAACCGATTTCACGCTT ACAATTTCTAGCCTCCAACCCGAAGATTTCGCGACGTACTACTGCCAACAATACCTT TACCATCCAGCCACATTTGGACAGGGCACGAAGGTTGAAATAAAAGGCGGAGGTGG ATCTGGCGGAGGAGGAAGTGGGGGTGGAGGTTCAGAAGTTCAGCTGGTTGAATCAG GCGGCGGACTTGTTCAGCCGGGCGGAAGCCTTCGGCTTAGCTGTGCTGCCAGTGGCT TCACATTCAGTGATAGCTGGATTCATTGGGTTCGCCAGGCACCAGGCAAAGGTTTGG AGTGGGTCGCCTGGATTAGTCCGTATGGGGGCTCCACCTACTACGCTGACTCAGTGA AAGGGCGGTTTACCATTAGTGCTGATACGTCCAAAAATACAGCTTACCTTCAGATGA ACTCTCTGAGGGCCGAAGATACTGCTGTGTACTACTGCGCTCGGAGACATTGGCCAG GAGGGTTCGATTACTGGGGGCAAGGCACTTTGGTGACAGTCAGTTCAGGTGGTTCCG GCAGCGCAGGAggtgggTTCCTGggaGGCGTAGACGGCGACAAAACTCACACATGCCCA CCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAA CCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGAC GTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGT GCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTG GTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTG CAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCA AAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTG ACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 GCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTC CCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGA GCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACA ACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAAggatcctctgggggaagtggaggta gcggtggttctgtgcccgataagacagtgcggtggtgcgccgtgtctgagcacgaggccaccaagtgccagagcttccgggaccacatg aagtccgtgatccccagcgacggccctagcgtggcctgtgtgaagaaggccagctacctggactgcatccgggccattgccgccaatga ggccgacgccgtgacactggatgccggcctggtgtacgatgcctacctggcccccaacaacctgaagcccgtggtggccgagttctacg gcagcaaagaggacccccagaccttctactacgccgtggccgtggtcaagaaggacagcggcttccagatgaaccagctgcggggcaa gaagtcctgtcacaccggcctgggcagaagcgccggctggaacatccccatcggcctgctgtactgcgatctgcccgagccccggaagc ctctggaaaaggccgtggccaacttcttcagcggcagctgcgccccttgtgctgacggaaccgacttcccccagctgtgtcagctgtgccc cggctgtggctgcagcaccctgaaccagtacttcggctacagcggcgccttcaagtgcctgaaggacggcgctggcgacgtggccttcgt gaagcacagcaccatcttcgagaacctggccaacaaggccgaccgggaccagtacgagctgctgtgcctggacaacaccagaaagccc gtggacgagtacaaggactgccacctcgcccaggtgccatctcacacagtggtggcccggtccatgggcggcaaagaggatctgatctg ggagctgctgaaccaggcccaggaacacttcggcaaggacaagagcaaagagttccagctgttcagcagcccccacggcaaggatctg ctgttcaaggacagcgcccacggctttctgaaggtgccccccagaatggacgccaagatgtacctgggctacgagtacgtgaccgccatc cggaacctgagagagggcacctgtcccgaggcccccaccgatgagtgcaagcccgtgaagtggtgcgccctgagccaccacgagcgg ctgaagtgcgacgagtggagcgtgaacagcgtgggcaagatcgagtgcgtgagcgccgagacaaccgaggactgtatcgccaagatca tgaacggcgaggccgatgccatgagcctggacggcggcttcgtgtacattgccggcaagtgcggcctggtgcctgtgctggccgagaac tacaacaagagcgacaactgcgaggatacccccgaggccggctactttgccatcgcagtcgtgaagaagtccgccagcgacctgacctg ggacaatctgaagggcaagaaaagctgccacaccgccgtgggaaggaccgccgggtggaatattcctatggggctgctgtacaacaaga tcaaccactgcagattcgacgagttcttcagcgagggctgcgctcccggcagcaagaaagacagcagcctgtgcaagctgtgcatgggca gcggcctgaacctgtgcgagcccaacaacaaagagggctactacggctacacaggggccttccggtgtctggtggagaagggggacgt ggcttttgtgaaacaccagaccgtgccccagaacaccggcggcaagaaccccgacccctgggccaagaacctgaacgagaaggactac gaactgctgtgtctcgacggcacccggaagccagtggaggaatacgccaactgtcacctggccagagcccccaatcacgccgtggtcac ccggaaggacaaagaggcctgcgtccacaagatcctgcggcagcagcagcacctgttcggcagcaacgtgaccgactgcagcggcaa cttctgcctgttcagaagcgagacaaaggacctcctgttccgggacgacaccgtgtgtctggccaagctgcacgaccggaacacctacga gaagtacctgggcgaggaatatgtgaaggccgtgggcaatctgcggaagtgcagcacctctagcctgctggaagcctgcacctttcgcag acctTAA

[0412] pDP86-PDL1-scfv-GFLG-fc-Tf Amino Acid Sequence (SEQ ID NO: 68)

[0413] MYRMQLLSCIALSLALVTNSDIQMTQSPSSLSASVGDRVTITCRASQDVSTA VAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQ QYLYHPATFGQGTKVEIKGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSC AASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNT AYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSSGGSGSAGGGFLGGV DGDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREP QVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGSSGGSGGSGGSVPDKTVRWCAVSEH EATKCQSFRDHMKSVIPSDGPSVACVKKASYLDCIRAIAANEADAVTLDAGLVYDAYLAPNNLK PVVAEFYGSKEDPQTFYYAVAVVKKDSGFQMNQLRGKKSCHTGLGRSAGWNIPIGLLYCDLPEP RKPLEKAVANFFSGSCAPCADGTDFPQLCQLCPGCGCSTLNQYFGYSGAFKCLKDGAGDVAFVK HSTIFENLANKADRDQYELLCLDNTRKPVDEYKDCHLAQVPSHTVVARSMGGKEDLIWELLNQA QEHFGKDKSKEFQLFSSPHGKDLLFKDSAHGFLKVPPRMDAKMYLGYEYVTAIRNLREGTCPEA PTDECKPVKWCALSHHERLKCDEWSVNSVGKIECVSAETTEDCIAKIMNGEADAMSLDGGFVYI AGKCGLVPVLAENYNKSDNCEDTPEAGYFAIAVVKKSASDLTWDNLKGKKSCHTAVGRTAGWNIDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 PMGLLYNKINHCRFDEFFSEGCAPGSKKDSSLCKLCMGSGLNLCEPNNKEGYYGYTGAFRCLVE KGDVAFVKHQTVPQNTGGKNPDPWAKNLNEKDYELLCLDGTRKPVEEYANCHLARAPNHAVVTR KDKEACVHKILRQQQHLFGSNVTDCSGNFCLFRSETKDLLFRDDTVCLAKLHDRNTYEKYLGEE YVKAVGNLRKCSTSSLLEACTFRRP*

[0414] pDP96-CD20_HC_Fc-N297G,S427C-Tf Nucleic Acid Sequence (SEQ ID NO: 69)

[0415] ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTC ACGAATTCGCAGATTGTTCTGAGCCAGTCCCCGGCAATCCTCTCTGCCAGCCCAGGC GAAAAGGTGACAATGACTTGCCGAGCGAGTTCCAGTGTCTCTTATATCCACTGGTTC CAGCAAAAGCCGGGAAGCAGCCCTAAACCATGGATATATGCAACGTCTAACCTGGC GAGCGGGGTCCCAGTGAGATTTTCCGGAAGCGGCAGCGGAACTAGTTACTCTTTGAC AATAAGCAGAGTGGAGGCTGAGGACGCTGCTACTTACTATTGCCAGCAATGGACGA GTAACCCGCCGACGTTTGGAGGTGGAACGAAGCTGGAGATTAAAGGTGGAGGTGGT TCTGGCGGAGGTGGTTCCGGTGGTGGTGGAAGTCAGGTGCAGCTCCAACAGCCTGG TGCCGAACTTGTCAAACCTGGGGCTAGTGTGAAGATGAGTTGCAAAGCTTCAGGGT ACACGTTTACGTCATACAACATGCATTGGGTAAAGCAAACACCAGGACGCGGCTTG GAATGGATCGGCGCGATATATCCAGGAAACGGTGACACTTCTTATAACCAGAAGTT CAAGGGGAAAGCTACTCTCACAGCGGACAAATCTTCTTCAACAGCGTATATGCAGTT GTCAAGCCTTACTAGCGAGGACAGTGCTGTTTATTACTGCGCCCGGTCCACCTATTA TGGGGGTGATTGGTACTTTAATGTTTGGGGCGCGGGTACTACCGTTACTGTGTCCGC GGGTGTAGACGGCAGCGGCAGCgacaaaactcacacatgcCCCC...

Claims

DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 CLAIMS What is claimed:

1. A fusion protein of Formula I: R1-R2-R3 (I): wherein: R1 is a protein of interest (POI) binder (POIB) or POI binding means for binding Apelin Receptor (APJ), Angiotensin II Receptor Type (AT1R), Relaxin Family Peptide Receptor 1 (RXFP1), C-C Chemokine Receptor Type 6 (CCR6), Adenosine A2A Receptor (A2AR), G Protein-Coupled Receptor 20 (GPR20), C-C Motif Chemokine Receptor 4 (CCR4), Protease- Activated Receptor 2 (PAR2), Frizzled Class Receptor 1 (FZD10), C-C Motif Chemokine Receptor 2 (CCR2), C-X-C Motif Chemokine Receptor 1 (CXCR1 / 1) or Complement Component 5a Receptor 1 (C5aR1); R2 is a linker of the formula R4-R5 or R5-R4, wherein: R4 is IgG Fc region; and R5 is an optional protease-sensitive linking means; and R3 is a TR binding means, wherein the TR binding means is non-competitive with endogenous transferrin for binding to the TR, and optionally, comprising a glycine-rich linker between R2 and R3.

2. The fusion protein of claim 1, wherein the POIB or POI binding means is for binding AT1R, APJ, RXFP1, CCR6, FZD10, and A2AR.

3. The fusion protein of claim 1, wherein the TR binding means is VHHA-12 (SEQ ID NO: 154), VHHA (SEQ ID NO: 146), VHHA-5 (SEQ ID NO: 155), VHHA-7 (SEQ ID NO: 156), or VHHB (SEQ ID NO: 148).

4. The fusion protein of claim 1, wherein the protease-sensitive linking means comprises a cathepsin-cleavable peptide.DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 5. The fusion protein of claim 4, wherein the cathepsin-cleavable peptide linker is selected from the group consisting of FK, VA, VK, SEQ ID NO: 7, 8, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144 and 145.

6. The fusion protein of claim 1, wherein the fusion protein of Formula I is a homodimer.

7. The fusion protein of claim 6, wherein the TR binding means within the homodimer has a binding affinity to TB (Kd) of about 0.001-50 nM.

8. The fusion protein of claim 1, wherein the fusion protein of Formula I is a heterodimer linked to a fusion protein of Formula II: R4’-R3’ (II): wherein: R4’ is IgG Fc region; and R3’ is a TR binding means; and optionally, wherein a linkage between R3’ and R4’ is a protease-sensitive linking means.

9. The fusion protein of claim 8, wherein the TR binding means within the heterodimer has a binding affinity to TB (Kd) of about 0.1-200 nM.

10. The fusion protein of claim 1, wherein the TR binding means comprises an antibody or polypeptide.

11. The fusion protein of claim 1, wherein the TR binding means is an antibody fragment having a heavy chain variable region.

12. The fusion protein of claim 1, wherein the TR binding means comprises: a. a VH CDR1 comprising the amino acid sequence of GTDFSINF (SEQ ID NO: 175), a VH CDR2 comprising the amino acid sequence of ITATGNT (SEQ IDDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 NO: 176), and a VH CDR3 comprising the amino acid sequence of YMLDK (SEQ ID NO: 177). b. a VH CDR1 comprising the amino acid sequence of GTDFSINF (SEQ ID NO: 175), a VH CDR2 comprising the amino acid sequence of FTATGNT (SEQ ID NO: 179), and a VH CDR3 comprising the amino acid sequence of AMLDK (SEQ ID NO: 180); or c. a VH CDR1 comprising the amino acid sequence of GTDFSINF (SEQ ID NO: 175), a VH CDR2 comprising the amino acid sequence of FTATGNT (SEQ ID NO: 179), and a VH CDR3 comprising the amino acid sequence of YMADK (SEQ ID NO: 183).

13. The fusion protein of claim 1, wherein the POIB or POI binding means comprises an antibody.

14. The fusion protein of claim 1, wherein the POIB or POI binding means binds to an extracellular domain of a transmembrane protein.

15. The fusion protein of claim 1, wherein the POIB or POI binding means binds to an extracellular domain of APJ, AT1R, RXFP1, CCR6, A2AR, GPR20, CCR4, PAR2, FZD1, CCR2, CXCR1 / 1 or C5aR1.

16. The fusion protein of claim 1, wherein the POIB or POI binding means binds AT1R and comprises a VH CDR1 comprising the amino acid sequence of GYIYSRY (SEQ ID NO: 187), a VH CDR2 comprising the amino acid sequence of SGGSS (SEQ ID NO: 188), and a VH CDR3 comprising the amino acid sequence of YKIDSNPRVY (SEQ ID NO: 189).

17. The fusion protein of claim 1, wherein the POIB or POI binding means binds APJ and comprises a VH CDR1 comprising the amino acid sequence of GSTYSSH (SEQ ID NO: 190), a VH CDR2 comprising the amino acid sequence of TRSRG (SEQ ID NO: 191), and a VH CDR3 comprising the amino acid sequence of VPRAGIESGAYCKWNMKDSGS (SEQ ID NO: 192).DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 18. The fusion protein of claim 1, wherein the POIB or POI binding means binds RXFP1 and comprises a VH CDR1 comprising the amino acid sequence of GNISRRV (SEQ ID NO: 193), a VH CDR2 comprising the amino acid sequence of DLGGN (SEQ ID NO: 194), and a VH CDR3 comprising the amino acid sequence of TYIDSDGYDYPHIY (SEQ ID NO: 195).

19. The fusion protein of claim 1, wherein the glycine-rich linker is selected from the group consisting of SEQ ID NO: 9, 10, 11, 12, 13, 14, 15 and 16.

20. A nucleic acid sequence encoding the fusion protein of claim 1.

21. A method for treating a subject that has a cancer, cardiovascular disease, kidney disease, neurological disease, respiratory disease or autoimmune disease associated with a GPCR, the method comprising administering the fusion protein of claim 1 to the subject.

22. The method of claim 21, wherein the subject has type 2 diabetes mellitus, Alzheimer’s disease, obesity and depression.

23. The fusion protein of any one of claim 1 for use in treating a cancer, cardiovascular disease, kidney disease, neurological disease, respiratory disease or autoimmune disease associated with a GPCR in a patient.

24. Use of the fusion protein of claim 1 for treating a subject having a cancer, cardiovascular disease, kidney disease, neurological disease, respiratory disease or autoimmune disease associated with a GPCR.

25. A homodimer of a fusion protein of Formula I: R1-R2-R3 (I): wherein: R1 is a of interest (POI) binder (POIB), or POI binding means for binding APJ, AT1R, RXFP1, CCR6, A2AR, GPR20, CCR4, PAR2, FZD1, CCR2, CXCR1 / 1 or C5aR1; R2 is a linker of the formula R4-R5 or R5-R4, wherein:DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 R4 is IgG Fc region; and R5 is an optional protease-sensitive linking means; and R3 is a TR binding means, wherein the TR binding means is non-competitive with endogenous transferrin for binding to the TR, and optionally, comprising a glycine-rich linker between R2 and R3 is a glycine-rich linker.

26. The homodimer of a fusion protein of claim 25, wherein the TR binding means is VHHA-5 (SEQ ID NO: 155), VHHA (SEQ ID NO: 146), VHHA-7 (SEQ ID NO: 156), VHHA-12 (SEQ ID NO: 154) or VHHB (SEQ ID NO: 148).

27. The homodimer of a fusion protein of claim 25, further comprising a disulfide bond between cysteine amino acids in R4 of separate fusion proteins.

28. A homodimer of a fusion protein of Formula III: R1-R6-R3 (III): wherein: R1 is a protein of interest (POI) binder (POIB), or POI binding means for binding APJ, AT1R, RXFP1, CCR6, A2AR, GPR20, CCR4, PAR2, FZD1, CCR2, CXCR1 / 1 or C5aR1; R6 is a dimerization means; and R3 is a TR binding means, wherein the TR binding means is non-competitive with endogenous transferrin for binding to the TR; wherein the homodimer optionally has a protease-sensitive linking means between R1 and R6.

29. The homodimer of a fusion protein of claim 28, wherein the TR binding means is VHHA-12 (SEQ ID NO: 154), VHHA (SEQ ID NO: 146), VHHA-5 (SEQ ID NO: 155), VHHA-7 (SEQ ID NO: 156), or VHHB (SEQ ID NO: 148).

30. A pharmaceutical composition, comprising the fusion protein or homodimer of a fusion protein of any one of claims 1, 6, 8, 25 or 28.DOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 31. A heterodimer of fusion proteins of Formula IV and V: R1-R2 (IV); R3-R2’ (V): wherein: R1 is at least one protein of interest (POI) binder (POIB), or POI binding means for binding APJ, AT1R, RXFP1, CCR6, A2AR, GPR20, CCR4, PAR2, FZD1, CCR2, CXCR1 / 1 or C5aR1; R2 and R2’ are linkers of the formula R4 or optionally R4-R5, wherein: R4 is an IgG Fc region; R5 is an optional protease-sensitive linking means; and R3 is at least one TR binding means, wherein the TR binding means is non-competitive with endogenous transferrin for binding to the TR.

32. The heterodimer of fusion proteins of claim 31, wherein the TR binding means is VHHA-12 (SEQ ID NO: 154), VHHA (SEQ ID NO: 146), VHHA-5 (SEQ ID NO: 155), VHHA-7 (SEQ ID NO: 156), or VHHB (SEQ ID NO: 148).

33. The heterodimer of fusion proteins of claim 31, wherein the R4 of separate fusion proteins have complementary knob and hole structures.

34. A heterodimer of fusion proteins of Formula I and VI: R1-R2-R3 (I); R2’ (VI): wherein: R1 is a protein of interest (POI) binder (POIB), or POI binding means for binding APJ, AT1R, RXFP1, CCR6, A2AR, GPR20, CCR4, PAR2, FZD1, CCR2, CXCR1 / 1 or C5aR1; R2 is a linker of the formula R4-R5 or R5-R4, wherein: R4 is an IgG Fc region; R5 is an optional protease-sensitive linking means; R3 is at least one TR binding means, wherein the TR binding means is non-competitive with endogenous transferrin for binding to the TR; andDOCKET NO: 5031461-000160-WO1 DATE OF FILING: 2025-03-17 R2’ is a linker of the formula R4 or optionally R4-R5.

35. The heterodimer of fusion proteins of claim 34, wherein the TR binding means is VHHA-12 (SEQ ID NO: 154), VHHA (SEQ ID NO: 146), VHHA-5 (SEQ ID NO: 155), VHHA-7 (SEQ ID NO: 156), or VHHB (SEQ ID NO: 148).

36. The heterodimer of fusion proteins of claim 34, wherein the R4 of separate fusion proteins have complementary knob and hole structures.

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