Coiled-coil immunoglobulin fusion protein and its composition

Abstract

Disclosed herein is an immunoglobulin fusion protein comprising a first antibody region, a first therapeutic agent, and a first connecting peptide; wherein the first therapeutic agent is attached to the first antibody region via the connecting peptide; and wherein the connecting peptide does not comprise a region having a beta strand secondary structure. The connecting peptide may comprise an extender peptide. The extender peptide may have an alpha helical secondary structure. The connecting peptide may comprise a connector peptide. The connector peptide may not comprise any secondary structure. Also disclosed herein are compositions comprising the immunoglobulin fusion protein and methods of using the immunoglobulin fusion protein to treat or prevent a disease or condition in a subject.

Description

[0001] This application is a divisional application of the Chinese patent application with application date of July 11, 2014, application number 201480050383.1, and invention name “Coiled-coil immunoglobulin fusion protein and its composition” (the application date of its corresponding PCT application is July 11, 2014, application number PCT / US2014 / 046419).

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit of U.S. Provisional Application No. 61 / 845,280, filed July 11, 2013, U.S. Provisional Application No. 61 / 845,287, filed July 11, 2013, U.S. Provisional Application No. 61 / 925,904, filed January 10, 2014, and U.S. Provisional Application No. 62 / 017,713, filed June 26, 2014, all of which are incorporated herein by reference in their entirety. Background Art

[0004] Antibodies are natural proteins formed by the vertebrate immune system in response to foreign substances (antigens), primarily to defend against infection. For more than a century, antibodies have been induced in animals under artificial conditions and harvested for treatment or diagnosis of disease conditions, or for biological research. Each individual antibody-producing cell produces a single type of antibody with a chemically defined composition, but antibodies obtained directly from animal serum in response to antigen inoculation actually contain the sum of different molecules (e.g., polyclonal antibodies) formed by the sum of individual antibody-producing cells.

[0005] Antibody fusion constructs can be used to improve the delivery of drugs or other agents to target cells, tissues, and tumors. Antibody fusion constructs can include chemical linkers to attach drugs or other agents to the antibody. Exemplary antibody fusion constructs and methods for producing antibody fusion constructs are disclosed in U.S. Patent Application Nos. 2006 / 0182751, 2007 / 0160617, and U.S. Patent No. 7,736,652.

[0006] Disclosed herein are novel immunoglobulin fusion proteins and methods for producing such immunoglobulin fusion proteins. Also disclosed herein are uses of the immunoglobulin fusion proteins in treating various diseases and conditions. Also disclosed herein are methods for extending the half-life of therapeutic agents. Summary of the Invention

[0007] In one aspect of the present disclosure, provided herein is an immunoglobulin fusion protein comprising a first antibody region, a first therapeutic agent, and a first connecting peptide; wherein the first therapeutic agent is attached to the first antibody region via the connecting peptide; and wherein the connecting peptide does not comprise a region having a beta strand secondary structure.

[0008] In one embodiment, the connecting peptide comprises a first extender peptide. The first extender peptide may comprise one or more regions having an alpha-helical secondary structure. In one example, the first extender peptide does not comprise more than 7 consecutive amino acids based on or derived from a bovine ultralong CDR3 amino acid sequence.

[0009] In one embodiment, the linking peptide comprises a first linking peptide. The first linking peptide may not comprise an α-helix or β-strand secondary structure. The first linking peptide may comprise from about 0 to about 50 amino acids.

[0010] In one embodiment, the connecting peptide comprises from about 0 to about 50 amino acids.The connecting peptide may comprise from about 4 to about 100 amino acids.

[0011] In one embodiment, the first connecting peptide is attached to a CDR of the first antibody region. In one embodiment, the first therapeutic peptide replaces one or more regions of the first antibody region. In another embodiment, the first connecting peptide replaces one or more regions of the first antibody region.

[0012] In one embodiment, the immunoglobulin fusion protein further comprises a second connecting peptide. In one embodiment, the second connecting peptide does not comprise a region having a β-strand secondary structure. The second connecting peptide may comprise a second extender peptide. The second extender peptide may comprise one or more regions having an α-helical secondary structure. The second connecting peptide may comprise a second connecting peptide. The second connecting peptide may not comprise an α-helical or β-strand secondary structure.

[0013] In one embodiment, the immunoglobulin fusion protein further comprises a second therapeutic agent.

[0014] In one embodiment, the first antibody region comprises an amino acid sequence based on or derived from any one of SEQ ID NOs: 19-36 and 271-273. In one embodiment, the first antibody region comprises an amino acid sequence that is at least about 50% identical and / or homologous to the amino acid sequence of any one of SEQ ID NOs: 19-36 and 271-273. In one embodiment, the first antibody region comprises an amino acid sequence that is at least about 80% identical and / or homologous to the amino acid sequence of any one of SEQ ID NOs: 19-36 and 271-273.

[0015] In one embodiment, the first antibody region comprises an amino acid sequence based on or derived from a trastuzumab immunoglobulin. In one embodiment, the first antibody region comprises an amino acid sequence that is at least about 50% identical and / or homologous to the amino acid sequence of the trastuzumab immunoglobulin. In one embodiment, the first antibody region comprises an amino acid sequence that is at least about 80% identical and / or homologous to the amino acid sequence of the trastuzumab immunoglobulin.

[0016] In one embodiment, the first antibody region comprises an amino acid sequence based on or derived from a palivizumab immunoglobulin. In one embodiment, the first antibody region comprises an amino acid sequence that is at least about 50% identical and / or homologous to the amino acid sequence of a palivizumab immunoglobulin. In one embodiment, the first antibody region comprises an amino acid sequence that is at least about 80% identical and / or homologous to the amino acid sequence of a palivizumab immunoglobulin.

[0017] In one embodiment, the immunoglobulin fusion protein further comprises a second antibody region. In one example, the first antibody region comprises a region of an antibody light chain, and the second antibody region comprises a region of an antibody heavy chain. In one example, the first antibody region comprises a region of an antibody heavy chain, and the second antibody region comprises a region of an antibody light chain. In one example, the first antibody region comprises a first region of an antibody light chain, and the second antibody region comprises a second region of an antibody light chain. In one example, the first antibody region comprises a first region of an antibody heavy chain, and the second antibody region comprises a second region of an antibody heavy chain. In one embodiment, the second antibody region comprises an amino acid sequence based on or derived from any one of SEQ ID NOs: 19-36 and 271-273. In one embodiment, the second antibody region comprises an amino acid sequence that is at least about 50% identical and / or homologous to the amino acid sequence of any one of SEQ ID NOs: 19-36 and 271-273. In one embodiment, the second antibody region comprises an amino acid sequence that is at least about 80% identical and / or homologous to the amino acid sequence of any one of SEQ ID NOs: 19-36 and 271-273.

[0018] In one embodiment, the second antibody region comprises an amino acid sequence based on or derived from a trastuzumab immunoglobulin. In one embodiment, the second antibody region comprises an amino acid sequence that is at least about 50% identical and / or homologous to the amino acid sequence of a trastuzumab immunoglobulin. In one embodiment, the second antibody region comprises an amino acid sequence that is at least about 80% identical and / or homologous to the amino acid sequence of a trastuzumab immunoglobulin. In one embodiment, the second antibody region comprises an amino acid sequence based on or derived from a palivizumab immunoglobulin. In one embodiment, the second antibody region comprises an amino acid sequence that is at least about 50% identical and / or homologous to the amino acid sequence of a palivizumab immunoglobulin. In one embodiment, the second antibody region comprises an amino acid sequence that is at least about 80% identical and / or homologous to the amino acid sequence of a palivizumab immunoglobulin.

[0019] In one embodiment, the first connecting peptide comprises an amino acid sequence based on or derived from any one of SEQ ID NOs: 144-185. In one embodiment, the first connecting peptide comprises an amino acid sequence that is at least about 50% identical and / or homologous to the amino acid sequence of any one of SEQ ID NOs: 144-185. In one embodiment, the first connecting peptide comprises an amino acid sequence that is at least about 80% identical and / or homologous to the amino acid sequence of any one of SEQ ID NOs: 144-185.

[0020] In one embodiment, the second connecting peptide comprises an amino acid sequence based on or derived from any one of SEQ ID NOs: 144-185. In one embodiment, the second connecting peptide comprises an amino acid sequence that is at least about 50% identical and / or homologous to the amino acid sequence of any one of SEQ ID NOs: 144-185. In one embodiment, the second connecting peptide comprises an amino acid sequence that is at least about 80% identical and / or homologous to the amino acid sequence of any one of SEQ ID NOs: 144-185.

[0021] In one embodiment, the first connecting peptide comprises a protease cleavage site. In one embodiment, the second connecting peptide comprises a protease cleavage site. In one example, the first therapeutic agent comprises an amino acid sequence configured to be recognized by a protease. In one example, the second therapeutic agent comprises an amino acid sequence configured to be recognized by a protease.

[0022] In one embodiment, the first connecting peptide comprises one or more extender peptides and one or more linker peptides. In one embodiment, the first connecting peptide comprises one or more extender peptides, one or more linker peptides and one or more protease cleavage sites. In one embodiment, the first connecting peptide comprises one or more extender peptides and one or more protease cleavage sites. In one embodiment, the first connecting peptide comprises one or more linker peptides and one or more protease cleavage sites.

[0023] In one embodiment, the second connecting peptide comprises one or more extender peptides and one or more linker peptides. In one embodiment, the second connecting peptide comprises one or more extender peptides, one or more linker peptides and one or more protease cleavage sites. In one embodiment, the second connecting peptide comprises one or more extender peptides and one or more protease cleavage sites. In one embodiment, the second connecting peptide comprises one or more linker peptides and one or more protease cleavage sites.

[0024] In one embodiment, the first therapeutic agent comprises an amino acid sequence based on or derived from any one of SEQ ID NOs: 227-267. In one embodiment, the first therapeutic agent comprises an amino acid sequence that is at least about 50% identical and / or homologous to an amino acid sequence of any one of SEQ ID NOs: 227-267. In one embodiment, the first therapeutic agent comprises an amino acid sequence that is at least about 80% identical and / or homologous to an amino acid sequence of any one of SEQ ID NOs: 227-267. In one embodiment, the first therapeutic agent comprises about 5 to about 1,000 amino acids, including about 5 to about 350 amino acids that are identical and / or homologous to any one of SEQ ID NOs: 227-267.

[0025] In one embodiment, the immunoglobulin fusion protein comprises an amino acid sequence based on or derived from any one of SEQ ID NOs: 68-99. In one embodiment, the immunoglobulin fusion protein comprises an amino acid sequence that is at least about 50% identical and / or homologous to the amino acid sequence of any one of SEQ ID NOs: 68-99. In one embodiment, the immunoglobulin fusion protein comprises an amino acid sequence that is at least about 80% identical and / or homologous to the amino acid sequence of any one of SEQ ID NOs: 68-99. In one embodiment, the immunoglobulin fusion protein comprises from about 5 to about 3,000 amino acids, of which from about 50 to about 700 amino acids are identical and / or homologous to any one of SEQ ID NOs: 68-99.

[0026] On the other hand, this paper provides a first genetic construct, which comprises a nucleic acid encoding an immunoglobulin fusion protein of any one of SEQ ID NOs: 68-99. In one embodiment, the first genetic construct comprises a nucleic acid derived from any one of SEQ ID NOs: 37-67. In one embodiment, the first genetic construct comprises a nucleic acid sequence that is at least about 50% identical and / or homologous to the nucleic acid sequence of any one of SEQ ID NOs: 37-67. In one embodiment, the first genetic construct comprises a nucleic acid sequence that is at least about 80% identical and / or homologous to the nucleic acid sequence of any one of SEQ ID NOs: 37-67.

[0027] On the other hand, the present invention provides a first expression vector comprising a first genetic construct comprising a nucleic acid encoding an immunoglobulin fusion protein of any one of SEQ ID NOs: 68-99. In one embodiment, the present invention provides a first expression vector comprising a first genetic construct comprising a nucleic acid derived from any one of SEQ ID NOs: 37-67. In one embodiment, the present invention provides a first expression vector comprising a first genetic construct comprising a nucleic acid sequence that is at least about 50% identical and / or homologous to a nucleic acid sequence of any one of SEQ ID NOs: 37-67. In one embodiment, the present invention provides a first expression vector comprising a first genetic construct comprising a nucleic acid sequence that is at least about 80% identical and / or homologous to a nucleic acid sequence of any one of SEQ ID NOs: 37-67. In one example, the present invention provides a mammalian expression host comprising the first expression vector. In one embodiment, provided herein is a method for producing an immunoglobulin fusion protein comprising (a) transiently transfecting a first expression vector in a mammalian cell culture, (b) culturing the cell culture in an expression medium at a controlled temperature and CO2 percentage, and (c) harvesting the secreted immunoglobulin fusion protein. In one embodiment, the method for producing an immunoglobulin fusion protein further comprises purifying the immunoglobulin fusion protein.

[0028] In another aspect, provided herein are methods of treating a disease or condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an immunoglobulin fusion protein comprising an amino acid sequence derived from any one of SEQ ID NOs 68-99. In another aspect, provided herein are methods of treating a disease or condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an immunoglobulin fusion protein comprising an amino acid sequence that is at least about 50% identical and / or homologous to any one of SEQ ID NOs 68-99. In another aspect, provided herein are methods of treating a disease or condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an immunoglobulin fusion protein comprising an amino acid sequence that is at least about 80% identical and / or homologous to any one of SEQ ID NOs 68-99.

[0029] In another aspect, provided herein are pharmaceutical compositions comprising an immunoglobulin fusion protein derived from any one of SEQ ID NOs: 68-99. In another aspect, provided herein are pharmaceutical compositions comprising an immunoglobulin fusion protein comprising an amino acid sequence that is at least about 50% identical and / or homologous to any one of SEQ ID NOs: 68-99. In another aspect, provided herein are pharmaceutical compositions comprising an immunoglobulin fusion protein comprising an amino acid sequence that is at least about 80% identical and / or homologous to any one of SEQ ID NOs: 68-99. In one embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

[0030] In one embodiment, the immunoglobulin fusion protein comprises an amino acid sequence based on or derived from any one of SEQ ID NOs: 122-143. In one embodiment, the immunoglobulin fusion protein comprises an amino acid sequence that is at least about 50% identical and / or homologous to the amino acid sequence of any one of SEQ ID NOs: 122-143. In one embodiment, the immunoglobulin fusion protein comprises an amino acid sequence that is at least about 80% identical and / or homologous to the amino acid sequence of any one of SEQ ID NOs: 122-143. In one embodiment, the first immunoglobulin fusion protein comprises about 5 to about 3,000 amino acids, which comprises about 50 to about 700 amino acids that are identical and / or homologous to any one of SEQ ID NOs: 122-143.

[0031] In another aspect, provided herein is a first genetic construct comprising a nucleic acid encoding an immunoglobulin fusion protein of any one of SEQ ID NOs: 122-143. In one embodiment, the first genetic construct comprises a nucleic acid derived from any one of SEQ ID NOs: 100-121. In one embodiment, the first genetic construct comprises a nucleic acid sequence that is at least about 50% identical and / or homologous to the nucleic acid sequence of any one of SEQ ID NOs: 100-121. In one embodiment, the first genetic construct comprises a nucleic acid sequence that is at least about 80% identical and / or homologous to the nucleic acid sequence of any one of SEQ ID NOs: 100-121.

[0032] In another aspect, the present invention provides a first expression vector comprising a first genetic construct comprising a nucleic acid encoding an immunoglobulin fusion protein of any one of SEQ ID NOs: 122-143. In one embodiment, the present invention provides a first expression vector comprising a first genetic construct comprising a nucleic acid derived from any one of SEQ ID NOs: 100-121. In one embodiment, the present invention provides a first expression vector comprising a first genetic construct comprising a nucleic acid sequence that is at least about 50% identical and / or homologous to a nucleic acid sequence of any one of SEQ ID NOs: 100-121. In one embodiment, the present invention provides a first expression vector comprising a first genetic construct comprising a nucleic acid sequence that is at least about 80% identical and / or homologous to a nucleic acid sequence of any one of SEQ ID NOs: 100-121. In one example, the present invention provides a mammalian expression host comprising the first expression vector. In one embodiment, provided herein is a method for producing an immunoglobulin fusion protein comprising (a) transiently transfecting a first expression vector in a mammalian cell culture, (b) growing the cell culture in an expression medium at a controlled temperature and CO2 percentage, and (c) harvesting the secreted immunoglobulin fusion protein. In one embodiment, the method for producing an immunoglobulin fusion protein further comprises purifying the immunoglobulin fusion protein.

[0033] In another aspect, provided herein are methods of treating a disease or condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an immunoglobulin fusion protein comprising an amino acid sequence derived from any one of SEQ ID NOs: 122-143. In another aspect, provided herein are methods of treating a disease or condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an immunoglobulin fusion protein comprising an amino acid sequence that is at least about 50% identical and / or homologous to any one of SEQ ID NOs: 122-143. In another aspect, provided herein are methods of treating a disease or condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an immunoglobulin fusion protein comprising an amino acid sequence that is at least about 80% identical and / or homologous to any one of SEQ ID NOs: 122-143.

[0034] In another aspect, provided herein are pharmaceutical compositions comprising an immunoglobulin fusion protein derived from any one of SEQ ID NOs: 122-143. In another aspect, provided herein are pharmaceutical compositions comprising an immunoglobulin fusion protein comprising an amino acid sequence that is at least about 50% identical and / or homologous to any one of SEQ ID NOs: 122-143. In another aspect, provided herein are pharmaceutical compositions comprising an immunoglobulin fusion protein comprising an amino acid sequence that is at least about 80% identical and / or homologous to any one of SEQ ID NOs: 122-143. In one embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

[0035] In one embodiment, the first therapeutic peptide is configured to treat neutropenia and / or neutropenia-related diseases. In one embodiment, the first therapeutic peptide is configured to treat diabetes and / or diabetes-related diseases. In one embodiment, the first therapeutic peptide is configured to treat obesity and / or obesity-related diseases. In one embodiment, the first therapeutic peptide is configured to treat autoimmune diseases and / or autoimmune-related diseases. In one embodiment, the first therapeutic peptide is configured to treat anemia and / or anemia-related diseases. In one embodiment, the first therapeutic peptide is configured to treat growth hormone deficiency and / or growth hormone-related diseases. In one embodiment, the first therapeutic peptide is configured to treat chronic obstructive pulmonary disease (COPD) and / or COPD-related diseases. In one embodiment, the first therapeutic peptide is configured to treat pain. In one embodiment, the first therapeutic peptide is configured to treat irritable bowel syndrome (IBS) and / or IBS-related diseases. In one embodiment, the first therapeutic peptide is configured to treat Crohn's disease and / or Crohn's disease-related diseases. In one embodiment, the first therapeutic peptide is configured to treat neutropenia and / or neutropenia-related diseases. In one embodiment, the first therapeutic peptide is configured to treat a metabolic disorder and / or a disease caused by the metabolic disorder. In one embodiment, the metabolic disorder includes lipodystrophy, diabetes, and hypertriglyceridemia. In one embodiment, the first therapeutic peptide is configured to treat short bowel syndrome and / or a disease associated with short bowel syndrome. In one embodiment, the first therapeutic peptide is configured to treat a patient with heart failure. In one embodiment, the first therapeutic peptide is configured to treat fibrosis and / or a disease associated with fibrosis.

[0036] In one embodiment, the immunoglobulin fusion protein is configured to treat neutropenia and / or neutropenia-related diseases. In one embodiment, the immunoglobulin fusion protein is configured to treat diabetes and / or diabetes-related diseases. In one embodiment, the immunoglobulin fusion protein is configured to treat obesity and / or obesity-related diseases. In one embodiment, the immunoglobulin fusion protein is configured to treat autoimmune diseases and / or autoimmune-related diseases. In one embodiment, the immunoglobulin fusion protein is configured to treat anemia and / or anemia-related diseases. In one embodiment, the immunoglobulin fusion protein is configured to treat growth hormone deficiency and / or growth hormone-related diseases. In one embodiment, the immunoglobulin fusion protein is configured to treat chronic obstructive pulmonary disease (COPD) and / or COPD-related diseases. In one embodiment, the immunoglobulin fusion protein is configured to treat pain. In one embodiment, the immunoglobulin fusion protein is configured to treat irritable bowel syndrome (IBS) and / or IBS-related diseases. In one embodiment, the immunoglobulin fusion protein is configured to treat Crohn's disease and / or Crohn's disease-related diseases. In one embodiment, the immunoglobulin fusion protein is configured to treat neutropenia and / or a disease associated with neutropenia. In one embodiment, the immunoglobulin fusion protein is configured to treat a metabolic disorder and / or a disease caused by the metabolic disorder. In one embodiment, the metabolic disorder includes lipodystrophy, diabetes, and hypertriglyceridemia. In one embodiment, the immunoglobulin fusion protein is configured to treat short bowel syndrome and / or a disease associated with short bowel syndrome. In one embodiment, the immunoglobulin fusion protein is configured to treat a patient with heart failure. In one embodiment, the immunoglobulin fusion protein is configured to treat fibrosis and / or a disease associated with fibrosis. In one embodiment, the first therapeutic peptide is configured to treat pulmonary hypertension, ventilator-induced immature lung damage, and / or lung transplant rejection. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] The above invention content and the following specific embodiments should be read in conjunction with the attached Figure 1 The present disclosure is better understood when read together. For the purpose of illustrating the present disclosure, presently preferred embodiments are shown in the accompanying drawings. However, it should be understood that the present disclosure is not limited to the exact arrangements, examples, and means shown. It is emphasized that, in accordance with common practice, the various features of the drawings are not to scale. On the contrary, the dimensions of the various features have been arbitrarily enlarged or reduced for the sake of clarity.

[0038] In some figures, trastuzumab is referred to as Herceptin. It should be understood that trastuzumab and Herceptin can be used interchangeably throughout the present disclosure. In some figures, immunoglobulin fusion proteins are described in the following order: antibody, coil or direct, therapeutic agent, and antibody region to which the therapeutic agent is attached; for example, trastuzumab-coiled hEPO (CDRH3). Immunoglobulin fusion proteins can be described in any other way, for example, trastuzumab-CDRH3-coiled-hEPO is the same fusion as trastuzumab-coiled hEPO (CDRH3). In some cases, antibodies are abbreviated in the figures, for example, bAb and BLVH12 are abbreviations for bovine antibodies. PBS is an abbreviation for phosphate-buffered saline. In some cases, hAb is an abbreviation for Herceptin or trastuzumab antibodies. In some cases, H2 is an abbreviation for CDRH2, H3 is an abbreviation for CDRH3, and L3 is an abbreviation for CDRL3. In some cases, CDRH3 and CDR3H represent complementarity determining region 3 of the heavy chain, CDRH2 and CDR2H represent complementarity determining region 2 of the heavy chain, and CDRL3 and CDR3L represent complementarity determining region 3 of the light chain.

[0039] Provided herein are immunoglobulin fusion proteins comprising the term "coiled," wherein in some cases, the immunoglobulin fusion proteins comprise at least one extender peptide comprising amino acids having an alpha-helical secondary structure. Provided herein are immunoglobulin fusion proteins comprising the term "direct," wherein in some cases, the immunoglobulin fusion proteins do not comprise an extender peptide.

[0040] The accompanying drawings include the following figures.

[0041] Figure 1 Schematic representations of various immunoglobulin fusion proteins with extender peptides comprising alpha-helical (eg, coiled) structures are depicted.

[0042] Figure 2 A-2G ​​depict schematic diagrams of various non-antibody regions.

[0043] Figure 3 Schematic representations of various immunoglobulin fusion proteins with extender peptides are depicted.

[0044] Figure 4 Depicted are SDS-PAGE gels of bovine-frizzled IgG, bovine-frizzled bGCSF IgG, trastuzumab IgG, and trastuzumab-frizzled bGCSF IgG.

[0045] Figure 5 Schematic representation of various immunoglobulin fusion proteins with therapeutic peptides directly inserted into the antibody region is depicted.

[0046] Figure 6 Depicted is a graph depicting the in vitro activity of bovine-frizzled IgG, bovine-frizzled bGCSF IgG, trastuzumab IgG, and trastuzumab-frizzled bGCSF IgG in mouse NFS-60 cells.

[0047] Figure 7 Graph depicting the binding affinity of trastuzumab-frizzled bGCSF IgG to the Her2 receptor.

[0048] Figure 8 Depicted are Western blots of BLV1H12-frizzled betatrophin IgG in the presence and absence of DTT.

[0049] Figure 9 Schematic representations of various immunoglobulin fusion proteins without extender peptides are depicted.

[0050] Figure 10 Depicted is SDS-PAGE of trastuzumab-direct bGCSF fusion protein in the presence and absence of DTT.

[0051] Figure 11 Graph depicting the in vitro activity of trastuzumab-direct bGCSF fusion protein and bGCSF in proliferating mouse NFS-60 cells.

[0052] Figure 12 Depicted is an SDS-PAGE gel of the Trastuzumab-Exendin-4 (CDRH3) fusion protein and the Trastuzumab-Exendin-4 fusion protein cleaved with Factor Xa to generate Trastuzumab-Exendin-4 RN.

[0053] Figure 13 Depicted are electrospray ionization mass spectrometry (ESI-MS) chromatograms of trastuzumab-exendin-4 (CDRH3) fusion protein treated with peptide N-glycosidase and DTT.

[0054] Figure 14 A-14B depicts ESI-MS chromatograms of trastuzumab-exendin-4 RN (CDRH3) fusion protein treated with peptide N-glycosidase and DTT, (A) N-terminal fragment, (B) C-terminal fragment.

[0055] Figure 15Depicted are graphs depicting the in vitro activities of Exendin-4 (Ex-4), trastuzumab, trastuzumab-Crisped Exendin-4 (CDRH3), and trastuzumab-Crisped Exendin-4 (CDRH3) RN in HEK 293 cells overexpressing the GLP-1R receptor and a cAMP-responsive element (CRE)-luciferase (Luc) reporter.

[0056] Figure 16 Graph depicting the in vitro activity of glucagon, Ex-4, trastuzumab, trastuzumab-CDRH3, and trastuzumab-CDRH3)RN in HEK 293 cells overexpressing the glucagon receptor (GCGR) and the CRE-Luc reporter.

[0057] Figure 17 A-17B depicts a graph of the pharmacokinetics of trastuzumab-Exendin-4 (CDRH3) IgG in mice following (A) intravenous and (B) subcutaneous injections.

[0058] Figure 18 A-18R depicts the pharmacodynamics of trastuzumab-CDRH3 IgG in mice at different time points.

[0059] Figure 19 A-19N depicts the pharmacodynamics of various concentrations of trastuzumab-Crisis Exendin-4 (CDRH3) IgG in mice at different time points.

[0060] Figure 20 Depicted are SDS-PAGE gels of trastuzumab-curled Moka IgG and trastuzumab-curled Vm24 IgG in the presence and absence of DTT.

[0061] Figure 21 Depicted is a graph depicting the in vitro activity of trastuzumab-frizzled Moka IgG and trastuzumab-frizzled Vm24 IgG on T cell activation in human peripheral blood mononuclear cells (PBMCs).

[0062] Figure 22 Depicted are SDS-PAGE of trastuzumab-frizzled hGCSF (CDRH2) and trastuzumab-frizzled hGCSF (CDRL3) in the presence and absence of DTT.

[0063] Figure 23A-23B depicts ESI-MS of (A) trastuzumab-frizzled hGCSF (CDRH2) treated with peptide N-glycosidase and DTT, and (B) trastuzumab-frizzled hGCSF (CDRL3) treated with peptide N-glycosidase and DTT.

[0064] Figure 24 Depicted are histograms of trastuzumab-frizzled hGCSF (CDRH2) and trastuzumab-frizzled hGCSF (CDRL3) binding to the HER2 receptor.

[0065] Figure 25 Graph depicting the in vitro activity of trastuzumab-frizzled hGCSF (CDRH2) and trastuzumab-frizzled hGCSF (CDRL3) in proliferating mouse NFS-60 cells.

[0066] Figure 26 Depicted is an SDS-PAGE gel of trastuzumab-curled hEPO (CDRH3) in the presence and absence of DTT.

[0067] Figure 27 Depicted is ESI-MS of trastuzumab-curled hEPO (CDRH3) IgG treated with peptide N-glycosidase and DTT.

[0068] Figure 28 Depicted is a histogram of trastuzumab-curled hEPO (CDRH3) binding to the HER2 receptor.

[0069] Figure 29 Graph depicting the in vitro activity of trastuzumab-frizzled hEPO (CDRH3) IgG in proliferating human TF-1 cells.

[0070] Figure 30 Depicted is an SDS-PAGE gel of double fusion trastuzumab-curled hEPO (CDRH3)-trastuzumab-curled hGCSF (CDRL3) IgG in the presence and absence of DTT.

[0071] Figure 31 A-31B depicts ESI-MS of the light chain (A) and heavy chain (B) of the dual fusion trastuzumab-crimped hEPO (CDRH3)-trastuzumab-crimped hGCSF (CDRL3) IgG treated with peptide N-glycosidase and DTT.

[0072] Figure 32 Depicted is a histogram of dual fusion trastuzumab-frizzled hEPO (CDRH3)-trastuzumab-frizzled hGCSF (CDRL3) IgG binding to the HER2 receptor.

[0073] Figure 33 Graph depicting the in vitro activity of dual fusion trastuzumab-crimped hEPO (CDRH3)-trastuzumab-crimped hGCSF (CDRL3) IgG in proliferating human TF-1 cells.

[0074] Figure 34 Graph depicting the in vitro activity of dual fusion trastuzumab-frizzled hEPO (CDRH3)-trastuzumab-frizzled hGCSF (CDRL3) IgG in proliferating mouse NFS-60 cells.

[0075] Figure 35 Depicted are SDS-PAGE gels of trastuzumab-curled hGH (CDRH3) IgG after purification, trastuzumab-direct fusion hGH (CDRH2), and trastuzumab-curled hGH (CDRH2) IgG in the presence and absence of DTT.

[0076] Figure 36 A-36B depicts the pharmacokinetics of trastuzumab-curled hGH (CDRH3) IgG in rats by (A) intravenous and (B) subcutaneous injections.

[0077] Figure 37 Depicted is a graph showing the percent body weight change in rats injected with vehicle, genotropin, trastuzumab, and trastuzumab-frizzled hGH (CDRH3) IgG (denoted as Ab-GH in the graph).

[0078] Figure 38 Depicted are SDS-PAGE gels of trastuzumab-curled hLeptin (CDRH2), trastuzumab-curled hLeptin (CDRH3), and trastuzumab-curled hLeptin (CDRL3) in the presence and absence of DTT.

[0079] Figure 39 A-39B depicts a graph of the in vitro activities of (A) hLeptin, trastuzumab-frizzled hLeptin (CDRH2), trastuzumab-frizzled hLeptin (CDRH3), and (B) hLeptin, trastuzumab-frizzled hLeptin (CDRL3) in activating human leptin receptor (LepR).

[0080] Figure 40 A-40C depicts histograms of SKBR3 cells binding to wild-type (wt) trastuzumab, trastuzumab-frizzled hLeptin (CDRH2), and trastuzumab-frizzled hLeptin (CDRH3).

[0081] Figure 41 Depicted is an SDS-PAGE gel of trastuzumab-curled elafin (CDRH3) in the presence and absence of DTT.

[0082] Figure 42 A-42B depicts a graph depicting the in vitro inhibitory activity of trastuzumab-frizzled elafin (CDRH3).

[0083] Figure 43 Depicted is an SDS-PAGE gel of trastuzumab-coiled GLP2 (CDRH3) in the presence and absence of DTT.

[0084] Figure 44 Depicted is an SDS-PAGE gel of trastuzumab-frizzledrelaxin (insulin c-peptide) (CDRH3) in the presence and absence of DTT.

[0085] Figure 45 Depicted is an SDS-PAGE gel of trastuzumab-frizzled relaxin (CDRH3) co-transfected with the cleavage enzyme prohormone convertase 2 (PC2) in the presence and absence of DTT.

[0086] Figure 46 Depicted is an SDS-PAGE gel of Trastuzumab-FrizzleRelaxin (XTEN35) (CDRH3) IgG with 6x HIS co-transfected with PC2 in the presence and absence of DTT. DETAILED DESCRIPTION

[0087] Disclosed herein are immunoglobulin fusion proteins and methods for producing such immunoglobulin fusion proteins. Disclosed herein are immunoglobulin fusion proteins comprising an antibody region and a non-antibody region, wherein the non-antibody region comprises: (a) a first extender peptide comprising at least one secondary structure; and (b) a therapeutic agent. The secondary structure may be an alpha helix. In some embodiments, the alpha helix is ​​configured to form a coiled coil. The therapeutic agent may be a therapeutic peptide. The non-antibody region may further comprise a linker. The linker may be a peptide linker. The peptide linker may not have a regular secondary structure. The non-antibody region may further comprise a proteolytic cleavage site. In some embodiments, the non-antibody region replaces at least a portion of the antibody region. In some embodiments, the extender peptide connects the therapeutic agent to the antibody region. In some cases, the non-antibody region connects to a CDR of an antibody. The CDR may be CDR1, CDR2, or CDR3. The CDR may be part of a light chain or a heavy chain. In exemplary embodiments, the first extender peptide is a connecting peptide or a portion of a connecting peptide. In some embodiments, the peptide linker is a connecting peptide or a portion of a connecting peptide. In some embodiments, the protease cleavage site is a linking peptide or a portion of a linking peptide.

[0088] Also disclosed herein are immunoglobulin fusion proteins comprising an antibody region and an extender fusion region, wherein the extender fusion region comprises: (a) a first extender peptide, wherein the first extender peptide comprises an amino acid sequence comprising an alpha-helix or coiled-coil secondary structure, and (b) a therapeutic agent. The therapeutic agent may be a therapeutic peptide. The extender fusion region may further comprise a linker. The linker may be a peptide linker. The peptide linker may not have a regular secondary structure. The extender fusion region may further comprise a proteolytic cleavage site. In some embodiments, the extender fusion region replaces at least a portion of the antibody region. In some embodiments, the extender peptide connects the therapeutic agent to the antibody region. In some cases, the extender fusion region is connected to a CDR of an antibody. The CDR may be CDR1, CDR2, or CDR3. The CDR may be part of a light chain or a heavy chain. In exemplary embodiments, the first extender peptide is a connecting peptide or a portion of a connecting peptide. In some embodiments, the peptide linker is a connecting peptide or a portion of a connecting peptide. In some embodiments, the protease cleavage site is a connecting peptide or a portion of a connecting peptide.

[0089] Also disclosed herein is an immunoglobulin fusion protein comprising an antibody region directly attached to a non-antibody region, wherein the non-antibody region comprises a therapeutic agent. In some cases, these immunoglobulin fusion proteins can be referred to as direct immunoglobulin fusion proteins. In some cases, the therapeutic agent is a therapeutic peptide. In some embodiments, without using a peptide comprising a secondary structure, the therapeutic agent is attached to the antibody region. In some embodiments, the therapeutic agent replaces at least a portion of the antibody region to which the therapeutic agent is attached. In some embodiments, the therapeutic peptide is attached to the antibody region by one or more connectors that do not comprise a regular secondary structure (e.g., do not comprise an α helix or a β chain). In some embodiments, the connector is a peptide connector. In some embodiments, the immunoglobulin fusion protein further comprises one or more protease cleavage sites. In some embodiments, the therapeutic agent is attached to the CDR of an antibody. The CDR can be CDR1, CDR2, or CDR3. The CDR can be a part of a light chain or a heavy chain. In exemplary embodiments, the peptide connector is a part of a connecting peptide or a connecting peptide. In some embodiments, the protease cleavage site is a part of a connecting peptide or a connecting peptide.

[0090] Also disclosed herein are immunoglobulin fusion proteins comprising an antibody region directly attached to an extension fusion region, wherein the extension fusion region comprises a therapeutic agent. In some cases, these immunoglobulin fusion proteins can be referred to as direct immunoglobulin fusion proteins. In some cases, the therapeutic agent is a therapeutic peptide. In some embodiments, the therapeutic agent is attached to the antibody region without using a peptide comprising a secondary structure. In some embodiments, the therapeutic agent replaces at least a portion of the antibody region to which the therapeutic agent is attached. In some embodiments, the therapeutic agent is attached to the antibody region by one or more connectors that do not comprise a regular secondary structure (e.g., do not comprise an α helix or a β chain). In some embodiments, the connector is a peptide connector. In some embodiments, the immunoglobulin fusion protein further comprises one or more protease cleavage sites. In some embodiments, the therapeutic agent is attached to the CDR of an antibody. The CDR can be CDR1, CDR2, or CDR3. The CDR can be a part of a light chain or a heavy chain. In exemplary embodiments, the peptide connector is a part of a connecting peptide or a connecting peptide. In some embodiments, the protease cleavage site is a part of a connecting peptide or a connecting peptide.

[0091] Also disclosed herein is an immunoglobulin fusion protein comprising a first antibody region, a first therapeutic agent, and a first connecting peptide; wherein the first therapeutic agent is attached to the first antibody region via the connecting peptide; and wherein the connecting peptide does not comprise a region having a beta-chain secondary structure. In an exemplary embodiment, the first therapeutic agent and the first connecting peptide are components of a non-antibody region. In an exemplary embodiment, the first therapeutic agent and the first connecting peptide are components of an extender fusion region. In some embodiments, the connecting peptide comprises one or more extender peptides. In some embodiments, the connecting peptide comprises one or more connecting peptides. In some embodiments, the connecting peptide comprises one or more protease cleavage sites. In some embodiments, the first connecting peptide comprises one or more extender peptides and one or more linker peptides. In some embodiments, the first connecting peptide comprises one or more extender peptides, one or more linker peptides, and one or more protease cleavage sites. In some embodiments, the first connecting peptide comprises one or more extender peptides and one or more protease cleavage sites. In some embodiments, the first connecting peptide comprises one or more linker peptides and one or more protease cleavage sites.

[0092] Also disclosed herein are immunoglobulin fusion proteins comprising (a) a non-antibody region and (b) an antibody region, wherein the non-antibody region replaces at least a portion of the antibody on which the antibody region is based or derived. The non-antibody region can replace at least a portion of the complementarity determining region. The non-antibody region can replace at least a portion of the variable domain. The non-antibody region can replace at least a portion of the constant domain. The non-antibody region can replace at least a portion of the heavy chain. The non-antibody region can replace at least a portion of the light chain. The non-antibody region can comprise a therapeutic peptide. The non-antibody region can comprise a linking peptide.

[0093] Also disclosed herein are immunoglobulin fusion proteins comprising (a) an extender fusion region and (b) an antibody region, wherein the extender fusion region replaces at least a portion of the antibody on which the antibody region is based or derived. The extender fusion region may replace at least a portion of the complementarity determining region. The extender fusion region may replace at least a portion of the variable domain. The extender fusion region may replace at least a portion of the constant domain. The extender fusion region may replace at least a portion of the heavy chain. The extender fusion region may replace at least a portion of the light chain. The extender fusion region may comprise a therapeutic peptide. The extender fusion region may comprise a linking peptide.

[0094] Also disclosed herein are dual fusion proteins comprising two or more therapeutic agents attached to one or more antibody regions or fragments thereof. At least one therapeutic agent may be inserted or attached to an antibody or fragment thereof. Two or more therapeutic agents may be inserted or attached to an antibody or fragment thereof. The therapeutic agent may replace at least a portion of the antibody or fragment thereof. In some cases, the dual fusion protein comprises two therapeutic agents attached to a heavy chain. In some cases, the dual fusion protein comprises two therapeutic agents attached to a light chain. In some cases, the dual fusion protein comprises one therapeutic agent attached to a heavy chain and another therapeutic agent attached to a light chain.

[0095] In some embodiments, the non-antibody region is an extender fusion region. In some cases, the extender fusion region comprises (a) a first extender peptide comprising at least one secondary structure, and (b) a therapeutic agent. The secondary structure can be an alpha helix. The secondary structure can be configured to form a coiled coil. The therapeutic agent can be a therapeutic peptide. In some embodiments, the non-antibody region comprises a connector. The connector may not have a regular secondary structure. In some embodiments, the non-antibody region comprises a protease cleavage site.

[0096] In some embodiments, immunoglobulin fusion proteins comprising an extender peptide (wherein the extender peptide forms an alpha helix and can be configured to form a coiled coil) are referred to as coiled-coil immunoglobulin fusion proteins. In some embodiments, immunoglobulin fusion proteins that do not comprise an extender peptide having a secondary structure are referred to as direct immunoglobulin fusion proteins.

[0097] The extender peptide may be based on or derived from an ultralong CDR3. The extender peptide may comprise 7 or fewer amino acids from an ultralong CDR3 sequence. Alternatively, or additionally, the extender peptide does not comprise an amino acid sequence based on or derived from an ultralong CDR3. The extender peptide may comprise one or more secondary structures. The one or more secondary structures may be alpha helices.

[0098] An exemplary immunoglobulin fusion protein comprising two coiled-coil-containing stretcher peptides (e.g., each stretcher peptide having an alpha-helical secondary structure) is depicted in Figure 1 In. Figure 1 As shown, an antibody region (110) comprising two immunoglobulin heavy chains (115, 120) and two immunoglobulin light chains (125, 130) is attached to a non-antibody region (135) comprising two extender peptides (140, 145) and a therapeutic agent (150) to produce an immunoglobulin fusion protein (160, 170, 180). Figure 1 As shown, the immunoglobulin fusion protein (160) comprises a non-antibody region of an immunoglobulin heavy chain attached to an antibody region. Figure 1 As shown, the immunoglobulin fusion protein (170) comprises a non-antibody region of an immunoglobulin light chain attached to an antibody region. Figure 1 As shown, the immunoglobulin fusion protein (180) comprises two non-antibody regions of two immunoglobulin chains attached to antibody regions. The two extender peptides can form a coiled coil. The two extender peptides can form an antiparallel coiled coil.

[0099] Figure 5 An exemplary direct immunoglobulin fusion protein is depicted in , in which a non-antibody region / stretcher fusion region (e.g., a therapeutic agent) is inserted directly into an antibody without the aid of a stretcher peptide having secondary structure. Figure 5 As shown, an antibody region (1010) comprising two immunoglobulin heavy chains (1015, 1020) and two immunoglobulin light chains (1025, 1030) is attached to a non-antibody region (1050) to produce an immunoglobulin fusion protein (1060, 1070, 1080). Figure 5 As shown, the immunoglobulin fusion protein (1060) comprises a non-antibody region of an immunoglobulin heavy chain attached to an antibody region. Figure 5 As shown, the immunoglobulin fusion protein (1070) comprises a non-antibody region of an immunoglobulin light chain attached to an antibody region. Figure 5 As shown, the immunoglobulin fusion protein (1080) comprises two non-antibody regions of two immunoglobulin chains attached to antibody regions.

[0100] Additional exemplary coiled-coil immunoglobulin fusion proteins are depicted in Figure 3 middle. Figure 3 Formula IA depicts an antibody region (A) comprising an antibody region attached to a fusion region of an extender. 1 ) of an immunoglobulin fusion protein, the extension fusion region comprising a region attached to a therapeutic agent (T 1 ) of the extended peptide (E 1 ).

[0101] Figure 3 Formula IIA depicts an antibody region comprising an antibody region attached to a fusion region of an extender (A 1 ) of an immunoglobulin fusion protein, the extension fusion region comprising a region attached to a therapeutic agent (T 1 ) two extension peptides (E 1 and E 2 ).

[0102] Figure 3 Formula IIIA depicts a system comprising two antibody regions (A 1 and A 2 ) immunoglobulin dual fusion protein. The immunoglobulin dual fusion protein may comprise (a) a first antibody region (A) attached to a first extension fusion region 1 ), the first extension fusion region comprises a first therapeutic agent (T 1 ) two extension peptides (E 1 and E 2 ); and (b) a second antibody region (A) attached to a second extension fusion region 2 ), the second extension fusion region comprises a second therapeutic agent (T 2 ) two extension peptides (E 3 and E 4 ).

[0103] Figure 3 Formula IVA depicts an antibody region (A) comprising an antibody region attached to a fusion region of an extender. 1 ) of an immunoglobulin fusion protein, the extension fusion region comprising a region attached to a therapeutic agent (T 1 ) linker (L 1 ), the linker and the therapeutic agent are located between two extended peptides (E 1 and E 2 )between.

[0104] Figure 3 Formula VA depicts an antibody region (A) comprising an antibody region attached to a fusion region of an extender. 1 ) of an immunoglobulin fusion protein, the extension fusion region comprising a region attached to a therapeutic agent (T 1 ) proteolytic cleavage site (P 1 ), the proteolytic cleavage site and the therapeutic agent are located in two extended peptides (E 1 and E 2)between. Figure 3 Formula VB depicts a cleaved version of Formula VA, in which the proteolytic cleavage site is cleaved by a protease, resulting in the release of one end of the therapeutic agent. Immunoglobulin fusion proteins that can be cleaved to release the amino terminus of the therapeutic agent are referred to as RN due to the release of the N-terminus. For example, trastuzumab-curled hGH RN indicates that upon proteolytic cleavage, the N-terminus of hGH is released.

[0105] Figure 3 Formula VIA depicts an antibody region (A) comprising an antibody region attached to a fusion region of an extender. 1 ) of an immunoglobulin fusion protein, the extension fusion region comprising a linker (L 1 ) and proteolytic cleavage sites (P 1 ) therapeutic agent (T 1 ), wherein the therapeutic agent, linker and proteolytic cleavage site are located between two extender peptides (E 1 and E 2 )between. Figure 3 Formula VIB depicts a cleaved version of Formula VIA, in which the proteolytic cleavage site is cleaved by a protease, resulting in the release of one end of the therapeutic agent. Immunoglobulin fusion proteins that can be cleaved to release the carboxyl terminus of the therapeutic agent are referred to as RCs due to the release of the C-terminus. For example, trastuzumab-curled hGH RC indicates that upon proteolytic cleavage, the C-terminus of hGH is released.

[0106] Figure 3 Formula VIIA depicts an antibody comprising two antibody regions (A 1 and A 2 ) immunoglobulin double fusion protein. The first antibody region (A 1 ) is attached to a first extension fusion region comprising a linker (L) at each end. 1 and L 2 ) therapeutic agent (T 1 ), the therapeutic agent and the linker are located between two extended peptides (E 1 and E 2 ). The second antibody region (A 2 ) is attached to a second stretcher fusion region comprising a region attached to a proteolytic cleavage site (P 1 ) therapeutic agent (T 2 The therapeutic agent and proteolytic cleavage site in the second extension fusion region are flanked by two linkers (L 3 and L 4 The second extension region is flanked by two extender peptides (E 1 and E 2 ).

[0107] Figure 3 Formula VIIIA depicts an antibody region (A) comprising an antibody region attached to a fusion region of an extender. 1 ) immunoglobulin fusion protein, the extender fusion region comprises two extender peptides (E 1 and E 2 ), two connectors (L 1 and L 2 ), two proteolytic cleavage sites (P 1 and P 2 ) and therapeutic agents (T 1 ). Figure 3 Formula VIIIB depicts a cleaved version of Formula VIIIA, wherein the proteolytic cleavage sites located at the N- and C-termini of the therapeutic agent are cleaved by a protease, which results in the release of the therapeutic agent from the immunoglobulin fusion protein.

[0108] Additional exemplary immunoglobulin fusion proteins without extender peptides (direct immunoglobulin fusion proteins) are depicted in Figure 9 middle. Figure 9 Formula IXA depicts a region comprising an antibody region (A 1 ) of an immunoglobulin fusion protein, wherein the non-antibody region comprises a therapeutic agent (T 1 ).

[0109] Figure 9 Formula XA depicts a region comprising an antibody region (A 1 ) immunoglobulin fusion protein, the non-antibody region comprising a region attached to a therapeutic agent (T 1 ) linker (L 1 ). Figure 9 Formula XIA depicts a region comprising two antibody regions (A 1 and A 2 ) immunoglobulin dual fusion protein. The immunoglobulin dual fusion protein may comprise (a) attached to a first therapeutic agent (T 1 ) of the first non-antibody region (A 1 ); and (b) attached to a cell comprising a second therapeutic agent (T 2 ) of the second non-antibody region (A 2 ).

[0110] Figure 9 Formula XIIA depicts a region comprising an antibody region (A 1 ) of an immunoglobulin fusion protein, wherein the non-antibody region comprises a linker (L 1 ), proteolytic cleavage site (P 1 ) and therapeutic agents (T 1), wherein the proteolytic cleavage site is located between the linker and the therapeutic agent. The proteolytic cleavage site in the second non-antibody region has been cleaved by a protease, resulting in the release of one end of the second therapeutic agent.

[0111] Figure 9 Formula XIIIA depicts a region comprising an antibody region (A 1 ) immunoglobulin fusion protein, the non-antibody region comprising a region attached to a therapeutic agent (T 1 ) proteolytic cleavage site (P 1 ). Figure 9 Formula XIIIB depicts a cleaved version of Formula XIIIA, wherein the proteolytic cleavage site is cleaved by a protease, which results in the release of one end of the therapeutic agent.

[0112] Figure 9 Formula XIVA depicts a region comprising an antibody region (A 1 ) of an immunoglobulin fusion protein, wherein the non-antibody region comprises a linker (L 1 ), therapeutic agents (T 1 ) and proteolytic cleavage sites (P 1 ), wherein the therapeutic agent is located between the linker and the proteolytic cleavage site. Figure 9 Formula XIVB depicts a cleaved version of Formula XIVA, wherein the proteolytic cleavage site is cleaved by a protease, resulting in the release of one end of the therapeutic agent.

[0113] Figure 9 Formula XVA depicts a polyclonal antibody comprising two antibody regions (A 1 and A 2 ) immunoglobulin double fusion protein. The first antibody region (A 1 ) is attached to a first non-antibody region comprising a linker (L 1 and L 2 ) therapeutic agent (T 1 ). Second antibody region (A 2 ) is attached to a second non-antibody region comprising a proteolytic cleavage site (P 1 ) of the second therapeutic agent (T 2 The therapeutic agent and the proteolytic cleavage site in the second non-antibody region are flanked by two linkers (L 3 and L 4 ). The proteolytic cleavage site in the second non-antibody region has been cleaved by a protease, resulting in the release of one end of the second therapeutic agent.

[0114] Figure 9 Formula XVIA depicts a region comprising an antibody region (A 1) immunoglobulin fusion protein, the non-antibody region comprises two linkers (L 1 and L 2 ), two proteolytic cleavage sites (P 1 and P 2 ) and therapeutic agents (T 1 ). Figure 9 Formula XVIB depicts a cleaved version of Formula XVIA, ​​wherein the proteolytic cleavage sites located at the N- and C-termini of the therapeutic agent are cleaved by a protease, which results in the release of the therapeutic agent from the immunoglobulin fusion protein.

[0115] Also disclosed herein is a method for treating a disease or condition of a subject in need. The method may include administering to the subject an immunoglobulin fusion protein comprising an antibody region attached to an extender fusion region, wherein the extender fusion region includes (a) an extender peptide and (b) a therapeutic agent, and wherein the extender peptide does not have a secondary structure comprising a beta chain. The method may include administering to the subject an immunoglobulin fusion protein comprising an antibody region attached to a non-immunoglobulin region, wherein the non-immunoglobulin region includes (a) an extender peptide and (b) a therapeutic agent, and wherein the extender peptide does not have a secondary structure comprising a beta chain. The method may include administering to the subject a direct immunoglobulin fusion protein comprising an antibody region attached to an extender fusion region, wherein the extender fusion region includes a therapeutic agent, and wherein the therapeutic agent is attached to the antibody region without using an extender peptide or a connecting peptide with a secondary structure. The method may comprise administering to a subject an immunoglobulin fusion protein comprising an antibody region attached to a non-immunoglobulin region, wherein the non-immunoglobulin region comprises a therapeutic agent, and wherein the therapeutic agent is attached to the antibody region without the use of a stretcher peptide or a linker peptide having a secondary structure. The method may comprise administering to a subject an immunoglobulin fusion protein comprising an antibody region attached to a therapeutic peptide via a linker peptide.

[0116] Also disclosed herein are methods for extending the half-life of therapeutic agents. The methods may include attaching the therapeutic agent to an antibody region. The methods may include attaching the therapeutic agent to the antibody region using one or more linker peptides that do not have a regular secondary structure. The methods may include attaching the therapeutic agent to the antibody region using one or more protease cleavage sites. The methods may include attaching the therapeutic agent to an extender fusion peptide. The methods may include attaching the therapeutic agent to the antibody region using an extender fusion peptide. The methods may include attaching the therapeutic agent to a linker peptide. The methods may include attaching the therapeutic agent to the antibody region using a linker peptide.

[0117] Also disclosed herein is a method for extending the half-life of a therapeutic agent. The method may include attaching an antibody region to the therapeutic agent to produce an immunoglobulin fusion protein. The method may further include attaching one or more linkers or proteolytic cleavage sites to the immunoglobulin fusion protein. The one or more linkers may be attached to the N- and / or C-terminus of the therapeutic agent. The one or more proteolytic cleavage sites may be attached to the N- and / or C-terminus of the therapeutic agent. The one or more proteolytic cleavage sites may be inserted into the therapeutic agent.

[0118] Also disclosed herein are methods for improving the delivery of therapeutic agents. The methods may include attaching a stretcher peptide to the therapeutic agent. The methods may further include attaching an antibody region to the stretcher peptide, the therapeutic agent, or the stretcher fusion peptide. The methods may include directly attaching the therapeutic peptide to the antibody region. The methods may include attaching a linker peptide to the therapeutic agent. The methods may further include attaching an antibody region to the linker peptide and the therapeutic agent.

[0119] Also disclosed herein are methods for improving the delivery of therapeutic agents. The method may include attaching an antibody region to the therapeutic agent to produce an immunoglobulin fusion protein. The method may further include attaching one or more linkers or proteolytic cleavage sites to the immunoglobulin fusion protein. The one or more linkers may be attached to the N- and / or C-termini of the therapeutic agent. The one or more proteolytic cleavage sites may be attached to the N- and / or C-termini of the therapeutic agent. The one or more proteolytic cleavage sites may be inserted into the therapeutic agent.

[0120] Before describing the present methods and compositions, it should be understood that the present invention is not limited to the specific methods or compositions described and therefore may of course be different. It should also be understood that the terms used herein are only used for the purpose of describing specific embodiments and are not intended to be limiting, as the scope of the present invention will only be limited by the appended claims. The examples are described to provide a complete disclosure and description of how to prepare and use the present invention for those of ordinary skill in the art and are not intended to limit the scope of the invention that the inventors regard as their inventions, nor are they intended to represent that the following experiments are all or only experiments performed. Efforts have been made to ensure accuracy with respect to the numbers used (e.g., amounts, temperatures, etc.), but some experimental errors and deviations should be taken into account. Unless otherwise noted, parts are parts by weight, molecular weight is weight average molecular weight, temperature is degrees Celsius, and pressure is atmospheric pressure or near atmospheric pressure.

[0121] The terms "homologous," "homology," or "percent homology," when used herein to describe an amino acid sequence or nucleic acid sequence relative to a reference sequence, can be determined using the formula described by Karlin and Altschul (Proc. Natl. Acad. Sci. USA 87:2264-2268, 1990, as modified in Proc. Natl. Acad. Sci. USA 90:5873-5877, 1993). Such a formula is incorporated into the Basic Alignment Search Tool (BLAST) program by Altschul et al. (J. Mol. Biol. 215:403-410, 1990). The percent homology of a sequence can be determined using the latest version of BLAST as of the filing date of this application.

[0122] When a range of values ​​is provided, it is understood that each intervening value (to one-tenth of the unit of the lower limit, unless the context clearly indicates otherwise) between the upper and lower limits of that range is also specifically disclosed. Each smaller range between any stated value or intervening value in a stated range and any other stated value or intervening value in that range is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included in or excluded from that range, and each range in which either, neither, or both limits are included in the smaller range is also encompassed within the invention, subject to any specifically excluded limits in the stated range. When a stated range includes one or two limits, ranges excluding either or both of those included limits are also encompassed within the invention.

[0123] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, some potential and preferred methods and materials are now described. All publications mentioned herein are hereby incorporated by reference to disclose and describe the methods and / or materials related to the cited publications. It is understood that in the event of a conflict, the present disclosure supersedes any disclosure of the incorporated publication.

[0124] It will be apparent to those skilled in the art after reading this disclosure that each individual embodiment described and illustrated herein has discrete components and features that can be readily separated or combined with the features of any other several embodiments without departing from the scope or spirit of the invention. Any recited method may be performed in the order of events recited or in any other order that is logically possible.

[0125] It must be noted that, as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a cell" includes a plurality of such cells, and reference to "the peptide" includes reference to one or more peptides and equivalents thereof, such as polypeptides known to those skilled in the art, and so forth.

[0126] As used herein, an amino acid sequence based on another amino acid sequence comprises one or more continuous amino acid portions of the other amino acid sequence. A continuous amino acid portion comprises any number of amino acids in the other amino acid sequence. The continuous amino acids can be 1-10%, 1-20%, 10-20%, 10-30%, 20-30%, 20-40%, 30-40%, 40-50%, 50-60%, 50-100%, 60-70%, 60-100%, 70-100%, 80-100%, 80-90%, 90-95%, 90-100% or 1-100% identical to any continuous amino acid region in the other amino acid sequence.

[0127] The publications discussed herein are provided solely as of the date of filing of the present application. Nothing herein should be construed as an admission that the present invention is not entitled to preempt such publications by virtue of prior invention. Furthermore, the publication dates provided may differ from the actual publication dates, which may require independent verification.

[0128] Immunoglobulin fusion proteins

[0129] The immunoglobulin fusion proteins disclosed herein may comprise one or more immunoglobulin domains. The immunoglobulin domain may be immunoglobulin A, immunoglobulin D, immunoglobulin E, immunoglobulin G, or immunoglobulin M. The immunoglobulin domain may be an immunoglobulin heavy chain region or a fragment thereof. The immunoglobulin domain may be derived from a mammalian antibody. Alternatively, the immunoglobulin domain may be derived from a chimeric antibody. The immunoglobulin domain may be derived from an engineered antibody or a recombinant antibody. The immunoglobulin domain may be derived from a humanized, human-engineered, or fully human antibody. The mammalian antibody may be a bovine antibody. The mammalian antibody may be a human antibody. The mammalian antibody may be a murine antibody. The mammalian antibody may be a non-human primate antibody.

[0130] The immunoglobulin fusion proteins disclosed herein may contain a therapeutic agent, wherein the therapeutic agent is a functional peptide. The immunoglobulin fusion protein may contain a functional peptide grafted into an antibody scaffold. The functional peptide may be a linear peptide. The functional peptide may be a modified cyclic peptide. The functional peptide may include a peptide modified to include a β-hairpin structure. The β-hairpin structure may be locked into the β-hairpin conformation by one or more bonds between two or more amino acid residues of the β-hairpin structure. The N-terminus and / or C-terminus of the functional peptide may be grafted into the stretcher fusion region of the immunoglobulin fusion protein. The N-terminus of the functional peptide may be grafted into the first stretcher peptide of the stretcher fusion region, and the C-terminus of the functional peptide may be grafted into the second stretcher peptide of the stretcher fusion region. The functional peptide may include a peptide modified to include a conformationally constrained peptide. The conformationally constrained peptide may have significantly improved binding affinity and / or specificity for the target relative to an endogenous or naturally occurring binding partner of the target. The endogenous or naturally occurring binding partner of the target may be a ligand or substrate of the target. As a non-limiting example, a conformationally constrained peptide may be a peptide comprising a β-hairpin structure. A conformationally constrained peptide may comprise a region that binds to a binding site of a target. The target may be a receptor. The target may be an enzyme. The binding site of a target may be a deep pocket of a ligand-binding domain or a substrate-binding domain. The functional peptide or a portion thereof may bind to a deep pocket of a ligand-binding domain or a substrate-binding domain, such that it blocks target ligand and / or substrate binding. The functional peptide or a portion thereof may bind to a deep pocket of a ligand-binding domain or a substrate-binding domain, such that it partially blocks target ligand and / or substrate binding. The functional peptide or a portion thereof may bind to a deep pocket of a ligand-binding domain or a substrate-binding domain, such that it completely blocks target ligand or substrate binding. The functional peptide or a portion thereof may bind to the surface of a ligand-binding domain or a substrate-binding domain. The functional peptide may be an agonist. The functional peptide may be an antagonist. The functional peptide may be an inhibitor. The functional peptide may be a ligand. The functional peptide may be a substrate.

[0131] The immunoglobulin fusion protein may comprise an amino acid sequence based on or derived from any one of SEQ ID NOs: 68-99 and 122-143. The immunoglobulin fusion protein may comprise an amino acid sequence that is at least about 50% homologous to any one of SEQ ID NOs: 68-99 and 122-143. The immunoglobulin fusion protein may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 97% homologous to any one of SEQ ID NOs: 68-99 and 122-143. The immunoglobulin fusion protein may comprise an amino acid sequence that is at least about 70% homologous to any one of SEQ ID NOs: 68-99 and 122-143. The immunoglobulin fusion protein may comprise an amino acid sequence that is at least about 80% homologous to any one of SEQ ID NOs: 68-99 and 122-143.

[0132] The immunoglobulin fusion protein may comprise an amino acid sequence comprising 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 or more amino acids based on or derived from any one of SEQ ID NOs: 68-99 and 122-143. The immunoglobulin fusion protein may comprise an amino acid sequence comprising 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 450, 500 or more amino acids based on or derived from any one of SEQ ID NOs: 68-99 and 122-143. The immunoglobulin fusion protein may comprise an amino acid sequence comprising 10 or more amino acids based on or derived from any one of SEQ ID NOs: 68-99 and 122-143. The immunoglobulin fusion protein may comprise an amino acid sequence comprising 50 or more amino acids based on or derived from any one of SEQ ID NOs: 68-99 and 122-143. The immunoglobulin fusion protein may comprise an amino acid sequence comprising 100 or more amino acids based on or derived from any one of SEQ ID NOs: 68-99 and 122-143. The immunoglobulin fusion protein may comprise an amino acid sequence comprising 200 or more amino acids based on or derived from any one of SEQ ID NOs: 68-99 and 122-143. The amino acids may be continuous. Alternatively, or additionally, the amino acids are discontinuous.

[0133] The immunoglobulin fusion protein can be encoded by a nucleotide sequence based on or derived from any one of SEQ ID NOs: 37-67 and 100-121. The immunoglobulin fusion protein can be encoded by a nucleotide sequence that is at least about 50% homologous to any one of SEQ ID NOs: 37-67 and 100-121. The immunoglobulin fusion protein can be encoded by a nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 97% homologous to any one of SEQ ID NOs: 37-67 and 100-121. The immunoglobulin fusion protein can be encoded by a nucleotide sequence that is at least about 70% homologous to any one of SEQ ID NOs: 37-67 and 100-121. The immunoglobulin fusion protein can be encoded by a nucleotide sequence that is at least about 80% homologous to any one of SEQ ID NOs: 37-67 and 100-121.

[0134] The immunoglobulin fusion protein can be encoded by a nucleotide sequence comprising 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 or more nucleotides based on or derived from any one of SEQ ID NOs: 37-67 and 100-121. The immunoglobulin fusion protein can be encoded by a nucleotide sequence comprising 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 450, 500 or more nucleotides based on or derived from any one of SEQ ID NOs: 37-67 and 100-121. The immunoglobulin fusion protein can be encoded by a nucleotide sequence comprising 600, 650, 700, 750, 800, 850, 900, 950, 1000 or more nucleotides based on or derived from any one of SEQ ID NOs: 37-67 and 100-121. The immunoglobulin fusion protein can be encoded by a nucleotide sequence comprising 1100, 1200, 1300, 1400, 1500 or more nucleotides based on or derived from any one of SEQ ID NOs: 37-67 and 100-121. The immunoglobulin fusion protein can be encoded by a nucleotide sequence comprising 100 or more nucleotides based on or derived from any one of SEQ ID NOs: 37-67 and 100-121. The immunoglobulin fusion protein can be encoded by a nucleotide sequence comprising 500 or more nucleotides based on or derived from any one of SEQ ID NOs: 37-67 and 100-121. The immunoglobulin fusion protein can be encoded by a nucleotide sequence comprising 1000 or more nucleotides based on or derived from any one of SEQ ID NOs: 37-67 and 100-121. The immunoglobulin fusion protein can be encoded by a nucleotide sequence comprising 1300 or more nucleotides based on or derived from any one of SEQ ID NOs: 37-67 and 100-121. The nucleotides can be contiguous. Alternatively, or additionally, the nucleotides are discontinuous.

[0135] The immunoglobulin fusion protein may further comprise one or more immunoglobulin light chains. The immunoglobulin fusion protein may comprise at least two immunoglobulin light chains. The immunoglobulin light chain may comprise one or more portions of an immunoglobulin light chain. The immunoglobulin light chain may be an immunoglobulin fusion light chain. The immunoglobulin fusion light chain comprises a therapeutic agent and an antibody region derived from an immunoglobulin light chain. The therapeutic agent may be attached to the antibody region via one or more linker peptides. The immunoglobulin light chain may comprise an amino acid sequence based on or derived from any one of SEQ ID NOs: 19-21, 28, 36, 68, 80, 94, 98, and 122. The immunoglobulin light chain may comprise an amino acid sequence that is at least about 50% homologous to any one of SEQ ID NOs: 19-21, 28, 36, 68, 80, 94, 98, and 122. The immunoglobulin light chain may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 97% homologous to any one of SEQ ID NOs: 19-21, 28, 36, 68, 80, 94, 98 and 122. The immunoglobulin light chain may comprise an amino acid sequence that is at least about 70% homologous to any one of SEQ ID NOs: 19-21, 28, 36, 68, 80, 94, 98 and 122. The immunoglobulin light chain may comprise an amino acid sequence that is at least about 80% homologous to any one of SEQ ID NOs: 19-21, 28, 36, 68, 80, 94, 98 and 122.

[0136] The immunoglobulin light chain may comprise an amino acid sequence comprising 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 or more amino acids based on or derived from any one of SEQ ID NOs: 19-21, 28, 36, 68, 80, 94, 98, and 122. The immunoglobulin light chain may comprise an amino acid sequence comprising 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 450, 500 or more amino acids based on or derived from any one of SEQ ID NOs: 19-21, 28, 36, 68, 80, 94, 98, and 122. The immunoglobulin light chain may comprise an amino acid sequence comprising 10 or more amino acids based on or derived from any one of SEQ ID NOs: 19-21, 28, 36, 68, 80, 94, 98, and 122. The immunoglobulin light chain may comprise an amino acid sequence comprising 50 or more amino acids based on or derived from any one of SEQ ID NOs: 19-21, 28, 36, 68, 80, 94, 98, and 122. The immunoglobulin light chain may comprise an amino acid sequence comprising 100 or more amino acids based on or derived from any one of SEQ ID NOs: 19-21, 28, 36, 68, 80, 94, 98, and 122. The immunoglobulin light chain may comprise an amino acid sequence comprising 200 or more amino acids based on or derived from any one of SEQ ID NOs: 19-21, 28, 36, 68, 80, 94, 98, and 122. The amino acids may be contiguous. Alternatively, or additionally, the amino acids are discontinuous.

[0137] The immunoglobulin light chain may be encoded by a nucleotide sequence based on or derived from any one of SEQ ID NOs: 1-3, 10, 18, 37, 49, 63, 67, and 100. The immunoglobulin light chain may be encoded by a nucleotide sequence that is at least about 50% homologous to any one of SEQ ID NOs: 1-3, 10, 18, 37, 49, 63, 67, and 100. The immunoglobulin light chain may be encoded by a nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 97% homologous to any one of SEQ ID NOs: 1-3, 10, 18, 37, 49, 63, 67, and 100. The immunoglobulin light chain may be encoded by a nucleotide sequence that is at least about 70% homologous to any one of SEQ ID NOs: 1-3, 10, 18, 37, 49, 63, 67, and 100. The immunoglobulin light chain can be encoded by a nucleotide sequence that is at least about 80% homologous to any one of SEQ ID NOs: 1-3, 10, 18, 37, 49, 63, 67, and 100.

[0138] The immunoglobulin light chain may be encoded by a nucleotide sequence comprising 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 or more nucleotides based on or derived from any one of SEQ ID NOs: 1-3, 10, 18, 37, 49, 63, 67, and 100. The immunoglobulin light chain may be encoded by a nucleotide sequence comprising 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 450, 500 or more nucleotides based on or derived from any one of SEQ ID NOs: 1-3, 10, 18, 37, 49, 63, 67, and 100. The immunoglobulin light chain may be encoded by a nucleotide sequence comprising 600, 650, 700, 750, 800, 850, 900, 950, 1000 or more nucleotides based on or derived from any one of SEQ ID NOs: 1-3, 10, 18, 37, 49, 63, 67, and 100. The immunoglobulin light chain may be encoded by a nucleotide sequence comprising 1100, 1200, 1300, 1400, 1500 or more nucleotides based on or derived from any one of SEQ ID NOs: 1-3, 10, 18, 37, 49, 63, 67, and 100. The immunoglobulin light chain may be encoded by a nucleotide sequence comprising 100 or more nucleotides based on or derived from any one of SEQ ID NOs: 1-3, 10, 18, 37, 49, 63, 67, and 100. The immunoglobulin light chain may be encoded by a nucleotide sequence comprising 500 or more nucleotides based on or derived from any one of SEQ ID NOs: 1-3, 10, 18, 37, 49, 63, 67, and 100. The immunoglobulin light chain may be encoded by a nucleotide sequence comprising 1000 or more nucleotides based on or derived from any one of SEQ ID NOs: 1-3, 10, 18, 37, 49, 63, 67, and 100. The immunoglobulin light chain may be encoded by a nucleotide sequence comprising 1300 or more nucleotides based on or derived from any one of SEQ ID NOs: 1-3, 10, 18, 37, 49, 63, 67, and 100. The nucleotides may be contiguous. Alternatively, or additionally, the nucleotides are discontinuous.

[0139] The immunoglobulin fusion protein may further comprise one or more immunoglobulin heavy chains. The immunoglobulin fusion protein may comprise at least two immunoglobulin heavy chains. The immunoglobulin heavy chain may comprise one or more portions of an immunoglobulin heavy chain. The immunoglobulin heavy chain may be an immunoglobulin fusion heavy chain. The immunoglobulin fusion heavy chain comprises a therapeutic agent and an antibody region derived from an immunoglobulin heavy chain. The therapeutic agent may be attached to the antibody region via one or more linker peptides. The immunoglobulin heavy chain may comprise an amino acid sequence based on or derived from any one of SEQ ID NOs: 22-27, 29-35, 69-79, 81-93, 95-97, 99, and 123-143. The immunoglobulin heavy chain may comprise an amino acid sequence that is at least about 50% homologous to any one of SEQ ID NOs: 22-27, 29-35, 69-79, 81-93, 95-97, 99, and 123-143. The immunoglobulin heavy chain may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 97% homologous to any one of SEQ ID NOs: 22-27, 29-35, 69-79, 81-93, 95-97, 99 and 123-143. The immunoglobulin heavy chain may comprise an amino acid sequence that is at least about 70% homologous to any one of SEQ ID NOs: 22-27, 29-35, 69-79, 81-93, 95-97, 99 and 123-143. The immunoglobulin heavy chain may comprise an amino acid sequence that is at least about 80% homologous to any one of SEQ ID NOs: 22-27, 29-35, 69-79, 81-93, 95-97, 99 and 123-143.

[0140] The immunoglobulin heavy chain may comprise an amino acid sequence comprising 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 or more amino acids based on or derived from any one of SEQ ID NOs: 22-27, 29-35, 69-79, 81-93, 95-97, 99, and 123-143. The immunoglobulin heavy chain may comprise an amino acid sequence comprising 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 450, 500 or more amino acids based on or derived from any one of SEQ ID NOs: 22-27, 29-35, 69-79, 81-93, 95-97, 99, and 123-143. The immunoglobulin heavy chain may comprise an amino acid sequence comprising 10 or more amino acids based on or derived from any one of SEQ ID NOs: 22-27, 29-35, 69-79, 81-93, 95-97, 99, and 123-143. The immunoglobulin heavy chain may comprise an amino acid sequence comprising 50 or more amino acids based on or derived from any one of SEQ ID NOs: 22-27, 29-35, 69-79, 81-93, 95-97, 99, and 123-143. The immunoglobulin heavy chain may comprise an amino acid sequence comprising 100 or more amino acids based on or derived from any one of SEQ ID NOs: 22-27, 29-35, 69-79, 81-93, 95-97, 99, and 123-143. The immunoglobulin heavy chain may comprise an amino acid sequence comprising 200 or more amino acids based on or derived from any one of SEQ ID NOs: 22-27, 29-35, 69-79, 81-93, 95-97, 99, and 123-143. The amino acids may be continuous. Alternatively, or additionally, the amino acids are discontinuous.

[0141] The immunoglobulin heavy chain can be encoded by a nucleotide sequence based on or derived from any one of SEQ ID NOs: 4-9, 11-17, 38-48, 50-62, 64-66, and 101-121. The immunoglobulin heavy chain can be encoded by a nucleotide sequence that is at least about 50% homologous to any one of SEQ ID NOs: 4-9, 11-17, 38-48, 50-62, 64-66, and 101-121. The immunoglobulin heavy chain can be encoded by a nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 97% homologous to any one of SEQ ID NOs: 4-9, 11-17, 38-48, 50-62, 64-66, and 101-121. The immunoglobulin heavy chain can be encoded by a nucleotide sequence that is at least about 70% homologous to any one of SEQ ID NOs: 4-9, 11-17, 38-48, 50-62, 64-66, and 101-121. The immunoglobulin heavy chain can be encoded by a nucleotide sequence that is at least about 80% homologous to any one of SEQ ID NOs: 4-9, 11-17, 38-48, 50-62, 64-66, and 101-121.

[0142] The immunoglobulin heavy chain can be encoded by a nucleotide sequence comprising 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 or more nucleotides based on or derived from any one of SEQ ID NOs: 4-9, 11-17, 38-48, 50-62, 64-66, and 101-121. The immunoglobulin heavy chain can be encoded by a nucleotide sequence comprising 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 450, 500 or more nucleotides based on or derived from any one of SEQ ID NOs: 4-9, 11-17, 38-48, 50-62, 64-66, and 101-121. The immunoglobulin heavy chain can be encoded by a nucleotide sequence comprising 600, 650, 700, 750, 800, 850, 900, 950, 1000 or more nucleotides based on or derived from any one of SEQ ID NOs: 4-9, 11-17, 38-48, 50-62, 64-66, and 101-121. The immunoglobulin heavy chain can be encoded by a nucleotide sequence comprising 1100, 1200, 1300, 1400, 1500 or more nucleotides based on or derived from any one of SEQ ID NOs: 4-9, 11-17, 38-48, 50-62, 64-66, and 101-121. The immunoglobulin heavy chain may be encoded by a nucleotide sequence comprising 100 or more nucleotides based on or derived from any one of SEQ ID NOs: 4-9, 11-17, 38-48, 50-62, 64-66, and 101-121. The immunoglobulin heavy chain may be encoded by a nucleotide sequence comprising 500 or more nucleotides based on or derived from any one of SEQ ID NOs: 4-9, 11-17, 38-48, 50-62, 64-66, and 101-121. The immunoglobulin heavy chain may be encoded by a nucleotide sequence comprising 1000 or more nucleotides based on or derived from any one of SEQ ID NOs: 4-9, 11-17, 38-48, 50-62, 64-66, and 101-121. The immunoglobulin heavy chain can be encoded by a nucleotide sequence comprising 1300 or more nucleotides based on or derived from any one of SEQ ID NOs: 4-9, 11-17, 38-48, 50-62, 64-66, and 101-121. The nucleotides can be continuous. Alternatively, or additionally, the nucleotides are discontinuous.

[0143] The immunoglobulin fusion protein can comprise (a) a first immunoglobulin fusion heavy chain comprising an amino acid sequence based on or derived from SEQ ID NOs: 69-79, 81-93, 95-97, 99, and 123-143; and (b) a first immunoglobulin light chain comprising an amino acid sequence based on or derived from SEQ ID NOs: 19-21, 28, and 36. The immunoglobulin fusion protein can comprise (a) a first immunoglobulin fusion heavy chain comprising an amino acid sequence at least about 50% identical to SEQ ID NOs: 69-79, 81-93, 95-97, 99, and 123-143; and (b) a first immunoglobulin light chain comprising an amino acid sequence at least about 50% identical to SEQ ID NOs: 19-21, 28, and 36. The first immunoglobulin fusion heavy chain can comprise an amino acid sequence that is at least about 60%, 70%, 75%, 80%, 90%, 95% or 97% identical to SEQ ID NOs: 69-79, 81-93, 95-97, 99 and 123-143. The first immunoglobulin light chain comprises an amino acid sequence that is at least about 60%, 70%, 75%, 80%, 90%, 95% or 97% identical to SEQ ID NOs: 19-21, 28 and 36.

[0144] The immunoglobulin fusion protein may comprise (a) a first immunoglobulin fusion heavy chain encoded by the nucleotide sequence of SEQ ID NOs: 38-48, 50-62, 64-66 and 101-121; and (b) a first immunoglobulin light chain encoded by the nucleotide sequence of SEQ ID NOs: 1-3, 10 and 18. The immunoglobulin fusion protein may comprise (a) a first immunoglobulin fusion heavy chain encoded by a nucleotide sequence that is at least 50% or more homologous to the nucleotide sequence of SEQ ID NOs: 38-48, 50-62, 64-66 and 101-121; and (b) a first immunoglobulin light chain encoded by a nucleotide sequence that is at least 50% or more homologous to the nucleotide sequence of SEQ ID NOs: 1-3, 10 and 18. The first immunoglobulin fusion heavy chain is encoded by a nucleotide sequence that is at least 60%, 70%, 75%, 80%, 90%, 95% or 97% or more homologous to the nucleotide sequences of SEQ ID NOs: 38-48, 50-62, 64-66 and 101-121. The first immunoglobulin light chain is encoded by a nucleotide sequence that is at least 60%, 70%, 75%, 80%, 90%, 95% or 97% or more homologous to the nucleotide sequences of SEQ ID NOs: 1-3, 10 and 18.

[0145] The immunoglobulin fusion protein can comprise (a) a first immunoglobulin heavy chain comprising an amino acid sequence based on or derived from SEQ ID NOs: 22-27 and 29-35; and (b) a first immunoglobulin fusion light chain comprising an amino acid sequence based on or derived from SEQ ID NOs: 68, 80, 94, 98, and 122. The immunoglobulin fusion protein can comprise (a) a first immunoglobulin heavy chain comprising an amino acid sequence at least about 50% identical to SEQ ID NOs: 22-27 and 29-35; and (b) a first immunoglobulin fusion light chain comprising an amino acid sequence at least about 50% identical to SEQ ID NOs: 68, 80, 94, 98, and 122. The first immunoglobulin heavy chain can comprise an amino acid sequence at least about 60%, 70%, 75%, 80%, 90%, 95%, or 97% identical to SEQ ID NOs: 22-27 and 29-35. The first immunoglobulin fusion light chain comprises an amino acid sequence that is at least about 60%, 70%, 75%, 80%, 90%, 95% or 97% identical to SEQ ID NO:68, 80, 94, 98 and 122.

[0146] The immunoglobulin fusion protein may comprise (a) a first immunoglobulin heavy chain encoded by the nucleotide sequence of SEQ ID NOs: 4-9 and 11-17; and (b) a first immunoglobulin fusion light chain encoded by the nucleotide sequence of SEQ ID NOs: 37, 49, 63, 67, and 100. The immunoglobulin fusion protein may comprise (a) a first immunoglobulin heavy chain encoded by a nucleotide sequence that is at least 50% or more homologous to the nucleotide sequence of SEQ ID NOs: 4-9 and 11-17; and (b) a first immunoglobulin fusion light chain encoded by a nucleotide sequence that is at least 50% or more homologous to the nucleotide sequence of SEQ ID NOs: 37, 49, 63, 67, and 100. The first immunoglobulin heavy chain is encoded by a nucleotide sequence that is at least 60%, 70%, 75%, 80%, 90%, 95%, or 97% or more homologous to the nucleotide sequence of SEQ ID NOs: 4-9 and 11-17. The first immunoglobulin fusion light chain is encoded by a nucleotide sequence that is at least 60%, 70%, 75%, 80%, 90%, 95% or 97% or more homologous to the nucleotide sequence of SEQ ID NO:37, 49, 63, 67 and 100.

[0147] The immunoglobulin fusion protein can comprise (a) a first immunoglobulin fusion heavy chain comprising an amino acid sequence based on or derived from SEQ ID NOs: 69-79, 81-93, 95-97, 99, and 123-143; and (b) a first immunoglobulin fusion light chain comprising an amino acid sequence based on or derived from SEQ ID NOs: 68, 80, 94, 98, and 122. The immunoglobulin fusion protein can comprise (a) a first immunoglobulin fusion heavy chain comprising an amino acid sequence at least about 50% identical to SEQ ID NOs: 69-79, 81-93, 95-97, 99, and 123-143; and (b) a first immunoglobulin fusion light chain comprising an amino acid sequence at least about 50% identical to SEQ ID NOs: 68, 80, 94, 98, and 122. The first immunoglobulin fusion heavy chain can comprise an amino acid sequence that is at least about 60%, 70%, 75%, 80%, 90%, 95% or 97% identical to SEQ ID NOs: 69-79, 81-93, 95-97, 99 and 123-143. The first immunoglobulin fusion light chain comprises an amino acid sequence that is at least about 60%, 70%, 75%, 80%, 90%, 95% or 97% identical to SEQ ID NOs: 68, 80, 94, 98 and 122.

[0148] The immunoglobulin fusion protein may comprise (a) a first immunoglobulin fusion heavy chain encoded by the nucleotide sequence of SEQ ID NOs: 38-48, 50-62, 64-66 and 101-121; and (b) a first immunoglobulin fusion light chain encoded by the nucleotide sequence of SEQ ID NOs: 37, 49, 63, 67 and 100. The immunoglobulin fusion protein may comprise (a) a first immunoglobulin fusion heavy chain encoded by a nucleotide sequence at least 50% or more homologous to the nucleotide sequence of SEQ ID NOs: 38-48, 50-62, 64-66 and 101-121; and (b) a first immunoglobulin fusion light chain encoded by a nucleotide sequence at least 50% or more homologous to the nucleotide sequence of SEQ ID NOs: 37, 49, 63, 67 and 100. The first immunoglobulin fusion heavy chain is encoded by a nucleotide sequence that is at least 60%, 70%, 75%, 80%, 90%, 95% or 97% or more homologous to the nucleotide sequence of SEQ ID NOs: 38-48, 50-62, 64-66 and 101-121. The first immunoglobulin fusion light chain is encoded by a nucleotide sequence that is at least 60%, 70%, 75%, 80%, 90%, 95% or 97% or more homologous to the nucleotide sequence of SEQ ID NOs: 37, 49, 63, 67 and 100.

[0149] Also disclosed herein is an immunoglobulin dual fusion protein comprising (a) an antibody region attached to a non-antibody region, wherein the non-antibody region comprises (i) a first extender peptide, wherein the first extender peptide comprises an amino acid sequence comprising an alpha-helical secondary structure, and wherein the first extender peptide does not comprise an ultralong CDR3, and (ii) a first therapeutic agent; and (b) a second therapeutic agent. Attachment of the antibody region to the non-antibody region may comprise inserting the non-antibody region into the antibody region. The first therapeutic agent and the second therapeutic agent may be the same. The first therapeutic agent and the second therapeutic agent may be different. The dual fusion protein may further comprise a second antibody region. The first and second therapeutic agents may be attached to the first antibody region. The first and second therapeutic agents may each be attached to the first and second antibody regions. The first and second antibody regions may be connected. The first and second antibody regions may be connected by one or more disulfide bonds. The first and second antibody regions may be part of an immunoglobulin light chain or heavy chain. The immunoglobulin dual fusion protein may further comprise one or more additional extender peptides. The immunoglobulin dual fusion protein may further comprise one or more linker peptides. The immunoglobulin dual fusion protein may further comprise one or more protease cleavage sites.

[0150] Alternatively, the immunoglobulin dual fusion protein comprises (a) an antibody region attached to a non-antibody region, wherein the non-antibody region comprises (i) a first extender peptide, wherein the first extender peptide comprises an amino acid sequence comprising an alpha-helical secondary structure, and wherein the extender peptide comprises 7 or fewer amino acids based on or derived from an ultralong CDR3, and (ii) a first therapeutic agent; and (b) a second therapeutic agent. The attachment of the antibody region to the non-antibody region may comprise inserting the non-antibody region into the antibody region. The first therapeutic agent and the second therapeutic agent may be the same. The first therapeutic agent and the second therapeutic agent may be different. The dual fusion protein may further comprise a second antibody region. The first and second therapeutic agents may be attached to the first antibody region. The first and second therapeutic agents may each be attached to the first and second antibody regions. The first and second antibody regions may be connected. The first and second antibody regions may be connected by one or more disulfide bonds. The first and second antibody regions may be part of an immunoglobulin light chain or heavy chain. The immunoglobulin dual fusion protein may further comprise one or more additional extender peptides. The immunoglobulin dual fusion protein may further comprise one or more connector peptides. The immunoglobulin dual fusion protein may further comprise one or more protease cleavage sites.

[0151] Alternatively, the immunoglobulin dual fusion protein comprises (a) an antibody region attached to an extender fusion region, wherein the extender fusion region comprises (i) a first extender peptide, wherein the first extender peptide comprises an amino acid sequence comprising an alpha-helical secondary structure, and wherein the first extender peptide does not comprise an ultralong CDR3, and (ii) a first therapeutic agent; and (b) a second therapeutic agent. Attachment of the antibody region to the extender fusion region may comprise inserting the extender fusion region into the antibody region. The first therapeutic agent and the second therapeutic agent may be the same. The first therapeutic agent and the second therapeutic agent may be different. The dual fusion protein may further comprise a second antibody region. The first and second therapeutic agents may be attached to the first antibody region. The first and second therapeutic agents may each be attached to the first and second antibody regions. The first and second antibody regions may be linked. The first and second antibody regions may be linked by one or more disulfide bonds. The first and second antibody regions may be part of an immunoglobulin light chain or heavy chain. The immunoglobulin dual fusion protein may further comprise one or more additional extender peptides. The immunoglobulin dual fusion protein may further comprise one or more linker peptides. The immunoglobulin dual fusion protein may further comprise one or more protease cleavage sites.

[0152] Alternatively, the immunoglobulin dual fusion protein comprises (a) an antibody region attached to an extender fusion region, wherein the extender fusion region comprises (i) a first extender peptide, wherein the first extender peptide comprises an amino acid sequence comprising an alpha-helical secondary structure, and wherein the extender peptide comprises 7 or fewer amino acids based on or derived from an ultralong CDR3, and (ii) a first therapeutic agent; and (b) a second therapeutic agent. Attachment of the antibody region to the extender fusion region may comprise inserting the extender fusion region into the antibody region. The first therapeutic agent and the second therapeutic agent may be the same. The first therapeutic agent and the second therapeutic agent may be different. The dual fusion protein may further comprise a second antibody region. The first and second therapeutic agents may be attached to the first antibody region. The first and second therapeutic agents may each be attached to the first and second antibody regions. The first and second antibody regions may be connected. The first and second antibody regions may be connected by one or more disulfide bonds. The first and second antibody regions may be part of an immunoglobulin light chain or heavy chain. The immunoglobulin dual fusion protein may further comprise one or more additional extender peptides. The immunoglobulin dual fusion protein may further comprise one or more linker peptides. The immunoglobulin dual fusion protein may further comprise one or more protease cleavage sites.

[0153] Alternatively, the immunoglobulin double fusion protein comprises (a) an antibody region attached to a non-antibody region, wherein the non-antibody region comprises (i) a first connecting peptide, wherein the first connecting peptide does not comprise an amino acid sequence containing an alpha helix or beta chain secondary structure, and (ii) a first therapeutic agent; and (b) a second therapeutic agent. The attachment of the antibody region to the non-antibody region may comprise inserting the non-antibody region into the antibody region. The first therapeutic agent and the second therapeutic agent may be the same. The first therapeutic agent and the second therapeutic agent may be different. The double fusion protein may further comprise a second antibody region. The first and second therapeutic agents may be attached to the first antibody region. The first and second therapeutic agents may each be attached to the first antibody region and the second antibody region. The first and second antibody regions may be connected. The first and second antibody regions may be connected by one or more disulfide bonds. The first and second antibody regions may be part of an immunoglobulin light chain or heavy chain. The immunoglobulin double fusion protein may further comprise one or more additional connector peptides. The immunoglobulin double fusion protein may further comprise one or more protease cleavage sites.

[0154] Alternatively, the immunoglobulin dual fusion protein comprises (a) an antibody region attached to an extender fusion region, wherein the extender fusion region comprises (i) a first connecting peptide, wherein the first connecting peptide does not comprise an amino acid sequence containing an alpha helix or beta chain secondary structure; and (ii) a first therapeutic agent; and (b) a second therapeutic agent. The attachment of the antibody region to the extender fusion region may comprise inserting the extender fusion region into the antibody region. The first therapeutic agent and the second therapeutic agent may be the same. The first therapeutic agent and the second therapeutic agent may be different. The dual fusion protein may further comprise a second antibody region. The first and second therapeutic agents may be attached to the first antibody region. The first and second therapeutic agents may each be attached to the first antibody region and the second antibody region. The first and second antibody regions may be connected. The first and second antibody regions may be connected by one or more disulfide bonds. The first and second antibody regions may be part of an immunoglobulin light chain or heavy chain. The immunoglobulin dual fusion protein may further comprise one or more additional connector peptides. The immunoglobulin dual fusion protein may further comprise one or more protease cleavage sites.

[0155] The dual fusion antibody may comprise (a) a first immunoglobulin fusion protein comprising a first antibody region attached to a first non-antibody region, wherein the first non-antibody region comprises (i) a first extender peptide, wherein the first extender peptide comprises an amino acid sequence containing an alpha helical secondary structure and (ii) a first therapeutic agent; and (b) a second immunoglobulin fusion protein comprising a second antibody region attached to a second non-antibody region, wherein the second non-antibody region comprises (i) a second extender peptide comprising at least one secondary structure and (ii) a second therapeutic agent. In some embodiments, the first extender peptide is a portion of a connecting peptide or a connecting peptide. In some embodiments, the second extender peptide is a portion of a connecting peptide or a connecting peptide. In some embodiments, the first extender peptide does not comprise amino acids having a beta chain secondary structure. In some embodiments, the second extender peptide does not comprise amino acids having a beta chain secondary structure. The dual fusion antibody may further comprise one or more peptide connectors. The dual fusion antibody may further comprise one or more protease cleavage sites.

[0156] The dual fusion antibody may comprise (a) a first immunoglobulin fusion protein comprising a first antibody region attached to a first non-antibody region, wherein the first non-antibody region comprises (i) a first extender peptide, wherein the first extender peptide comprises an amino acid sequence comprising an alpha helical secondary structure and (ii) a first therapeutic agent; and (b) a second immunoglobulin fusion protein comprising a second antibody region attached to the extender fusion region, wherein the extender fusion region comprises (i) a second extender peptide comprising at least one secondary structure and (ii) a second therapeutic agent. In some embodiments, the first extender peptide is a portion of a connecting peptide or a connecting peptide. In some embodiments, the second extender peptide is a portion of a connecting peptide or a connecting peptide. The dual fusion antibody may further comprise one or more peptide connectors. The dual fusion antibody may further comprise one or more protease cleavage sites. In some embodiments, the first extender peptide does not comprise amino acids having a beta chain secondary structure. In some embodiments, the second extender peptide does not comprise amino acids having a beta chain secondary structure.

[0157] The dual fusion antibody may comprise (a) a first immunoglobulin fusion protein comprising a first antibody region attached to a first extender fusion region, wherein the first extender fusion region comprises (i) a first extender peptide, wherein the first extender peptide comprises an amino acid sequence comprising an alpha helical secondary structure and (ii) a first therapeutic agent; and (b) a second immunoglobulin fusion protein comprising a second antibody region attached to a non-antibody region, wherein the non-antibody region comprises (i) a second extender peptide comprising at least one secondary structure and (ii) a second therapeutic agent. In some embodiments, the first extender peptide is a connecting peptide or a portion of a connecting peptide. In some embodiments, the second extender peptide is a connecting peptide or a portion of a connecting peptide. The dual fusion antibody may further comprise one or more peptide connectors. The dual fusion antibody may further comprise one or more protease cleavage sites. In some embodiments, the first extender peptide does not comprise amino acids having a beta chain secondary structure. In some embodiments, the second extender peptide does not comprise amino acids having a beta chain secondary structure.

[0158] The dual fusion antibody may comprise (a) a first immunoglobulin fusion protein comprising a first antibody region attached to a first extender fusion region, wherein the first extender fusion region comprises (i) a first extender peptide, wherein the first extender peptide comprises an amino acid sequence comprising an alpha helical secondary structure and (ii) a first therapeutic agent; and (b) a second immunoglobulin fusion protein comprising a second antibody region attached to a second extender fusion region, wherein the extender fusion region comprises (i) a second extender peptide comprising at least one secondary structure and (ii) a second therapeutic agent. In some embodiments, the first extender peptide is a connecting peptide or a portion of a connecting peptide. In some embodiments, the second extender peptide is a connecting peptide or a portion of a connecting peptide. The dual fusion antibody may further comprise one or more peptide connectors. The dual fusion antibody may further comprise one or more protease cleavage sites. In some embodiments, the first extender peptide does not comprise amino acids having a beta chain secondary structure. In some embodiments, the second extender peptide does not comprise amino acids having a beta chain secondary structure.

[0159] The double fusion antibody may comprise (a) a first immunoglobulin fusion protein comprising a first antibody region attached to a first non-antibody region, wherein the first non-antibody region comprises (i) a first peptide connector, wherein the first peptide connector does not comprise an amino acid sequence containing an α-helix or β-chain secondary structure and (ii) a first therapeutic agent; and (b) a second immunoglobulin fusion protein comprising a second antibody region attached to a second non-antibody region, wherein the second non-antibody region comprises (i) a second peptide connector, wherein the second peptide connector does not comprise an amino acid sequence containing an α-helix or β-chain secondary structure and (ii) a second therapeutic agent. In some embodiments, the first connector peptide is a portion of a connecting peptide or a connecting peptide. In some embodiments, the second connector peptide is a portion of a connecting peptide or a connecting peptide. The double fusion antibody may further comprise one or more protease cleavage sites.

[0160] The double fusion antibody may comprise (a) a first immunoglobulin fusion protein comprising a first antibody region attached to a first non-antibody region, wherein the first non-antibody region comprises (i) a first peptide connector, wherein the first peptide connector does not comprise an amino acid sequence containing an α-helix or β-chain secondary structure and (ii) a first therapeutic agent; and (b) a second immunoglobulin fusion protein comprising a second antibody region attached to an extender fusion region, wherein the extender fusion region comprises (i) a second peptide connector, wherein the second peptide connector does not comprise an amino acid sequence containing an α-helix or β-chain secondary structure and (ii) a second therapeutic agent. In some embodiments, the first connector peptide is a portion of a connecting peptide or a connecting peptide. In some embodiments, the second connector peptide is a portion of a connecting peptide or a connecting peptide. The double fusion antibody may further comprise one or more protease cleavage sites.

[0161] The dual fusion antibody may comprise (a) a first immunoglobulin fusion protein comprising a first antibody region attached to a first extension fusion region, wherein the first extension fusion region comprises (i) a first peptide connector, wherein the first peptide connector does not comprise an amino acid sequence containing an α-helix or β-chain secondary structure and (ii) a first therapeutic agent; and (b) a second immunoglobulin fusion protein comprising a second antibody region attached to a non-antibody region, wherein the non-antibody region comprises (i) a second peptide connector, wherein the second peptide connector does not comprise an amino acid sequence containing an α-helix or β-chain secondary structure and (ii) a second therapeutic agent. In some embodiments, the first connector peptide is a connecting peptide or a portion of a connecting peptide. In some embodiments, the second connector peptide is a connecting peptide or a portion of a connecting peptide. The dual fusion antibody may further comprise one or more protease cleavage sites.

[0162] The dual fusion antibody may comprise (a) a first immunoglobulin fusion protein comprising a first antibody region attached to a first extension fusion region, wherein the first extension fusion region comprises (i) a first peptide connector, wherein the first peptide connector does not comprise an amino acid sequence containing an alpha helix or beta chain secondary structure and (ii) a first therapeutic agent; and (b) a second immunoglobulin fusion protein comprising a second antibody region attached to a second extension fusion region, wherein the extension fusion region comprises (i) a second peptide connector, wherein the second peptide connector does not comprise an amino acid sequence containing an alpha helix or beta chain secondary structure and (ii) a second therapeutic agent. In some embodiments, the first connector peptide is a connecting peptide or a portion of a connecting peptide. In some embodiments, the second connector peptide is a connecting peptide or a portion of a connecting peptide. The dual fusion antibody may further comprise one or more protease cleavage sites.

[0163] The dual fusion antibody may comprise (a) a first immunoglobulin fusion protein comprising a first antibody region attached to a first non-antibody region, wherein the first non-antibody region comprises (i) a first peptide connector, wherein the first peptide connector does not comprise an amino acid sequence containing an α-helix or β-chain secondary structure and (ii) a first therapeutic agent; and (b) a second immunoglobulin fusion protein comprising a second antibody region attached to a second non-antibody region, wherein the second non-antibody region comprises (i) an extender peptide comprising at least one secondary structure and (ii) a second therapeutic agent. In some embodiments, the extender peptide comprises amino acids having an α-helix secondary structure. In some embodiments, the extender peptide does not comprise amino acids having a β-chain secondary structure. In some embodiments, the extender peptide is a portion of a connecting peptide or a connecting peptide. In some embodiments, the peptide connector is a portion of a connecting peptide or a connecting peptide. The dual fusion antibody may further comprise one or more protease cleavage sites. The dual fusion antibody may further comprise one or more additional connectors. The second non-antibody region of the second immunoglobulin fusion protein may further comprise one or more additional extender peptides.

[0164] The dual fusion antibody may comprise (a) a first immunoglobulin fusion protein comprising a first antibody region attached to a first non-antibody region, wherein the first non-antibody region comprises (i) a first peptide connector, wherein the first peptide connector does not comprise an amino acid sequence containing an alpha helix or beta chain secondary structure and (ii) a first therapeutic agent; and (b) a second immunoglobulin fusion protein comprising a second antibody region attached to an extender fusion region, wherein the extender fusion region comprises (i) an extender peptide comprising at least one secondary structure and (ii) a second therapeutic agent. In some embodiments, the extender peptide comprises amino acids having an alpha helix secondary structure. In some embodiments, the extender peptide does not comprise amino acids having a beta chain secondary structure. In some embodiments, the extender peptide is a connecting peptide or a portion of a connecting peptide. In some embodiments, the peptide connector is a connecting peptide or a portion of a connecting peptide. The dual fusion antibody may further comprise one or more protease cleavage sites. The dual fusion antibody may further comprise one or more additional connectors. The extender fusion region of the second immunoglobulin fusion protein may further comprise one or more additional extender peptides.

[0165] The dual fusion antibody may comprise (a) a first immunoglobulin fusion protein comprising a first antibody region attached to a first extension fusion region, wherein the first extension fusion region comprises (i) a first peptide connector, wherein the first peptide connector does not comprise an amino acid sequence containing an alpha helix or beta chain secondary structure and (ii) a first therapeutic agent; and (b) a second immunoglobulin fusion protein comprising a second antibody region attached to a non-antibody region, wherein the non-antibody region comprises (i) an extension peptide comprising at least one secondary structure and (ii) a second therapeutic agent. In some embodiments, the extension peptide comprises amino acids having an alpha helix secondary structure. In some embodiments, the extension peptide does not comprise amino acids having a beta chain secondary structure. In some embodiments, the extension peptide is a connecting peptide or a portion of a connecting peptide. In some embodiments, the peptide connector is a connecting peptide or a portion of a connecting peptide. The dual fusion antibody may further comprise one or more protease cleavage sites. The dual fusion antibody may further comprise one or more additional connectors. The non-antibody region of the second immunoglobulin fusion protein may further comprise one or more additional extension peptides.

[0166] The dual fusion antibody may comprise (a) a first immunoglobulin fusion protein comprising a first antibody region attached to a first extension fusion region, wherein the first extension fusion region comprises (i) a first peptide linker, wherein the first peptide linker does not comprise an amino acid sequence comprising an alpha helix or beta chain secondary structure and (ii) a first therapeutic agent; and (b) a second immunoglobulin fusion protein comprising a second antibody region attached to a second extension fusion region, wherein the extension fusion region comprises (i) a second extension peptide comprising at least one secondary structure and (ii) a second therapeutic agent. In some embodiments, the extension peptide comprises amino acids having an alpha helix secondary structure. In some embodiments, the extension peptide does not comprise amino acids having a beta chain secondary structure. In some embodiments, the extension peptide is a connecting peptide or a portion of a connecting peptide. In some embodiments, the peptide linker is a connecting peptide or a portion of a connecting peptide. The dual fusion antibody may further comprise one or more protease cleavage sites. The dual fusion antibody may further comprise one or more additional linkers. The second extension fusion region of the second immunoglobulin fusion protein may further comprise one or more additional extension peptides.

[0167] The dual fusion antibody may comprise (a) a first immunoglobulin fusion protein comprising a first antibody region attached to a first non-antibody region, wherein the first non-antibody region comprises (i) a first extender peptide, wherein the first extender peptide comprises an amino acid sequence containing an α-helical secondary structure and (ii) a first therapeutic agent; and (b) a second immunoglobulin fusion protein comprising a second antibody region attached to a second non-antibody region, wherein the second non-antibody region comprises (i) a peptide connector, wherein the peptide connector does not comprise an amino acid sequence containing an α-helical or β-chain secondary structure and (ii) a second therapeutic agent. In some embodiments, the extender peptide does not comprise an amino acid having a β-chain secondary structure. In some embodiments, the extender peptide is a connecting peptide or a portion of a connecting peptide. In some embodiments, the peptide connector is a connecting peptide or a portion of a connecting peptide. The dual fusion antibody may further comprise one or more protease cleavage sites. The dual fusion antibody may further comprise one or more additional connectors. The first non-antibody region of the first immunoglobulin fusion protein may further comprise one or more additional extender peptides.

[0168] The dual fusion antibody may comprise (a) a first immunoglobulin fusion protein comprising a first antibody region attached to a first non-antibody region, wherein the first non-antibody region comprises (i) a first extender peptide, wherein the first extender peptide comprises an amino acid sequence containing an alpha helical secondary structure and (ii) a first therapeutic agent; and (b) a second immunoglobulin fusion protein comprising a second antibody region attached to the extender fusion region, wherein the extender fusion region comprises (i) a peptide connector, wherein the second peptide connector does not comprise an amino acid sequence containing an alpha helical or beta chain secondary structure and (ii) a second therapeutic agent. In some embodiments, the extender peptide does not comprise an amino acid having a beta chain secondary structure. In some embodiments, the extender peptide is a connecting peptide or a portion of a connecting peptide. In some embodiments, the peptide connector is a connecting peptide or a portion of a connecting peptide. The dual fusion antibody may further comprise one or more protease cleavage sites. The dual fusion antibody may further comprise one or more additional connectors. The first non-antibody region of the first immunoglobulin fusion protein may further comprise one or more additional extender peptides.

[0169] The dual fusion antibody may comprise (a) a first immunoglobulin fusion protein comprising a first antibody region attached to a first extender fusion region, wherein the first extender fusion region comprises (i) a first extender peptide, wherein the first extender peptide comprises an amino acid sequence comprising an alpha helical secondary structure and (ii) a first therapeutic agent; and (b) a second immunoglobulin fusion protein comprising a second antibody region attached to a non-antibody region, wherein the non-antibody region comprises (i) a peptide connector, wherein the peptide connector does not comprise an amino acid sequence comprising an alpha helical or beta chain secondary structure and (ii) a second therapeutic agent. In some embodiments, the extender peptide does not comprise an amino acid having a beta chain secondary structure. In some embodiments, the extender peptide is a connecting peptide or a portion of a connecting peptide. In some embodiments, the peptide connector is a connecting peptide or a portion of a connecting peptide. The dual fusion antibody may further comprise one or more protease cleavage sites. The dual fusion antibody may further comprise one or more additional connectors. The first extender fusion region of the first immunoglobulin fusion protein may further comprise one or more additional extender peptides.

[0170] The dual fusion antibody may comprise (a) a first immunoglobulin fusion protein comprising a first antibody region attached to a first extension fusion region, wherein the first extension fusion region comprises (i) a first extension peptide, wherein the first extension peptide comprises an amino acid sequence containing an alpha helical secondary structure and (ii) a first therapeutic agent; and (b) a second immunoglobulin fusion protein comprising a second antibody region attached to a second extension fusion region, wherein the extension fusion region comprises (i) a peptide connector, wherein the peptide connector does not comprise an amino acid sequence containing an alpha helical or beta chain secondary structure and (ii) a second therapeutic agent. In some embodiments, the extension peptide does not comprise an amino acid having a beta chain secondary structure. In some embodiments, the extension peptide is a connecting peptide or a portion of a connecting peptide. In some embodiments, the peptide connector is a connecting peptide or a portion of a connecting peptide. The dual fusion antibody may further comprise one or more protease cleavage sites. The dual fusion antibody may further comprise one or more additional connectors. The first extension fusion region of the first immunoglobulin fusion protein may further comprise one or more additional extension peptides.

[0171] The first therapeutic agent and the second therapeutic agent may be the same. The first therapeutic agent and the second therapeutic agent may be different. The immunoglobulin dual fusion protein may further comprise one or more additional therapeutic agents. The two or more therapeutic agents may be the same. Alternatively, or additionally, the two or more therapeutic agents may be different. The first therapeutic agent may be a therapeutic peptide. The second therapeutic agent may be a therapeutic peptide. One or more of the additional therapeutic agents may be a therapeutic peptide. The first therapeutic agent may comprise a therapeutic peptide. The second therapeutic agent may comprise a therapeutic peptide. One or more of the additional therapeutic agents may comprise one or more therapeutic peptides. The therapeutic agent may comprise one or more therapeutic peptides or regions of therapeutic peptides. The therapeutic agent may comprise, for example, a first therapeutic peptide or a portion thereof, an internal peptide, and a second therapeutic peptide or a portion thereof. The internal peptide may comprise, for example, a protease cleavage site or an affinity tag, such as a histidine tag (6X HIS). The internal peptide may comprise, for example, another therapeutic peptide or a portion thereof. For example, the therapeutic agent may comprise the first portion of the first therapeutic peptide, the first portion of the second therapeutic peptide, and the second portion of the first therapeutic peptide.

[0172] The first antibody region and the second antibody region can be the same. For example, the first antibody region and the second antibody region comprise immunoglobulin heavy chains. Alternatively, the first antibody region and the second antibody region can comprise immunoglobulin light chains. The first antibody region and the second antibody region can be different. For example, the first antibody region comprises immunoglobulin heavy chains and the second antibody region comprises immunoglobulin light chains, or vice versa. The immunoglobulin dual fusion protein can further comprise one or more additional antibody regions. The two or more antibody regions can be the same. Alternatively, or additionally, the two or more antibody regions can be different.

[0173] The immunoglobulin dual fusion protein may further comprise one or more extender peptides. The one or more extender peptides may be identical. Alternatively, or additionally, the one or more extender peptides may be different. In some embodiments, the extender peptide comprises 7 or fewer amino acids based on or derived from an ultralong CDR3.

[0174] The immunoglobulin double fusion protein may further comprise one or more additional antibody regions. The two or more antibody regions may be the same. Alternatively, or additionally, the two or more antibody regions are different.

[0175] The immunoglobulin dual fusion protein may further comprise one or more linkers. The immunoglobulin dual fusion protein may further comprise two or more linkers. The two or more linkers may be the same. Alternatively, or additionally, the two or more linkers are different.

[0176] The immunoglobulin dual fusion protein may further comprise one or more proteolytic cleavage sites. The immunoglobulin dual fusion protein may further comprise two or more proteolytic cleavage sites. The two or more proteolytic cleavage sites may be the same. Alternatively, or additionally, the two or more proteolytic cleavage sites are different.

[0177] The immunoglobulin dual fusion protein may further comprise one or more therapeutic agents containing internal peptides. The internal peptide may comprise an affinity tag or label, such as a HHHHHH (6X) histidine tail. The internal peptide may comprise a portion of a therapeutic peptide.

[0178] Exemplary immunoglobulin dual fusion proteins are depicted in Figure 3 Formula IIIA and Formula VIIA. Figure 8 The immunoglobulin dual fusion protein may comprise (a) a first antibody region (A) attached to a first extension fusion region. 1 ), the first stretcher fusion region comprises two stretcher peptides (E 1 , E 2 ) of the first therapeutic agent (T 1 ); and (b) a second antibody region (A) attached to a second extension fusion region 2 ), the second stretcher fusion region comprises two stretcher peptides (E 3 , E 4 ) of the second therapeutic agent (T 2). The immunoglobulin dual fusion protein may further comprise one or more linkers and one or more proteolytic cleavage sites. The one or more proteolytic cleavage sites may be attached to the N- and / or C-termini of the therapeutic agent. Proteolytic cleavage of the proteolytic cleavage site may release the N- and / or C-termini of the therapeutic agent from the immunoglobulin fusion protein. Figure 3 Formula VIIA depicts an exemplary immunoglobulin dual fusion protein, wherein the second therapeutic agent (T 2 ) has been released from the N-terminus.

[0179] Antibody region

[0180] The immunoglobulin fusion proteins disclosed herein comprise one or more antibody regions. An antibody region may comprise an immunoglobulin or a fragment thereof. The antibody region may comprise at least a portion of an immunoglobulin heavy chain, an immunoglobulin light chain, or a combination thereof. An antibody region may comprise two or more immunoglobulin chains or portions thereof. An antibody region may comprise three or more immunoglobulin chains or portions thereof. An antibody region may comprise four or more immunoglobulin chains or portions thereof. An antibody region may comprise five or more immunoglobulin chains or portions thereof. An antibody region may comprise two immunoglobulin heavy chains and two immunoglobulin light chains.

[0181] The antibody region may comprise an entire immunoglobulin molecule or any polypeptide comprising an immunoglobulin fragment, including but not limited to a heavy chain, a light chain, a variable domain, a constant domain, a complementarity determining region (CDR), a framework region, a fragment antigen (Fab) binding region, Fab', F(ab')2, F(ab')3, Fab', a fragment crystallizable (Fc) region, a single-chain variable fragment (scFV), a di-scFv, a single-domain immunoglobulin, a trifunctional immunoglobulin, a chemically linked F(ab')2, and any combination thereof. The immunoglobulin region may comprise one or more mutations. The Fc region may be a mutated Fc region. The mutated Fc region may comprise one or more mutations that eliminate the antibody-dependent cell-mediated cytotoxicity (ADCC) effect of the Fc region. The mutated Fc region may comprise about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 1 to about 10, about 1 to about 20, or about 1 to about 30 mutations.

[0182] In some embodiments, an immunoglobulin heavy chain may comprise an entire heavy chain or a portion of a heavy chain. For example, a variable domain derived from a heavy chain or a region thereof may be referred to as a heavy chain or a region of a heavy chain. In some embodiments, an immunoglobulin light chain may comprise an entire light chain or a portion of a light chain. For example, a variable domain derived from a light chain or a region thereof may be referred to as a light chain or a region of a light chain. Single-domain immunoglobulins include, but are not limited to, a single monomeric variable immunoglobulin domain, for example, a shark variable novel antigen receptor immunoglobulin fragment (VNAR).

[0183] The immunoglobulin can be derived from any type known to those skilled in the art, including but not limited to IgA, IgD, IgE, IgG, IgM, IgY, IgW. The antibody region can comprise one or more units, including but not limited to 1, 2, 3, 4 and 5 units. Functional units can include but are not limited to non-antibody regions, heavy chains, light chains, variable domains, constant domains, complementary determining regions (CDRs), framework regions, fragment antigen binding (Fab) regions, Fab', F(ab')2, F(ab')3, Fab', fragment crystallizable (Fc) regions, single-chain variable fragments (scFV), di-scFv, single-domain immunoglobulins, trifunctional immunoglobulins, chemically linked F(ab')2 and any combination or fragment thereof. The non-antibody region includes but is not limited to carbohydrates, lipids, small molecules and therapeutic peptides. The antibody region can comprise one or more units connected by one or more disulfide bonds. The antibody region can comprise one or more units connected by a peptide connector, such as an scFv immunoglobulin. The immunoglobulin can be a recombinant immunoglobulin, including an immunoglobulin with amino acid mutations, substitutions and / or deletions. The immunoglobulin may be a recombinant immunoglobulin comprising chemical modifications.The immunoglobulin may comprise the whole or part of an immunoglobulin-drug conjugate.

[0184] The antibody region may comprise at least a portion of an immunoglobulin heavy chain. The antibody region may comprise one or more immunoglobulin heavy chains or portions thereof. The antibody region may comprise two or more immunoglobulin heavy chains or portions thereof. The antibody region may comprise an amino acid sequence that is at least about 50% homologous to an immunoglobulin heavy chain. The antibody region may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 95% or 97% or more homologous to an immunoglobulin heavy chain. The antibody region may comprise an amino acid sequence that is at least about 70% homologous to an immunoglobulin heavy chain. The antibody region may comprise an amino acid sequence that is at least about 80% homologous to an immunoglobulin heavy chain. The antibody region may comprise an amino acid sequence that is at least about 90% homologous to an immunoglobulin heavy chain. The immunoglobulin heavy chain may comprise amino acids based on or derived from any one of SEQ ID NOs: 22-27 and 29-35. In some embodiments, the antibody region comprises an amino acid sequence that is at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 97% homologous to the amino acid sequence of any one of SEQ ID NOs: 22-27 and 29-35. In some embodiments, the antibody region comprises an amino acid sequence that is at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 97% identical to the amino acid sequence of any one of SEQ ID NOs: 22-27 and 29-35.

[0185] The antibody region may comprise an amino acid sequence comprising 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90 or more amino acids of an immunoglobulin heavy chain. The antibody region may comprise an amino acid sequence comprising 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900 or more amino acids of an immunoglobulin heavy chain. The amino acids may be continuous. Alternatively, or additionally, the amino acids are discontinuous.

[0186] The immunoglobulin heavy chain can be encoded by a nucleotide sequence based on or derived from SEQ ID NOs: 4-9 and 11-17. The immunoglobulin heavy chain can be encoded by a nucleotide sequence that is at least about 50% homologous to SEQ ID NOs: 4-9 and 11-17. The immunoglobulin heavy chain can be encoded by a nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 95% or 97% or more homologous to SEQ ID NOs: 4-9 and 11-17. The immunoglobulin heavy chain can be encoded by a nucleotide sequence that is at least about 75% homologous to SEQ ID NOs: 4-9 and 11-17. The immunoglobulin heavy chain can be encoded by a nucleotide sequence that is at least about 85% homologous to SEQ ID NOs: 4-9 and 11-17. In some embodiments, the antibody region is encoded by a nucleotide sequence that is at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 97% homologous to the nucleotide sequence of any one of SEQ ID NOs: 4-9 and 11-17. In some embodiments, the antibody region is encoded by a nucleotide sequence that is at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 97% identical to the nucleotide sequence of any one of SEQ ID NOs: 4-9 and 11-17.

[0187] The antibody region may comprise at least a portion of an immunoglobulin light chain. The antibody region may comprise one or more immunoglobulin light chains or portions thereof. The antibody region may comprise two or more immunoglobulin light chains or portions thereof. The antibody region may comprise an amino acid sequence that is at least about 50% homologous to an immunoglobulin light chain. The antibody region may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 95% or 97% or more homologous to an immunoglobulin light chain. The antibody region may comprise an amino acid sequence that is at least about 70% homologous to an immunoglobulin light chain. The antibody region may comprise an amino acid sequence that is at least about 80% homologous to an immunoglobulin light chain. The antibody region may comprise an amino acid sequence that is at least about 90% homologous to an immunoglobulin light chain. The immunoglobulin light chain may comprise amino acids based on or derived from any one of SEQ ID NOs: 19-21, 28 and 36. In some embodiments, the antibody region comprises an amino acid sequence that is at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 97% homologous to the amino acid sequence of any one of SEQ ID NOs: 19-21, 28, and 36. In some embodiments, the antibody region comprises an amino acid sequence that is at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 97% identical to the amino acid sequence of any one of SEQ ID NOs: 19-21, 28, and 36.

[0188] The antibody region may comprise an amino acid sequence comprising 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90 or more amino acids of an immunoglobulin light chain. The antibody region may comprise an amino acid sequence comprising 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900 or more amino acids of an immunoglobulin light chain. The amino acids may be continuous. Alternatively, or additionally, the amino acids are discontinuous.

[0189] The immunoglobulin light chain can be encoded by a nucleotide sequence based on or derived from SEQ ID NOs: 1-3, 10, and 18. The immunoglobulin light chain can be encoded by a nucleotide sequence that is at least about 50% homologous to SEQ ID NOs: 1-3, 10, and 18. The immunoglobulin light chain can be encoded by a nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 95%, or 97% or more homologous to SEQ ID NOs: 1-3, 10, and 18. The immunoglobulin light chain can be encoded by a nucleotide sequence that is at least about 75% homologous to SEQ ID NOs: 1-3, 10, and 18. The immunoglobulin light chain can be encoded by a nucleotide sequence that is at least about 85% homologous to SEQ ID NOs: 1-3, 10, and 18. In some embodiments, the antibody region is encoded by a nucleotide sequence that is at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 97% homologous to the nucleotide sequence of any one of SEQ ID NOs: 1-3, 10, and 18. In some embodiments, the antibody region is encoded by a nucleotide sequence that is at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 97% identical to the nucleotide sequence of any one of SEQ ID NOs: 1-3, 10, and 18.

[0190] The antibody region may comprise at least a portion of a variable domain. The antibody region may comprise one or more variable domains or portions thereof. The antibody region may comprise 2, 3, 4, 5 or more variable domains or portions thereof. The antibody region may comprise an amino acid sequence of 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 140, 160, 180, 200, 225, 250, 275, 300, 350, 400, 500 or more amino acids based on or derived from the amino acid sequence of one or more variable domains. The amino acids may be continuous. The amino acids may be discontinuous.

[0191] The antibody region may comprise at least a portion of a constant domain. The antibody region may comprise one or more constant domains or portions thereof. The antibody region may comprise 2, 3, 4, 5, 6, 7, 8, 9, 10 or more constant domains or portions thereof. The antibody region may comprise an amino acid sequence of 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 140, 160, 180, 200, 225, 250, 275, 300, 350, 400, 500, 600, 700, 800, 900, 1000, 1200, 1400 or more amino acids based on or derived from the amino acid sequence of one or more constant domains. The amino acid may be continuous. The amino acid may be discontinuous.

[0192] The antibody region may comprise at least a portion of a complementary determining region (CDR). The antibody region may comprise one or more complementary determining regions (CDRs) or portions thereof. The antibody region may comprise 2, 3, 4, 5 or more complementary determining regions (CDRs) or portions thereof. The antibody region may comprise 6, 7, 8 or more complementary determining regions (CDRs) or portions thereof. The antibody region may comprise four or more complementary determining regions (CDRs) or portions thereof. The antibody region may comprise 9, 10, 11 or more complementary determining regions (CDRs) or portions thereof. The one or more CDRs may be CDR1, CDR2, CDR3 or a combination thereof. The one or more CDRs may be CDR1. The one or more CDRs may be CDR2. The one or more CDRs may be CDR3. The CDRs may be heavy chain CDRs. The one or more CDRs may be light chain CDRs.

[0193] The antibody region may comprise an amino acid sequence containing 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acids based on or derived from the amino acid sequence of the CDR. The antibody region may comprise an amino acid sequence containing 3 or more amino acids based on or derived from the amino acid sequence of the CDR. The antibody region may comprise an amino acid sequence containing 5 or more amino acids based on or derived from the amino acid sequence of the CDR. The antibody region may comprise an amino acid sequence containing 10 or more amino acids based on or derived from the amino acid sequence of the CDR. The amino acids may be continuous. The amino acids may be discontinuous.

[0194] The antibody region may be based on or derived from at least a portion of an anti-T cell receptor immunoglobulin.The antibody region may be based on or derived from at least a portion of an anti-B cell receptor immunoglobulin.

[0195] The antibody region may be based on or derived from at least a portion of an anti-T cell co-receptor immunoglobulin. The antibody region may be based on or derived from at least a portion of an anti-CD3 immunoglobulin. The antibody region may be based on or derived from an anti-CD3 immunoglobulin. The anti-CD3 immunoglobulin may be UCHT1. The antibody region may be based on or derived from at least a portion of a Fab fragment of an anti-CD3 immunoglobulin. The antibody region may be based on or derived from an immunoglobulin fragment of an anti-CD3 immunoglobulin.

[0196] The antibody region may be based on or derived from an immunoglobulin or immunoglobulin fragment that binds to at least a portion of a receptor on a cell. The antibody region may be based on or derived from an immunoglobulin or immunoglobulin fragment that binds to at least a portion of a co-receptor on a cell. The antibody region may be based on or derived from an immunoglobulin or immunoglobulin fragment that binds to at least a portion of an antigen or a cell surface marker on a cell. The cell may be a hematopoietic cell. The hematopoietic cell may be a bone marrow cell. The bone marrow cell may be an erythrocyte, a thrombocyte, a neutrophil, a monocyte, a macrophage, an eosinophil, a basophil, or a mast cell. The hematopoietic cell may be a lymphoid cell. The lymphoid cell may be a B cell, a T cell, or a NK cell. The hematopoietic cell may be a leukocyte. The hematopoietic cell may be a lymphocyte.

[0197] The antibody region can be based on or derived from an immunoglobulin or immunoglobulin fragment that binds to at least a portion of a receptor on a T cell. The receptor can be a T cell receptor (TCR). The TCR can include TCR alpha, TCR beta, TCR gamma and / or TCR delta. The receptor can be a T cell receptor zeta.

[0198] The antibody region can be based on or derived from an immunoglobulin or immunoglobulin fragment that binds to at least a portion of a receptor on a lymphocyte, B cell, macrophage, monocyte, neutrophil and / or NK cell. The receptor can be an Fc receptor. The Fc receptor can be an Fc-γ receptor, an Fc-α receptor and / or an Fc-ε receptor. Fc-γ receptors include but are not limited to FcγRI (CD64), FcγRIIA (CD32), FcγRIIB (CD32), FcγRIIIA (CD16a) and FcγRIIIB (CD16b). Fc-α receptors include but are not limited to FcαRI. Fc-ε receptors include but are not limited to FcεRI and FcεRII. The receptor can be CD89 (Fc fragment of IgA receptor or FCAR).

[0199] The antibody region may be based on or derived from an immunoglobulin or immunoglobulin fragment that binds to at least a portion of a co-receptor on a T cell. The co-receptor may be CD3, CD4, and / or CD8. The antibody region may be based on or derived from an immunoglobulin fragment that binds to a CD3 co-receptor. The CD3 co-receptor may include CD3-γ, CD3-Δ, and / or CD3-ε. CD8 may comprise CD8-α and / or CD8-β chains.

[0200] In some embodiments, the antibody region is not specific for a mammalian target. In some embodiments, the immunoglobulin is an antiviral immunoglobulin. In some embodiments, the immunoglobulin is an antibacterial immunoglobulin. In some embodiments, the immunoglobulin is an antiparasitic immunoglobulin. In some embodiments, the immunoglobulin is an antifungal immunoglobulin. In some embodiments, the antibody region is derived from an immunoglobulin vaccine.

[0201] In some embodiments, the antibody region is based on or derived from an immunoglobulin including, but not limited to, actoxumab, bezlotoxumab, CR6261, ebalizumab, ifengolmab, ivavirumab, fovelumab, fulavirumab, ibalizumab (TMB-355, TNX-355), rivimab, motavirumab, nebacilumab, paxilumab, palivizumab, panocurumab, ravirumab, ravirumab, regavir, sevirumab (MSL-109), suvizumab, tefizumab, tuvirumab, and urovituximab.

[0202] In some embodiments, the antibody region is based on or derived from an immunoglobulin against Clostridium difficile, Orthomyxoviruses (Influenzavirus A, Influenzavirus B, Influenzavirus C, Isavirus, Thogotovirus), Escherichia coli, Candida, Rabies, Human Immunodeficiency Virus, Hepatitis, Staphylococcus, Respiratory Syncytial Virus, Pseudomonas aeruginosa, Bacillus anthracis, Cytomegalovirus, or Staphylococcus aureus.

[0203] Antibody region can be based on or derived from antiviral immunoglobulin. Antiviral immunoglobulin can be directed against the epitope of viral protein. Antibacterial immunoglobulin can target one or more viruses, including but not limited to adenovirus (Adenoviruses), herpesvirus (Herpesviruses), poxvirus (Poxviruses), parvovirus (Parvoviruses), reovirus (Reoviruses), picornavirus (Picornaviruses), togavirus (Togaviruses), orthomyxovirus, rhabdovirus (Rhabdoviruses), retrovirus (Retroviruses) and hepadnavirus (Hepadnaviruses). Viral protein can be from respiratory syncytial virus. Viral protein can be the F protein of respiratory syncytial virus. The epitope can be in the A antigen site of F protein. Antiviral immunoglobulin can be based on or derived from palivizumab. Immunoglobulin can be based on or derived from antiviral vaccine. The antiviral immunoglobulin may be based on or derived from elvirumab, flavuzumab, rivuzumab, ramucirumab, regavir, sevirumab, tuvirumab, fovirizumab, motavizumab, palivizumab, and / or suvizumab.

[0204] The antibody region may be based on or derived from antiviral immunoglobulin G. The antibody region may comprise at least a portion of antiviral immunoglobulin G. The antibody region may comprise an amino acid sequence that is at least about 50% homologous to at least a portion of antiviral immunoglobulin G. The antibody region may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 95% or 97% or more homologous to at least a portion of antiviral immunoglobulin G. The antibody region may comprise an amino acid sequence that is at least about 70% homologous to at least a portion of antiviral immunoglobulin G. The antibody region may comprise an amino acid sequence that is at least about 80% homologous to at least a portion of antiviral immunoglobulin G. In some embodiments, the antibody region comprises an amino acid sequence based on or derived from antiviral immunoglobulin M.

[0205] The antibody region may comprise an amino acid sequence of 10, 20, 30, 40, 50, 60, 70, 80, 90 or more amino acids of an antiviral immunoglobulin G sequence. The antibody region may comprise an amino acid sequence of 100, 200, 300, 400, 500, 600, 700, 800, 900 or more amino acids of an antiviral immunoglobulin G sequence. The antibody region may comprise an amino acid sequence of 50 or more amino acids of an antiviral immunoglobulin G sequence. The antibody region may comprise an amino acid sequence of 100 or more amino acids of an antiviral immunoglobulin G sequence. The antibody region may comprise an amino acid sequence of 200 or more amino acids of an antiviral immunoglobulin G sequence.

[0206] The antibody region may be based on or derived from a palivizumab immunoglobulin. The antibody region may comprise at least a portion of a palivizumab immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 50% homologous to at least a portion of a palivizumab immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 95% or 97% or more homologous to at least a portion of a palivizumab immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 70% homologous to at least a portion of a palivizumab immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 80% homologous to at least a portion of a palivizumab immunoglobulin.

[0207] The antibody region may comprise an amino acid sequence comprising 10, 20, 30, 40, 50, 60, 70, 80, 90 or more amino acids of a palivizumab immunoglobulin sequence. The antibody region may comprise an amino acid sequence comprising 100, 200, 300, 400, 500, 600, 700, 800, 900 or more amino acids of a palivizumab immunoglobulin sequence. The antibody region may comprise an amino acid sequence comprising 50 or more amino acids of a palivizumab immunoglobulin sequence. The antibody region may comprise an amino acid sequence comprising 100 or more amino acids of a palivizumab immunoglobulin sequence. The antibody region may comprise an amino acid sequence comprising 200 or more amino acids of a palivizumab immunoglobulin sequence.

[0208] The antibody region can be based on or derived from ividavir, fulavirumab, rivimab, revimab, regavir, sevirumab, tuvirumab, panvizumab, motavizumab, palivizumab and / or suvizumab immunoglobulins. The antibody region can comprise at least a portion of ividavir, fulavirumab, rivimab, revimab, regavir, sevirumab, tuvirumab, panvizumab, motavizumab, palivizumab and / or suvizumab immunoglobulins. The antibody region can comprise an amino acid sequence that is at least about 50% homologous to at least a portion of ividavir, fulavirumab, rivimab, revimab, regavir, sevirumab, tuvirumab, panvizumab, motavizumab, palivizumab and / or suvizumab immunoglobulins. The antibody region may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 95% or 97% or more homologous to at least a portion of an ividavir, flavavirumab, rividarumab, revidarumab, regavir, sevirumab, tuvirumab, fovirizumab, motavizumab, palivizumab and / or suvizumab immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 70% homologous to at least a portion of an ividavir, flavavirumab, rividarumab, revidarumab, regavir, sevirumab, tuvirumab, fovirizumab, motavizumab, palivizumab and / or suvizumab immunoglobulin. The antibody region can comprise an amino acid sequence that is at least about 80% homologous to at least a portion of the elvirumab, flavuzumab, rivuzumab, ramucirumab, regavir, sevirumab, tuvirumab, panavizumab, motavizumab, palivizumab, and / or suvizumab immunoglobulin.

[0209] The antibody region may comprise an amino acid sequence comprising 10, 20, 30, 40, 50, 60, 70, 80, 90 or more amino acids of the immunoglobulin sequence of ivavirmab, fulavirmab, rivavirmab, revavirmab, regavir, sevimab, tuviromab, panvizumab, motavizumab, palivizumab and / or suvizumab. The antibody region may comprise an amino acid sequence comprising 100, 200, 300, 400, 500, 600, 700, 800, 900 or more amino acids of the immunoglobulin sequence of ivavirmab, fulavirmab, rivavirmab, revavirmab, regavir, sevimab, tuviromab, panvizumab, motavizumab, palivizumab and / or suvizumab. The antibody region may comprise an amino acid sequence comprising 50 or more amino acids of the immunoglobulin sequence of ividavir, flavavirumab, rividarumab, revidarumab, regavir, sevirumab, tuvirumab, panvizumab, motavizumab, palivizumab and / or suvizumab. The antibody region may comprise an amino acid sequence comprising 100 or more amino acids of the immunoglobulin sequence of ividavir, flavavirumab, rividarumab, revidarumab, regavir, sevirumab, tuvirumab, panvizumab, motavizumab, palivizumab and / or suvizumab. The antibody region may comprise an amino acid sequence of 200 or more amino acids comprising the ivavirumab, flavuzumab, rivavirumab, ramavirumab, regavirumab, sevirumab, tuvirumab, fovirizumab, motavirumab, palivizumab, and / or suvizumab immunoglobulin sequence.

[0210] The antibody region may be based on or derived from an antibacterial immunoglobulin. The antibacterial immunoglobulin may be directed against an epitope of a bacterial protein. The antibacterial immunoglobulin can target bacteria including, but not limited to, Acetobacteraurantius, Agrobacterium radiobacter, Anaplasma phagocytophilum, Azorhizobium caulinodans, Bacillus anthracis, Bacillus brevis, Bacillus cereus, Bacillus subtilis, Bacteroides fragilis, Bacteroides gingivalis, Bacteroides melaninogenicus, Bartonella quintana, Bordetella bronchiseptica, Bordetella pertussis, Borrelia burgdorferi), Brucella abortus, Brucella melitensis, Brucella suis, Burkholderia mallei, Burkholderia pseudomallei, Burkholderia cepacia, Calymmatobacterium granulomatis, Campylobacter coli, Campylobacter fetus, Campylobacter jejuni, Helicobacter pylori, Chlamydia trachomatis, Chlamydophila pneumoniae, Chlamydophila psittaci, Clostridium botulinum, Clostridium difficile, Corynebacteriumdiphtheriae), Corynebacterium fusiforme, Coxiella burnetii, Enterobacter cloacae, Enterococcus faecalis, Enterococcus faecium, Enterococcus galllinarum, Enterococcus maloratus, Escherichia coli, Francisella tularensis, Fusobacterium nucleatum, Gardnerella vaginalis, Haemophilus influenzae, Haemophilus parainfluenzae, Haemophilus pertussis, Gardnerella vaginalis, Helicobacter pylori, Klebsiella pneumoniae pneumoniae), Lactobacillus acidophilus, Lactococcus lactis, Legionella pneumophila, Listeria monocytogenes, Methanobacterium extroquens, Microbacterium multiforme, Micrococcus luteus, Moraxella catarrhalis, Mycobacterium phlei, Mycobacterium smegmatis, Mycobacterium tuberculosis, Mycoplasma genitalium, Mycoplasma hominis, Mycoplasma pneumoniae, Neisseria gonorrhoeae, Neisseria meningitidis meningitidis), Pasteurella multocidamultocida), Pasteurella tularensis, Peptostreptococcus, Porphyromonas gingivalis, Prevotella melaninogenica, Pseudomonas aeruginosa, Rhizobium radiobacter, Rickettsia rickettsii, Rothia dentocariosa, Salmonella enteritidis, Salmonella typhi, Salmonella typhimurium, Shigella dysenteriae, Staphylococcus aureus, Staphylococcus epidermidis, Stenotrophomonas maltophilia maltophilia, Streptococcus pneumoniae, Streptococcus pyogenes, Treponema allidum, Treponema denticola, Vibrio cholerae, Vibrio comma, Vibrio parahaemolyticus, Vibrio vulnificus, Yersinia enterocolitica, and Yersinia pseudotuberculosis. The immunoglobulins can be based on or derived from bacterial vaccines. The antiviral immunoglobulins can be based on or derived from nebacumab, panocumab, raxicumab, ebumab, paxilumab, and / or tefizumab.

[0211] The antibody region may be based on or derived from antibacterial immunoglobulin G. The antibody region may comprise at least a portion of antibacterial immunoglobulin G. The antibody region may comprise an amino acid sequence that is at least about 50% homologous to at least a portion of antibacterial immunoglobulin G. The antibody region may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 95% or 97% or more homologous to at least a portion of antibacterial immunoglobulin G. The antibody region may comprise an amino acid sequence that is at least about 70% homologous to at least a portion of antibacterial immunoglobulin G. The antibody region may comprise an amino acid sequence that is at least about 80% homologous to at least a portion of antibacterial immunoglobulin G. In some embodiments, the antibody region comprises an amino acid sequence based on or derived from antiviral immunoglobulin M.

[0212] The antibody region may comprise an amino acid sequence of 10, 20, 30, 40, 50, 60, 70, 80, 90 or more amino acids of an antibacterial immunoglobulin G sequence. The antibody region may comprise an amino acid sequence of 100, 200, 300, 400, 500, 600, 700, 800, 900 or more amino acids of an antibacterial immunoglobulin G sequence. The antibody region may comprise an amino acid sequence of 50 or more amino acids of an antibacterial immunoglobulin G sequence. The antibody region may comprise an amino acid sequence of 100 or more amino acids of an antibacterial immunoglobulin G sequence. The antibody region may comprise an amino acid sequence of 200 or more amino acids of an antibacterial immunoglobulin G sequence.

[0213] The antibody region may be based on or derived from a nebacumab, panocumab, raxicumab, ebumab, paliximumab, and / or tefizumab immunoglobulin. The antibody region may comprise at least a portion of a nebacumab, panocumab, raxicumab, ebumab, paliximumab, and / or tefizumab immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 50% homologous to at least a portion of a nebacumab, panocumab, raxicumab, ebumab, paliximumab, and / or tefizumab immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 95%, or 97% or more homologous to at least a portion of a nebacumab, panocumab, raxicumab, ebumab, paliximumab, and / or tefizumab immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 70% homologous to at least a portion of a nebacumab, panocumab, raxicumab, ebumab, paliximumab, and / or tefizumab immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 80% homologous to at least a portion of a nebacumab, panocumab, raxicumab, ebumab, paliximumab, and / or tefizumab immunoglobulin.

[0214] The antibody region may comprise an amino acid sequence comprising 10, 20, 30, 40, 50, 60, 70, 80, 90 or more amino acids of the immunoglobulin sequence of nebacumab, panocumab, raxicumab, ebumab, paliximumab and / or tefizumab. The antibody region may comprise an amino acid sequence comprising 100, 200, 300, 400, 500, 600, 700, 800, 900 or more amino acids of the immunoglobulin sequence of nebacumab, panocumab, raxicumab, ebumab, paliximumab and / or tefizumab. The antibody region may comprise an amino acid sequence comprising 50 or more amino acids of the immunoglobulin sequence of nebacumab, panocumab, raxicumab, ebumab, paliximumab and / or tefizumab. The antibody region may comprise an amino acid sequence of 100 or more amino acids comprising the immunoglobulin sequence of nebacumab, panocumab, raxicumab, ebumab, paxilumab, and / or teficillin. The antibody region may comprise an amino acid sequence of 200 or more amino acids comprising the immunoglobulin sequence of nebacumab, panocumab, raxicumab, ebumab, paxilumab, and / or teficillin.

[0215] The antibody region may be based on or derived from an antiparasitic immunoglobulin. The antiparasitic immunoglobulin may be directed against an epitope of a parasite protein. Antiparasitic immunoglobulins can target parasites or parasite proteins, including but not limited to the parasites Acanthamoeba, Balamuthia mandrillaris, Babesia (B. divergens, B. bigemina, B. equi, B. microfti, B. duncani, Balantidium coli, Blastocystis, Cryptosporidium, Dientamoeba fragilis, Entamoeba histolytica, Giardia lamblia, Isospora belli, Leishmania, Naegleria fowleri, and fowleri), Plasmodium falciparum, Plasmodium vivax, Plasmodium ovale curtisi, Plasmodium ovale wallikeri, Plasmodium malariae, Plasmodium knowlesi, Rhinosporidium seeberi, Sarcocystis bovihominis, Sarcocystis suihominis, Toxoplasma gondii, Trichomonas vaginalis, Trypanosoma brucei, Trypanosoma cruzi, Cestoda, Taenia multiceps), Diphyllobothrium latum, Echinococcus granulosus, Echinococcus multilocularis, Echinococcus wollerivogeli), Echinococcus oligarthrus, Hymenolepis nana, Hymenolepis diminuta, Taenia saginata, Taenia solium, Bertiella mucronata, Bertiella studeri, Spirometra erinacei ieuropaei, Clonorchis sinensis; Clonorchis viverrini, Dicrocoelium dendriticum, Fasciola hepatica, Fasciola gigantica, Fasciolopsis brucei buski), Gnathostomaspinigerum, Gnathostoma hispidum, Metagonimus yokogawai, Opisthorchis viverrini, Opisthorchis felineus, Clonorchis sinensis, Paragonimus westermani; Paragonimus africanus; Paragonimus caliensis; Paragonimus kellicotti; Paragonimus skrjabini; Paragonimus uterobilateralis; Schistosoma sp., Schistosoma mansoni, Schistosoma haematobium haematobium), Schistosoma japonicum, Schistosoma mekongi, Echinostomaechinatum, Trichobilharzia regenti, Schistosomatidae, Ancylostoma duodenale, Necatoramericanus), Angiostrongylus costaricensis, Anisakis, Ascaris species (Ascaris lumbricoides), Baylisascaris procyonis, Brugia malayi, Brugia timori, Dioctophyme renale, Dracunculus medinensis, Enterobius vermicularis, Enterobius gregorii, Halicephalobus gingivalis, Loa filaria, Mansonella streptocerca, Onchocerca volvulus, Strongyloides stercoralis, Thelazia californiensis), Thelazia callipaeda, Toxocara canis, Toxocaracati, Trichinella spiralis, Trichinella britovi, Trichinella nelsoni, Trichinella nativa, Trichuris trichiura, Trichuris vulpis, Wuchereria bancrofti, Archiacanthocephala, Moniliformis moniliformis, Linguatula serrata, Oestroidea, Calliphoridae, Sarcophagidae, Tunga penetrans, Dermatobia hominis), Ixodidae, Argasidae, Cimex lectularius, Pediculus humanus, Pediculus humanuscorporis), Pthirus pubis, Demodex folliculorum / brevis / canis, Sarcoptes scabiei, Cochliomyia hominivorax, and Pulex irritans.

[0216] The antibody region may be based on or derived from antiparasitic immunoglobulin G. The antibody region may comprise at least a portion of antiparasitic immunoglobulin G. The antibody region may comprise an amino acid sequence that is at least about 50% homologous to at least a portion of antiparasitic immunoglobulin G. The antibody region may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 95% or 97% or more homologous to at least a portion of antiparasitic immunoglobulin G. The antibody region may comprise an amino acid sequence that is at least about 70% homologous to at least a portion of antiparasitic immunoglobulin G. The antibody region may comprise an amino acid sequence that is at least about 80% homologous to at least a portion of antiparasitic immunoglobulin G. In some embodiments, the antibody region comprises an amino acid sequence based on or derived from antiparasitic immunoglobulin M.

[0217] The antibody region may comprise an amino acid sequence of 10, 20, 30, 40, 50, 60, 70, 80, 90 or more amino acids of an antiparasitic immunoglobulin G sequence. The antibody region may comprise an amino acid sequence of 100, 200, 300, 400, 500, 600, 700, 800, 900 or more amino acids of an antiparasitic immunoglobulin G sequence. The antibody region may comprise an amino acid sequence of 50 or more amino acids of an antiparasitic immunoglobulin G sequence. The antibody region may comprise an amino acid sequence of 100 or more amino acids of an antiparasitic immunoglobulin G sequence. The antibody region may comprise an amino acid sequence of 200 or more amino acids of an antiparasitic immunoglobulin G sequence.

[0218] The antibody region may be based on or derived from an antifungal immunoglobulin. The antibacterial immunoglobulin may be directed against an epitope of a fungal protein. The antifungal immunoglobulins can target fungi or fungal proteins, including but not limited to Cryptococcus neoformans, Cryptococcus gattii, Candida albicans, Candida tropicalis, Candida stellatoidea, Candida glabrata, Candida krusei, Candida parapsilosis, Candida guilliermondii, Candida viswanathii, Candida lusitaniae, Rhodotorula mucilaginosa, Schizosaccharomyces pombe, Saccharomyces cerevisiae, Brettanomyces bruxellensis, Candida stellata), Schizosaccharomyces pombe, Torulaspora delbrueckii, Zygosaccharomyces bailii, Yarrowia lipolytica, Saccharomyces exiguus, and Pichia pastoris. The antifungal immunoglobulin may be based on or derived from ifingucomab.

[0219] The antibody region may be based on or derived from antifungal immunoglobulin G. The antibody region may comprise at least a portion of antifungal immunoglobulin G. The antibody region may comprise an amino acid sequence that is at least about 50% homologous to at least a portion of antifungal immunoglobulin G. The antibody region may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 95%, or 97% or more homologous to at least a portion of antifungal immunoglobulin G. The antibody region may comprise an amino acid sequence that is at least about 70% homologous to at least a portion of antifungal immunoglobulin G. The antibody region may comprise an amino acid sequence that is at least about 80% homologous to at least a portion of antifungal immunoglobulin G. In some embodiments, the antibody region comprises an amino acid sequence based on or derived from antifungal immunoglobulin M.

[0220] The antibody region may comprise an amino acid sequence comprising 10, 20, 30, 40, 50, 60, 70, 80, 90 or more amino acids of an antifungal immunoglobulin G sequence. The antibody region may comprise an amino acid sequence comprising 100, 200, 300, 400, 500, 600, 700, 800, 900 or more amino acids of an antifungal immunoglobulin G sequence. The antibody region may comprise an amino acid sequence comprising 50 or more amino acids of an antifungal immunoglobulin G sequence. The antibody region may comprise an amino acid sequence comprising 100 or more amino acids of an antifungal immunoglobulin G sequence. The antibody region may comprise an amino acid sequence comprising 200 or more amino acids of an antifungal immunoglobulin G sequence.

[0221] The antibody region may be based on or derived from an ifenggu monoclonal antibody immunoglobulin. The antibody region may comprise at least a portion of an ifenggu monoclonal antibody immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 50% homologous to at least a portion of an ifenggu monoclonal antibody immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 95% or 97% or more homologous to at least a portion of an ifenggu monoclonal antibody immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 70% homologous to at least a portion of an ifenggu monoclonal antibody immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 80% homologous to at least a portion of an ifenggu monoclonal antibody immunoglobulin.

[0222] The antibody region may comprise an amino acid sequence of 10, 20, 30, 40, 50, 60, 70, 80, 90 or more amino acids of the immunoglobulin sequence of Ifinggu monoclonal antibody. The antibody region may comprise an amino acid sequence of 100, 200, 300, 400, 500, 600, 700, 800, 900 or more amino acids of the immunoglobulin sequence of Ifinggu monoclonal antibody. The antibody region may comprise an amino acid sequence of 50 or more amino acids of the immunoglobulin sequence of Ifinggu monoclonal antibody. The antibody region may comprise an amino acid sequence of 100 or more amino acids of the immunoglobulin sequence of Ifinggu monoclonal antibody. The antibody region may comprise an amino acid sequence of 200 or more amino acids of the immunoglobulin sequence of Ifinggu monoclonal antibody.

[0223] The antibody region may be based on or derived from the trastuzumab immunoglobulin G immunoglobulin. The antibody region may comprise at least a portion of the trastuzumab immunoglobulin G immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 50% homologous to at least a portion of the trastuzumab immunoglobulin G immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 95%, or 97% or more homologous to at least a portion of the trastuzumab immunoglobulin G immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 70% homologous to at least a portion of the trastuzumab immunoglobulin G immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 80% homologous to at least a portion of the trastuzumab immunoglobulin G immunoglobulin. The antibody region may comprise a mutated trastuzumab antibody. The antibody region may comprise a trastuzumab antibody comprising seven mutations in the IgG1 heavy chain. The antibody region may comprise a trastuzumab antibody comprising three mutations in an IgG4 heavy chain.

[0224] The antibody region may comprise an amino acid sequence comprising 10, 20, 30, 40, 50, 60, 70, 80, 90 or more amino acids of the trastuzumab immunoglobulin G immunoglobulin sequence. The antibody region may comprise an amino acid sequence comprising 100, 200, 300, 400, 500, 600, 700, 800, 900 or more amino acids of the trastuzumab immunoglobulin G immunoglobulin sequence. The antibody region may comprise an amino acid sequence comprising 50 or more amino acids of the trastuzumab immunoglobulin G immunoglobulin sequence. The antibody region may comprise an amino acid sequence comprising 100 or more amino acids of the trastuzumab immunoglobulin G immunoglobulin sequence. The antibody region may comprise an amino acid sequence comprising 200 or more amino acids of the trastuzumab immunoglobulin G immunoglobulin sequence.

[0225] The antibody region may be based on or derived from an anti-Her2 immunoglobulin. The antibody region may comprise at least a portion of an anti-Her2 immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 50% homologous to at least a portion of an anti-Her2 immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 95% or 97% or higher homologous to at least a portion of an anti-Her2 immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 70% homologous to at least a portion of an anti-Her2 immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 80% homologous to at least a portion of an anti-Her2 immunoglobulin.

[0226] The antibody region may comprise an amino acid sequence of 10, 20, 30, 40, 50, 60, 70, 80, 90 or more amino acids of an anti-Her2 immunoglobulin sequence. The antibody region may comprise an amino acid sequence of 100, 200, 300, 400, 500, 600, 700, 800, 900 or more amino acids of an anti-Her2 immunoglobulin sequence. The antibody region may comprise an amino acid sequence of 50 or more amino acids of an anti-Her2 immunoglobulin sequence. The antibody region may comprise an amino acid sequence of 100 or more amino acids of an anti-Her2 immunoglobulin sequence. The antibody region may comprise an amino acid sequence of 200 or more amino acids of an anti-Her2 immunoglobulin sequence.

[0227] The antibody region may be based on or derived from an anti-CD47 immunoglobulin. The antibody region may comprise at least a portion of an anti-CD47 immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 50% homologous to at least a portion of an anti-CD47 immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 95%, or 97% or more homologous to at least a portion of an anti-CD47 immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 70% homologous to at least a portion of an anti-CD47 immunoglobulin. The antibody region may comprise an amino acid sequence that is at least about 80% homologous to at least a portion of an anti-CD47 immunoglobulin.

[0228] The antibody region may comprise an amino acid sequence comprising 10, 20, 30, 40, 50, 60, 70, 80, 90 or more amino acids of an anti-CD47 immunoglobulin sequence. The antibody region may comprise an amino acid sequence comprising 100, 200, 300, 400, 500, 600, 700, 800, 900 or more amino acids of an anti-CD47 immunoglobulin sequence. The antibody region may comprise an amino acid sequence comprising 50 or more amino acids of an anti-CD47 immunoglobulin sequence. The antibody region may comprise an amino acid sequence comprising 100 or more amino acids of an anti-CD47 immunoglobulin sequence. The antibody region may comprise an amino acid sequence comprising 200 or more amino acids of an anti-CD47 immunoglobulin sequence.

[0229] The antibody region can be based on or derived from an anti-cancer immunoglobulin. Examples of anti-cancer immunoglobulins include, but are not limited to, abciximab, adalimumab, alemtuzumab, basiliximab, belimumab, bevacizumab, brentuximab, canakinumab, certolizumab pegol, cetuximab, daclizumab, denosumab, eculizumab, efalizumab, gemtuzumab, golimumab, ibritumomab tiuxetan, infliximab, ipilimumab, muromonab-cd3, natalizumab, ofatumumab, omalizumab, palivizumab, panitumumab, ranibizumab, rituximab, tocilizumab, tositumomab, and trastuzumab.

[0230] The antibody region may comprise at least a portion of a human immunoglobulin. The antibody region may comprise at least a portion of a humanized immunoglobulin. The antibody region may comprise at least a portion of a chimeric immunoglobulin. The antibody region may be based on or derived from a human immunoglobulin. The antibody region may be based on or derived from a humanized immunoglobulin. The antibody region may be based on or derived from a chimeric immunoglobulin. The antibody region may be based on or derived from a monoclonal immunoglobulin. The antibody region may be based on or derived from a polyclonal immunoglobulin. The antibody region may comprise at least a portion of an immunoglobulin from a mammal, bird, reptile, amphibian, or a combination thereof. The mammal may be a human. The mammal may be a non-human primate. The mammal may be a dog, cat, sheep, goat, cow, rabbit, or mouse.

[0231] The antibody region can comprise a sequence based on or derived from one or more immunoglobulins and / or immunoglobulin fragment sequences. The antibody region can comprise a sequence that is at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher homologous to a sequence based on or derived from one or more immunoglobulins and / or immunoglobulin fragment sequences. The antibody region can comprise a sequence that is at least about 70% homologous to a sequence based on or derived from one or more immunoglobulins and / or immunoglobulin fragment sequences. The antibody region can comprise a sequence that is at least about 80% homologous to a sequence based on or derived from one or more immunoglobulins and / or immunoglobulin fragment sequences. The antibody region can comprise a sequence that is at least about 90% homologous to a sequence based on or derived from one or more immunoglobulins and / or immunoglobulin fragment sequences. The antibody region can comprise a sequence that is at least about 95% homologous to a sequence based on or derived from one or more immunoglobulins and / or immunoglobulin fragment sequences. The sequence may be a peptide sequence. The sequence may be a nucleotide sequence.

[0232] The antibody region may comprise a peptide sequence that differs from a peptide sequence based on or derived from one or more immunoglobulins and / or immunoglobulin fragments by less than or equal to about 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, 20, 17, 15, 12, 10, 8, 6, 5, 4 or fewer amino acids. The antibody region may comprise a peptide sequence that differs from a peptide sequence based on or derived from one or more immunoglobulins and / or immunoglobulin fragments by less than or equal to about 4 or fewer amino acids. The antibody region may comprise a peptide sequence that differs from a peptide sequence based on or derived from one or more immunoglobulins and / or immunoglobulin fragments by less than or equal to about 3 or fewer amino acids. The antibody region may comprise a peptide sequence that differs from a peptide sequence based on or derived from one or more immunoglobulins and / or immunoglobulin fragments by less than or equal to about 2 or fewer amino acids. The antibody region may comprise a peptide sequence that is different from a peptide sequence based on or derived from one or more immunoglobulins and / or immunoglobulin fragments by less than or equal to about 1 or less amino acid. The amino acids may be continuous, discontinuous, or a combination thereof. For example, the antibody region may comprise a peptide sequence that is different from a peptide sequence based on or derived from one or more immunoglobulins and / or immunoglobulin fragments by less than about 3 consecutive amino acids. Alternatively, or additionally, the antibody region may comprise a peptide sequence that is different from a peptide sequence based on or derived from one or more immunoglobulins and / or immunoglobulin fragments by less than about 2 discontinuous amino acids. In another example, the antibody region may comprise a peptide sequence that is different from a peptide sequence based on or derived from one or more immunoglobulins and / or immunoglobulin fragments by less than about 5 amino acids, wherein 2 amino acids are continuous and 2 amino acids are discontinuous.

[0233] The antibody region can comprise a nucleotide sequence that is less than or equal to about 500, 400, 300, 200, 100, 90, 80, 70, 60, 50, 40, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4 or more nucleotides or base pairs different from the nucleotide sequence based on or derived from one or more antibodies and / or immunoglobulin fragments. The antibody region can comprise a nucleotide sequence that is less than or equal to about 15 or more nucleotides or base pairs different from the nucleotide sequence based on or derived from one or more immunoglobulins and / or immunoglobulin fragments. The antibody region can comprise a nucleotide sequence that is less than or equal to about 12 or more nucleotides or base pairs different from the nucleotide sequence based on or derived from one or more immunoglobulins and / or immunoglobulin fragments. The antibody region may comprise a nucleotide sequence that differs from a nucleotide sequence based on or derived from one or more immunoglobulins and / or immunoglobulin fragments in less than or equal to about 9 or fewer nucleotides or base pairs. The antibody region may comprise a nucleotide sequence that differs from a nucleotide sequence based on or derived from one or more immunoglobulins and / or immunoglobulin fragments in less than or equal to about 6 or fewer nucleotides or base pairs. The antibody region may comprise a nucleotide sequence that differs from a nucleotide sequence based on or derived from one or more immunoglobulins and / or immunoglobulin fragments in less than or equal to about 4 or fewer nucleotides or base pairs. The antibody region may comprise a nucleotide sequence that differs from a nucleotide sequence based on or derived from one or more immunoglobulins and / or immunoglobulin fragments in less than or equal to about 3 or fewer nucleotides or base pairs. The antibody region may comprise a nucleotide sequence that differs from a nucleotide sequence based on or derived from one or more immunoglobulins and / or immunoglobulin fragments in less than or equal to about 2 or fewer nucleotides or base pairs. The antibody region can include a nucleotide sequence that is less than or equal to about 1 or less nucleotides or base pairs based on or derived from a nucleotide sequence of one or more immunoglobulins and / or immunoglobulin fragments. The nucleotides or base pairs can be continuous, discontinuous, or a combination thereof. For example, the antibody region can include a nucleotide sequence that is less than about 3 continuous nucleotides or base pairs based on or derived from a nucleotide sequence of one or more immunoglobulins and / or immunoglobulin fragments. Alternatively, or in addition, the antibody region can include a nucleotide sequence that is less than about 2 discontinuous nucleotides or base pairs based on or derived from a nucleotide sequence of one or more immunoglobulins and / or immunoglobulin fragments. In another example, the antibody region can include a nucleotide sequence that is less than about 5 nucleotides or base pairs based on or derived from a nucleotide sequence of one or more immunoglobulins and / or immunoglobulin fragments, wherein 2 nucleotides or base pairs are continuous and 2 nucleotides or base pairs are discontinuous.

[0234] The peptide sequence of the antibody region may differ from the peptide sequence of the immunoglobulin or immunoglobulin fragment on which it is based and / or derived from by one or more amino acid substitutions. The peptide sequence of the antibody region may differ from the peptide sequence of the immunoglobulin or immunoglobulin fragment on which it is based and / or derived from by two or more amino acid substitutions. The peptide sequence of the antibody region may differ from the peptide sequence of the immunoglobulin or immunoglobulin fragment on which it is based and / or derived from by three or more amino acid substitutions. The peptide sequence of the antibody region may differ from the peptide sequence of the immunoglobulin or immunoglobulin fragment on which it is based and / or derived from by four or more amino acid substitutions. The peptide sequence of the antibody region may differ from the peptide sequence of the immunoglobulin or immunoglobulin fragment on which it is based and / or derived from by five or more amino acid substitutions. The peptide sequence of the antibody region may differ from the peptide sequence of the immunoglobulin or immunoglobulin fragment on which it is based and / or derived from by six or more amino acid substitutions. The peptide sequence of the antibody region may differ from the peptide sequence of the immunoglobulin or immunoglobulin fragment on which it is based and / or derived by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 15, 17, 20, 25 or more amino acid substitutions. The peptide sequence of the antibody region may differ from the peptide sequence of the immunoglobulin or immunoglobulin fragment on which it is based and / or derived by about 20-30, 30-40, 40-50, 50-60, 60-70, 80-90, 90-100, 100-150, 150-200, 200-300 or more amino acid substitutions.

[0235] The nucleotide sequence of an antibody region may differ from the nucleotide sequence of the immunoglobulin or immunoglobulin fragment on which it is based and / or derived from by one or more nucleotide and / or base pair substitutions. The nucleotide sequence of an antibody region may differ from the nucleotide sequence of the immunoglobulin or immunoglobulin fragment on which it is based and / or derived from by two or more nucleotide and / or base pair substitutions. The nucleotide sequence of an antibody region may differ from the nucleotide sequence of the immunoglobulin or immunoglobulin fragment on which it is based and / or derived from by three or more nucleotide and / or base pair substitutions. The nucleotide sequence of an antibody region may differ from the nucleotide sequence of the immunoglobulin or immunoglobulin fragment on which it is based and / or derived from by four or more nucleotide and / or base pair substitutions. The nucleotide sequence of an antibody region may differ from the nucleotide sequence of the immunoglobulin or immunoglobulin fragment on which it is based and / or derived from by five or more nucleotide and / or base pair substitutions. The nucleotide sequence of an antibody region may differ from the nucleotide sequence of the immunoglobulin or immunoglobulin fragment on which it is based and / or derived from by six or more nucleotide and / or base pair substitutions. The nucleotide sequence of an antibody region may differ from the nucleotide sequence of the immunoglobulin or immunoglobulin fragment on which it is based and / or derived from by nine or more nucleotide and / or base pair substitutions. The nucleotide sequence of an antibody region may differ from the nucleotide sequence of the immunoglobulin or immunoglobulin fragment on which it is based and / or derived from by twelve or more nucleotide and / or base pair substitutions. The nucleotide sequence of an antibody region may differ from the nucleotide sequence of the immunoglobulin or immunoglobulin fragment on which it is based and / or derived from by fifteen or more nucleotide and / or base pair substitutions. The nucleotide sequence of an antibody region may differ from the nucleotide sequence of the immunoglobulin or immunoglobulin fragment on which it is based and / or derived from by eighteen or more nucleotide and / or base pair substitutions. The nucleotide sequence of the antibody region may differ from the nucleotide sequence of the immunoglobulin or immunoglobulin fragment on which it is based and / or derived by 20, 22, 24, 25, 27, 30 or more nucleotide and / or base pair substitutions. The nucleotide sequence of the antibody region may differ from the nucleotide sequence of the immunoglobulin or immunoglobulin fragment on which it is based and / or derived by about 30-40, 40-50, 50-60, 60-70, 70-80, 80-90, 90-100, 100-200, 200-300, 300-400 or more nucleotide and / or base pair substitutions.

[0236] The antibody region may comprise at least about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 or more amino acids. The antibody region may comprise at least about 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625, 650, 675, 700 or more amino acids. The antibody region may comprise at least about 100 amino acids. The antibody region may comprise at least about 200 amino acids. The antibody region may comprise at least about 400 amino acids. The antibody region may comprise at least about 500 amino acids. The antibody region may comprise at least about 600 amino acids.

[0237] The antibody region may comprise less than about 2000, 1900, 1800, 1700, 1600, 1500, 1400, 1300, 1200, or 1100 amino acids. The antibody region may comprise less than about 1000, 950, 900, 850, 800, 750, or 700 amino acids. The antibody region may comprise less than about 1500 amino acids. The antibody region may comprise less than about 1000 amino acids. The antibody region may comprise less than about 800 amino acids. The antibody region may comprise less than about 700 amino acids.

[0238] The immunoglobulin fusion protein may further comprise an antibody region comprising 30 or fewer consecutive amino acids of a complementary determining region 3 (CDR3). The antibody region may comprise 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1 or fewer consecutive amino acids of CDR3. The antibody region may comprise 15 or fewer consecutive amino acids of CDR3. The antibody region may comprise 14 or fewer consecutive amino acids of CDR3. The antibody region may comprise 13 or fewer consecutive amino acids of CDR3. The antibody region may comprise 12 or fewer consecutive amino acids of CDR3. The antibody region may comprise 11 or fewer consecutive amino acids of CDR3. The antibody region may comprise 10 or fewer consecutive amino acids of CDR3. The antibody region may comprise 9 or fewer consecutive amino acids of CDR3. The antibody region may comprise 8 or fewer consecutive amino acids of CDR3. The antibody region may comprise 7 or fewer consecutive amino acids of CDR3. The antibody region may comprise 6 or fewer consecutive amino acids of CDR3. The antibody region may comprise 5 or fewer consecutive amino acids of CDR3. The antibody region may comprise 4 or fewer consecutive amino acids of CDR3. The antibody region may comprise 3 or fewer consecutive amino acids of CDR3. The antibody region may comprise 2 or fewer consecutive amino acids of CDR3. The antibody region may comprise 1 or fewer consecutive amino acids of CDR3. In some cases, the antibody region does not contain CDR3.

[0239] The immunoglobulin fusion protein may comprise a first antibody region comprising 6 or fewer consecutive amino acids of a complementary determining region 3 (CDR3). The first antibody region may comprise 5 or fewer consecutive amino acids of a CDR3. The first antibody region may comprise 4 or fewer consecutive amino acids of a CDR3. The first antibody region may comprise 3 or fewer consecutive amino acids of a CDR3. The first antibody region may comprise 2 or fewer consecutive amino acids of a CDR3. The first antibody region may comprise 1 or fewer consecutive amino acids of a CDR3. In some cases, the first antibody region does not contain a CDR3.

[0240] The immunoglobulin fusion protein may further comprise a second antibody region comprising 30 or fewer consecutive amino acids of a complementary determining region 3 (CDR3). The second antibody region may comprise 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1 or fewer consecutive amino acids of CDR3. The second antibody region may comprise 15 or fewer consecutive amino acids of CDR3. The second antibody region may comprise 14 or fewer consecutive amino acids of CDR3. The second antibody region may comprise 13 or fewer consecutive amino acids of CDR3. The second antibody region may comprise 12 or fewer consecutive amino acids of CDR3. The second antibody region may comprise 11 or fewer consecutive amino acids of CDR3. The second antibody region may comprise 10 or fewer consecutive amino acids of CDR3. The second antibody region may comprise 9 or fewer consecutive amino acids of CDR3. The second antibody region may comprise 8 or fewer consecutive amino acids of CDR3. The second antibody region may comprise 7 or fewer consecutive amino acids of CDR3. The second antibody region may comprise 6 or fewer consecutive amino acids of CDR3. The second antibody region may comprise 5 or fewer consecutive amino acids of CDR3. The second antibody region may comprise 4 or fewer consecutive amino acids of CDR3. The second antibody region may comprise 3 or fewer consecutive amino acids of CDR3. The second antibody region may comprise 2 or fewer consecutive amino acids of CDR3. The second antibody region may comprise 1 or fewer consecutive amino acids of CDR3. In some cases, the second antibody region does not contain CDR3.

[0241] The antibody region may comprise an amino acid sequence based on or derived from any one of SEQ ID NOs: 19-36 and 271-273. The antibody region may comprise an amino acid sequence that is at least about 50% homologous to any one of SEQ ID NOs: 19-36 and 271-273. The antibody region may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 97% homologous to any one of SEQ ID NOs: 19-36 and 271-273. The antibody region may comprise an amino acid sequence that is at least about 70% homologous to any one of SEQ ID NOs: 19-36 and 271-273. The antibody region may comprise an amino acid sequence that is at least about 80% homologous to any one of SEQ ID NOs: 19-36 and 271-273. The antibody region may comprise an amino acid sequence that is at least about 50% identical to any one of SEQ ID NOs: 19-36 and 271-273. The antibody region may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 97% identical to any one of SEQ ID NOs: 19-36 and 271-273. The antibody region may comprise an amino acid sequence that is at least about 70% identical to any one of SEQ ID NOs: 19-36 and 271-273. The antibody region may comprise an amino acid sequence that is at least about 80% identical to any one of SEQ ID NOs: 19-36 and 271-273. The antibody region may comprise an amino acid sequence that is 100% identical to any one of SEQ ID NOs: 19-36 and 271-273. In some embodiments, the antibody region comprises an amino acid sequence that is at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 97% homologous to the amino acid sequence of any one of SEQ ID NOs: 19-36 and 271-273. In some embodiments, the antibody region comprises an amino acid sequence that is at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 97% identical to the amino acid sequence of any one of SEQ ID NOs: 19-36 and 271-273.

[0242] The antibody region may comprise an amino acid sequence comprising 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 or more amino acids based on or derived from any one of SEQ ID NOs: 19-36 and 271-273. The antibody region may comprise an amino acid sequence comprising 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 450, 500 or more amino acids based on or derived from any one of SEQ ID NOs: 19-36 and 271-273. The antibody region may comprise an amino acid sequence comprising 10 or more amino acids based on or derived from any one of SEQ ID NOs: 19-36 and 271-273. The antibody region may comprise an amino acid sequence containing 50 or more amino acids based on or derived from any one of SEQ ID NOs: 19-36 and 271-273. The antibody region may comprise an amino acid sequence containing 100 or more amino acids based on or derived from any one of SEQ ID NOs: 19-36 and 271-273. The antibody region may comprise an amino acid sequence containing 200 or more amino acids based on or derived from any one of SEQ ID NOs: 19-36 and 271-273. The amino acids may be continuous. Alternatively, or additionally, the amino acids may be discontinuous. In some embodiments, the antibody region may comprise an amino acid derived from any one of SEQ ID NOs: 19-36 and 271-273 and an amino acid not derived from any one of SEQ ID NOs: 19-36 and 271-273. In some embodiments, the antibody region may comprise amino acids derived from one or more of SEQ ID NOs: 19-36 and 271-273 and amino acids not derived from any of SEQ ID NOs: 19-36 and 271-273. In some embodiments, the antibody region comprises amino acids derived from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more of SEQ ID NOs: 19-36 and 271-273.

[0243] The antibody region may be encoded by a nucleotide sequence based on or derived from any one of SEQ ID NOs: 1-18 and 268-270. The antibody region may be encoded by a nucleotide sequence that is at least about 50% homologous to any one of SEQ ID NOs: 1-18 and 268-270. The antibody region may be encoded by a nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 97% homologous to any one of SEQ ID NOs: 1-18 and 268-270. The antibody region may be encoded by a nucleotide sequence that is at least about 70% homologous to any one of SEQ ID NOs: 1-18 and 268-270. The antibody region may be encoded by a nucleotide sequence that is at least about 80% homologous to any one of SEQ ID NOs: 1-18 and 268-270. The antibody region may be encoded by a nucleotide sequence that is at least about 50% identical to any one of SEQ ID NOs: 1-18 and 268-270. The antibody region may be encoded by a nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 97% identical to any one of SEQ ID NOs: 1-18 and 268-270. The antibody region may be encoded by a nucleotide sequence that is at least about 70% identical to any one of SEQ ID NOs: 1-18 and 268-270. The antibody region may be encoded by a nucleotide sequence that is at least about 80% identical to any one of SEQ ID NOs: 1-18 and 268-270. The antibody region may be encoded by a nucleotide sequence that is 100% identical to any one of SEQ ID NOs: 1-18 and 268-270.

[0244] The antibody region may be encoded by a nucleotide sequence comprising 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 or more nucleotides based on or derived from any one of SEQ ID NOs: 1-18 and 268-270. The antibody region may be encoded by a nucleotide sequence comprising 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 450, 500 or more nucleotides based on or derived from any one of SEQ ID NOs: 1-18 and 268-270. The antibody region can be encoded by a nucleotide sequence comprising 600, 650, 700, 750, 800, 850, 900, 950, 1000 or more nucleotides based on or derived from any one of SEQ ID NO:1-18 and 268-270. The antibody region can be encoded by a nucleotide sequence comprising 1100, 1200, 1300, 1400, 1500 or more nucleotides based on or derived from any one of SEQ ID NO:1-18 and 268-270. The antibody region can be encoded by a nucleotide sequence comprising 100 or more nucleotides based on or derived from any one of SEQ ID NO:1-18 and 268-270. The antibody region can be encoded by a nucleotide sequence comprising 500 or more nucleotides based on or derived from any one of SEQ ID NO:1-18 and 268-270. The antibody region can be encoded by a nucleotide sequence comprising 1000 or more nucleotides based on or derived from any one of SEQ ID NO:1-18 and 268-270. The antibody region can be encoded by a nucleotide sequence comprising 1300 or more nucleotides based on or derived from any one of SEQ ID NO:1-18 and 268-270. The nucleotides can be continuous. In some embodiments, the antibody region is encoded by a nucleotide sequence comprising nucleotides derived from any one of SEQ ID NO:1-18 and 268-270 and non-nucleotides derived from any one of SEQ ID NO:1-18 and 268-270. In some embodiments, the antibody region is encoded by a nucleotide sequence comprising nucleotides derived from one or more of SEQ ID NO:1-18 and 268-270 and non-nucleotides derived from any one of SEQ ID NO:1-18 and 268-270. In some embodiments, the antibody region is encoded by a nucleotide sequence derived from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more of SEQ ID NOs: 1-18 and 268-270.

[0245] Non-antibody region

[0246] The immunoglobulin fusion proteins disclosed herein may comprise one or more non-antibody regions. The immunoglobulin fusion proteins disclosed herein may comprise two or more non-antibody regions. The immunoglobulin fusion proteins disclosed herein may comprise 3, 4, 5, 6, 7, 8, 9, 10 or more non-antibody regions.

[0247] The two or more non-antibody regions may be attached to one or more antibody regions. The two or more non-antibody regions may be attached to two or more antibody regions. The two or more non-antibody regions may be attached to one or more immunoglobulin chains. The two or more non-antibody regions may be attached to two or more immunoglobulin chains. The two or more non-antibody regions may be attached to one or more subunits within one or more antibody regions. The two or more non-antibody regions may be attached to two or more subunits within one or more antibody regions.

[0248] The non-antibody region may comprise one or more therapeutic agents. The non-antibody region may comprise two or more therapeutic agents. The non-antibody region may comprise 3, 4, 5, 6, 7 or more therapeutic agents. The therapeutic agents may be different. The therapeutic agents may be the same.

[0249] The non-antibody region may comprise one or more extender peptides. The non-antibody region may comprise two or more extender peptides. The non-antibody region may comprise 3, 4, 5, 6, 7 or more extender peptides. The extender peptides may be different. The extender peptides may be the same. In some embodiments, the extender peptide comprises an amino acid sequence having an alpha helical secondary structure. In some embodiments, the extender peptide does not comprise amino acids having a beta chain secondary structure. In some embodiments, the extender fusion region comprises two extender peptides, wherein the two extender peptides are configured to form a coiled coil. In some cases, the non-antibody region does not comprise an extender peptide. The extender peptide can directly connect the therapeutic peptide to the antibody region.

[0250] The non-antibody region may comprise one or more linkers. The non-antibody region may comprise two or more linkers. The non-antibody region may comprise 3, 4, 5, 6, 7 or more linkers. The linkers may be different. The linkers may be the same. The linker may directly connect the therapeutic agent to the antibody region. The linker may connect the therapeutic peptide to the extender peptide. In some cases, the non-antibody region does not comprise a linker. In some embodiments, the linker peptide does not comprise amino acids having alpha-helical or beta-strand secondary structures.

[0251] The non-antibody region may comprise one or more protease cleavage sites. The non-antibody region may comprise two or more protease cleavage sites. The cleavage sites may be different. The cleavage sites may be the same. The cleavage site may directly connect the therapeutic agent to the antibody region. The cleavage site may connect the therapeutic agent to a linker peptide. The cleavage site may connect the therapeutic agent to an extender peptide. In some embodiments, the therapeutic agent comprises a protease cleavage site. In some cases, the non-antibody region does not comprise a protease cleavage site.

[0252] The extender fusion region may comprise one or more linker peptides. The linker peptide may comprise an extender peptide. The linker peptide may comprise a linker peptide. The linker peptide may comprise a protease cleavage site. The linker peptide may comprise any sequence of amino acids configured to link the therapeutic agent to the antibody region.

[0253] The non-antibody region can be inserted into the antibody region. The insertion of the non-antibody region into the antibody region may include the removal or deletion of a portion of the antibody on which the antibody region is based or derived. The non-antibody region may replace at least a portion of the heavy chain. The non-antibody region may replace at least a portion of the light chain. The non-antibody region may replace at least a portion of the V region. The non-antibody region may replace at least a portion of the D region. The non-antibody region may replace at least a portion of the J region. The non-antibody region may replace at least a portion of the variable region. The non-antibody region may replace at least a portion of the constant region. The non-antibody region may replace at least a portion of the complementary determining region (CDR). The non-antibody region may replace at least a portion of CDR1. The non-antibody region may replace at least a portion of CDR2. The non-antibody region may replace at least a portion of CDR3. The non-antibody region can replace at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more of an antibody or portion thereof. For example, a non-antibody region can replace at least about 50% of a CDR. A non-antibody region can replace at least about 70% of a CDR. A non-antibody region can replace at least about 80% of a CDR. A non-antibody region can replace at least about 90% of a CDR. A non-antibody region can replace at least about 95% of a CDR.

[0254] The non-antibody region may comprise (a) one or more extender peptides; (b) one or more therapeutic agents; (c) optionally, one or more linkers; and (d) optionally, one or more proteolytic cleavage sites. In some embodiments, the one or more extender peptides comprise an amino acid sequence having an alpha helical secondary structure. In some cases, an immunoglobulin fusion protein comprising an antibody region and a non-antibody region (wherein the non-antibody region comprises one or more extender peptides comprising amino acids having an alpha helical secondary structure) is referred to as a coiled immunoglobulin fusion protein.

[0255] The non-antibody region may comprise (a) one or more linker peptides; (b) one or more therapeutic agents; and (c) optionally, one or more proteolytic cleavage sites. In some embodiments, the one or more linker peptides do not comprise an amino acid sequence having an α-helix or β-chain secondary structure. In some cases, an immunoglobulin fusion protein comprising an antibody region and a non-antibody region (wherein one or more linker peptides do not comprise an amino acid sequence having an α-helix or β-chain secondary structure) is referred to as a direct immunoglobulin fusion protein.

[0256] In some embodiments, the non-antibody region is an extender fusion region.

[0257] Extension fusion zone

[0258] The immunoglobulin fusion proteins disclosed herein may comprise one or more extender fusion regions. The immunoglobulin fusion proteins may comprise two or more extender fusion regions. The immunoglobulin fusion proteins may comprise 3, 4, 5, 6, 7, 8, 9, 10 or more extender fusion regions.

[0259] The two or more extension fusion regions may be attached to one or more antibody regions. The two or more extension fusion regions may be attached to two or more antibody regions. The two or more extension fusion regions may be attached to one or more immunoglobulin chains. The two or more extension fusion regions may be attached to two or more immunoglobulin chains. The two or more extension fusion regions may be attached to one or more subunits within one or more antibody regions. The two or more extension fusion regions may be attached to two or more subunits within one or more antibody regions.

[0260] The extender fusion region may comprise one or more extender peptides. The extender fusion region may comprise two or more extender peptides. The extender fusion region may comprise 3, 4, 5, 6 or more extender peptides. The extender peptides may be different. The extender peptides may be identical. In some embodiments, the extender peptide comprises an amino acid sequence having an alpha helical secondary structure. In some embodiments, the extender peptide does not comprise amino acids having a beta chain secondary structure. In some embodiments, the extender fusion region comprises two extender peptides, wherein the two extender peptides are configured to form a coiled coil. In some cases, the extender fusion region does not comprise an extender peptide. In some embodiments, the extender peptide directly connects the therapeutic agent to the antibody region.

[0261] The extension fusion region may contain one or more therapeutic agents. The extension fusion region may contain two or more therapeutic agents. The extension fusion region may contain 3, 4, 5, 6, 7 or more therapeutic agents. The therapeutic agents may be different. The therapeutic agents may be the same.

[0262] The stretcher fusion region may comprise one or more linkers. The stretcher fusion region may comprise two or more linkers. The stretcher fusion region may comprise 3, 4, 5, 6, 7, or more linkers. The linkers may be different. The linkers may be identical. The linker may connect the therapeutic agent to the stretcher peptide. The linker may directly connect the therapeutic agent to the antibody region. In some cases, the stretcher fusion region does not comprise a linker. In some embodiments, the linker peptide does not comprise amino acids having alpha-helical or beta-strand secondary structures.

[0263] The extender fusion region may comprise one or more protease cleavage sites. The extender fusion region may comprise two or more protease cleavage sites. The cleavage sites may be different. The cleavage sites may be the same. The cleavage site may directly connect the therapeutic agent to the antibody region. The cleavage site may connect the therapeutic agent to the extender peptide. The cleavage site may connect the therapeutic agent to the linker peptide. In some cases, the extender fusion region does not comprise a protease cleavage site.

[0264] The extender fusion region may comprise one or more linker peptides. The linker peptide may comprise an extender peptide. The linker peptide may comprise a linker peptide. The linker peptide may comprise a protease cleavage site. The linker peptide may comprise any sequence of amino acids configured to link the therapeutic agent and the antibody region.

[0265] The immunoglobulin fusion proteins disclosed herein may comprise an antibody region attached to an extender fusion region. The extender fusion region may be attached to the N-terminus, C-terminus, or both the N-terminus and the C-terminus of the antibody region. The antibody region may be directly attached to the extender fusion region. Alternatively, or additionally, the antibody region may be indirectly attached to a non-antibody sequence. The attachment of the extender fusion region to the antibody region may comprise covalent attachment. The attachment may comprise fusion of the extender fusion region to the antibody region. The attachment may comprise chemical conjugation.

[0266] Alternatively, or additionally, attachment includes insertion of an extender fusion region into an antibody region. The extender fusion region may be inserted into the heavy chain of the antibody region. The extender fusion region may be inserted into the light chain of the antibody region. The extender fusion region may be inserted into the variable domain of the antibody region. The extender fusion region may be inserted into the constant domain of the antibody region. The extender fusion region may be inserted into the complementary determining region (CDR) of the antibody region.

[0267] The extension fusion region can replace at least a portion of the antibody on which the antibody region is based or derived from. The extension fusion region can replace at least a portion of the heavy chain of the antibody on which the antibody region is based or derived from. The extension fusion region can replace at least a portion of the light chain of the antibody on which the antibody region is based or derived from. The extension fusion region can replace at least a portion of the variable domain of the antibody on which the antibody region is based or derived from. The extension fusion region can replace at least a portion of the variable domain of the antibody on which the antibody region is based or derived from. The extension fusion region can replace at least a portion of the complementary determining region (CDR) of the antibody on which the antibody region is based or derived from. The extension fusion region can replace at least a portion of the CDR1, CDR2, CDR3, or a combination thereof of the antibody on which the antibody or its fragment is based or derived from. The extension fusion region can replace at least a portion of the CDR3 of the antibody on which the antibody region is based or derived from.

[0268] The extension fusion region can replace at least about 1, 2, 3, 4, 5, 6, 7, 8, 9 or more amino acids of the antibody on which the antibody region is based or derived from. The extension fusion region can replace at least about 1 or more amino acids of the antibody on which the antibody region is based or derived from. The extension fusion region can replace at least about 3 or more amino acids of the antibody on which the antibody region is based or derived from. The extension fusion region can replace at least about 5 or more amino acids of the antibody on which the antibody region is based or derived from.

[0269] The extension fusion region may comprise at least about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100 or more amino acids. The extension fusion region may comprise at least about 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1000, 1500, 2000 or more amino acids. The extension fusion region may comprise at least about 10 or more amino acids. The extension fusion region may comprise at least about 25 or more amino acids. The extension fusion region may comprise at least about 50 or more amino acids. The extension fusion region may comprise at least about 75 or more amino acids. The extension fusion region may comprise at least about 100 or more amino acids.

[0270] The extension fusion region may comprise less than about 2000, 1500, 1000, 900, 800, 700, 600, or 500 amino acids. The extension fusion region may comprise less than about 450, 400, 350, 300, 275, 250, 225, 200, 175, 150, 125, 100, 90, 80, 70, 60, or 50 amino acids. The extension fusion region may comprise less than about 400 amino acids. The extension fusion region may comprise less than about 300 amino acids. The extension fusion region may comprise less than about 250 amino acids.

[0271] The stretch fusion region may comprise about 10 to about 1000 amino acids. The stretch fusion region may comprise about 10 to about 500 amino acids. The stretch fusion region may comprise about 10 to about 400 amino acids. The stretch fusion region may comprise about 10 to about 300 amino acids. The stretch fusion region may comprise about 10 to about 250 amino acids. The stretch fusion region may comprise about 20 to about 500 amino acids. The stretch fusion region may comprise about 20 to about 400 amino acids. The stretch fusion region may comprise about 20 to about 300 amino acids.

[0272] The stretcher fusion region may comprise (a) one or more stretcher peptides; (b) one or more therapeutic agents; (c) optionally, one or more linkers; and (d) optionally, one or more proteolytic cleavage sites. Exemplary stretcher fusion regions are Figure 2 AG is shown. For example, Figure 2 As shown in Figure A, the extender fusion region comprises an extender peptide (210) and a therapeutic agent (220). Figure 2 As shown in B, the stretcher fusion region comprises two stretcher peptides (210, 230) and a therapeutic agent (220). Figure 2 As shown in Figure C, the extender fusion region comprises an extender peptide (210) and a therapeutic agent (220) connected by a linker (240). Figure 2 As shown in Figure D, the extender fusion region comprises an extender peptide (210) and a therapeutic agent (220) flanked by two linkers (240, 250). Figure 2 As shown in E, the extender fusion region includes an extender peptide (210), a therapeutic agent (220), and a proteolytic cleavage site (260), wherein the proteolytic cleavage site (260) is inserted between the extender peptide and the therapeutic agent. Figure 2 As shown in Figure F, the extender fusion region comprises two extender peptides (210, 230), two linkers (240, 250) and a therapeutic agent (220). Figure 2 As shown in Figure G, the extender fusion region comprises two extender peptides (210, 230), two linkers (240, 250), a proteolytic cleavage site (260) and a therapeutic agent (220).

[0273] The stretcher fusion region may comprise (a) a first stretcher peptide, wherein the first stretcher peptide comprises (i) an amino acid sequence comprising an alpha-helical secondary structure; and (ii) 7 or fewer amino acids based on or derived from an ultralong CDR3; and (b) a therapeutic agent. The stretcher fusion region may further comprise one or more additional stretcher peptides comprising at least one secondary structure. The stretcher fusion region may further comprise one or more linkers. The stretcher fusion region may comprise one or more proteolytic cleavage sites.

[0274] The stretcher fusion region may comprise (a) a first stretcher peptide, wherein the first stretcher peptide comprises (i) an amino acid sequence comprising an alpha-helical secondary structure; and (ii) an amino acid sequence that does not comprise an ultralong CDR3; and (b) a first therapeutic agent. The stretcher fusion region may further comprise one or more additional stretcher peptides comprising at least one secondary structure. The stretcher fusion region may further comprise one or more linkers. The stretcher fusion region may comprise one or more proteolytic cleavage sites.

[0275] The stretcher fusion region may comprise (a) one or more stretcher peptides; (b) one or more therapeutic agents; (c) optionally, one or more linkers; and (d) optionally, one or more proteolytic cleavage sites. In some embodiments, the one or more stretcher peptides comprise an amino acid sequence having an alpha-helical secondary structure. In some cases, an immunoglobulin fusion protein comprising an antibody region and a stretcher fusion region (wherein the stretcher fusion region comprises one or more stretcher peptides comprising amino acids having an alpha-helical secondary structure) is referred to as a coiled immunoglobulin fusion protein.

[0276] The extender fusion region may comprise (a) one or more linker peptides; (b) one or more therapeutic agents; and (c) optionally, one or more proteolytic cleavage sites. In some embodiments, the one or more linker peptides do not comprise an amino acid sequence having an α-helix or β-chain secondary structure. In some cases, an immunoglobulin fusion protein comprising an antibody region and an extender fusion region (wherein the one or more linker peptides do not comprise an amino acid sequence having an α-helix or β-chain secondary structure) is referred to as a direct immunoglobulin fusion protein.

[0277] In some embodiments, the stretcher fusion region does not comprise amino acids based on or derived from an antibody. In some cases, the stretcher fusion region is a non-antibody region.

[0278] Extender peptide

[0279] The immunoglobulin fusion proteins disclosed herein may comprise one or more extender peptides. The immunoglobulin fusion proteins disclosed herein may comprise two or more extender peptides. The one or more extender peptides may be attached to the N-terminus, C-terminus, or both the N-terminus and the C-terminus of the therapeutic agent. The one or more extender peptides may be attached to each end of the therapeutic agent. The one or more extender peptides may be attached to different ends of the therapeutic agent.

[0280] The stretcher fusion region of the immunoglobulin fusion proteins disclosed herein may comprise one or more stretcher peptides. The stretcher fusion region may comprise two or more stretcher peptides. The stretcher fusion region may comprise three or more stretcher peptides. The stretcher fusion region may comprise four or more stretcher peptides. The stretcher fusion region may comprise five or more stretcher peptides. The stretcher fusion region may comprise a first stretcher peptide and a second stretcher peptide.

[0281] The stretch peptide may comprise one or more secondary structures. The stretch peptide may comprise two or more secondary structures. The stretch peptide may comprise 3, 4, 5, 6, 7 or more secondary structures. The two or more stretch peptides may comprise one or more secondary structures. The two or more stretch peptides may comprise two or more secondary structures. The two or more stretch peptides may comprise 3, 4, 5, 6, 7 or more secondary structures. Each stretch peptide may comprise at least one secondary structure. The secondary structures of the two or more stretch peptides may be the same. Alternatively, the secondary structures of the two or more stretch peptides may be different.

[0282] Alternatively, or in addition, the one or more secondary structures may comprise one or more alpha helices. The stretcher peptide may comprise two or more alpha helices. For example, a first stretcher peptide comprises a first alpha helix and a second stretcher peptide comprises a second alpha helix. The stretcher peptide may comprise 3, 4, 5, 6, 7, or more alpha helices. The two or more alpha helices may be antiparallel. The two or more alpha helices may be parallel. The two or more alpha helices may form one or more coiled-coil domains.

[0283] The one or more extender peptides may comprise at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acids. The one or more extender peptides may comprise at least about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more amino acids. The one or more extender peptides may comprise at least about 35, 40, 45, 50, 60, 70, 80, 90, 100 or more amino acids.

[0284] The one or more stretch peptides may comprise less than about 100 amino acids. The one or more stretch peptides may comprise less than about 95, 90, 85, 80, 75, 70, 65, 60, 55, or 50 amino acids. The one or more stretch peptides may comprise less than about 90 amino acids. The one or more stretch peptides may comprise less than about 80 amino acids. The one or more stretch peptides may comprise less than about 70 amino acids.

[0285] The two or more stretch peptides can be the same length. For example, the first stretch peptide and the second stretch peptide are the same length. Alternatively, the two or more stretch peptides are different lengths. In another example, the first stretch peptide and the second stretch peptide are different lengths. The lengths of the two or more stretch peptides can differ by at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more amino acids. The lengths of the two or more stretch peptides can differ by at least about 1 or more amino acids. The lengths of the two or more stretch peptides can differ by at least about 3 or more amino acids. The lengths of the two or more stretch peptides can differ by at least about 5 or more amino acids.

[0286] The extender peptide may be adjacent to an antibody region. The extender peptide may be attached to the N-terminus, C-terminus, or N-terminus and C-terminus of the antibody region. The extender peptide may be adjacent to a non-antibody region. The extender peptide may be attached to the N-terminus, C-terminus, or N-terminus and C-terminus of the non-antibody region. The extender peptide may be adjacent to a therapeutic agent. The extender peptide may be attached to the N-terminus, C-terminus, or N-terminus and C-terminus of the therapeutic agent. The extender peptide may be adjacent to a connector. The extender peptide may be attached to the N-terminus, C-terminus, or N-terminus and C-terminus of the connector. The extender peptide may be adjacent to a proteolytic cleavage site. The extender peptide may be attached to the N-terminus, C-terminus, or N-terminus and C-terminus of the proteolytic cleavage site.

[0287] The extender peptide can connect the therapeutic agent to the antibody region. The extender peptide can be between the antibody region and the therapeutic agent, connector and / or proteolytic cleavage site. The extender peptide can be between two or more antibody regions, therapeutic agents, connectors, proteolytic cleavage sites, or a combination thereof. The extender peptide can be at the N-terminus of the antibody region, therapeutic agent, connector, proteolytic cleavage site, or a combination thereof. The extender peptide can be at the C-terminus of the antibody region, therapeutic agent, connector, proteolytic cleavage site, or a combination thereof.

[0288] The extender peptide may comprise an amino acid sequence based on or derived from any one of SEQ ID NOs: 144-175. The extender peptide may comprise an amino acid sequence that is at least about 50% homologous to an amino acid sequence based on or derived from any one of SEQ ID NOs: 144-175. The extender peptide may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97% or more homologous to an amino acid sequence based on or derived from any one of SEQ ID NOs: 144-175. The extender peptide may comprise an amino acid sequence that is at least about 70% homologous to an amino acid sequence based on or derived from any one of SEQ ID NOs: 144-175. The extender peptide may comprise an amino acid sequence that is at least about 80% homologous to an amino acid sequence based on or derived from any one of SEQ ID NOs: 144-175. The extender peptide may comprise an amino acid sequence that is at least about 85% homologous to an amino acid sequence based on or derived from any one of SEQ ID NOs: 144-175.

[0289] The first extender peptide may comprise an amino acid sequence based on or derived from any one of SEQ ID NOs: 144-175. The first extender peptide may comprise an amino acid sequence that is at least about 50% homologous to an amino acid sequence based on or derived from any one of SEQ ID NOs: 144-175. The first extender peptide may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97% or more homologous to an amino acid sequence based on or derived from any one of SEQ ID NOs: 144-175. The first extender peptide may comprise an amino acid sequence that is at least about 75% homologous to an amino acid sequence based on or derived from any one of SEQ ID NOs: 144-175. The first extender peptide may comprise an amino acid sequence that is at least about 80% homologous to an amino acid sequence based on or derived from any one of SEQ ID NOs: 144-175.

[0290] The first extender peptide may comprise an amino acid sequence based on or derived from any one of SEQ ID NOs: 144-153. The first extender peptide may comprise an amino acid sequence that is at least about 50% homologous to an amino acid sequence based on or derived from any one of SEQ ID NOs: 144-153. The first extender peptide may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97% or more homologous to an amino acid sequence based on or derived from any one of SEQ ID NOs: 144-153. The first extender peptide may comprise an amino acid sequence that is at least about 75% homologous to an amino acid sequence based on or derived from any one of SEQ ID NOs: 144-153. The first extender peptide may comprise an amino acid sequence that is at least about 80% homologous to an amino acid sequence based on or derived from any one of SEQ ID NOs: 144-153.

[0291] The second extender peptide may comprise an amino acid sequence based on or derived from any one of SEQ ID NOs: 144-175. The second extender peptide may comprise an amino acid sequence that is at least about 50% homologous to an amino acid sequence based on or derived from any one of SEQ ID NOs: 144-175. The second extender peptide may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97% or more homologous to an amino acid sequence based on or derived from any one of SEQ ID NOs: 144-175. The second extender peptide may comprise an amino acid sequence that is at least about 70% homologous to an amino acid sequence based on or derived from any one of SEQ ID NOs: 144-175. The second extender peptide may comprise an amino acid sequence that is at least about 80% homologous to an amino acid sequence based on or derived from any one of SEQ ID NOs: 144-175.

[0292] The second extender peptide may comprise an amino acid sequence based on or derived from any one of SEQ ID NOs: 154-163. The second extender peptide may comprise an amino acid sequence that is at least about 50% homologous to an amino acid sequence based on or derived from any one of SEQ ID NOs: 154-163. The second extender peptide may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97% or more homologous to an amino acid sequence based on or derived from any one of SEQ ID NOs: 154-163. The second extender peptide may comprise an amino acid sequence that is at least about 70% homologous to an amino acid sequence based on or derived from any one of SEQ ID NOs: 154-163. The second extender peptide may comprise an amino acid sequence that is at least about 80% homologous to an amino acid sequence based on or derived from any one of SEQ ID NOs: 154-163.

[0293] The extender peptides disclosed herein can be based on or derived from a CDR3. The CDR3 can be an ultralong CDR3. "Ultralong CDR3" or "ultralong CDR3 sequence" used interchangeably herein can include a CDR3 that is not derived from a human antibody sequence. The length of an ultralong CDR3 can be 35 amino acids or longer, e.g., 40 amino acids or longer, 45 amino acids or longer, 50 amino acids or longer, 55 amino acids or longer, or 60 amino acids or longer. An ultralong CDR3 can be a heavy chain CDR3 (CDR-H3 or CDRH3). An ultralong CDR3 can include a sequence derived from or based on a ruminant (e.g., bovine) sequence. An ultralong CDR3 can include one or more cysteine ​​motifs. An ultralong CDR3 can include at least 3 or more cysteine ​​residues, e.g., 4 or more cysteine ​​residues, 6 or more cysteine ​​residues, or 8 or more cysteine ​​residues. Additional details on ultralong CDR3 sequences can be found in Saini SS et al. (Exceptionally long CDR3H region with multiple cysteine ​​residues in functional bovine IgM antibodies, European Journal of Immunology, 1999), Zhang Y et al. (Functional antibody CDR3 fusion proteins with enhanced pharmacological properties, Angew Chem Int Ed Engl, 2013), Wang F et al. (Reshaping antibody diversity, Cell, 2013), and U.S. Patent No. 6,740,747.

[0294] The stretch peptide may comprise 7 or fewer amino acids based on or derived from a CDR. The stretch peptide may comprise 6, 5, 4, 3, 2, 1, or fewer amino acids based on or derived from a CDR. The amino acids may be contiguous. The amino acids may be discontinuous. The CDR may be CDR1. The CDR may be CDR2. The CDR may be CDR3. The CDR may be an ultralong CDR.

[0295] The stretch peptide may be based on or derived from a CDR, wherein the CDR is not an ultralong CDR3. The stretch peptide may comprise 10 or fewer amino acids based on or derived from a CDR3. The stretch peptide may comprise 9, 8, 7, 6, 5, 4, 3, 2, 1 or fewer amino acids based on or derived from a CDR3. The stretch peptide may comprise 8 or fewer amino acids based on or derived from a CDR3. The stretch peptide may comprise 7 or fewer amino acids based on or derived from a CDR3. The stretch peptide may comprise 5 or fewer amino acids based on or derived from a CDR3.

[0296] The extender peptide may comprise an amino acid sequence that is less than about 50% identical to the amino acid sequence comprising the ultralong CDR3. The extender peptide may comprise an amino acid sequence that is less than about 45%, 40%, 35%, 30%, 25%, 20%, 25%, or 10% identical to the amino acid sequence comprising the ultralong CDR3. The extender peptide may comprise an amino acid sequence that is less than about 30% identical to the amino acid sequence comprising the ultralong CDR3. The extender peptide may comprise an amino acid sequence that is less than about 25% identical to the amino acid sequence comprising the ultralong CDR3. The extender peptide may comprise an amino acid sequence that is less than about 20% identical to the amino acid sequence comprising the ultralong CDR3.

[0297] The extender peptide may comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more amino acids attached to or inserted into the portion of the extender peptide based on the ultralong CDR3. The extender peptide may comprise 1 or more amino acids attached to or inserted into the portion of the extender peptide based on the ultralong CDR3. The extender peptide may comprise 3 or more amino acids attached to or inserted into the portion of the extender peptide based on the ultralong CDR3. The extender peptide may comprise 5 or more amino acids attached to or inserted into the portion of the extender peptide based on the ultralong CDR3. The two or more amino acids attached to or inserted into the ultralong CDR3 may be continuous. Alternatively, or additionally, the two or more amino acids attached to or inserted into the ultralong CDR3 are discontinuous.

[0298] The extender peptide may comprise 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10 or fewer amino acids attached to or inserted into the portion of the extender peptide based on the ultralong CDR3. The extender peptide may comprise 20 or fewer amino acids attached to or inserted into the portion of the extender peptide based on the ultralong CDR3. The extender peptide may comprise 15 or fewer amino acids attached to or inserted into the portion of the extender peptide based on the ultralong CDR3. The extender peptide may comprise 10 or fewer amino acids attached to or inserted into the portion of the extender peptide based on the ultralong CDR3. The amino acids attached to or inserted into the ultralong CDR3 may be continuous. Alternatively, or additionally, the amino acids attached to or inserted into the ultralong CDR3 are discontinuous.

[0299] The extender peptide may comprise the sequence 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 (SEQ ID NO: 144). In some embodiments, the first extender peptide comprises the sequence X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 (SEQ ID NO: 144). In some cases, the first extender peptide is located between the amino terminus of the therapeutic agent and the antibody region. 1 -X 14 Can be independently selected from positively charged amino acids or hydrophobic amino acids. 1 -X 14 Can be independently selected from the group consisting of alanine (A), asparagine (N), isoleucine (I), leucine (L), valine (V), glutamine (Q), glutamic acid (E) and lysine (K). 1 -X 14 Can be independently selected from: alanine (A), leucine (L), and lysine (K). Alanine can constitute at least about 30% of the total amino acid composition. Alanine can constitute less than about 70% of the total amino acid composition. Leucine can constitute at least about 20% of the total amino acid composition. Leucine can constitute less than about 50% of the total amino acid composition. Lysine can constitute at least about 20% of the total amino acid composition. Lysine can constitute less than about 50% of the total amino acid composition. Hydrophobic amino acids can constitute at least about 50% of the total amino acid composition. Hydrophobic amino acids can constitute at least about 60% of the total amino acid composition. Hydrophobic amino acids can constitute at least about 70% of the total amino acid composition. Hydrophobic amino acids can constitute less than about 90% of the total amino acid composition.

[0300] The extender peptide may comprise the sequence (X 1 X 2 X 3 X4 X 5 X 6 X 7 ) n (SEQ ID NO. 145). In some embodiments, the first extender peptide comprises the sequence (X 1 X 2 X 3 X 4 X 5 X 6 X 7 ) n (SEQ ID NO. 145). In some cases, the first extender peptide is located between the amino terminus of the therapeutic agent and the antibody region. In some embodiments, n is between about 1 and about 10. In some embodiments, n is between about 1 and about 5. In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more. n can be about 1 to about 3. X 1 -X 7 Can be independently selected from positively charged amino acids or hydrophobic amino acids. 1 -X 7 Can be independently selected from: alanine (A), asparagine (N), isoleucine (I), leucine (L), valine (V), glutamine (Q), glutamic acid (E), and lysine (K). Alanine (A) can constitute at least about 30% of the total amino acid composition. Alanine (A) can constitute less than about 70% of the total amino acid composition. Leucine can constitute at least about 20% of the total amino acid composition. Leucine can constitute less than about 50% of the total amino acid composition. Lysine can constitute at least about 20% of the total amino acid composition. Lysine can constitute less than about 50% of the total amino acid composition. Asparagine can constitute about 50% of the total amino acid composition. Isoleucine can constitute about 50% of the total amino acid composition. Valine can constitute about 50% of the total amino acid composition. Glutamine can constitute about 50% of the total amino acid composition. Glutamic acid can constitute about 50% of the total amino acid composition. Hydrophobic amino acids can constitute at least about 50% of the total amino acid composition. Hydrophobic amino acids can constitute at least about 60% of the total amino acid composition. Hydrophobic amino acids can constitute at least about 70% of the total amino acid composition. Hydrophobic amino acids may constitute less than about 90% of the total amino acid composition.

[0301] The extender peptide may comprise the sequence X a X b X c X d (X 1 X 2 X 3 X 4 X 5 X 6 X 7 ) n(SEQ ID NO: 148). In some embodiments, the first extender peptide comprises the sequence X a X b X c X d (X 1 X 2 X 3 X 4 X 5 X 6 X 7 ) n (SEQ ID NO: 146). In some cases, the first extender peptide is located between the amino terminus of the therapeutic agent and the antibody region. In some embodiments, n is between about 1 and about 10. In some embodiments, n is between about 1 and about 5. In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more. n can be about 1 to about 3. X a 、X b and X d Can be independently selected from hydrophobic amino acids. c Can be polar, uncharged amino acids. X a 、X b and X d Can be the same amino acid. a 、X b and X d Can be different amino acids. X 1 -X 7 Can be independently selected from positively charged amino acids or hydrophobic amino acids. 1 -X 7 Can be independently selected from the group consisting of alanine (A), asparagine (N), isoleucine (I), leucine (L), valine (V), glutamine (Q), glutamic acid (E) and lysine (K). 1 -X 7 Can be independently selected from: A, L, and K. A can constitute at least about 30% of the total amino acid composition. A can constitute less than about 70% of the total amino acid composition. L can constitute at least about 20% of the total amino acid composition. L can constitute less than about 50% of the total amino acid composition. K can constitute at least about 20% of the total amino acid composition. K can constitute less than about 50% of the total amino acid composition. Hydrophobic amino acids can constitute at least about 50% of the total amino acid composition. Hydrophobic amino acids can constitute at least about 60% of the total amino acid composition. Hydrophobic amino acids can constitute at least about 70% of the total amino acid composition. Hydrophobic amino acids can constitute less than about 90% of the total amino acid composition. In some embodiments, X a is glycine (G). In some embodiments, X b is G. In some embodiments, X d It is glycine.a 、X b and X d It can be glycine (G). X c It may be serine (S).

[0302] The extender peptide may comprise the sequence X a X b X c X d (AKLAALK) n (SEQ ID NO. 147). In some embodiments, the first extender peptide comprises the sequence X a X b X c X d (AKLAALK) n (SEQ ID NO. 147). In some cases, the first extender peptide is located between the amino terminus of the therapeutic agent and the antibody region. In some embodiments, n is between about 1 and about 10. In some embodiments, n is between about 1 and about 5. In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more. n can be about 1 to about 3. X a 、X b and X d Can be independently selected from hydrophobic amino acids. c Can be polar, uncharged amino acids. X a 、X b and X d Can be the same amino acid. a 、X b and X d Can be different amino acids. In some embodiments, X a is glycine (G). In some embodiments, X b In some embodiments, X d It is glycine. a 、X b and X d It can be glycine (G). X c It may be serine (S).

[0303] The extender peptide may comprise the sequence (AKLAALK) n (SEQ ID NO. 148). In some embodiments, the first extender peptide comprises the sequence (AKLAALK) n(SEQ ID NO. 148). In some cases, the first extender peptide is located between the amino terminus of the therapeutic agent and the antibody region. In some embodiments, n is between about 1 and about 10. In some embodiments, n is between about 1 and about 5. In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more. n can be between about 1 and about 3.

[0304] The extender peptide may comprise the sequence GGSG(AKLAALK) n (SEQ ID NO: 149). In some embodiments, the first extender peptide comprises the sequence GGSG(AKLAALK) n (SEQ ID NO: 149). In some cases, the first extender peptide is located between the amino terminus of the therapeutic agent and the antibody region. In some embodiments, n is between about 1 and about 10. In some embodiments, n is between about 1 and about 5. In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more. n can be about 1 to about 3.

[0305] The extender peptide may comprise the sequence X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 (SEQ ID NO: 154); wherein X 1 -X 14 are independently selected from negatively charged amino acids or hydrophobic amino acids. In some embodiments, the second extender peptide comprises the sequence X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 (SEQ ID NO: 154); wherein X 1 -X 14are independently selected from negatively charged amino acids or hydrophobic amino acids. In some cases, the second extender peptide is located between the carboxyl terminus of the therapeutic agent and the antibody region. In some embodiments, n is between about 1 and about 10. In some embodiments, n is between about 1 and about 5. In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more. n can be from about 1 to about 3. In some embodiments, X 1 -X 14 Independently selected from: alanine (A), leucine (L) and glutamic acid (E). In one embodiment, A constitutes at least about 30% of the total amino acid composition. In one embodiment, A constitutes less than about 70% of the total amino acid composition. In one embodiment, L constitutes at least about 20% of the total amino acid composition. In one embodiment, L constitutes less than about 50% of the total amino acid composition. In one embodiment, E constitutes at least about 20% of the total amino acid composition. In one embodiment, E constitutes less than about 50% of the total amino acid composition. In one embodiment, hydrophobic amino acids constitute at least about 50% of the total amino acid composition. In one embodiment, hydrophobic amino acids constitute at least about 60% of the total amino acid composition. In one embodiment, hydrophobic amino acids constitute at least about 70% of the total amino acid composition. In one embodiment, hydrophobic amino acids constitute less than about 90% of the total amino acid composition.

[0306] The second extender peptide may comprise the sequence (X 1 X 2 X 3 X 4 X 5 X 6 X 7 ) n (SEQ ID NO: 155). In some embodiments, the second extender peptide comprises the sequence (X 1 X 2 X 3 X 4 X 5 X 6 X 7 ) n (SEQ ID NO: 155). In some cases, the second extender peptide is located between the carboxyl terminus of the therapeutic agent and the antibody region. In some embodiments, n is between about 1 and about 10. In some embodiments, n is between about 1 and about 5. In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more. n can be about 1 to about 3. X 1 -X 7 Can be independently selected from positively charged amino acids or hydrophobic amino acids. 1 -X 7Can be independently selected from: alanine (A), asparagine (N), isoleucine (I), leucine (L), valine (V), glutamine (Q), glutamic acid (E), and lysine (K). Alanine (A) can constitute at least about 30% of the total amino acid composition. Alanine (A) can constitute less than about 70% of the total amino acid composition. Leucine can constitute at least about 20% of the total amino acid composition. Leucine can constitute less than about 50% of the total amino acid composition. Lysine can constitute at least about 20% of the total amino acid composition. Lysine can constitute less than about 50% of the total amino acid composition. Asparagine can constitute about 50% of the total amino acid composition. Isoleucine can constitute about 50% of the total amino acid composition. Valine can constitute about 50% of the total amino acid composition. Glutamine can constitute about 50% of the total amino acid composition. Glutamic acid can constitute about 50% of the total amino acid composition. Hydrophobic amino acids can constitute at least about 50% of the total amino acid composition. Hydrophobic amino acids can constitute at least about 60% of the total amino acid composition. Hydrophobic amino acids can constitute at least about 70% of the total amino acid composition. Hydrophobic amino acids may constitute less than about 90% of the total amino acid composition.

[0307] The extender peptide may comprise the sequence (X 1 X 2 X 3 X 4 X 5 X 6 X 7 ) n X a X b X c X d (SEQ ID NO: 158). In some embodiments, the second extender peptide comprises the sequence (X 1 X 2 X 3 X 4 X 5 X 6 X 7 ) n X a X b X c X d (SEQ ID NO: 156). In some cases, the second extender peptide is located between the carboxyl terminus of the therapeutic agent and the antibody region. In some embodiments, n is between about 1 and about 10. In some embodiments, n is between about 1 and about 5. In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more. n can be from about 1 to about 3. X 1 -X 7 Can be independently selected from positively charged amino acids or hydrophobic amino acids. 1 -X 7Can be independently selected from: alanine (A), leucine (L), and lysine (K). A can constitute at least about 30% of the total amino acid composition. A can constitute less than about 70% of the total amino acid composition. L can constitute at least about 20% of the total amino acid composition. L can constitute less than about 50% of the total amino acid composition. K can constitute at least about 20% of the total amino acid composition. K can constitute less than about 50% of the total amino acid composition. Hydrophobic amino acids can constitute at least about 50% of the total amino acid composition. Hydrophobic amino acids can constitute at least about 60% of the total amino acid composition. Hydrophobic amino acids can constitute at least about 70% of the total amino acid composition. Hydrophobic amino acids can constitute less than about 90% of the total amino acid composition. X a 、X b and X d Can be independently selected from hydrophobic amino acids. c Can be polar, uncharged amino acids. X a 、X b and X d Can be the same amino acid. a 、X b and X d Can be different amino acids. In some embodiments, X a is glycine (G). In some embodiments, X b In some embodiments, X d It is glycine. a 、X b and X d It can be glycine (G). X c It may be serine (S).

[0308] The extender peptide may comprise the sequence (ELAALEA) n X a X b X c X d (SEQ ID NO: 157). In some embodiments, the second extender peptide comprises the sequence (ELAALEA) n X a X b X c X d (SEQ ID NO: 157). In some cases, the second extender peptide is located between the carboxyl terminus of the therapeutic agent and the antibody region. In some embodiments, n is between about 1 and about 10. In some embodiments, n is between about 1 and about 5. In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more. n can be about 1 to about 3. X a 、X b and X d Can be independently selected from hydrophobic amino acids.c Can be polar, uncharged amino acids. X a 、X b and X d Can be the same amino acid. a 、X b and X d Can be different amino acids. In some embodiments, X a is glycine (G). In some embodiments, X b In some embodiments, X d It is glycine. a 、X b and X d It can be glycine (G). X c It may be serine (S).

[0309] The extender peptide may comprise the sequence (ELAALEA) n (SEQ ID NO: 158). In some embodiments, the second extender peptide comprises the sequence (ELAALEA) n (SEQ ID NO: 158). In some cases, the second extender peptide is located between the carboxyl terminus of the therapeutic agent and the antibody region. In some embodiments, n is between about 1 and about 10. In some embodiments, n is between about 1 and about 5. In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more. n can be from about 1 to about 3.

[0310] The extender peptide may comprise the sequence (ELAALEA) n GGSG (SEQ ID NO: 159). In some embodiments, the second extender peptide comprises the sequence (ELAALEA) n GGSG (SEQ ID NO: 159). In some cases, the second extender peptide is located between the carboxyl terminus of the therapeutic agent and the antibody region. In some embodiments, n is between about 1 and about 10. In some embodiments, n is between about 1 and about 5. In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more. n can be about 1 to about 3.

[0311] The immunoglobulin fusion protein may comprise (a) a first extender peptide comprising an amino acid sequence based on or derived from SEQ ID NO: 151; and (b) a second extender peptide comprising an amino acid sequence based on or derived from SEQ ID NO: 161. The immunoglobulin fusion protein may comprise (a) a first extender peptide comprising an amino acid sequence that is at least about 50% homologous to the amino acid sequence of SEQ ID NO: 151; and (b) a second extender peptide comprising an amino acid sequence that is at least about 50% homologous to the amino acid sequence of SEQ ID NO: 161. The first extender peptide may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more homologous to the amino acid sequence of SEQ ID NO: 151. The second extender peptide may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more homologous to the amino acid sequence of SEQ ID NO: 161. The first extender peptide may comprise an amino acid sequence comprising 3, 4, 5, 6, 7 or more amino acids based on or derived from the amino acid sequence of SEQ ID NO: 151. The first extender peptide may comprise an amino acid sequence comprising 5 or more amino acids based on or derived from the amino acid sequence of SEQ ID NO: 151. The second extender peptide may comprise an amino acid sequence comprising 3, 4, 5, 6, 7 or more amino acids based on or derived from the amino acid sequence of SEQ ID NO: 161. The second extender peptide may comprise an amino acid sequence comprising 5 or more amino acids based on or derived from the amino acid sequence of SEQ ID NO: 161.

[0312] Aliphatic amino acids may constitute at least about 20% of the total amino acids of the stretch peptide. Aliphatic amino acids may constitute at least about 22%, 25%, 27%, 30%, 32%, 35%, 37%, 40%, 42%, 45% or more of the total amino acids of the stretch peptide. Aliphatic amino acids may constitute at least about 22% of the total amino acids of the stretch peptide. Aliphatic amino acids may constitute at least about 27% of the total amino acids of the stretch peptide.

[0313] Aliphatic amino acids may constitute less than about 50% of the total amino acids of the stretcher peptide. Aliphatic amino acids may constitute less than about 47%, 45%, 43%, 40%, 38%, 35%, 33% or 30% of the total amino acids of the stretcher peptide.

[0314] Aliphatic amino acids may constitute about 20% to about 45% of the total amino acids of the stretcher peptide. Aliphatic amino acids may constitute about 23% to about 45% of the total amino acids of the stretcher peptide. Aliphatic amino acids may constitute about 23% to about 40% of the total amino acids of the stretcher peptide.

[0315] Aromatic amino acids may constitute less than about 20% of the total amino acids of the extended peptide. Aromatic amino acids may constitute less than about 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11% or 10% of the total amino acids of the extended peptide. Aromatic amino acids may constitute 0% to about 20% of the total amino acids of the extended peptide.

[0316] Non-polar amino acids may constitute at least about 30% of the total amino acids of the stretcher peptide. Non-polar amino acids may constitute at least about 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39% or 40% of the total amino acids of the stretcher peptide. Non-polar amino acids may constitute at least about 32% of the total amino acids of the stretcher peptide.

[0317] Non-polar amino acids may constitute less than about 80% of the total amino acids of the stretcher peptide.Non-polar amino acids may constitute less than about 77%, 75%, 72%, 70%, 69% or 68% of the total amino acids of the stretcher peptide.

[0318] Non-polar amino acids may constitute about 35% to about 80% of the total amino acids of the stretcher peptide. Non-polar amino acids may constitute about 38% to about 80% of the total amino acids of the stretcher peptide. Non-polar amino acids may constitute about 38% to about 75% of the total amino acids of the stretcher peptide. Non-polar amino acids may constitute about 35% to about 70% of the total amino acids of the stretcher peptide.

[0319] Polar amino acids may constitute at least about 20% of the total amino acids of the stretcher peptide. Polar amino acids may constitute at least about 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35% or more of the total amino acids of the stretcher peptide. Polar amino acids may constitute at least about 23% of the total amino acids of the stretcher peptide.

[0320] Polar amino acids may constitute less than about 80% of the total amino acids of the stretcher peptide. Polar amino acids may constitute less than about 77%, 75%, 72%, 70%, 69% or 68% of the total amino acids of the stretcher peptide. Polar amino acids may constitute less than about 77% of the total amino acids of the stretcher peptide. Polar amino acids may constitute less than about 75% of the total amino acids of the stretcher peptide. Polar amino acids may constitute less than about 72% of the total amino acids of the stretcher peptide.

[0321] Polar amino acids may constitute about 25% to about 70% of the total amino acids of the stretcher peptide. Polar amino acids may constitute about 27% to about 70% of the total amino acids of the stretcher peptide. Polar amino acids may constitute about 30% to about 70% of the total amino acids of the stretcher peptide.

[0322] Alternatively, the immunoglobulin fusion proteins disclosed herein do not comprise an extender peptide.

[0323] therapeutic agents

[0324] The immunoglobulin fusion proteins disclosed herein may comprise one or more therapeutic agents. The therapeutic agent may be a peptide. The therapeutic agent may be a small molecule. The immunoglobulin fusion proteins disclosed herein may comprise two or more therapeutic agents. The immunoglobulin fusion proteins disclosed herein may comprise 3, 4, 5, 6, or more therapeutic agents. The two or more therapeutic agents may be the same. The two or more therapeutic agents may be different.

[0325] The therapeutic agent may comprise any secondary structure, such as an alpha helix or a beta chain, or may not comprise a regular secondary structure. The therapeutic agent may comprise an amino acid with one or more modifications, including but not limited to: myristoylation, palmitoylation, prenylation, glycosylphosphatidylinositolization (glypiation), lipidation, acylation, acetylation, alkylation, methylation, glycosylation, malonylation, hydroxylation, iodination, nucleotide addition, oxidation, phosphorylation, adenylylation, propionylation, succinylation, sulfation, selenylation, biotinylation, pegylation, deimination, deamidation, eliminylation, and carbamylation. The therapeutic agent may comprise one or more amino acids conjugated to one or more small molecules (e.g., drugs). In some embodiments, the therapeutic agent comprises one or more non-natural amino acids. In some embodiments, the therapeutic agent comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30, 40, 50 or more unnatural amino acids. In some embodiments, the therapeutic agent comprises one or more amino acid substitutions. In some embodiments, the therapeutic agent comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30, 40, 50 or more amino acid substitutions.

[0326] The therapeutic agent can be inserted into the immunoglobulin region. The insertion of the therapeutic agent into the immunoglobulin region can include the removal or deletion of a portion of the immunoglobulin on which the immunoglobulin region is based or derived. The therapeutic agent can replace at least a portion of the heavy chain. The therapeutic agent can replace at least a portion of the light chain. The therapeutic agent can replace at least a portion of the variable domain. The therapeutic agent can replace at least a portion of the constant domain. The therapeutic agent can replace at least a portion of the complementary determining region (CDR). The therapeutic agent can replace at least a portion of CDR1. The therapeutic agent can replace at least a portion of CDR2. The therapeutic agent can replace at least a portion of CDR3. The therapeutic agent can replace at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more of the immunoglobulin or portion thereof.

[0327] The one or more therapeutic agents may be based on or derived from a protein. The protein may be a growth factor, cytokine, hormone, or toxin. The growth factor may be GCSF, GMCSF, GDF11, or FGF21. The GCSF may be bovine GCSF. The GCSF may be human GCSF. The GMCSF may be bovine GMCSF or human GMCSF. The FGF21 may be bovine FGF21. The FGF21 may be human FGF21. The protein may be elastin. The protein may be a peptidase inhibitor. The protease may be skin-derived anti-leukocyte protease (SKALP).

[0328] The cytokine may be an interferon or an interleukin. The cytokine may be stromal cell-derived factor 1 (SDF-1). The interferon may be interferon-β. The interferon may be interferon-α. The interleukin may be interleukin 11 (IL-11). The interleukin may be interleukin 8 (IL-8) or interleukin 21 (IL-21).

[0329] The hormone can be exendin-4, GLP-1, relaxin, oxyntomodulin, hLeptin (human leptin), betatrophin, bovine growth hormone (bGH), human growth hormone (hGH), erythropoietin (EPO), or parathyroid hormone. The hormone can be somatostatin. The parathyroid hormone can be human parathyroid hormone. The erythropoietin can be human erythropoietin.

[0330] The toxin may be Moka1, VM24, or Mamba1. The toxin may be ziconotide or chlorotoxin. In one embodiment, the toxin is mu-SLPTX-Ssm6a (Ssam6).

[0331] The protein may be angiopoietin-like 3 (ANGPTL3). The angiopoietin-like 3 may be human angiopoietin-like 3.

[0332] The therapeutic agent may be glucagon-like peptide 2 (GLP2).

[0333] In some embodiments, the therapeutic agent is a glucagon analog.

[0334] In some embodiments, the therapeutic agent is a dual agonist.

[0335] In some embodiments, one or more regions of the therapeutic agent are configured to treat diabetes and / or conditions associated with diabetes. In some embodiments, 2, 3, 4, 5 or more regions of the therapeutic agent are configured to treat diabetes and / or conditions associated with diabetes. Diabetes may include type 1 diabetes, type 2 diabetes, gestational diabetes, and prediabetes. In some embodiments, one or more regions of the therapeutic agent are configured to treat obesity and / or conditions associated with obesity. In some embodiments, 2, 3, 4, 5 or more regions of the therapeutic agent are configured to treat obesity and / or conditions associated with obesity. Conditions may include complications and diseases. Examples of conditions associated with diabetes include, but are not limited to, diabetic retinopathy, diabetic nephropathy, diabetic heart disease, diabetic foot disease, diabetic neuropathy, macroangiopathy, diabetic cardiomyopathy, infection, and diabetic ketoacidosis. Diabetic neuropathy may include, but is not limited to, symmetrical polyneuropathy, autonomic neuropathy, radiculopathy, cranial neuropathy, and mononeuropathy. Obesity-related conditions include, but are not limited to, heart disease, stroke, high blood pressure, diabetes, osteoarthritis, gout, sleep apnea, asthma, gallbladder disease, gallstones, abnormal blood lipids (e.g., abnormal LDL and HDL cholesterol levels), obesity hypoventilation syndrome, reproductive problems, liver steatosis, and mental health conditions.

[0336] In some embodiments, one or more regions of the therapeutic agent are glucagon-like protein-1 (GLP-1) receptor agonists or formulations thereof. In some embodiments, one or more regions of the therapeutic agent are incretin mimetics. In some embodiments, one or more regions of the therapeutic agent comprise an amino acid sequence based on or derived from the amino acid sequence of exendin-4, exenatide, or a synthetic thereof. In some embodiments, one or more regions of the therapeutic agent are glucagon analogs or formulations thereof. In some embodiments, one or more regions of the therapeutic agent comprise an amino acid sequence based on or derived from the amino acid sequence of insulin. In some embodiments, one or more regions of the therapeutic agent are bispecific. In some embodiments, the therapeutic agent is specific for the GLP-1 receptor and the glucagon receptor. In some embodiments, one or more regions of the therapeutic agent comprise an amino acid sequence based on or derived from the amino acid sequence of oxyntomodulin.

[0337] In some embodiments, one or more regions of the therapeutic agent are configured to treat short bowel syndrome and / or conditions associated with short bowel syndrome. In some embodiments, 2, 3, 4, 5 or more regions of the therapeutic agent are configured to treat short bowel syndrome and / or conditions associated with short bowel syndrome. Conditions associated with short bowel syndrome may include, but are not limited to, bacterial overgrowth in the small intestine, metabolic acidosis, gallstones, kidney stones, malnutrition, osteomalacia, intestinal failure, and weight loss. In some embodiments, one or more regions of the therapeutic agent are configured to treat inflammatory bowel disease and / or conditions associated with inflammatory bowel disease. In some embodiments, 2, 3, 4, 5 or more regions of the therapeutic agent are configured to treat inflammatory bowel disease and / or conditions associated with inflammatory bowel disease. Inflammatory bowel disease and / or inflammatory bowel disease-associated conditions include, but are not limited to, ulcerative colitis, Crohn's disease, collagenous colitis, lymphocytic colitis, ischemic colitis, diversion colitis, Behcet's disease, indeterminate colitis, anemia, arthritis, pyoderma gangrenosum, primary sclerosing cholangitis, euthyroid syndrome; and abdominal pain, vomiting, diarrhea, rectal bleeding, internal cramping or muscle cramps, and weight loss in individuals with inflammatory bowel disease.

[0338] In some embodiments, one or more regions of the therapeutic agent comprise an amino acid sequence based on or derived from the amino acid sequence of glucagon, a glucagon analog, a glucagon-like peptide, and / or a glucagon-like peptide analog. In some embodiments, one or more regions of the therapeutic agent comprise an amino acid sequence based on or derived from the amino acid sequence of glucagon-like peptide-2 (GLP2).

[0339] In some embodiments, one or more regions of a therapeutic agent are configured to treat an autoimmune disease and / or an autoimmune disease-related condition. In some embodiments, 2, 3, 4, 5 or more regions of a therapeutic agent are configured to treat an autoimmune disease and / or an autoimmune disease-related condition. Autoimmune diseases and / or autoimmune disease-related conditions may include, but are not limited to, acute disseminated encephalomyelitis, alopecia areata, antiphospholipid syndrome, autoimmune cardiomyopathy, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inner ear disease, autoimmune lymphoproliferative syndrome, autoimmune peripheral neuropathy, autoimmune pancreatitis, autoimmune polyendocrine disease syndrome, autoimmune progesterone dermatitis, autoimmune thrombocytopenic purpura, autoimmune urticaria, autoimmune uveitis, Behçet's disease, celiac disease, cold agglutinin disease, Crohn's disease, dermatomyositis, type 1 diabetes mellitus, eosinophilic fasciitis, gastrointestinal pemphigoid, Goodpasture's syndrome, Graves' disease, Guillain-Barre syndrome, Hashimoto's encephalopathy, Hashimoto's thyroiditis, and other autoimmune conditions. thyroiditis), idiopathic thrombocytopenic purpura, lupus erythematosus, Miller Fisher syndrome, mixed connective tissue disease, multiple sclerosis, myasthenia gravis, narcolepsy, pemphigus vulgaris, pernicious anemia, polymyositis, primary biliary cirrhosis, psoriasis, psoriatic arthritis, relapsing polychondritis, rheumatoid arthritis, rheumatic fever, Sjögren's syndrome, temporal arteritis, transverse myelitis, ulcerative colitis, undifferentiated connective tissue disease, vasculitis, and Wegener's granulomatosis.

[0340] In some embodiments, one or more regions of the therapeutic agent comprise an amino acid sequence based on or derived from an amino acid sequence that binds to a potassium channel. In some embodiments, one or more regions of the therapeutic agent comprise an amino acid sequence based on or derived from the amino acid sequence of Mokatoxin-1 (Moka).

[0341] In some embodiments, one or more regions of therapeutic agent are configured to treat pain. In some embodiments, 2, 3, 4, 5 or more regions of therapeutic agent are configured to treat pain.

[0342] In some embodiments, one or more regions of the therapeutic agent comprise an amino acid sequence based on or derived from an amino acid sequence that is a neurotoxin. In some embodiments, one or more regions of the therapeutic agent comprise an amino acid sequence based on or derived from the amino acid sequence of the neurotoxin mu-SLPTX-Ssm6a (Ssam6). In some embodiments, one or more regions of the therapeutic agent comprise an amino acid sequence based on or derived from the amino acid sequence of mambalign-1.

[0343] In some embodiments, one or more regions of the therapeutic agent are configured to treat heart failure and / or fibrosis. In some embodiments, one or more regions of the therapeutic agent are configured to treat conditions associated with heart failure and / or fibrosis. In some embodiments, 2, 3, 4, 5 or more regions of the therapeutic agent are configured to treat heart failure and / or fibrosis. In some embodiments, 2, 3, 4, 5 or more regions of the therapeutic agent are configured to treat conditions associated with heart failure and / or fibrosis. In some embodiments, 2, 3, 4, 5 or more regions of the therapeutic agent are configured to treat conditions associated with heart failure and / or fibrosis. Conditions associated with heart failure may include coronary heart disease, hypertension, diabetes, cardiomyopathy, valvular heart disease, arrhythmia, congenital heart defect, obstructive sleep apnea, myocarditis, hyperthyroidism, hypothyroidism, emphysema, hemochromatosis and amyloidosis. Heart failure may be left-sided heart failure, right-sided heart failure, systolic heart failure and diastolic heart failure. Fibrosis can include, but is not limited to, pulmonary fibrosis, idiopathic pulmonary fibrosis, cystic fibrosis, cirrhosis, endomyocardial fibrosis, myocardial infarction, atrial fibrosis, mediastinal fibrosis, myelofibrosis, retroperitoneal fibrosis, progressive massive fibrosis, nephrogenic systemic fibrosis, Crohn's disease, keloids, scleroderma / systemic sclerosis, arthrofibrosis, Peyronie's disease, Dupuytren's contracture, and adhesive capsulitis.

[0344] In some embodiments, one or more regions of the therapeutic agent comprise an amino acid sequence based on or derived from an amino acid sequence belonging to the insulin superfamily. In some embodiments, one or more regions of the therapeutic agent comprise an amino acid sequence based on or derived from the amino acid sequence of insulin.

[0345] In some embodiments, the amino acids of the therapeutic agent are based on or derived in whole or in part from any one of SEQ ID NOs: 227-267. The therapeutic agent may comprise an amino acid sequence that is at least about 50% homologous to any one of SEQ ID NOs: 227-267. The therapeutic agent may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 97% homologous to any one of SEQ ID NOs: 227-267. The therapeutic agent may comprise an amino acid sequence that is at least about 70% homologous to any one of SEQ ID NOs: 227-267. The therapeutic agent may comprise an amino acid sequence that is at least about 80% homologous to any one of SEQ ID NOs: 227-267. The therapeutic agent may comprise an amino acid sequence that is at least about 50% identical to any one of SEQ ID NOs: 227-267. The therapeutic agent may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 97% identical to any one of SEQ ID NOs: 227-267. The therapeutic agent may comprise an amino acid sequence that is at least about 70% identical to any one of SEQ ID NOs: 227-267. The therapeutic agent may comprise an amino acid sequence that is at least about 80% identical to any one of SEQ ID NOs: 227-267. The therapeutic agent may comprise an amino acid sequence that is 100% identical to any one of SEQ ID NOs: 227-267. In some embodiments, the therapeutic agent comprises an amino acid sequence that is at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 97% homologous to the amino acid sequence of any one of SEQ ID NOs: 227-267. In some embodiments, the therapeutic agent comprises an amino acid sequence that is at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 97% identical to the amino acid sequence of any one of SEQ ID NOs: 227-267. In some embodiments, the therapeutic agent comprises an amino acid sequence that is 100% identical to the amino acid sequence of any one of SEQ ID NOs: 227-267.

[0346] The therapeutic agent may comprise an amino acid sequence comprising 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 or more amino acids based on or derived from any one of SEQ ID NOs: 227-267. The therapeutic agent may comprise an amino acid sequence comprising 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 450, 500 or more amino acids based on or derived from any one of SEQ ID NOs: 227-267. The therapeutic agent may comprise an amino acid sequence comprising 10 or more amino acids based on or derived from any one of SEQ ID NOs: 227-267. The therapeutic agent may comprise an amino acid sequence comprising 50 or more amino acids based on or derived from any one of SEQ ID NOs: 227-267. The therapeutic agent may comprise an amino acid sequence comprising 100 or more amino acids based on or derived from any one of SEQ ID NOs: 227-267. The therapeutic agent may comprise an amino acid sequence comprising 200 or more amino acids based on or derived from any one of SEQ ID NOs: 227-267. The amino acids may be continuous. Alternatively, or additionally, the amino acids are discontinuous. In some embodiments, the therapeutic agent may comprise amino acids derived from any one of SEQ ID NOs: 227-267 and amino acids not derived from any one of SEQ ID NOs: 227-267. In some embodiments, the therapeutic agent may comprise amino acids derived from one or more of SEQ ID NOs: 227-267 and amino acids not derived from any one of SEQ ID NOs: 227-267. In some embodiments, the therapeutic agent comprises amino acids derived from 1, 2, 3, or 4 of SEQ ID NOs: 227-267.

[0347] The therapeutic agent may be encoded by a nucleic acid sequence based on or derived from any one of SEQ ID NOs: 186-226. The therapeutic agent may be encoded by a nucleic acid sequence that is at least about 50% homologous to any one of SEQ ID NOs: 186-226. The therapeutic agent may be encoded by a nucleic acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more homologous to any one of SEQ ID NOs: 186-226. The therapeutic agent may be encoded by a nucleic acid sequence that is at least about 70% homologous to any one of SEQ ID NOs: 186-226. The therapeutic agent may be encoded by a nucleic acid sequence that is at least about 80% homologous to any one of SEQ ID NOs: 186-226.

[0348] The therapeutic agent may comprise a protease cleavage site. The protease cleavage site may be inserted into the therapeutic agent. In some embodiments, the therapeutic agent comprises a first therapeutic agent region and a second therapeutic agent region. In some embodiments, the therapeutic agent comprises a protease cleavage site disposed between the first therapeutic agent region and the second therapeutic agent region. In some embodiments, the first therapeutic agent region and the second therapeutic agent region are derived from the same protein or amino acid sequence group. In some embodiments, the first therapeutic agent region and the second therapeutic agent region are derived from different protein or amino acid sequence groups. One or more protease cleavage sites may be attached to the N-terminus, C-terminus, or both the N-terminus and the C-terminus of the therapeutic agent region.

[0349] The therapeutic agent may comprise one or more internal connector peptides. The therapeutic agent may comprise two or more internal connector peptides. The therapeutic agent may comprise 3, 4, 5, 6, 7 or more internal connector peptides. The connector peptides may be different. The connector peptides may be the same. The connector peptides may be inserted within the therapeutic agent. In some embodiments, the therapeutic agent comprises a first therapeutic region, a second therapeutic region, and one or more connector peptides located between the first therapeutic region and the second therapeutic region. The one or more connector peptides may be attached to the N-terminus, C-terminus, or both the N-terminus and C-terminus of the therapeutic agent region. In some embodiments, the connector peptide is a protease cleavage site. In some embodiments, the connector peptide is a label, such as an affinity tag. An example of an affinity tag is a 6X (HHHHHH) histidine tag. In some embodiments, the internal connector comprises amino acids having a repeating sequence. In some embodiments, the internal connector has 2, 3, 4, 5, 6, 7, 8, 9, 10 or more repeating sequences. In some embodiments, the internal connector is hypoimmunogenic. In some embodiments, the internal connector is biodegradable.

[0350] The therapeutic agent can be inserted into the antibody region. The insertion of the therapeutic agent into the antibody region may include the removal or deletion of one or more amino acids from the antibody region.

[0351] In some embodiments, the immunoglobulin fusion protein comprises one or more extender peptides. The one or more extender peptides can be attached to the N-terminus, the C-terminus, or both the N-terminus and the C-terminus of the therapeutic agent.

[0352] In some embodiments, the immunoglobulin fusion protein comprises one or more linker peptides. The one or more linker peptides can be attached to the N-terminus, the C-terminus, or both the N-terminus and the C-terminus of the therapeutic agent.

[0353] In some embodiments, the immunoglobulin fusion protein comprises one or more proteolytic cleavage sites. The one or more proteolytic cleavage sites can be attached to the N-terminus, the C-terminus, or both the N-terminus and the C-terminus of the therapeutic agent.

[0354] In some embodiments, the therapeutic agent can be linked to the antibody region without the aid of a stretcher peptide.The therapeutic agent can be linked to the antibody via one or more linkers.

[0355] Connector

[0356] The immunoglobulin fusion protein, antibody region, non-antibody region, and / or extension fusion region may further comprise one or more linkers. The immunoglobulin fusion protein, antibody region, non-antibody region, and / or extension fusion region may further comprise 2, 3, 4, 5, 6, 7, 8, 9, 10, or more linkers. The extension fusion region may further comprise one or more linkers. The extension fusion region may further comprise 2, 3, 4, 5, 6, 7, 8, 9, 10, or more linkers.

[0357] The one or more linkers are attached to the N-terminus, C-terminus, or both of the therapeutic agent. The one or more linkers are attached to the N-terminus, C-terminus, or both of the stretcher peptide. The one or more linkers are attached to the N-terminus, C-terminus, or both of the stretcher peptide. The one or more linkers are attached to the N-terminus, C-terminus, or both of the proteolytic cleavage site. The one or more linkers can be attached to the therapeutic agent, the stretcher peptide, the proteolytic cleavage site, the stretcher fusion region, the antibody region, or a combination thereof.

[0358] In some embodiments, the linker peptide is a connecting peptide or a portion of a connecting peptide.

[0359] The one or more linkers may comprise the sequence (X e X f X g X h ) n (SEQ ID NO: 176). In one embodiment, n is between about 1 and about 20. In one embodiment, n is any one of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. In one embodiment, n is between about 1 and about 10. In one embodiment, n is between about 1 and about 5. In one embodiment, n is between about 1 and about 3. In one embodiment, X e 、X f and X g Independently selected from hydrophobic amino acids. hIt can be a polar, uncharged amino acid. The linker sequence can also contain one or more cysteine ​​(C) residues. The one or more cysteine ​​residues are at the N-terminus, the C-terminus, or a combination thereof. The linker peptide can contain the sequence CX e X f X g X h (SEQ ID NO: 177). In one embodiment, X e 、X f and X g Independently selected from hydrophobic amino acids. h Can be a polar, uncharged amino acid. The linker peptide may comprise the sequence X e X f X g X h C (SEQ ID NO: 178). In one embodiment, X e 、X f and X g Independently selected from hydrophobic amino acids. h May be a polar, uncharged amino acid.

[0360] The one or more linkers may comprise the sequence (GGGGS) n (SEQ ID NO: 179). In one embodiment, n is between about 1 and about 20. In one embodiment, n is any one of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. In one embodiment, n is between about 1 and about 10. In one embodiment, n is between about 1 and about 5. In one embodiment, n is between about 1 and about 3.

[0361] The one or more linkers may comprise an amino acid sequence selected from any one of SEQ ID NOs: 176-181. The one or more linkers may comprise an amino acid sequence that is at least about 50% homologous to any one of SEQ ID NOs: 176-181. The one or more linkers may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more homologous to any one of SEQ ID NOs: 176-181. The one or more linkers may comprise an amino acid sequence that is at least about 70% homologous to any one of SEQ ID NOs: 176-181. The one or more linkers may comprise an amino acid sequence that is at least about 80% homologous to any one of SEQ ID NOs: 176-181.

[0362] Proteolytic cleavage site

[0363] The immunoglobulin fusion proteins disclosed herein may further comprise one or more proteolytic cleavage sites. The immunoglobulin fusion proteins disclosed herein may further comprise two or more proteolytic cleavage sites. The immunoglobulin fusion proteins disclosed herein may further comprise three or more proteolytic cleavage sites. The immunoglobulin fusion proteins disclosed herein may further comprise four, five, six, seven or more proteolytic cleavage sites. The therapeutic agents disclosed herein may further comprise one or more proteolytic cleavage sites.

[0364] The immunoglobulin fusion protein may comprise a sequence having one or more cleavage sites between the antibody region and the non-antibody region. The immunoglobulin fusion protein may comprise a sequence having one or more cleavage sites between the antibody region and the extension fusion region. In some embodiments, the proteolytic cleavage site is or is part of a connecting peptide.

[0365] The one or more proteolytic cleavage sites may be attached to the N-terminus, C-terminus, or both of the therapeutic peptide. The one or more proteolytic cleavage sites may be attached to the N-terminus, C-terminus, or both of the extender peptide. The one or more proteolytic cleavage sites may be attached to the N-terminus, C-terminus, or both of the extender peptide. The one or more proteolytic cleavage sites may be attached to the N-terminus, C-terminus, or both of the linker peptide. The one or more proteolytic cleavage sites may be attached to the therapeutic agent, the extender peptide, the linker, the extender fusion region, the immunoglobulin region, the non-immunoglobulin region, or a combination thereof.

[0366] Digestion of the proteolytic cleavage site can result in release of the N- or C-terminus of the therapeutic agent from the immunoglobulin fusion protein. The proteolytic cleavage site can be at the N- and C-termini of the therapeutic agent. Digestion of the proteolytic cleavage site can result in release of the therapeutic agent from the immunoglobulin fusion protein.

[0367] Alternatively, or additionally, the proteolytic cleavage site is located within the amino acid sequence of the therapeutic agent, the stretcher peptide, the antibody region, or a combination thereof. The therapeutic agent may comprise one or more proteolytic cleavage sites within the amino acid sequence. For example, SEQ ID NO: 89 discloses a relaxin protein comprising two internal proteolytic cleavage sites. Digestion of the proteolytic cleavage sites within the relaxin protein can result in the release of internal fragments of the relaxin protein.

[0368] The two or more proteolytic cleavage sites can surround the therapeutic agent, the extender peptide, the linker, the antibody region, or a combination thereof. Digestion of the proteolytic cleavage site can result in the release of the peptide fragment located between the two or more proteolytic cleavage sites. For example, the proteolytic cleavage sites can flank the therapeutic agent-linker peptide. Digestion of the proteolytic cleavage site can result in the release of the therapeutic agent-linker.

[0369] The proteolytic cleavage site can be recognized by one or more polymerases. The one or more polymerases can be serine proteases, threonine proteases, cysteine ​​proteases, aspartic proteases, glutamic proteases, metalloproteinases, exopeptidases, endopeptidases or a combination thereof. The protease can be selected from Factor VII or Factor Xa. Other examples of the protease include, but are not limited to, aminopeptidase, carboxypeptidase, trypsin, chymotrypsin, pepsin, papain and elastase. The protease can be prohormone convertase 2 (PC2).

[0370] The one or more proteolytic cleavage sites may comprise an amino acid sequence selected from any one of SEQ ID NOs: 182-185. The one or more proteolytic cleavage sites may comprise an amino acid sequence that is at least about 50% homologous to any one of SEQ ID NOs: 182-185. The one or more proteolytic cleavage sites may comprise an amino acid sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more homologous to any one of SEQ ID NOs: 182-185. The one or more proteolytic cleavage sites may comprise an amino acid sequence that is at least about 70% homologous to any one of SEQ ID NOs: 182-185. The one or more proteolytic cleavage sites may comprise an amino acid sequence that is at least about 80% homologous to any one of SEQ ID NOs: 182-185.

[0371] Vectors, host cells and recombination methods

[0372] Immunoglobulin fusion proteins as disclosed herein can be expressed by recombinant methods. Generally, the nucleic acid encoding the immunoglobulin fusion protein can be separated and inserted into a reproducible vector for further cloning (amplification of DNA) or expression. The DNA encoding the immunoglobulin fusion protein can be prepared by PCR amplification and sequenced using conventional procedures (for example, by adopting oligonucleotide probes that can be specifically bound to the nucleotides encoding the immunoglobulin fusion protein). In an exemplary embodiment, the nucleic acid encoding the immunoglobulin fusion protein is amplified by PCR, digested with restriction enzymes and gel purified. The digested nucleic acid can be inserted into a reproducible vector. The reproducible vector containing the immunoglobulin fusion protein inserted by digestion can be transformed or transduced into a host cell for further cloning (amplification of DNA) or expression. The host cell can be a prokaryotic cell or a eukaryotic cell.

[0373] The polynucleotide sequence of the polypeptide component (e.g., antibody region, extension peptide, therapeutic agent) encoding the immunoglobulin fusion protein can be obtained by PCR amplification. The polynucleotide sequence can be isolated from the cell containing the nucleic acid encoding the polypeptide component and sequenced. Alternatively, or additionally, a nucleotide synthesizer or PCR technology can be used to synthesize the polynucleotide. Once obtained, the sequence encoding the polypeptide component can be inserted into a recombinant vector that can replicate and express the heterologous polynucleotide in a prokaryotic and / or eukaryotic host.

[0374] In addition, phage vectors containing replicons and control sequences compatible with the host microorganism can be used as transformation vectors relevant to these hosts. For example, phage such as λGEM TM -11 can be used in the preparation of recombinant vectors that can be used to transform susceptible host cells (such as Escherichia coli LE392).

[0375] The immunoglobulin fusion protein can be expressed intracellularly (eg, cytoplasmically) or extracellularly (eg, secreted). For extracellular expression, the vector can contain a secretion signal that enables the immunoglobulin fusion protein to be transferred outside the cell.

[0376] Suitable host cells for cloning or expressing vectors encoding immunoglobulin fusion proteins include prokaryotes or eukaryotic cells. The host cell can be a eukaryotic cell. Examples of eukaryotic cells include, but are not limited to, human embryonic kidney (HEK) cells, Chinese hamster ovary (CHO) cells, fungi, yeast, invertebrate cells (e.g., plant cells and insect cells), lymphoid cells (e.g., YO, NSO, Sp20 cells). Other examples of suitable mammalian host cell lines are monkey kidney CV1 lines (COS-7) transformed by SV40; baby hamster kidney cells (BHK); mouse Sertoli cells; monkey kidney cells (CV1); African green monkey kidney cells (VERO-76); human cervical cancer cells (HELA); canine kidney cells (MDCK); buffalo rat liver cells (BRL 3A); human lung cells (W138); human liver cells (Hep G2); mouse mammary tumor (MMT 060562); TR1 cells; MRC5 cells; and FS4 cells. The host cell can be a prokaryotic cell (eg, E. coli).

[0377] Host cells can be transformed with vectors containing nucleotides encoding immunoglobulin fusion proteins. Transformed host cells can be cultivated in culture medium. Culture medium can be supplemented with one or more agents to induce promoters, select transformants or amplify, or express genes encoding the desired sequence. Methods for transforming host cells are known in the art and may include electroporation, calcium chloride, or polyethylene glycol / DMSO.

[0378] Alternatively, host cells can be transfected or transduced with a vector containing nucleotides encoding the immunoglobulin fusion protein. The transfected or transduced host cells can be cultured in a culture medium. The culture medium can be supplemented with one or more agents for inducing a promoter, selecting transfected or transduced cells, or expressing a gene encoding a desired sequence.

[0379] Host cells can be transfected or transduced with a vector comprising nucleotides encoding one or more proteases. A vector comprising a protease can be co-transfected with a vector encoding any immunoglobulin fusion protein disclosed herein. Proteases include Factor Xa and PC2.

[0380] The expressed immunoglobulin fusion protein can be secreted into the periplasm of the host cell and therefrom recovered, or transported to the culture medium. Recovering protein from the periplasm can include destroying the host cell. The destruction of the host cell can include osmotic shock, ultrasonic treatment or cracking. Centrifugation or filtration can be used to remove cell debris or whole cells. For example, the immunoglobulin fusion protein can be further purified by affinity resin chromatography.

[0381] Alternatively, the immunoglobulin fusion protein secreted into the culture medium can be isolated in the culture medium. Cells can be removed from the culture and a culture supernatant can be generated that is filtered and concentrated for further protein purification. The expressed polynucleotides can be further isolated and identified using known methods such as polyacrylamide gel electrophoresis (PAGE) and Western blotting.

[0382] Can carry out the production of immunoglobulin fusion protein in a large number by fermentation process.Multiple large-scale fed-batch fermentation processes can be used for the production of recombinant protein.Large-scale fermentation has a throughput of at least 1000 liters, preferably about 1,000 to 100,000 liters of throughput.These fermentor tanks utilize agitator impellers to distribute oxygen and nutrients, particularly glucose (preferred carbon source / energy source).Small-scale fermentation generally refers to the fermentation in the fermentor tank that volume capacity is no more than about 100 liters, and can be about 1 liter to about 100 liters scope.

[0383] In the fermentation process, induction of protein expression is usually initiated after the cells have grown to a desired density (e.g., OD550 of about 180-220) under suitable conditions, at which stage the cells are in the early stationary phase. As known in the art and as described herein, a variety of inducers can be used, depending on the vector construct employed. Cells can be grown for a shorter period of time before induction. Although longer or shorter induction times can be employed, cells are typically induced for about 12-50 hours.

[0384] In order to improve the production yield and quality of the immunoglobulin fusion proteins disclosed herein, multiple fermentation conditions can be modified. For example, to improve the correct assembly and folding of the secreted immunoglobulin fusion protein polypeptide, additional overexpression chaperone proteins such as Dsb proteins (DsbA, DsbB, DsbC, DsbD and / or DsbG) or FkpA (a peptidyl prolyl cis, trans-isomerase with chaperone activity) can be used to co-transform host prokaryotic cells. Chaperone proteins have been shown to promote the correct folding and solubility of heterologous proteins produced in bacterial host cells.

[0385] In order to minimize the proteolysis of the expressed heterologous proteins (particularly those that are proteolytically sensitive), certain host strains lacking proteolytic enzymes can be used in the present invention. For example, the host bacterial strain can be modified to produce genetic mutations in the genes encoding known bacterial proteases (such as protease III, OmpT, DegP, Tsp, protease I, protease Mi, protease V, protease VI, and combinations thereof). Some E. coli strains lacking protease are available.

[0386] Standard protein purification methods known in the art can be employed. The following procedures are exemplary of suitable purification procedures: fractionation on an immunoaffinity column or an ion exchange column, ethanol precipitation, reversed-phase HPLC, chromatography on silica or a cation exchange resin such as DEAE, chromatofocusing, SDS-PAGE, ammonium sulfate precipitation, hydroxyapatite chromatography, gel electrophoresis, dialysis, and affinity chromatography and gel filtration using, for example, Sephadex G-75.

[0387] Commercially available protein concentration filters such as Amicon or Millipore can be used. Ultrafiltration unit to concentrate immunoglobulin fusion proteins.

[0388] A protease inhibitor or cocktail of protease inhibitors may be included in any of the foregoing steps to inhibit proteolysis of the immunoglobulin fusion protein.

[0389] In some cases, the immunoglobulin fusion protein may not have biological activity after separation. A variety of methods for "refolding" or converting the polypeptide into its tertiary structure and generating disulfide bonds can be used to restore biological activity. Such methods include exposing the dissolved polypeptide to a pH generally greater than 7 and in the presence of a specific concentration of a chaotropic agent. The choice of chaotropic agent is very similar to that used for inclusion body dissolution, but the chaotropic agent is usually used at a lower concentration and does not have to be the same as the chaotropic agent used for dissolution. In most cases, the refolding / oxidation solution will also contain a reducing agent or a specific ratio of a reducing agent plus its oxidized form to generate a specific redox potential, thereby allowing disulfide shuffling to occur during the formation of cysteine ​​bridges in the protein. Some commonly used redox pairs include cysteine / cystamine, glutathione (GSH) / disulfide diGSH, copper chloride, dithiothreitol (DTT) / dithiane DTT, and 2-mercaptoethanol (bME) / disulfide-b(ME). In many cases, a cosolvent can be used to increase the efficiency of refolding, and common reagents used for this purpose include glycerol, polyethylene glycols of various molecular weights, arginine, etc.

[0390] Composition

[0391] Disclosed herein are compositions comprising the immunoglobulin fusion proteins and / or immunoglobulin fusion protein components disclosed herein. The composition may comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more immunoglobulin fusion proteins. These immunoglobulin fusion proteins may be different. Alternatively, these immunoglobulin fusion proteins may be the same or similar. These immunoglobulin fusion proteins may comprise different antibody regions, extender fusion regions, extender peptides, therapeutic agents, or combinations thereof.

[0392] The composition may also comprise one or more pharmaceutically acceptable salts, excipients or carriers. Pharmaceutically acceptable salts, excipients or carriers used in the pharmaceutical compositions of the present invention include carriers, excipients, diluents, antioxidants, preservatives, colorants, flavorings and diluents, emulsifiers, suspending agents, solvents, fillers, swelling agents, buffers, delivery vehicles, tonicity agents, cosolvents, wetting agents, complexing agents, buffers, antimicrobial agents and surfactants.

[0393] Neutral buffered saline or saline mixed with serum albumin is an exemplary suitable carrier. The pharmaceutical composition may include antioxidants such as ascorbic acid; low molecular weight polypeptides; proteins such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinyl pyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides and other carbohydrates, including glucose, mannose or dextrin; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and / or non-ionic surfactants such as Tween, pluronics or polyethylene glycol (PEG). Also by way of example, suitable tonicity enhancers include alkali metal halides (preferably sodium chloride or potassium chloride), mannitol, sorbitol, and the like. Suitable preservatives include benzalkonium chloride, thimerosal, phenylethyl alcohol, methylparaben, propylparaben, chlorhexidine, sorbic acid, and the like. Hydrogen peroxide can also be used as a preservative. Suitable cosolvents include glycerol, propylene glycol, and PEG. Suitable complexing agents include caffeine, polyvinyl pyrrolidone, beta-cyclodextrin, or hydroxy-propyl-beta-cyclodextrin. Suitable surfactants or wetting agents include sorbitan esters, polysorbates such as polysorbate 80, tromethamine, lecithin, cholesterol, tyloxapal, and the like. The buffer can be a conventional buffer such as acetate, borate, citrate, phosphate, bicarbonate, or Tris-HCl. Acetate buffer can be approximately pH 4-5.5, while Tris buffer can be approximately pH 7-8.5. Additional agents are described in Remington's Pharmaceutical Sciences, 18th Edition, A.R. Gennaro, ed., Mack Publishing Company, 1990.

[0394] The composition can be in liquid form or in a lyophilized or freeze-dried form and can include one or more lyoprotectants, excipients, surfactants, high molecular weight structural additives and / or bulking agents (see, for example, U.S. Patents 6,685,940, 6,566,329 and 6,372,716). In one embodiment, a lyoprotectant is included that is a non-reducing sugar such as sucrose, lactose or trehalose. The amount of lyoprotectant typically included is such that after reconstitution, the resulting formulation will be isotonic, although hypertonic or slightly hypotonic formulations may also be suitable. In addition, the amount of lyoprotectant should be sufficient to prevent degradation and / or aggregation of the protein to an unacceptable degree upon lyophilization. Exemplary lyoprotectant concentrations of sugars (e.g., sucrose, lactose, trehalose) in pre-lyophilized formulations are from about 10 mM to about 400 mM. In another embodiment, surfactants are included, for example, nonionic surfactants and ionic surfactants such as polysorbates (e.g., polysorbate 20, polysorbate 80); poloxamers (e.g., poloxamer 188); poly(ethylene glycol) phenyl ethers (e.g., Triton); sodium dodecyl sulfate (SDS); sodium lauryl sulfate; sodium octyl glucoside (sodium octyl glycoside); glycoside); lauryl-, myristyl-, linoleyl-, or stearyl-sulfobetaine; lauryl-, myristyl-, linoleyl-, or stearyl-sarcosine; linoleyl-, myristyl-, or cetyl-betaine; lauramidopropyl-, cocamidopropyl-, linoleamidopropyl-, myristamidopropyl-, palmitamidopropyl-, or isostearamidopropyl-betaine (e.g., lauramidopropyl); myristamidopropyl-, palmitamidopropyl-, or isostearamidopropyl-dimethylamine; sodium methyl co...

Claims

1. A nucleic acid encoding an immunoglobulin fusion protein, wherein the immunoglobulin fusion protein comprises a first extension fusion region located in the complementarity determining region 3 (CDR3) of a first antibody region, the first extension fusion region comprising a therapeutic agent located between a first extension peptide and a second extension peptide configured to form an antiparallel coiled coil, the first extension peptide comprising GGSGAKLAALKAKLAALK (SEQ ID NO: 151), and the second extension peptide comprising ELAALEAELAALEAGGSG (SEQ ID NO: 161), wherein the nucleic acid comprises a nucleotide sequence comprising 50 or more nucleotides based on or derived from one of SEQ ID NOs: 37-56 and 58-67.

2. The nucleic acid of claim 1, further comprising a second therapeutic agent.

3. The nucleic acid of claim 1, wherein the antibody from which the first antibody region is derived comprises an amino acid sequence that is at least 90% identical to any one of SEQ ID NOs: 19-36 and 271-273.

4. The nucleic acid of claim 1, wherein the antibody from which the first antibody region is derived comprises an amino acid sequence that is at least 90% identical to a trastuzumab immunoglobulin.

5. The nucleic acid of claim 1, wherein the antibody from which the first antibody region is derived comprises an amino acid sequence that is at least 90% identical to the palivizumab immunoglobulin. The nucleic acid of claim 1 , further comprising a second antibody region.

7. A nucleic acid encoding any one of the immunoglobulin fusion proteins of SEQ ID NOs: 68-99.

8. A nucleic acid comprising a sequence that is at least 90% identical to any one of SEQ ID NOs:37-56 and 58-67, wherein the nucleic acid sequence encodes an immunoglobulin fusion protein comprising a first extender fusion region located in the complementarity determining region 3 (CDR3) of a first antibody region, the first extender fusion region comprising a therapeutic agent located between a first extender peptide and a second extender peptide configured to form an antiparallel coiled coil, the first extender peptide comprising GGSGAKLAALKAKLAALK (SEQ ID NO: 151), and the second extender peptide comprising ELAALEAELAALEAGGSG (SEQ ID NO: 161).

9. A vector comprising the nucleic acid according to any one of claims 1 to 8.

10. A host cell comprising the vector according to claim 9.

11. A host cell comprising the nucleic acid according to any one of claims 1 to 8.

12. Use of the nucleic acid according to any one of claims 1 to 8 in the preparation of a medicament for treating an autoimmune disease, wherein the autoimmune disease is multiple sclerosis.

13. Use of the nucleic acid according to any one of claims 1 to 8 in the preparation of a medicament for treating a central nervous system disorder, wherein the central nervous system disorder is Alzheimer's disease.

14. Use of the nucleic acid according to any one of claims 1 to 8 in the preparation of a medicament for treating a central nervous system disorder, wherein the central nervous system disorder is Parkinson's disease.

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