Liquid formulation and use thereof

Abstract

The present application relates to a liquid formulation comprising an oil phase solvent system and a protein, wherein the oil phase solvent system comprises an oil phase solvent, the mass fraction of which is about 50% or more.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biological medicine, in particular to a liquid preparation and application thereof. BACKGROUND

[0002] Tumor is a disease that seriously threatens human health. In recent years, immunotherapy as a new therapy has shown great potential in tumor treatment. Cytokine is a very important immune signal in the body. Cytokine fusion protein technology is another hot direction of current tumor immunotherapy. This method is based on the fact that these cytokines have the same or related functional activity and different target points. Two or more cytokines are fused together by genetic engineering technology. However, the effect of tumor treatment by using cytokine fusion protein technology is still not satisfactory, and there are many needs for improvement. SUMMARY

[0003] The present application provides a liquid preparation, which comprises an oil phase solvent system and a protein, wherein the oil phase solvent system comprises an oil phase solvent, and the mass fraction of the oil phase solvent is about 50% or more.

[0004] In some embodiments, the oil phase solvent is selected from the group consisting of glycerol, propylene glycol, polyethylene glycol, polyvinylpyrrolidone, mannitol, sorbitol, polyoxypropylene and tromethamine.

[0005] In some embodiments, the mass fraction of the oil phase solvent is about 60% or more.

[0006] In some embodiments, the mass fraction of the oil phase solvent is about 70% or more.

[0007] In some embodiments, the mass fraction of the protein is about 1% to about 50%.

[0008] In some embodiments, the protein comprises a cytokine and / or an antibody.

[0009] In some embodiments, the cytokine comprises two or more selected from the group consisting of IL12, IL2, GMCSF, IL7, IL15, IL21 and FLT3L.

[0010] In some embodiments, the cytokine is derived from a mammal.

[0011] In some embodiments, the protein comprises a fusion protein, and the fusion protein comprises at least two cytokines selected from the group consisting of IL12, IL2, GMCSF, IL7, IL15, IL21 and FLT3L.

[0012] In certain embodiments, the protein comprises a fusion protein, and the fusion protein comprises any one group of cytokines selected from the group consisting of:

[0013] a) IL12, IL2 and GMSCF;

[0014] b) IL12, IL7 and GMSCF;

[0015] c) IL12, IL15 and GMSCF;

[0016] d) IL12, IL21 and GMSCF;

[0017] e) IL12, IL2 and FLT3L;

[0018] f) IL12, IL7 and FLT3L;

[0019] g) IL12, IL15 and FLT3L; and

[0020] h) IL12, IL21 and FLT3L.

[0021] In certain embodiments, the protein further comprises a targeting moiety.

[0022] In certain embodiments, the targeting moiety is capable of specifically recognizing and / or binding to a tumor-associated antigen.

[0023] In certain embodiments, the tumor-associated antigen is selected from the group consisting of EDB domain of fibronectin, EDA domain of fibronectin, and necrotic regions.

[0024] In certain embodiments, the targeting moiety comprises an antibody or an antigen-binding fragment thereof.

[0025] In certain embodiments, the targeting moiety comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-15.

[0026] In certain embodiments, the protein comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-67.

[0027] In certain embodiments, the protein is a single chain protein.

[0028] In certain embodiments, the single chain protein comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-51.

[0029] In certain embodiments, the protein is a dimer consisting of a first polypeptide chain and a second polypeptide chain, wherein the first polypeptide chain is different from the second polypeptide chain.

[0030] In some embodiments, the first polypeptide chain comprises IL12a and the second polypeptide chain comprises IL12b.

[0031] In some embodiments, IL2 or a functional fragment thereof is in the first polypeptide chain or in the second polypeptide chain, GMCSF or a functional fragment thereof is in the first polypeptide chain or in the second polypeptide chain, and the one or more targeting moieties are each independently in the first polypeptide chain or in the second polypeptide chain.

[0032] In some embodiments, in the first polypeptide chain, the IL2 or a functional fragment thereof, the IL12a or a functional fragment thereof, and the GMCSF or a functional fragment thereof are contained in this order from the N-terminus to the C-terminus.

[0033] In some embodiments, in the first polypeptide chain, the targeting moiety, the IL12a or a functional fragment thereof, the IL2 or a functional fragment thereof, and the GMCSF or a functional fragment thereof are contained in this order from the N-terminus to the C-terminus.

[0034] In some embodiments, in the second polypeptide chain, the IL12b or a functional fragment thereof and the targeting moiety are contained in this order from the N-terminus to the C-terminus.

[0035] In some embodiments, wherein,

[0036] a) the first polypeptide chain comprises the amino acid sequence set forth in SEQ ID NO: 53 and the second polypeptide chain comprises the amino acid sequence set forth in SEQ ID NO: 52;

[0037] b) the first polypeptide chain comprises the amino acid sequence set forth in SEQ ID NO: 55 and the second polypeptide chain comprises the amino acid sequence set forth in SEQ ID NO: 54;

[0038] c) the first polypeptide chain comprises the amino acid sequence set forth in SEQ ID NO: 56 and the second polypeptide chain comprises the amino acid sequence set forth in SEQ ID NO: 57;

[0039] d) the first polypeptide chain comprises the amino acid sequence set forth in SEQ ID NO: 58 and the second polypeptide chain comprises the amino acid sequence set forth in SEQ ID NO: 59;

[0040] e) the first polypeptide chain comprises the amino acid sequence set forth in SEQ ID NO: 60 and the second polypeptide chain comprises the amino acid sequence set forth in SEQ ID NO: 61;

[0041] f) the first polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 62 and the second polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 63;

[0042] g) the first polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 64 and the second polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 65;

[0043] h) the first polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 66 and the second polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 67.

[0044] In another aspect, the present application also provides a medicament comprising the liquid preparation described herein.

[0045] In certain embodiments, the liquid preparation described herein is formulated as an injection.

[0046] In certain embodiments, the medicament described herein further comprises a diluent, wherein the diluent and the liquid preparation do not mix with each other.

[0047] In another aspect, the present application also provides use of the liquid preparation described herein in the manufacture of a medicament for treating a tumor.

[0048] In certain embodiments, the tumor comprises lung cancer.

[0049] In certain embodiments, the liquid preparation described herein is used for treating a tumor.

[0050] In another aspect, the present application also provides a method for treating a tumor, comprising administering to a subject in need thereof the liquid preparation described herein.

[0051] In certain embodiments, the method of administration is intratumoral injection.

[0052] In another aspect, the present application also provides an excipient for preparing the liquid preparation described herein, comprising an oil phase solvent, wherein the mass fraction of the oil phase solvent is about 50% or more.

[0053] Other aspects and advantages of the present application will be readily appreciated from the detailed description that follows, when considered in connection with the accompanying drawings. The detailed description and drawings show only exemplary embodiments of the present application. As will be readily appreciated, the present application makes it possible to make modifications to the disclosed specific embodiments without departing from the spirit and scope of the application. Accordingly, the drawings and description are merely exemplary and not limiting. BRIEF DESCRIPTION OF DRAWINGS

[0054] The specific features of the invention to which this application relates are set forth in the claims appended hereto. The features and advantages of the invention to which this application relates are better understood with reference to the detailed description of the example embodiments and the accompanying drawings. A brief description of the drawings is as follows:

[0055] Figure 1 To show the effect of glycerol on tumor growth in mice;

[0056] Figure 2 To show the effect of the liquid formulation described in this application on tumor growth in mice;

[0057] Figure 3 To show the effect of the liquid formulation described in this application on tumor growth in mice;

[0058] Figure 4 To show the effect of the liquid formulation described in this application on tumor growth in mice;

[0059] Figure 5 To show the effect of the liquid formulation described in this application on tumor growth in mice;

[0060] Figure 6 To show the effect of the liquid formulation described in this application on tumor growth in mice. DETAILED DESCRIPTION

[0061] The embodiments of the invention will be described in relation to specific examples, which will enable a person skilled in the art to readily understand other advantages and effects of the invention disclosed in this specification.

[0062] The present application is further described below: In the present application, unless otherwise specified, the scientific and technical terms used herein have the meanings commonly understood by one of ordinary skill in the art. Also, the terms and techniques related to protein and nucleic acid chemistry, molecular biology, cell and tissue culture, microbiology, immunology, and the like, and the laboratory procedures set forth herein are those commonly used by those skilled in the art. In addition, for better understanding of the present application, the definitions and explanations of the related terms are provided below.

[0063] In the present application, the term "liquid formulation" generally refers to a pharmaceutical composition in the form of a liquid, which generally comprises an active ingredient and a pharmaceutically acceptable carrier or excipient. For example, in the present application, the liquid formulation can include an oil phase solvent system and a protein, wherein the oil phase solvent system includes an oil phase solvent having a mass fraction of about 50% or more. The oil phase solvent can include glycerol, propylene glycol, and / or polyethylene glycol. In the present application, the protein can be dispersed or dissolved in the oil phase solvent to prepare the liquid formulation described in the present application.

[0064] In the present application, the term "protein" can be considered as belonging to "cytokine fusion protein", which generally refers to a fusion protein obtained by fusing two or more cytokines together through genetic recombination technology. It has both the unique biological activity of its constituent factors or significantly improves some of its activities, and it can also play a complex biological function that single cytokine simple combination does not have through the complementary and synergistic effect of biological activity, and it can even produce some new structures and biological functions.

[0065] In the present application, the terms "IL12", "IL12a", "IL12b", "IL2", "GMCSF", "IL7", "IL15", "IL21", "FLT3L" can be considered as belonging to "cytokine". The "cytokine" generally refers to a class of small molecular proteins with a wide range of biological activities synthesized and secreted by immune cells (such as mononuclear, macrophages, T cells, B cells, NK cells, etc.) and some non-immune cells (for example, endothelial cells, epidermal cells, fibroblasts, etc.) stimulated. The cytokine has an important regulatory effect on cell-cell interaction, cell growth and differentiation. In the present application, the cytokine can be selected from one or more of the following group: interleukin (IL), FMS-related tyrosine kinase 3 ligand (FLT3L) and colony stimulating factor (Colony Stimulating Factor, CSF). The interleukin generally refers to a cytokine produced by lymphocytes, monocytes or other non-mononuclear cells. In the present application, the interleukin can be selected from one or more of the following group: IL12, IL2, IL7, IL15, IL21. In the present application, the colony stimulating factor generally refers to a cytokine that can stimulate different hematopoietic stem cells to form cell colonies in semi-solid culture medium. In the present application, the colony stimulating factor can be granulocyte macrophage colony stimulating factor (Granulocyte Macrophage Colony Stimulating Factor, GMCSF).

[0066] In the present application, the term "IL12" generally refers to interleukin-12, which plays an important regulatory role in cell-cell interactions, immunomodulation, hematopoiesis, and inflammatory processes. The molecule of IL12 is generally a heterodimer, which generally includes two subunits, a p40 subunit (40 kd) and a p35 subunit (35 kd), which are connected together by a disulfide bond. In the present application, IL12 containing the p35 subunit (35 kd) can be denoted as IL12a, and IL12 containing the p40 subunit (40 kd) can be denoted as IL12b. For example, the p35 subunit in IL12 derived from a mouse (mIL12) can comprise an amino acid sequence as set forth in SEQ ID NO: 16, and the p40 subunit can comprise an amino acid sequence as set forth in SEQ ID NO: 17. For another example, the p35 subunit in IL12 derived from a human (hIL12) can comprise an amino acid sequence as set forth in SEQ ID NO: 18, and the p40 subunit can comprise an amino acid sequence as set forth in SEQ ID NO: 19.

[0067] In the present application, the term "IL2" generally refers to interleukin-2, which plays an important regulatory role in cell-cell interactions, immunomodulation, hematopoiesis, and inflammatory processes. For example, IL2 derived from a mouse (mIL2) can comprise an amino acid sequence as set forth in SEQ ID NO: 20. For another example, IL2 derived from a human (hIL2) can comprise an amino acid sequence as set forth in SEQ ID NO: 21.

[0068] In the present application, the term "IL15" generally refers to interleukin-15, which plays an important regulatory role in cell-cell interactions, immunomodulation, hematopoiesis, and inflammatory processes. For example, IL15 derived from a mouse (mIL15) can comprise an amino acid sequence as set forth in SEQ ID NO: 22. For another example, IL15 derived from a human (hIL15) can comprise an amino acid sequence as set forth in SEQ ID NO: 23.

[0069] In the present application, the term "IL7" generally refers to interleukin-7, which plays an important regulatory role in cell-cell interactions, immunomodulation, hematopoiesis, and inflammatory processes. For example, IL7 derived from a mouse (mIL7) can comprise an amino acid sequence as set forth in SEQ ID NO: 24. For another example, IL7 derived from a human (hIL7) can comprise an amino acid sequence as set forth in SEQ ID NO: 25.

[0070] In the present application, the term "IL21" generally refers to interleukin-21, which plays an important regulatory role in cell-cell interactions, immune regulation, hematopoiesis, and inflammatory processes. For example, IL21 derived from mouse (mIL21) can comprise an amino acid sequence as set forth in SEQ ID NO: 26. For another example, IL21 derived from human (hIL21) can comprise an amino acid sequence as set forth in SEQ ID NO: 27.

[0071] In the present application, the term "FLT3L" generally refers to FMS-related tyrosine kinase 3 ligand, which can regulate the proliferation and differentiation of non-erythroid hematopoietic stem cells, promote the proliferation, differentiation and maturation of pre-B lymphocytes, dendritic cells, NK cells, cytotoxic T lymphocytes, and has important anti-tumor effect. For example, FLT3L derived from mouse (mFLT3L) can comprise an amino acid sequence as set forth in SEQ ID NO: 28. For another example, FLT3L derived from human (hFLT3L) can comprise an amino acid sequence as set forth in SEQ ID NO: 29.

[0072] In the present application, the term "GMCSF" generally refers to granulocyte macrophage colony-stimulating factor. The GMCSF can have 4 alpha-helix bundle structure. For example, GMCSF derived from mouse (mGMCSF) can comprise an amino acid sequence as set forth in SEQ ID NO: 30. For another example, GMCSF derived from human (hGMCSF) can comprise an amino acid sequence as set forth in SEQ ID NO: 31.

[0073] In the present application, the term "antibody" generally refers to an immunoglobulin or a fragment or a derivative thereof, and encompasses any polypeptide that comprises an antigen binding site, whether produced in vitro or in vivo. The term includes, but is not limited to, polyclonal, monoclonal, monospecific, multispecific, non-specific, humanized, single-chain, chimeric, synthetic, recombinant, hybrid, mutated, and grafted antibodies. Unless otherwise modified by the term "intact", such as in "intact antibody", for the purposes of the present application, the term "antibody" also includes antibody fragments, such as Fab, F(ab')2, Fv, scFv, Fd, dAb, and other antibody fragments that retain antigen binding function (i.e., specifically bind to, for example, OX40 or PD-L1). Typically, such fragments should include an antigen binding domain.

[0074] The basic 4-chain antibody unit is a heterotetrameric glycoprotein composed of two identical light (L) chains and two identical heavy (H) chains. IgM antibodies comprise 5 of the basic heterotetramer units along with an additional polypeptide called J chain, and contain 10 antigen binding sites. IgA antibodies, which are present as a dimer of 4 chain units, can also contain J chain. For both IgG and IgA, the 4 chain unit is generally about 150,000 Daltons. Each L chain is linked to a H chain by one covalent disulfide bond, while each H chain is inter-connected to another H chain by one or more disulfide bonds depending on the H chain isotype. Each H and L chain also has regularly spaced intrachain disulfide bridges. Each H chain has at the N-terminus a variable region (VH) followed by three constant domains (CH) for each of the α and γ chains, and four CH domains for μ and ε isotypes. Each L chain has a variable region (VL) at its N-terminus, and a constant domain at its other end. The VL is aligned with the VHand the CL is aligned with a first constant domain of a heavy chain (CHI). Particular amino acid residues of each of these domains have been described as forming an interface between the light chain and heavy chain variable domains. The VHand VLpair together to form a single antigen binding site. For the structure and properties of the different classes of antibodies, see, e.g., Basic and Clinical Immunology, 8th Edition, Daniel P. Sties, Abba I. Terr and Tristram G. Parsolw (eds), Appleton & Lange, Norwalk, Conn., 1994, page 71 and Chapter 6. The L chain from any vertebrate species can be assigned to one of two clearly distinct types, called kappa and lambda, based on the amino acid sequences of their constant domains. Depending on their heavy chain constant domain (CH) amino acid sequence, immunoglobulins can be assigned to different classes or isotypes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, with several of these having subclasses (e.g., IgG1, IgG2A, IgG2B, IgG3, IgG4, IgA1, and IgA2). The γ and α classes are further divided based on relatively small differences in the CH sequence and function into subclasses, e.g., humans express the following subclasses: IgG1, IgG2A, IgG2B, IgG3, IgG4, IgA1, and IgA2.

[0075] In the present application, the term "targeting moiety" generally refers to a class of moieties that act on certain specific tissues, cells. For example, the targeting moiety can specifically target a tumor-associated antigen. In the present application, the targeting moiety includes an antibody or an antigen-binding fragment thereof.

[0076] The term "specifically recognizes and / or binds" as used herein generally refers to a measurable and reproducible interaction, such as binding, between a target and an antibody, which can be determined in the presence of a heterogeneous population of molecules (including biological molecules). For example, an antibody that specifically binds a target (which can be an epitope) is one that binds that target with greater affinity, avidity, more readily, and / or for longer duration than it binds other targets. In one embodiment, the extent of binding of an antibody to an unrelated, non-target, is less than about 10% of the binding of the antibody to the target, as measured, e.g., by radioimmunoassay (RIA). In certain embodiments, an antibody that specifically binds a target has a dissociation constant (KD) of <1 x 10 -6 M, <1 x 10 -7 M, <1 x 10 -8 M, <1 x 10 - 9 M, or <1 x 10 -10 M. In certain embodiments, an antibody specifically binds an epitope on a protein that is conserved among proteins from different species. In another embodiment, specific binding can include, but does not require, exclusive binding.

[0077] In the present application, the term "tumor-associated antigen" (TAA) generally refers to an antigenic molecule present on a tumor cell or a normal cell. The tumor-associated antigen can include an embryonic protein, a glycoprotein antigen, and a squamous cell antigen. The tumor-associated antigen can be selected from the group consisting of an EDB domain of fibronectin, an EDA domain of fibronectin, and necrotic regions of cells.

[0078] In the present application, the term "antigen-binding fragment" generally refers to a fragment having antigen-binding activity. In the present application, the antigen-binding fragment can be selected from the group consisting of Fab, Fab', F(ab')2, F(ab)2, dAb, an isolated complementarity determining region (CDR), Fv, and scFv.

[0079] In the present application, the term "single-chain protein" generally refers to a polypeptide of primary structure consisting of an uninterrupted sequence of one continuous amino acid residue. For example, in the present application, the single-chain protein can comprise an amino acid sequence as set forth in any one of the group consisting of SEQ ID NOs: 32-51.

[0080] In the present application, the term "dimer" generally refers to a macromolecular complex formed from two monomeric units, which are usually non-covalently bonded. Each monomeric unit can be a macromolecule, such as a polypeptide chain or a polynucleotide. For example, in the present application, the protein can be a dimer consisting of a first polypeptide chain and a second polypeptide chain.

[0081] In the present application, the term "polypeptide chain" generally refers to a macromolecule comprising two or more covalently linked peptides. The peptides within a polypeptide chain can be connected to each other by a peptide bond. Each polypeptide chain can comprise an N-terminal or amino-terminal end and a C-terminal or carboxyl-terminal end.

[0082] In the present application, the term "functional fragment" generally refers to a fragment that retains a certain specific function, for example, an IL12a functional fragment refers to a fragment that retains the function of IL12a. For example, an IL12a functional fragment can be IL12a, fragment (GenBank: AIC49052.1). For another example, an IL12b functional fragment can be IL12b, fragment (GenBank: AIC54621.1).

[0083] In the present application, the term "injection" generally refers to a sterile solution (including emulsions and suspensions) of a drug prepared for infusion into the body and sterile powders or concentrated solutions prepared for the extemporaneous preparation of a solution or suspension for injection. For example, an injection can be used to introduce a drug into the body by intravenous injection or subcutaneous injection. In the present application, the liquid preparation can be formulated as an injection.

[0084] In the present application, the term "diluent" generally refers to a solvent used to dilute a substance, for example, a solvent that reduces the concentration of a substance. In the present application, the drug can also contain a diluent that does not mix with the liquid preparation.

[0085] In the present application, the term "tumor" generally refers to a neoplasm or solid lesion formed by abnormal cell growth. In the present application, a tumor can be a solid tumor or a blood tumor. For example, a tumor can include lung cancer.

[0086] In the present application, the term "subject" generally refers to a human or non-human animal, including but not limited to a cat, dog, horse, pig, cow, sheep, rabbit, mouse, rat, or monkey.

[0087] In the present application, the term "administration" generally refers to a method of giving a subject (e.g., a patient) a dose of a liquid preparation or a drug. Administration can be performed by any suitable means, including parenterally, intrapulmonarily, and intranasally, as well as intralesionally if local treatment is desired. Parenteral infusions include, for example, intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. The drug can be administered by any suitable route, for example, by injection (such as intravenous or subcutaneous injection), depending in part on whether the administration is short-term or long-term. Various dosing schedules are contemplated herein, including but not limited to a single administration or multiple administrations at various time points, bolus administration, and pulse infusion. For example, in the present application, the administration can be an intratumoral injection. The "intratumoral injection" generally refers to an injection of a dose of a liquid preparation or a drug into the interior of a tumor.

[0088] In the present application, the term "adjuvant" generally refers to an auxiliary material, e.g., a material that aids in the preparation of a certain substance. For example, in the present application, an adjuvant used in the preparation of the liquid formulation can include an oil phase solvent, which can be present in a mass fraction of about 50% or more.

[0089] In the present application, the term "comprising" generally means including, but not limited to.

[0090] In the present application, the term "about" generally means a variation of 0.5-10% above or below the specified value, e.g., a variation of 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% above or below the specified value.

[0091] Liquid formulations

[0092] In one aspect, the present application provides a liquid formulation, which can include an oil phase solvent system and a protein, wherein the oil phase solvent system can include an oil phase solvent, which can be present in a mass fraction of about 50% or more.

[0093] In addition, in some embodiments, the oil phase solvent can be present in a mass fraction of about 60% or more. For example, in other embodiments, the oil phase solvent can be present in a mass fraction of about 70% or more. For example, the oil phase solvent can be present in a mass fraction of about 50% to about 60%, about 60% to about 70%, about 70% to about 80%, about 80% to about 90%, about 90% to about 95%, about 50% to about 70%, about 50% to about 80%, about 50% to about 90%, about 50% to about 95%, about 60% to about 80%, about 60% to about 90%, about 60% to about 95%, about 70% to about 90%, about 70% to about 95%, or about 80% to about 95%.

[0094] In the present application, the oil phase solvent can be selected from the group consisting of glycerol, propylene glycol, polyethylene glycol, polyvinylpyrrolidone, mannitol, sorbitol, polyoxypropylene, and tromethamine.

[0095] In the present application, the oil phase solvent can be glycerol, and the mass fraction of the glycerol can be about 50% or more. In some embodiments, the mass fraction of the glycerol can be about 60% or more. For example, in other embodiments, the mass fraction of the glycerol can be about 70% or more. For example, the mass fraction of the glycerol can be about 50% to about 60%, about 60% to about 70%, about 70% to about 80%, about 80% to about 90%, about 90% to about 95%, about 50% to about 70%, about 50% to about 80%, about 50% to about 90%, about 50% to about 95%, about 60% to about 80%, about 60% to about 90%, about 60% to about 95%, about 70% to about 90%, about 70% to about 95%, or about 80% to about 95%.

[0096] In the present application, the mass fraction of the protein can be about 1% to about 50%, for example, the mass fraction of the protein can be about 1% to about 10%, about 1% to about 20%, about 1% to about 30%, about 1% to about 40%, about 1% to about 50%, 5% to about 10%, about 5% to about 20%, about 5% to about 30%, about 5% to about 40%, about 5% to about 50%, about 10% to about 20%, about 10% to about 30%, about 10% to about 40%, about 10% to about 50%, about 20% to about 30%, about 20% to about 40%, about 20% to about 50%, about 30% to about 40%, about 30% to about 50%, about 40% to about 50%, or about 45% to about 50%.

[0097] In some embodiments, the liquid formulation described in the present application can be prepared by dissolving or dispersing the protein in the oil phase solvent system (e.g., in the oil phase solvent).

[0098] In the present application, the oil phase solvent and the protein can be mutually soluble or mutually insoluble.

[0099] In the present application, the oil phase solvent and the protein have a synergistic effect, thereby enhancing the effect of the liquid formulation described in the present application in treating tumors.

[0100] In the present application, the liquid formulation described in the present application can be used for treating tumors, in which the oil phase solvent and the protein can synergistically act on each other, thereby enhancing the effect of the liquid formulation described in the present application in treating tumors, for example, the effect of the liquid formulation described in the present application in treating tumors is significantly and enhanced compared to the oil phase solvent or the protein alone. For example, a protein solution alone generally cannot induce tumor regression, but using the liquid formulation described in the present application can induce tumor regression.

[0101] In the present application, the liquid formulation can be used to inhibit tumor growth. For example, the liquid formulation of the present application can inhibit or delay the development or progression of a disease, can reduce the size of a tumor (even substantially eliminate the tumor) by promoting the expression of cytokines, and / or can reduce and / or stabilize the disease state.

[0102] Further, it is also noted that the liquid formulation of the present application, in addition to including an oil phase solvent system and a protein, can also include an amino acid, which can be any one or more of glycine, glutamine, asparagine, arginine, or lysine. The liquid formulation of the present application can also include an antimicrobial agent or an antioxidant. The antioxidant can be any one or more of ascorbic acid, sodium sulfite, or sodium bisulfite. The liquid formulation of the present application can also include a buffering agent, which can be any one or more of borate, bicarbonate, Tris-HCl, citrate, phosphate, or other organic acids. The liquid formulation of the present application can also include a bulking agent, such as mannitol or glycine. The liquid formulation of the present application can also include a chelating agent, such as ethylenediaminetetraacetic acid (EDTA. The liquid formulation of the present application can also include a complexing agent, such as any one or more of caffeine, polyvinylpyrrolidone, beta-cyclodextrin, or hydroxypropyl-beta-cyclodextrin. The liquid formulation of the present application can also include a carbohydrate, such as dextrose, mannose, or dextrin. The liquid formulation of the present application can also include a preservative, such as any one or more of benzalkonium chloride, benzoic acid, salicylic acid, thimerosal, phenethyl alcohol, methylparaben, propylparaben, chlorhexidine, sorbic acid, or hydrogen peroxide.

[0103] Proteins

[0104] In the present application, the protein can include a cytokine and / or an antibody.

[0105] In the present application, the antibody can be selected from one or more of a monoclonal antibody, a polyclonal antibody, an antibody fragment, a chimeric antibody, a bispecific antibody, a heteroconjugate antibody, a single chain (ScFv), a fusion protein comprising an antibody portion (e.g., a domain antibody), a humanized antibody, and any other modified configuration of the immunoglobulin molecule that comprises an antigen recognition site of the required specificity. Among these, the antibody fragment can be selected from one or more of a Fab, a Fab', a F(ab')2, a Fv, and a Fc. The fusion protein comprising an antibody portion can be a domain antibody.

[0106] In the present application, the antibody can also include a glycosylation variant of an antibody, an amino acid sequence variant of an antibody, and a covalently modified antibody. The antibody can be a canine, a rat, a human, or any other source, such as a chimeric or humanized antibody.

[0107] In the present application, the cytokine can be any one of IL12, IL2, GMCSF, IL7, IL15, IL21 and FLT3L.

[0108] In the present application, the cytokine can also include two or more selected from the group consisting of IL12, IL2, GMCSF, IL7, IL15, IL21 and FLT3L. Wherein the cytokine can be derived from a mammal. The mammal can be a human or a mouse. For example, the amino acid sequence of IL12a derived from a mouse (denoted as mIL12a) can be as shown in SEQ ID NO. 16, the amino acid sequence of IL12b derived from a mouse (denoted as mIL12b) can be as shown in SEQ ID NO. 17, the amino acid sequence of IL2 derived from a mouse (denoted as mIL2) can be as shown in SEQ ID NO. 20, the amino acid sequence of GMCSF derived from a mouse (denoted as mGMCSF) can be as shown in SEQ ID NO. 30, the amino acid sequence of IL7 derived from a mouse (denoted as mIL7) can be as shown in SEQ ID NO. 24, the amino acid sequence of IL15 derived from a mouse (denoted as mIL15) can be as shown in SEQ ID NO. 22, the amino acid sequence of IL21 derived from a mouse (denoted as mIL21) can be as shown in SEQ ID NO. 26, and the amino acid sequence of FLT3L derived from a mouse (denoted as mFLT3L) can be as shown in SEQ ID NO. 28. For another example, the amino acid sequence of IL12a derived from a human (denoted as hIL12a) can be as shown in SEQ ID NO. 18, the amino acid sequence of IL12b derived from a human (denoted as hIL12b) can be as shown in SEQ ID NO. 19, the amino acid sequence of IL2 derived from a human (denoted as hIL2) can be as shown in SEQ ID NO. 21, the amino acid sequence of GMCSF derived from a human (denoted as hGMCSF) can be as shown in SEQ ID NO. 31, the amino acid sequence of IL7 derived from a human (denoted as hIL7) can be as shown in SEQ ID NO. 25, the amino acid sequence of IL15 derived from a human (denoted as hIL15) can be as shown in SEQ ID NO. 23, the amino acid sequence of IL21 derived from a human (denoted as hIL21) can be as shown in SEQ ID NO. 27, and the amino acid sequence of FLT3L derived from a human (denoted as hFLT3L) can be as shown in SEQ ID NO. 29.

[0109] In the present application, the protein can include a fusion protein that fuses two or more of the cytokines, i.e., IL12, IL2, IL7, IL15, IL21, FLT3L and GMCSF, together by genetic recombination technology. The protein can have both the unique biological activity of the constituent factors and can exert biological functions that single cytokines do not have through the complementary and synergistic effects of biological activity, and can even produce some new structures and biological functions. For example, in the present application, the protein can include a fusion protein, and the fusion protein can include any group of cytokines selected from the following: a) IL12, IL2 and GMSCF; b) IL12, IL7 and GMSCF; c) IL12, IL15 and GMSCF; d) IL12, IL21 and GMSCF; e) IL12, IL2 and FLT3L; f) IL12, IL7 and FLT3L; g) IL12, IL15 and FLT3L; and h) IL12, IL21 and FLT3L.

[0110] In the present application, the protein can also include a targeting moiety, and the number of targeting moieties can be one or more. The targeting moieties can be the same or different. The targeting moieties can specifically recognize and / or bind to tumor-associated antigens. The tumor-associated antigens can be selected from the group consisting of the EDB domain of fibronectin, the EDA domain of fibronectin and necrotic regions. The targeting moieties can include antibodies or antigen-binding fragments thereof.

[0111] In the present application, the antigen-binding fragment can be selected from the group consisting of Fab, Fab', F(ab')2, F(ab)2, dAb, isolated complementarity determining region CDR, Fv and scFv. In some embodiments, the antigen-binding fragment can be scFv.

[0112] In the present application, the targeting moiety can include any one of the amino acid sequences set forth in SEQ ID NOs: 1-15.

[0113] For example, the targeting moiety of the protein can be selected from the group consisting of L19V L (the amino acid sequence of which can be as set forth in SEQ ID NO. 10), L19V H (the amino acid sequence of which can be as set forth in SEQ ID NO. 11), F8V L (the amino acid sequence of which can be as set forth in SEQ ID NO. 12), F8V H (the amino acid sequence of which can be as set forth in SEQ ID NO. 13), NHS76V L(amino acid sequence of which can be as shown in SEQ ID NO. 14) and NHS76V H (amino acid sequence of which can be as shown in SEQ ID NO. 15).

[0114] It should be noted that in the present application, the cytokines or the cytokines and the targeting moieties can be connected by a linker. The linker can be a connecting peptide. In the present application, the linker can comprise an amino acid sequence as shown in any one of SEQ ID NO: 73-76.

[0115] For example, the cytokines can be connected by the linker. In the present application, the IL12a, IL12b, IL2, IL7, IL15, IL21, FLT3L and GMCSF can be connected by the connecting peptide. For example, the connecting peptide can comprise an amino acid sequence as shown in SEQ ID NO. 73 or SEQ ID NO. 75.

[0116] For example, the cytokines and the targeting moieties can be connected by the linker. In the present application, the targeting moieties and IL12a, IL12b, IL2, IL7, IL15, IL21, FLT3L and GMCSF can be connected by the connecting peptide. For example, the connecting peptide can comprise an amino acid sequence as shown in any one of SEQ ID NO: 73-76.

[0117] In the present application, the protein can comprise an amino acid sequence as shown in any one of SEQ ID NO: 32-67.

[0118] For example, in the present application, the protein can be a single chain protein, wherein the single chain protein can comprise an amino acid sequence as shown in any one of SEQ ID NO: 32-51.

[0119] For example, the structure of the single chain protein can be that the C-terminus of mIL12b and the N-terminus of mIL12a are fused, the C-terminus of mIL12a and the N-terminus of mIL2 are fused, and the C-terminus of mIL2 and the N-terminus of mGMCSF are fused, thereby forming a mIL12b-mIL12a-mIL2-mGMCSF single chain protein, denoted as mIL12bIL12aIL2GMCSF (amino acid sequence of which can be as shown in SEQ ID NO. 32).

[0120] For example, the structure of the single-chain protein can be that the C-terminus of mIL12b and the N-terminus of mIL12a are fused, the C-terminus of mIL12a and the N-terminus of mIL7 are fused, and the C-terminus of mIL7 and the N-terminus of mFLT3L are fused, thereby forming a mIL12b-mIL12a-mIL7-mFLT3L single-chain protein, denoted as mIL12bIL12aIL7FLT3L (the amino acid sequence of which can be as shown in SEQ ID NO. 37).

[0121] For example, the structure of the single-chain protein can be that the C-terminus of mIL12b and the N-terminus of mIL12a are fused, the C-terminus of mIL12a and the N-terminus of mIL7 are fused, and the C-terminus of mIL7 and the N-terminus of mFLT3L are fused, thereby forming a mIL12b-mIL12a-mIL7-mFLT3L single-chain protein, denoted as mIL12bIL12aIL7FLT3L (the amino acid sequence of which can be as shown in SEQ ID NO. 37).

[0122] For example, the structure of the single-chain protein can be that the C-terminus of mIL12b and the N-terminus of mIL12a are fused, the C-terminus of mIL12a and the N-terminus of mIL7 are fused, and the C-terminus of mIL7 and the N-terminus of mFLT3L are fused, thereby forming a mIL12b-mIL12a-mIL7-mFLT3L single-chain protein, denoted as mIL12bIL12aIL7FLT3L (the amino acid sequence of which can be as shown in SEQ ID NO. 37).

[0123] For example, the structure of the single-chain protein can be that the C-terminus of mIL12b and the N-terminus of mIL12a are fused, the C-terminus of mIL12a and the N-terminus of mIL7 are fused, and the C-terminus of mIL7 and the N-terminus of mFLT3L are fused, thereby forming a mIL12b-mIL12a-mIL7-mFLT3L single-chain protein, denoted as mIL12bIL12aIL7FLT3L (the amino acid sequence of which can be as shown in SEQ ID NO. 37).

[0124] For example, the structure of the single-chain protein can be that the C-terminus of mIL12b and the N-terminus of mIL12a are fused, the C-terminus of mIL12a and the N-terminus of mIL7 are fused, and the C-terminus of mIL7 and the N-terminus of mFLT3L are fused, thereby forming a mIL12b-mIL12a-mIL7-mFLT3L single-chain protein, denoted as mIL12bIL12aIL7FLT3L (the amino acid sequence of which can be as shown in SEQ ID NO. 37).

[0125] For example, the structure of the single-chain protein can be that the C-terminus of mIL12b and the N-terminus of mIL12a are fused, the C-terminus of mIL12a and the N-terminus of mIL21 are fused, and the C-terminus of mIL21 and the N-terminus of mFLT3L are fused, thereby forming a mIL12b-mIL12a-mIL21-mFLT3L single-chain protein, denoted as mIL12bIL12aIL21FLT3L (the amino acid sequence of which can be as shown in SEQ ID NO. 39).

[0126] For example, the structure of the single-chain protein can be that the C-terminus of mIL12b and the N-terminus of mIL12a are fused, the C-terminus of mIL12a and the N-terminus of mIL21 are fused, and the C-terminus of mIL21 and the N-terminus of mFLT3L are fused, thereby forming a mIL12b-mIL12a-mIL21-mFLT3L single-chain protein, denoted as mIL12bIL12aIL21FLT3L (the amino acid sequence of which can be as shown in SEQ ID NO. 39).

[0127] For example, the structure of the single-chain protein can be that the C-terminus of hIL12b and the N-terminus of hIL12a are fused, the C-terminus of hIL12a and the N-terminus of hIL2 are fused, and the C-terminus of hIL2 and the N-terminus of hGMCSF are fused, thereby forming a hIL12b-hIL12a-hIL2-hGMCSF single-chain protein, denoted as hIL12bIL12aIL2GMCSF (the amino acid sequence of which can be as shown in SEQ ID NO. 40).

[0128] For example, the structure of the single-chain protein can be that the C-terminus of hIL12b and the N-terminus of hIL12a are fused, the C-terminus of hIL12a and the N-terminus of hIL7 are fused, and the C-terminus of hIL7 and the N-terminus of hGMCSF are fused, thereby forming a hIL12b-hIL12a-hIL7-hGMCSF single-chain protein, denoted as hIL12bIL12aIL7GMCSF (the amino acid sequence of which can be as shown in SEQ ID NO. 41).

[0129] For example, the structure of the single-chain protein can be that the C-terminus of hIL12b and the N-terminus of hIL12a are fused, the C-terminus of hIL12a and the N-terminus of hIL15 are fused, and the C-terminus of hIL15 and the N-terminus of hGMCSF are fused, thereby forming a hIL12b-hIL12a-hIL15-hGMCSF single-chain protein, denoted as hIL12bIL12aIL15GMCSF (the amino acid sequence of which can be as shown in SEQ ID NO. 42).

[0130] For example, the structure of the single-chain protein can be that the C-terminus of hIL12b and the N-terminus of hIL12a are fused, the C-terminus of hIL12a and the N-terminus of hIL21 are fused, and the C-terminus of hIL21 and the N-terminus of hGMCSF are fused, thereby forming a hIL12b-hIL12a-hIL21-hGMCSF single-chain protein, denoted as hIL12bIL12aIL21GMCSF (the amino acid sequence of which can be as shown in SEQ ID NO. 43).

[0131] For example, the structure of the single-chain protein can be that the C-terminus of hIL12b and the N-terminus of hIL12a are fused, the C-terminus of hIL12a and the N-terminus of hIL2 are fused, and the C-terminus of hIL2 and the N-terminus of hFLT3L are fused, thereby forming a hIL12b-hIL12a-hIL2-hFLT3L single-chain protein, denoted as hIL12bIL12aIL2FLT3L (the amino acid sequence of which can be as shown in SEQ ID NO. 44).

[0132] For example, the structure of the single-chain protein can be that the C-terminus of hIL12b and the N-terminus of hIL12a are fused, the C-terminus of hIL12a and the N-terminus of hIL7 are fused, and the C-terminus of hIL7 and the N-terminus of hFLT3L are fused, thereby forming a hIL12b-hIL12a-hIL7-hFLT3L single-chain protein, denoted as hIL12bIL12aIL7FLT3L (the amino acid sequence of which can be as shown in SEQ ID NO. 45).

[0133] For example, the structure of the single-chain protein can be that the C-terminus of hIL12b and the N-terminus of hIL12a are fused, the C-terminus of hIL12a and the N-terminus of hIL15 are fused, and the C-terminus of hIL15 and the N-terminus of hFLT3L are fused, thereby forming a hIL12b-hIL12a-hIL15-hFLT3L single-chain protein, denoted as hIL12bIL12aIL15FLT3L (the amino acid sequence of which can be as shown in SEQ ID NO. 46).

[0134] For example, the structure of the single-chain protein can be that the C-terminus of hIL12b and the N-terminus of hIL12a are fused, the C-terminus of hIL12a and the N-terminus of hIL21 are fused, and the C-terminus of hIL21 and the N-terminus of hFLT3L are fused, thereby forming a hIL12b-hIL12a-hIL21-hFLT3L single-chain protein, denoted as hIL12bIL12aIL21FLT3L (the amino acid sequence of which can be as shown in SEQ ID NO. 47).

[0135] For example, the structure of the single-chain protein can be that the C-terminus of mIL12b is fused with the N-terminus of mIL12a, the C-terminus of mIL12a is fused with the N-terminus of mIL2, the C-terminus of mIL2 is fused with the N-terminus of NHS76V H , the C-terminus of NHS76V H is fused with the N-terminus of F8V L , the C-terminus of F8V L is fused with the N-terminus of F8V H , the C-terminus of F8V H is fused with the N-terminus of NHS76V L , the C-terminus of NHS76V L is fused with the N-terminus of mGMCSF, thereby forming a mIL12b-mIL12a-mIL2-NHS76V H -F8V L -F8V H -NHS76V L -mGMCSF single-chain protein. It should be noted that the linker of the single-chain protein contains a thrombin cleavage site, and the name of the single-chain protein is represented by mIL12bIL12aIL2DiaNHS76F8GMCSF-Thr (the amino acid sequence of which can be shown as SEQ ID NO. 48).

[0136] For example, the structure of the single-chain protein can be that the C-terminus of mIL12b is fused with the N-terminus of mIL12a, the C-terminus of mIL12a is fused with the N-terminus of mIL2, the C-terminus of mIL2 is fused with the N-terminus of NHS76V H , the C-terminus of NHS76V H is fused with the N-terminus of F8V L , the C-terminus of F8V L is fused with the N-terminus of F8V H , the C-terminus of F8V H is fused with the N-terminus of NHS76V L , the C-terminus of NHS76V L is fused with the N-terminus of hGMCSF, thereby forming a hIL12b-hIL12a-hIL2-NHS76V H -F8V L -F8V H -NHS76V L -hGMCSF single-chain protein. It should be noted that the linker of the single-chain protein contains a thrombin cleavage site, and the name of the single-chain protein is represented by hIL12bIL12aIL2DiaNHS76F8GMCSF-Thr (the amino acid sequence of which can be shown as SEQ ID NO. 49).

[0137] For example, the structure of the single-chain protein can be that the C-terminus of mIL12b is fused with the N-terminus of mIL12a, the C-terminus of mIL12a is fused with the N-terminus of mIL2, the C-terminus of mIL2 is fused with the N-terminus of F8V H , the C-terminus of F8V H is fused with the N-terminus of F8V L , the C-terminus of F8V L is fused with the N-terminus of F8V H , the C-terminus of F8V H is fused with the N-terminus of F8V L , and the C-terminus of F8V L is fused with the N-terminus of mGMCSF, thereby forming a mIL12b-mIL12a-mIL2-F8V H -F8V L -F8V H -F8V L -mGMCSF single-chain protein, which is denoted as mIL12bIL12aIL2DiaF8GMCSF (the amino acid sequence of which can be shown as SEQ ID NO. 50).

[0138] For example, the structure of the single-chain protein can be that the C-terminus of hIL12b is fused with the N-terminus of hIL12a, the C-terminus of hIL12a is fused with the N-terminus of hIL2, the C-terminus of hIL2 is fused with the N-terminus of F8V H , the C-terminus of F8V H is fused with the N-terminus of F8V L , the C-terminus of F8V L is fused with the N-terminus of F8V H , the C-terminus of F8V H is fused with the N-terminus of F8V L , and the C-terminus of F8V L is fused with the N-terminus of hGMCSF, thereby forming a hIL12b-hIL12a-hIL2-F8V H -F8V L -F8V H -F8V L -hGMCSF single-chain protein, which is denoted as hIL12bIL12aIL2DiaF8GMCSF (the amino acid sequence of which can be shown as SEQ ID NO. 51).

[0139] For example, in the present application, the protein can also be a dimer composed of a first polypeptide chain and a second polypeptide chain, wherein the first polypeptide chain is different from the second polypeptide chain. The first polypeptide chain can comprise IL12a, and the second polypeptide chain can comprise IL12b.

[0140] In the present application, the IL2 or a functional fragment thereof can be located in the first polypeptide chain or in the second polypeptide chain, the GMCSF or a functional fragment thereof can be located in the first polypeptide chain or in the second polypeptide chain, and the one or more targeting moieties can each independently be located in the first polypeptide chain or in the second polypeptide chain.

[0141] In the present application, in the first polypeptide chain, the IL2 or a functional fragment thereof, the IL12a or a functional fragment thereof, and the GMCSF or a functional fragment thereof can be contained in this order from the N-terminus to the C-terminus.

[0142] In the present application, in the first polypeptide chain, the targeting moiety, the IL12a or a functional fragment thereof, the IL2 or a functional fragment thereof, and the GMCSF or a functional fragment thereof can be contained in this order from the N-terminus to the C-terminus.

[0143] In the present application, in the second polypeptide chain, the IL12b or a functional fragment thereof and the targeting moiety can be contained in this order from the N-terminus to the C-terminus.

[0144] In certain embodiments, in the dimer described in the present application, the first polypeptide chain can comprise the amino acid sequence set forth in SEQ ID NO: 53 and the second polypeptide chain can comprise the amino acid sequence set forth in SEQ ID NO: 52.

[0145] In certain embodiments, in the dimer described in the present application, the first polypeptide chain can comprise the amino acid sequence set forth in SEQ ID NO: 55 and the second polypeptide chain can comprise the amino acid sequence set forth in SEQ ID NO: 54;

[0146] In certain embodiments, in the dimer described in the present application, the first polypeptide chain can comprise the amino acid sequence set forth in SEQ ID NO: 56 and the second polypeptide chain can comprise the amino acid sequence set forth in SEQ ID NO: 57;

[0147] In certain embodiments, in the dimer described in the present application, the first polypeptide chain can comprise the amino acid sequence set forth in SEQ ID NO: 58 and the second polypeptide chain can comprise the amino acid sequence set forth in SEQ ID NO: 59;

[0148] In certain embodiments, in the dimer described in the present application, the first polypeptide chain can comprise the amino acid sequence set forth in SEQ ID NO: 60 and the second polypeptide chain can comprise the amino acid sequence set forth in SEQ ID NO: 61;

[0149] In some embodiments, in the dimer described herein, the first polypeptide chain can comprise the amino acid sequence set forth in SEQ ID NO: 62 and the second polypeptide chain can comprise the amino acid sequence set forth in SEQ ID NO: 63.

[0150] In some embodiments, in the dimer described herein, the first polypeptide chain can comprise the amino acid sequence set forth in SEQ ID NO: 64 and the second polypeptide chain can comprise the amino acid sequence set forth in SEQ ID NO: 65.

[0151] In some embodiments, in the dimer described herein, the first polypeptide chain can comprise the amino acid sequence set forth in SEQ ID NO: 66 and the second polypeptide chain can comprise the amino acid sequence set forth in SEQ ID NO: 67.

[0152] For example, in the dimer, the C-terminus of mIL12b can be fused with the N-terminus of F8V H , the C-terminus of F8V H may be fused with the N-terminus of F8V L to form the second polypeptide chain (the amino acid sequence of which can be set forth in SEQ ID NO. 52), and the C-terminus of F8V H may be fused with the N-terminus of F8V L , the C-terminus of F8V L may be fused with the N-terminus of mIL12a, the C-terminus of mIL12a can be fused with the N-terminus of mIL2, the C-terminus of mIL2 can be fused with the N-terminus of mGMCSF to form the first polypeptide chain (the amino acid sequence of which can be set forth in SEQ ID NO. 53), thereby forming the mIL12b-F8V H -F8V L -F8V H -F8V L -mIL12a-mIL2-mGMCSF dimer, denoted as mIL12b-scF8-scF8-IL12a-IL2-GMCSF.

[0153] For example, in the dimer, the C-terminus of hIL12b can be fused with the N-terminus of NHS76V H , the C-terminus of NHS76V H may be fused with the N-terminus of NHS76V L to form the second polypeptide chain (the amino acid sequence of which can be set forth in SEQ ID NO. 54), and the C-terminus of NHS76V H may be fused with the N-terminus of NHS76V L , the C-terminus of NHS76V LThe C-terminus of hIL12a can be fused with the N-terminus of hIL2, the C-terminus of hIL2 can be fused with the N-terminus of hGMCSF to form a first polypeptide chain (the amino acid sequence of which can be as shown in SEQ ID NO. 55), thereby forming hIL12b-NHS76V H -NHS76V L -NHS76V H -NHS76V L -hIL12a-hIL2-hGMCSF dimer, denoted as hIL12b-scNHS76-scNHS76IL12aIL2GMCSF.

[0154] For example, in the dimer, the C-terminus of mIL12a and the N-terminus of mIL2 can be fused to form a mIL12a-mIL2 first polypeptide chain (the sequence of which is shown in SEQ ID NO. 56), and the C-terminus of mIL12b and the N-terminus of mGMCSF can be fused to form a mIL12b-mGMCSF second polypeptide chain (the sequence of which is shown in SEQ ID NO. 57), thereby forming a mIL12a-mIL2-mIL12b-mGMCSF dimer, denoted as mIL12aIL2IL12bGMCSF.

[0155] For example, in the dimer, the C-terminus of mIL12a and the N-terminus of mIL7 can be fused to form a mIL12a-mIL7 first polypeptide chain (the sequence of which is shown in SEQ ID NO. 58), and the C-terminus of mIL12b and the N-terminus of mGMCSF can be fused to form a mIL12b-mGMCSF second polypeptide chain (the sequence of which is shown in SEQ ID NO. 59), thereby forming a mIL12a-mIL7-mIL12b-mGMCSF dimer, denoted as mIL12aIL7IL12bGMCSF.

[0156] For example, in the dimer, the C-terminus of mIL12a and the N-terminus of mIL21 can be fused to form a mIL12a-mIL21 first polypeptide chain (the sequence of which is shown in SEQ ID NO. 60), and the C-terminus of mIL12b and the N-terminus of mGMCSF can be fused to form a mIL12b-mGMCSF second polypeptide chain (the sequence of which is shown in SEQ ID NO. 61), thereby forming a mIL12a-mIL21-mIL12b-mGMCSF dimer, denoted as mIL12aIL21IL12bGMCSF.

[0157] For example, in the dimer, the C-terminus of hIL12a and the N-terminus of hIL2 can be fused to form a hIL12a-hIL2 first polypeptide chain (the sequence is shown as SEQ ID NO. 62), and the C-terminus of hIL12b and the N-terminus of hGMCSF can be fused to form a hIL12b-hGMCSF second polypeptide chain (the sequence is shown as SEQ ID NO. 63), thereby forming a hIL12a-hIL2-hIL12b-hGMCSF dimer, denoted as hIL12aIL2IL12bGMCSF.

[0158] For example, in the dimer, the C-terminus of hIL12a and the N-terminus of hIL7 can be fused to form a hIL12a-hIL7 first polypeptide chain (the sequence is shown as SEQ ID NO. 64), and the C-terminus of hIL12b and the N-terminus of hGMCSF can be fused to form a hIL12b-hGMCSF second polypeptide chain (the sequence is shown as SEQ ID NO. 65), thereby forming a hIL12a-hIL7-hIL12b-hGMCSF dimer, denoted as hIL12aIL7IL12bGMCSF.

[0159] For example, in the dimer, the C-terminus of hIL12a and the N-terminus of hIL21 can be fused to form a hIL12a-hIL21 first polypeptide chain (the sequence is shown as SEQ ID NO. 66), and the C-terminus of hIL12b and the N-terminus of hGMCSF can be fused to form a hIL12b-hGMCSF second polypeptide chain (the sequence is shown as SEQ ID NO. 67), thereby forming a hIL12a-hIL21-hIL12b-hGMCSF dimer, denoted as hIL12aIL21IL12bGMCSF.

[0160] The protein, polypeptide and / or amino acid sequence involved in the present application should also be understood to at least include the range of variants or homologues having the same or similar functions as the said protein or polypeptide.

[0161] In the present application, the variant can be a protein or polypeptide that has one or more amino acids substituted, deleted, or added in the amino acid sequence of the protein and / or the polypeptide (e.g., the protein molecule). For example, the functional variant can comprise a protein or polypeptide that has an amino acid alteration by at least 1, for example, 1-30, 1-20, or 1-10, and further for example, 1, 2, 3, 4, or 5 amino acid substitutions, deletions, and / or insertions. The functional variant can substantially maintain the biological properties of the protein or the polypeptide before the alteration (e.g., substitution, deletion, or addition). For example, the functional variant can maintain at least 60%, 70%, 80%, 90%, or 100% of the biological activity of the protein or the polypeptide before the alteration.

[0162] In the present application, the homolog can be a protein or polypeptide that has at least about 80% (e.g., at least about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more) sequence homology to the amino acid sequence of the protein and / or the polypeptide (e.g., the protein molecule).

[0163] In the present application, the homology generally refers to the similarity, analogy or correlation between two or more sequences. The "percent sequence homology" can be calculated by comparing two sequences to be aligned in a comparison window, determining the number of positions at which the identical nucleic acid base (e.g., A, T, C, G, I) or identical amino acid residue (e.g., Ala, Pro, Ser, Thr, Gly, Val, Leu, lie, Phe, Tyr, Trp, Lys, Arg, His, Asp, Glu, Asn, Gin, Cys and Met) occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the comparison window (i.e., the window size), and multiplying the result by 100 to yield the percent sequence homology. The alignment for determining percent sequence homology can be achieved in various ways known in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms needed to achieve the maximum alignment over the full length or a specified region of a sequence being compared. The homology can also be determined by FASTA and BLAST. The description of the FASTA algorithm can be found in W. R. Pearson and D. J. Lipman, "Improved tools for biological sequence comparison", Proc. Natl. Acad. Sci., 85:2444-2448, 1988; and D. J. Lipman and W. R. Pearson, "Fast

[0164] Drugs, uses, methods of treating tumors

[0165] In another aspect, the present application also provides a medicament comprising the liquid formulation described in the present application. Wherein the liquid formulation can be formulated as an injection, so that the liquid formulation described in the present application can be used for intravenous injection or subcutaneous injection.

[0166] In certain embodiments, the medicament described in the present application can be formulated for oral administration, intravenous administration, intramuscular administration, in situ administration at the site of a tumor, inhalation, rectal administration, vaginal administration, transdermal administration or administration through a subcutaneous depot.

[0167] In the present application, the medicament described in the present application can also comprise a diluent, wherein the diluent and the liquid preparation described in the present application do not mix with each other, so that the diluent can be used to dilute the liquid preparation described in the present application when different concentrations of the liquid preparation described in the present application are required.

[0168] In some embodiments, the medicament described in the present application can also comprise a pharmaceutically acceptable carrier. For example, the pharmaceutically acceptable carrier can include buffers, antioxidants, preservatives, low molecular weight polypeptides, proteins, hydrophilic polymers, amino acids, sugars, chelating agents, counterions, metal complexes, and / or nonionic surfactants, etc. For example, the pharmaceutically acceptable carrier can include excipients, for example, the excipients can be selected from the group consisting of starch, dextrin, sucrose, magnesium stearate, calcium sulfate, carboxymethylcellulose, talc, calcium alginate gel, chitosan, and nanomicrospheres, etc. For example, the pharmaceutically acceptable carrier can also be selected from the group consisting of pH adjusters, osmotic pressure adjusters, solubilizers, and bacteriostatic agents.

[0169] In another aspect, the present application also provides the use of the liquid preparation described in the present application in the preparation of a medicament for treating tumors. Wherein the tumors can include lung cancer.

[0170] In another aspect, the present application also provides a method for treating tumors, which comprises administering the liquid preparation described in the present application to a subject in need thereof.

[0171] Wherein the administration method can be intratumoral injection, for example, injecting the liquid preparation described in the present application into the interior of a tumor. In some embodiments, the administration method can also be oral administration, intravenous administration, intramuscular administration, in situ administration at the tumor site, inhalation, rectal administration, vaginal administration, transdermal administration, or administration through a subcutaneous depot.

[0172] In addition, in some embodiments, the dose level of the liquid preparation described in the present application administered can vary depending on the molecule delivered, the route of administration, and the size (weight, body surface, or organ size) and / or physical condition (age and general health) of the patient.

[0173] In another aspect, the present application provides a liquid preparation or a medicament for treating tumors. Wherein the tumors can include lung cancer.

[0174] In another aspect, the present application also provides an excipient for preparing the liquid formulation described in the present application, which comprises an oil phase solvent, and the mass fraction of the oil phase solvent is about 50% or more, for example, the mass fraction of the oil phase solvent in the excipient can be about 50% to about 60%, about 60% to about 70%, about 70% to about 80%, about 80% to about 90%, about 90% to about 95%, about 50% to about 70%, about 50% to about 80%, about 50% to about 90%, about 50% to about 95%, about 60% to about 80%, about 60% to about 90%, about 60% to about 95%, about 70% to about 90%, about 70% to about 95%, or about 80% to about 95%, so that the liquid formulation described in the present application can be easily prepared.

[0175] Examples

[0176] The following examples are put forth merely to illustrate the liquid formulation, medicaments and uses of the present application, and are not intended to limit the scope of the present application, nor are they intended to represent that the experiments were carried out in the exact and only manner described. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees Centigrade, and pressure is at or near atmospheric. Standard abbreviations can be used, e.g., bp, base pair(s); kb, kilobase(s); pl, picoliter(s); s or sec, second(s); min, minute(s); h or hr, hour(s); aa, amino acid; nt, nucleotide; iv, intravenous; i.m., intramuscular; i.p., intraperitoneal; s.c., subcutaneous; and the like.

[0177] Reagents: DMEM medium, 1640 medium, fetal bovine serum were purchased from lifetechnologies company; cell culture bottles, culture plates were purchased from Corning company; doxycycline (DOX) was purchased from Shanghai shenguo biological engineering co., LTD; puromycin, blasticidin were purchased from Chemicon company; restriction endonuclease was purchased from Takara and NEB company; ligase was purchased from NEB company; DNA polymerase was purchased from Takara company; plasmid extraction kit, gel recovery kit were purchased from OmegaBiotech company; primer synthesis was completed by Shanghai shenguo biological engineering co., LTD; gene synthesis was completed by Nanjing Kingsway company; ELISA kit was purchased from Doctor Deer company.

[0178] Example 1 Expression of mIL12bIL12aIL2 GMCSF protein

[0179] 1.1 Construction of expression vector

[0180] mIL12bIL12aIL2GMCSF protein with a secretion signal peptide at the front end and 6*His added at the rear end for purification, the DNA sequence corresponding to the mIL12bIL12aIL2GMCSF protein gene was synthesized, and the sequence was mutated at the BamHI or XhoI site with a degenerate codon, and the front and rear ends of the synthesized sequence were provided with BamHI and XhoI enzyme digestion sites, respectively. The synthesized plasmid with the target gene was digested with the following system: 5 μg of plasmid, 4 μl of enzyme digestion buffer, 1 μl of BamHI and 1 μl of XhoI, and water to a total volume of 40 μl, and incubated at 37°C for 12 hours. The EP tube was removed, 4.4 μl of 10x loading buffer was added, and electrophoresis was performed on a 1% agarose gel. After electrophoresis, the mIL12bIL12aIL2GMCSF protein gene fragment was recovered and used as needed.

[0181] The amino acid sequence of the mIL12bIL12aIL2GMCSF protein is shown in SEQ ID NO. 32, and the nucleotide sequence encoding the mIL12bIL12aIL2GMCSF is shown in SEQ ID NO. 68.

[0182] The pLentis-CMV-MCS-IRES-PURO vector was digested in an EP tube with the following system: 2 μg of pLentis-CMV-MCS-IRES-PURO vector plasmid, 3 μl of enzyme digestion buffer, 1 μl of BamHI and 1 μl of XhoI, and water to a total volume of 30 μl, and incubated at 37°C for 12 hours. The EP tube was removed, 3.3 μl of 10x loading buffer was added, and electrophoresis was performed on a 1% agarose gel. After electrophoresis, the pLentis-CMV-MCS-IRES-PURO vector fragment was recovered and used as needed.

[0183] The mIL12bIL12aIL2GMCSF and pLentis-CMV-MCS-IRES-PURO were ligated with the following system: 2 μl of pLentis-CMV-MCS-IRES-PURO vector fragment, 2 μl of gene fragment, 1 μl of ligase buffer, 0.5 μl of T4 DNA ligase, and 4.5 μl of water. Incubate at room temperature for 4 hours. Then the ligation system is transformed into E. coli competent cells. The next day, colonies were picked from the transformed plate and incubated in LB medium at 37°C overnight. The plasmid was extracted from the cultured bacteria using a plasmid extraction kit, and the gene fragment was identified by enzyme digestion to determine whether it was successfully ligated into the vector. The correct vector was then sequenced to confirm successful construction. The expression vector pLentis-CMV-mIL12bIL12aIL2GMCSF-IRES-PURO was obtained.

[0184] 1.2 Preparation of expression virus

[0185] 1) Digest the cultured 293FT cells, count and then seed 3x10 6 cells / well into 10cm culture dish, the culture volume is 10ml.

[0186] 2) The next day evening, observe the cell status, if the cell status is good, then proceed to transfection. Add chloroquine to the final concentration of 25μM in the culture plate, take a test tube, add sterile water and the following plasmids (pMD2.G 6μg + pSPAX2 15μg + expression vector obtained in the above 1.1 embodiment 20μg), the total volume is 1045μl, then add 2M CaCl2155μl, mix well, finally add 1200μl 2xHBS, dropwise add and oscillate, after the dropwise addition is completed, quickly add the mixture to the cell culture well, mix gently.

[0187] 3) The next morning, observe the cell status, replace the culture medium with 10ml fresh DMEM culture medium.

[0188] 4) The next morning, observe the cell status, and collect the supernatant in the culture dish, filter with 0.45μm filter, then place in a high-speed centrifuge tube, centrifuge at 50000g for 2 hours, carefully discard the supernatant, try to absorb the liquid with absorbent paper, then resuspend the precipitate with 200μl HBSS, dissolve for 2 hours, then aliquot into small tubes, and store at -70℃.

[0189] 1.3 Preparation of expression cells

[0190] Digest the cultured 293A cells, seed 10 5 cells / well into 6-well plates, the culture volume is 1ml. After 24 hours, add 10μl of virus expressing the above-mentioned target gene (i.e. the virus obtained in Example 1.2), continue to culture in the incubator for 24 hours, then discard the supernatant and replace it with fresh culture medium for continued culture. When the cells are fully grown, they are transferred to culture bottles, 3μg / ml puromycin is added to the final concentration, and the culture medium is replaced every two days to maintain the concentration of puromycin. After one week of screening, the surviving cells are stable expression of the protein, named 293A-mIL12bIL12aIL2GMCSF.

[0191] 1.4 Protein expression and purification

[0192] The constructed cells expressing mIL12bIL12aIL2GMCSF (293A-mIL12bIL12aIL2GMCSF) were passaged into 15cm culture dishes. After the cells reached confluence, the culture medium was replaced with 30ml CDM4HEK293, and the cells were cultured for another 5 days. The supernatant was then collected, filtered through a 0.45μm filter, and concentrated using a 50kd AMICON ULTRA-15 ultrafiltration tube. The concentrated protein solution was purified using nickel chelate magnetic beads (purchased from Beaver Biotechnology Co., Ltd.) according to the instructions. The purified protein solution was then ultrafiltered using an AMICON ULTRA-0.5 ultrafiltration tube, and the buffer was replaced with PBS. Finally, the protein concentration was detected using an IL12p70 ELISA kit. After adjusting the protein concentration to 2μg / μl with PBS, the protein solution was aliquoted and stored at -20℃.

[0193] Example 2 Expression of protein molecules mIL12bIL12aIL2DiaNHS76F8GMCSF-Thr

[0194] 2.1 Constructing the expression vector

[0195] The mIL12bIL12aIL2DiaNHS76F8GMCSF-Thr protein, with a secretion signal peptide at its anterior end and a 6*His peptide added to its posterior end for purification, was synthesized. The corresponding DNA sequence was synthesized, with BamHI or XhoI sites mutated using degenerate codons. The synthesized sequence contained BamHI and XhoI restriction sites at its anterior and posterior ends, respectively. The resulting plasmid containing the target gene was digested using the following mixture: 5 μg plasmid, 4 μl digestion buffer, 1 μl BamHI, and 1 μl XhoI, with water added to a total volume of 40 μl. The mixture was incubated at 37°C for 12 hours. The EP tube was then removed, and 4.4 μl of 10× loading buffer was added. Electrophoresis was performed on a 1% agarose gel. After electrophoresis, the mIL12bIL12aIL2DiaNHS76F8GMCSF-Thr protein gene fragment was recovered for later use. It should be noted that the linker of the mIL12bIL12aIL2DiaNHS76F8GMCSF-Thr protein contains a thrombin cleavage site.

[0196] The amino acid sequence of the mIL12bIL12aIL2DiaNHS76F8GMCSF-Thr protein is shown in SEQ ID NO.48, and the nucleotide sequence encoding the mIL12bIL12aIL2DiaNHS76F8GMCSF-Thr is shown in SEQ ID NO.69.

[0197] Enzymatic digestion of the pLentis-CMV-MCS-IRES-PURO vector in an EP tube, the system is as follows: 2 μg of pLentis-CMV-MCS-IRES-PURO vector plasmid, 3 μl of enzyme digestion buffer, 1 μl of BamHI and 1 μl of XhoI, add water to a total volume of 30 μl, 37°C for 12 hours. Take out the EP tube, add 3.3 μl of 10×loading buffer, and perform electrophoresis with a 1% agarose gel. After electrophoresis, recover the pLentis-CMV-MCS-IRES-PURO vector fragment for use.

[0198] Connect mIL12bIL12aIL2DiaNHS76F8GMCSF-Thr and pLentis-CMV-MCS-IRES-PURO, the system is as follows, 2 μl of pLentis-CMV-MCS-IRES-PURO vector fragment, 2 μl of gene fragment, 1 μl of ligase buffer, 0.5 μl of T4 DNA ligase and 4.5 μl of water. Place at room temperature for 4 hours. Then perform E. coli competent transformation of the ligation system. The next day, pick colonies from the transformed plate and incubate in LB medium at 37°C overnight. Use a plasmid extraction kit to extract plasmids from the cultured bacteria. Identify whether the fragment has been successfully connected to the vector by enzyme digestion, then sequence the correct vector to determine whether the construction is successful. Obtain the expression vector pLentis-CMV-mIL12bIL12aIL2DiaNHS76F8GMCSF-Thr-IRES-PURO.

[0199] 2.2 Preparation of expression virus

[0200] 1) Digest the cultured 293FT cells, count and then plate 3×10 6 cells / well in a 10 cm culture dish with a culture volume of 10 ml.

[0201] 2) The next night, observe the cell state, and if the cell state is good, proceed with transfection. Add chloroquine to a final concentration of 25 μM in the culture plate, take a test tube, add sterile water and the following plasmids (pMD2.G 6 μg + pSPAX2 15 μg + expression vector obtained in Example 2.1 20 μg), a total volume of 1045 μl, then add 2M CaCl2155 μl, mix well, finally add 1200 μl of 2×HBS, dropwise and oscillate, after the dropwise addition is completed, quickly add the mixture to the cell culture well and mix gently.

[0202] 3) The next morning, observe the cell state and replace the culture medium with 10 ml of fresh DMEM medium.

[0203] 4) On the fifth morning, observe the state of the cells, and collect the supernatant in the culture dish, filter it with a 0.45 μm filter, then place it in a high-speed centrifuge tube, centrifuge at 50000g for 2 hours, carefully discard the supernatant, try to absorb the liquid with a water-absorbing paper, then resuspend the precipitate with 200 μl of HBSS, dissolve for 2 hours, then aliquot into small tubes, and store at -70°C.

[0204] 2.3 Preparation of expression cells

[0205] Digest the cultured 293A cells, seed 10 5 cells / well into a 6-well plate, with a culture volume of 1 ml. After 24 hours, add 10 μl of virus expressing the above-mentioned target gene (i.e., the virus obtained in Example 2.2), continue to culture in the incubator for 24 hours, then discard the supernatant and replace it with fresh culture medium for continued culture. After the cells have grown, they are subcultured into culture bottles, and 3 μg / ml of puromycin is added, and the culture medium is replaced every two days, and the concentration of puromycin is maintained. After one week of screening, the surviving cells are cells that stably express the protein, and are named 293A-mIL12bIL12aIL2DiaNHS76F8GMCSF-Thr.

[0206] 2.4 Protein expression and purification

[0207] Subculture the constructed cells 293A-mIL12bIL12aIL2DiaNHS76F8GMCSF-Thr expressing mIL12bIL12aIL2DiaNHS76F8GMCSF-Thr into a 15 cm culture dish, and after the cells have grown, replace the culture medium with 30 ml of CDM4HEK293, continue to culture for 5 days, then collect the supernatant, filter it with a 0.45 μm filter, and then concentrate it with a 50 kd AMICON ULTRA-15 ultrafiltration tube. The obtained concentrated protein solution is purified with nickel-chelated magnetic beads (purchased from Beaver Biosciences Technology Co., Ltd.), and the operation procedure is performed according to the instructions. The obtained purified protein solution is ultrafiltered with an AMICON ULTRA-0.5 ultrafiltration tube, the buffer is replaced with PBS, and finally the obtained protein solution is detected for protein concentration with an IL12p70 ELISA kit. After adjusting the protein concentration to 2 μg / μl with PBS, the protein is aliquoted and stored at -20°C.

[0208] Example 3 Expression of mIL12bIL12aIL7GMCSF protein

[0209] 3.1 Construction of expression vector

[0210] mIL12bIL12aIL7GMCSF protein with a secretion signal peptide at the front end and 6*His added at the rear end for purification, the DNA sequence corresponding to the mIL12bIL12aIL7GMCSF protein gene was synthesized, and the sequence was mutated at the BamHI or XhoI site with a degenerate codon, and the front and rear ends of the synthesized sequence were provided with BamHI and XhoI enzyme digestion sites, respectively. The synthesized plasmid with the target gene was digested with the following system: 5 μg of plasmid, 4 μl of digestion buffer, 1 μl of BamHI and 1 μl of XhoI, and water to a total volume of 40 μl, 37°C for 12 hours. Take out the EP tube, add 4.4 μl of 10x loading buffer, and perform electrophoresis on a 1% agarose gel. After electrophoresis, the mIL12bIL12aIL7GMCSF protein gene fragment was recovered and used.

[0211] The amino acid sequence of the mIL12bIL12aIL7GMCSF protein is shown in SEQ ID NO. 33, and the nucleotide sequence encoding the mIL12bIL12aIL7GMCSF is shown in SEQ ID NO. 70.

[0212] The pLentis-CMV-MCS-IRES-PURO vector was digested in an EP tube with the following system: 2 μg of pLentis-CMV-MCS-IRES-PURO vector plasmid, 3 μl of digestion buffer, 1 μl of BamHI and 1 μl of XhoI, and water to a total volume of 30 μl, 37°C for 12 hours. Take out the EP tube, add 3.3 μl of 10x loading buffer, and perform electrophoresis on a 1% agarose gel. After electrophoresis, the pLentis-CMV-MCS-IRES-PURO vector fragment was recovered and used.

[0213] The mIL12bIL12aIL7GMCSF and pLentis-CMV-MCS-IRES-PURO were ligated with the following system: 2 μl of pLentis-CMV-MCS-IRES-PURO vector fragment, 2 μl of gene fragment, 1 μl of ligase buffer, 0.5 μl of T4 DNA ligase, and 4.5 μl of water. Incubate at room temperature for 4 hours. Then the ligation system was transformed into E. coli competent cells. The next day, colonies were picked from the transformed plate and incubated in LB medium at 37°C overnight. The plasmid was extracted from the cultured bacteria using a plasmid extraction kit, and the gene fragment was identified by enzyme digestion to determine whether it was successfully ligated into the vector. The correct vector was then sequenced to confirm successful construction. The expression vector pLentis-CMV-mIL12bIL12aIL7GMCSF-IRES-PURO was obtained.

[0214] 3.2 Preparation of expression virus

[0215] 1) Digest the cultured 293FT cells, count and then seed 3x10 6 cells / well into 10cm culture dish, the culture volume is 10ml.

[0216] 2) The next day evening, observe the cell status, if the cell status is good, then proceed to transfection. Add chloroquine to the final concentration of 25μM in the culture plate, take a test tube, add sterile water and the following plasmids (pMD2.G 6μg + pSPAX2 15μg + expression vector obtained in the above 3.1 embodiment 20μg), the total volume is 1045μl, then add 2M CaCl2155μl, mix well, finally add 1200μl 2xHBS, dropwise add and oscillate, after the dropwise addition is completed, quickly add the mixture to the cell culture well, mix gently.

[0217] 3) The next morning, observe the cell status, replace the culture medium with 10ml fresh DMEM culture medium.

[0218] 4) The next morning, observe the cell status, and collect the supernatant in the culture dish, filter with 0.45μm filter, then place in a high-speed centrifuge tube, centrifuge at 50000g for 2 hours, carefully discard the supernatant, try to absorb the liquid with absorbent paper, then resuspend the precipitate with 200μl HBSS, dissolve for 2 hours, then aliquot into small tubes, and store at -70℃.

[0219] 3.3 Preparation of expression cells

[0220] Digest the cultured 293A cells, seed 10 5 cells / well into 6-well plates, the culture volume is 1ml. After 24 hours, add 10μl of virus expressing the above-mentioned target gene (i.e. the virus obtained in Example 3.2), continue to culture in the incubator for 24 hours, then discard the supernatant and replace it with fresh culture medium for continued culture. When the cells are fully grown, they are transferred to culture bottles, and 3μg / ml puromycin is added to the final concentration, and the culture medium is replaced every two days to maintain the concentration of puromycin. After one week of screening, the surviving cells are stable expression of the protein, named 293A-mIL12bIL12aIL7GMCSF.

[0221] 3.4 Protein expression and purification

[0222] The constructed cell 293A-mIL12bIL12aIL7GMCSF expressing mIL12bIL12aIL7GMCSF was subcultured into a 15 cm culture dish. After the cells grew to fullness, the culture medium was replaced with 30 ml CDM4HEK293, and the cells were cultured for another 5 days. Then the supernatant was collected, filtered through a 0.45 μm filter, concentrated by 50 kd AMICON ULTRA-15 ultrafiltration, and the obtained concentrated protein solution was purified by nickel chelate magnetic beads (purchased from Beaver Bioscience Co., Ltd.). The operation procedure was performed according to the instructions. The obtained purified protein solution was ultrafiltered by an AMICON ULTRA-0.5 ultrafiltration tube, the buffer was replaced with PBS, and finally the protein solution was detected for protein concentration by an IL12p70 ELISA kit. After the protein concentration was adjusted to 2 μg / μl with PBS, the protein solution was aliquoted and stored at -20°C.

[0223] Example 4 Expression of mIL12bIL12aIL21GMCSF Protein

[0224] 4.1 Construction of expression vector

[0225] mIL12bIL12aIL21GMCSF protein with a secretory signal peptide at the front end and 6*His added at the rear end for purification. The DNA sequence corresponding to the mIL12bIL12aIL21GMCSF protein gene was synthesized. The sequence was mutated at the BamHI or XhoI site with a degenerate codon. The front and rear ends of the synthesized sequence were provided with BamHI and XhoI enzyme digestion sites, respectively. The synthesized plasmid with the target gene was digested by the following system: 5 μg plasmid, 4 μl enzyme digestion buffer, 1 μl BamHI, and 1 μl XhoI, and water was added to a total volume of 40 μl. The system was incubated at 37°C for 12 hours. The EP tube was taken out, 4.4 μl of 10x loading buffer was added, and electrophoresis was performed on a 1% agarose gel. After electrophoresis, the mIL12bIL12aIL21GMCSF protein gene fragment was recovered for use.

[0226] The amino acid sequence of the mIL12bIL12aIL21GMCSF protein is shown in SEQ ID NO. 35, and the nucleotide sequence encoding the mIL12bIL12aIL21GMCSF is shown in SEQ ID NO. 71.

[0227] Enzymatic digestion of the EP tube vector pLentis-CMV-MCS-IRES-PURO, the system is as follows: 2 μg pLentis-CMV-MCS-IRES-PURO vector plasmid, 3 μl enzyme digestion buffer, 1 μl BamHI and 1 μl XhoI, add water to a total volume of 30 μl, 37°C for 12 hours. Take out the EP tube, add 3.3 μl 10×loading buffer, and perform electrophoresis with 1% agarose gel. After electrophoresis, the pLentis-CMV-MCS-IRES-PURO vector fragment is recovered for use.

[0228] Connect mIL12bIL12aIL21GMCSF and pLentis-CMV-MCS-IRES-PURO, the system is as follows, 2 μl pLentis-CMV-MCS-IRES-PURO vector fragment, 2 μl gene fragment, 1 μl ligase buffer, 0.5 μl T4 DNA ligase and water 4.5 μl. Place at room temperature for 4 hours. Then the ligation system is transformed into E. coli competent. The next day, colonies are picked from the transformed plate and placed in LB medium at 37°C in a shaker overnight. The plasmid is extracted from the cultured bacteria using a plasmid extraction kit, and the gene fragment is identified by enzyme digestion to determine whether it is successfully connected to the vector. Then the correct vector is sequenced to determine the success of construction. The expression vector pLentis-CMV-mIL12bIL12aIL21GMCSF-IRES-PURO is obtained.

[0229] 4.2 Preparation of expression virus

[0230] 1) Digest the cultured 293FT cells, count and then 3×10 6 cells / well are plated in a 10 cm culture dish with a culture volume of 10 ml.

[0231] 2) The next night, observe the cell state, and if the cell state is good, proceed with the transfection. Add chloroquine to a final concentration of 25 μM in the culture plate, take a test tube, add sterile water and the following plasmids (pMD2.G 6 μg + pSPAX2 15 μg + expression vector obtained in Example 4.1 above 20 μg), the total volume is 1045 μl, then add 2M CaCl2155 μl, mix well, finally add 1200 μl 2×HBS, dropwise add and oscillate, after the dropwise addition is completed, quickly add the mixture to the cell culture well and mix gently.

[0232] 3) The next morning, observe the cell state and replace the culture medium with 10 ml of fresh DMEM medium.

[0233] 4) On the fifth morning, observe the state of the cells, and collect the supernatant in the culture dish, filter it with a 0.45 μm filter, then place it in a high-speed centrifuge tube, centrifuge at 50000g for 2 hours, carefully discard the supernatant, try to absorb the liquid with a water-absorbing paper, then resuspend the precipitate with 200 μl of HBSS, dissolve for 2 hours, then aliquot into small tubes, and store at -70°C.

[0234] 4.3 Preparation of expression cells

[0235] Digest the cultured 293A cells, seed 10 5 cells / well into a 6-well plate, with a culture volume of 1 ml. After 24 hours, add 10 μl of virus expressing the above-mentioned target gene (i.e., the virus obtained in Example 4.2), continue to culture in the incubator for 24 hours, then discard the supernatant and replace it with fresh culture medium for continued culture. After the cells have grown, they are subcultured into culture bottles, and 3 μg / ml of puromycin is added, and the culture medium is replaced every two days, and the concentration of puromycin is maintained. After one week of screening, the surviving cells are cells that stably express the protein, and are named 293A-mIL12bIL12aIL21GMCSF.

[0236] 4.4 Protein expression and purification

[0237] Subculture the constructed 293A-mIL12bIL12aIL21GMCSF cells expressing mIL12bIL12aIL21GMCSF into a 15 cm culture dish, and after the cells have grown, replace the culture medium with 30 ml of CDM4HEK293, continue to culture for 5 days, then collect the supernatant, filter it with a 0.45 μm filter, and then concentrate it with a 50 kd AMICON ULTRA-15 ultrafiltration tube. The obtained concentrated protein solution is purified with nickel chelate magnetic beads (purchased from Beaver Biosciences Technology Co., Ltd.), and the operation procedure is performed according to the instructions. The obtained purified protein solution is ultrafiltered with an AMICON ULTRA-0.5 ultrafiltration tube, the buffer is replaced with PBS, and finally the obtained protein solution is detected for protein concentration with an IL12p70 ELISA kit. After adjusting the protein concentration to 2 μg / μl with PBS, the protein solution is aliquoted and stored at -20°C.

[0238] Example 5 Expression of mIL12aIL2IL12bGMCSF protein

[0239] 5.1 Construction of expression vector

[0240] The mIL12aIL2IL12bGMCSF protein is a heterodimer composed of mIL12aIL2 and mIL12bGMCSF, wherein 6*His is added at the rear end of mIL12aIL2 for purification. The DNA sequence corresponding to the mIL12aIL2IL12bGMCSF protein gene is synthesized, a T2A cleavage peptide is added between the two peptide chains to form a heterodimer after translation, and the sequence is mutated at the BamHI or XhoI site with a degenerate codon. The synthesized sequence has BamHI and XhoI enzyme digestion sites at the front and rear ends, respectively. The plasmid with the target gene is digested by enzymes as follows: 5 μg of plasmid, 4 μl of enzyme digestion buffer, 1 μl of BamHI, and 1 μl of XhoI. Add water to a total volume of 40 μl, and incubate at 37°C for 12 hours. Remove the EP tube, add 4.4 μl of 10× loading buffer, and perform electrophoresis on a 1% agarose gel. Recover the mIL12aIL2IL12bGMCSF protein gene fragment after electrophoresis for later use.

[0241] The amino acid sequence of the second polypeptide chain of the mIL12aIL2IL12bGMCSF protein is shown in SEQ ID NO. 57, the amino acid sequence of the first polypeptide chain is shown in SEQ ID NO. 56, and the nucleotide sequence encoding the mIL12aIL2IL12bGMCSF is shown in SEQ ID NO. 72.

[0242] The pLentis-CMV-MCS-IRES-PURO vector is digested in an EP tube as follows: 2 μg of pLentis-CMV-MCS-IRES-PURO vector plasmid, 3 μl of enzyme digestion buffer, 1 μl of BamHI, and 1 μl of XhoI. Add water to a total volume of 30 μl, and incubate at 37°C for 12 hours. Remove the EP tube, add 3.3 μl of 10× loading buffer, and perform electrophoresis on a 1% agarose gel. Recover the pLentis-CMV-MCS-IRES-PURO vector fragment after electrophoresis for later use.

[0243] Connect mIL12aIL2IL12bGMCSF and pLentis-CMV-MCS-IRES-PURO, system as follows, 2 μl pLentis-CMV-MCS-IRES-PURO vector fragment, 2 μl gene fragment, 1 μl ligase buffer, 0.5 μl T4 DNA ligase and water 4.5 μl. Place in room temperature connection 4 hours. Then the ligation system is transformed into E. coli competent. The next day from the transformed plate pick colonies, placed in LB medium 37 degrees in the incubator overnight culture, using the plasmid extraction kit from the cultured bacteria to extract plasmid, by enzyme cutting to identify whether the gene fragment is successfully connected into the vector, then the correct vector sequencing, determine the success of the construction. Obtain expression vector pLentis-CMV-mIL12aIL2IL12bGMCSF-IRES-PURO.

[0244] 5.2 Preparation of expression virus

[0245] 1) Digest the cultured 293FT cells, count 3x10 6 cells / well in a 10 cm culture dish, culture volume is 10 ml.

[0246] 2) The next night, observe the cell state, if the cell state is good, proceed to transfection. Add chloroquine to the final concentration of 25 μM in the culture plate, take a test tube, add sterile water and the following plasmid (pMD2.G 6 μg + pSPAX2 15 μg + expression vector obtained in the above 5.1 example 20 μg), the total volume is 1045 μl, then add 2M CaCl2 155 μl, mix, finally add 1200 μl 2xHBS, dropwise add and oscillate, after the dropwise addition, quickly add the mixture to the cell culture well, gently shake and mix.

[0247] 3) The next morning, observe the cell state, replace the culture medium with 10 ml of fresh DMEM medium.

[0248] 4) The next morning, observe the cell state, and collect the supernatant in the culture dish, filter with a 0.45 μm filter, then place in a high-speed centrifuge tube, 50000g centrifuge for 2 hours, carefully discard the supernatant, try to absorb the liquid with a water-absorbing paper, then resuspend the precipitate with 200 μl HBSS, dissolve for 2 hours, then aliquot into small tubes, -70°C.

[0249] 5.3 Preparation of expression cells

[0250] Digest the cultured 293A cells, 10 5Cells were seeded at 1 cell / well in 6-well plates with 1 ml culture volume. After 24 hours, 10 μl of virus expressing the above-mentioned target gene (i.e., the virus obtained in Example 5.2) was added, and after 24 hours of further culture in the incubator, the supernatant was discarded and replaced with fresh culture medium for continued culture. After the cells grew to confluence, they were subcultured into culture bottles, and 3 μg / ml puromycin was added to the culture medium. The culture medium was replaced every two days, and the concentration of puromycin was maintained. After one week of selection, the surviving cells were stable cells expressing the protein, and were named 293A-mIL12aIL2IL12bGMCSF.

[0251] 5.4 Protein expression and purification

[0252] The constructed 293A-mIL12aIL2IL12bGMCSF cells expressing mIL12aIL2IL12bGMCSF were subcultured into 15 cm culture dishes. After the cells grew to confluence, the culture medium was replaced with 30 ml CDM4HEK293, and the cells were cultured for 5 days. The supernatant was then collected, filtered through a 0.45 μm filter, and concentrated by ultrafiltration using a 50 kd AMICON ULTRA-15. The concentrated protein solution was purified using nickel-chelated magnetic beads (purchased from Beagle Biotechnology Co., Ltd.), according to the manufacturer's instructions. The purified protein solution was further ultrafiltrated using an AMICON ULTRA-0.5 ultrafiltration tube, and the buffer was replaced with PBS. The final protein solution was detected for protein concentration using an IL12p70 ELISA kit. After the protein concentration was adjusted to 0.2 μg / μl using PBS, the solution was aliquoted and stored at -20 °C.

[0253] Example 6 Effect of glycerol injection on tumor growth in mice

[0254] 5 x 10 5 mouse lung cancer cells (LLC) were injected subcutaneously into the right side of the body of C57BL / 6 mice. When the long diameter of the tumor reached 7-10 mm, treatment was initiated. PBS was used to prepare 50%, 60%, and 70% glycerol solutions, respectively. The prepared glycerol solutions were drawn into a 29G insulin syringe, and slowly injected into the tumor at a volume of 150 μl. After injection, the needle was retained for a short time to reduce the overflow of the solution. The mice were returned to the cage, and the survival of the mice was recorded. Mice injected with PBS only were used as a control. The results are shown in Figure 1 Table 1. Survival of mice after injection of glycerol solution

[0255] Example 7 Effect of mIL12bIL12aIL2GMCSF injection on tumor growth in mice

[0256] 5 x 10 5 The mice were injected with 5 x 10

[0257] A series of liquid formulations of the present application (i.e. mIL12bIL12aIL2GMCSF glycerol injection) with different glycerol mass percentages were prepared using the protein solution prepared in Example 1. Specifically, 50 μl of the protein solution prepared in Example 1 was added to 50 μl of glycerol, and mixed quickly with a syringe to avoid the formation of bubbles, to obtain a liquid formulation with a glycerol concentration of 50%. Similarly, 50 μl of the protein solution prepared in Example 1 was added to 75 μl of glycerol to obtain a liquid formulation with a glycerol concentration of 60%, and 50 μl of the protein solution prepared in Example 1 was added to 117 μl of glycerol to obtain a liquid formulation with a glycerol concentration of 70%. The preparation of the control formulation was as follows: 50 μl of the protein solution prepared in Example 1 was added to 117 μl of PBS, and mixed quickly with a syringe to avoid the formation of bubbles, to obtain the control formulation 1. The liquid formulation of the present application or the control formulation 1 was drawn up using a 29G insulin syringe, and injected slowly into the tumor. After injection, the needle was left in place for a short time to reduce the overflow of the solution. The mice were returned to their cages, and the survival of the mice was recorded. The results are shown in Figure 2, where PBS indicates the control group. As can be seen, all of the mice injected with the liquid formulation of the present application (i.e. mIL12bIL12aIL2GMCSF glycerol injection) survived, indicating that the glycerol solution and the protein molecules produced a synergistic effect, thereby inhibiting the growth of the tumor. Figure 2

[0258] Example 8 Effect of mIL12bIL12aIL2DiaNHS76F8GMCSF-Thr injection on the growth of tumors in mice

[0259] 5 x 10 5 The mice were injected with 5 x 10

[0260] ​A series of liquid formulations of the present application (i.e., mIL12bIL12aIL2DiaNHS76F8GMCSF-Thr glycerol injection) with different glycerol mass percentages were prepared using the protein solution prepared in Example 2. Specifically, 50 μl of the protein solution prepared in Example 2 was added to 50 μl of glycerol, and mixed quickly with a syringe tip to avoid the formation of bubbles, to obtain a liquid formulation with a glycerol concentration of 50%. Similarly, 50 μl of the protein solution prepared in Example 2 was added to 75 μl of glycerol to obtain a liquid formulation with a glycerol concentration of 60%, and 50 μl of the protein solution prepared in Example 2 was added to 117 μl of glycerol to obtain a liquid formulation with a glycerol concentration of 70%. The preparation of the control comparative formulation was as follows: 50 μl of the protein solution prepared in Example 2 was added to 117 μl of PBS, and mixed quickly with a syringe tip to avoid the formation of bubbles, to obtain Comparative Formulation 2. The prepared liquid formulation of the present application (i.e., mIL12bIL12aIL2DiaNHS76F8GMCSF-Thr glycerol injection) or Comparative Formulation 2 was drawn up using a 29G insulin syringe, and injected slowly into the tumor. After injection, the needle was left in place for a short period of time to reduce the overflow of the solution. The mice were returned to their cages after injection, and the survival of the mice was recorded. The results are shown in Figure 2, where PBS indicates the control group. As can be seen, all of the mice injected with the liquid formulation of the present application, mIL12bIL12aIL2DiaNHS76F8GMCSF-Thr glycerol injection, survived, indicating that the glycerol solution and the protein molecules produced a synergistic effect, thereby jointly inhibiting the growth of the tumor. Figure 3

[0261] Example 9 Effect of mIL12bIL12aIL7GMCSF injection on the growth of tumors in mice

[0262] 5 x 10 5 The digested and cultured mouse lung cancer cells (LLC) were injected subcutaneously into the right side of the body of C57BL / 6 mice, and treatment was started when the long diameter of the tumor reached 7-10 mm.

[0263] 50 μl of the protein solution prepared in Example 3 was added to 80 μl of glycerol, and mixed quickly with a syringe tip to avoid the formation of bubbles, to obtain the prepared liquid formulation of the present application. The preparation of the control comparative formulation was as follows: 50 μl of the protein solution prepared in Example 3 was added to 80 μl of PBS, and mixed quickly with a syringe tip to avoid the formation of bubbles, to obtain Comparative Formulation 3. The prepared liquid formulation of the present application or Comparative Formulation 3 was drawn up using a 29G insulin syringe, and injected slowly into the tumor. After injection, the needle was left in place for a short period of time to reduce the overflow of the solution. The mice were returned to their cages after injection, and the survival of the mice was recorded. The results are shown in Figure 3, where PBS indicates the control group. As can be seen, all of the mice injected with the liquid formulation of the present application survived, indicating that the glycerol solution and the protein molecules produced a synergistic effect, thereby jointly inhibiting the growth of the tumor. Figure 4 ​As shown in the figure, PBS represents the control group. It can be seen that after injection of the liquid preparation of the present application (i.e. mIL12bIL12aIL7GMCSF glycerol injection), all mice survived, indicating that the glycerol solution and the protein molecules produced a synergistic effect, thereby jointly inhibiting the growth of the tumor.

[0264] Example 10 Effect of mIL12bIL12aIL21GMCSF injection on tumor growth in mice

[0265] 5 x 10 5 The digested and cultured mouse lung cancer cells (LLC) were injected subcutaneously into the right side of the body of C57BL / 6 mice, and treatment was started when the long diameter of the tumor reached 7-10 mm.

[0266] 50 μl of the protein solution prepared in Example 4 was taken and then added to 80 μl of glycerol, and mixed quickly with a syringe head to avoid the generation of bubbles, thereby obtaining the prepared liquid preparation of the present application. The preparation method of the control comparative preparation was as follows: 50 μl of the protein solution prepared in Example 4 was taken and then added to 80 μl of PBS, and mixed quickly with a syringe head to avoid the generation of bubbles, thereby obtaining the prepared comparative preparation 4. The prepared liquid preparation of the present application or comparative preparation 4 was taken with a 29G insulin syringe, and slowly injected into the tumor, and the needle was retained for a short time after injection to reduce the overflow of the solution. The mice were returned to the cage after injection, and the survival of the mice was recorded. The results are shown in Figure 5 As shown in the figure, PBS represents the control group. It can be seen that after injection of the liquid preparation of the present application (i.e. mIL12bIL12aIL7GMCSF glycerol injection), all mice survived, indicating that the glycerol solution and the protein molecules produced a synergistic effect, thereby jointly inhibiting the growth of the tumor.

[0267] Example 11 Effect of mIL12aIL2IL12bGMCSF injection on tumor growth in mice

[0268] 5 x 10 5 The digested and cultured mouse lung cancer cells (LLC) were injected subcutaneously into the right side of the body of C57BL / 6 mice, and treatment was started when the long diameter of the tumor reached 7-10 mm.

[0269] The protein solution prepared in Example 5 was taken 50 μl, then added to 80 μl of glycerol, and mixed quickly with a syringe head to avoid the generation of bubbles, to obtain the prepared liquid preparation of the present application. The preparation method of the control comparative preparation was as follows: the protein solution prepared in Example 5 was taken 50 μl, then added to 80 μl of PBS, and mixed quickly with a syringe head to avoid the generation of bubbles, to obtain the prepared comparative preparation 5. The prepared liquid preparation of the present application or comparative preparation 5 was taken with a 29G insulin syringe, and slowly injected into the tumor, and the needle was retained for a short time after injection to reduce the overflow of the solution. The mice were put back into the cage after injection, and the survival of the mice was recorded. The results are shown in Figure 1, wherein PBS represents the control group. It can be seen that after the injection of the liquid preparation of the present application (i.e. mIL12aIL2IL12bGMCSF glycerol injection), all the mice survived, indicating that the glycerol solution and the protein molecules produced a synergistic effect, thereby jointly inhibiting the growth of the tumor. Figure 6 Liquid formulations Proteins Drugs, uses, methods of treating tumors

[0270] The foregoing detailed description is provided by way of explanation and example only, and is not intended to limit the scope of the appended claims. Various modifications to the embodiments described herein will be apparent to those of ordinary skill in the art, and are intended to be within the scope of the claims and their equivalents. SEQUENCE LISTING <110> Zhang Jin-yu <120> A liquid preparation and its use <130> 0069-PA-007 <160> 76 <170> PatentIn version 3.5 <210> 1 <211> 469 <212> PRT <213> Artificial Sequence <220> <223> DiaL19 <400> 1 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Phe 20 25 30 ​Ser Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser lie Ser Gly Ser Ser Gly Thr Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Pro Phe Pro Tyr Phe Asp Tyr Trp Gly Gin Gly Thr Leu Val 100 105 110 Thr Val Gly Gly Ser Gly Gly Glu lie Val Leu Thr Gin Ser Pro Gly 115 120 125 Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala 130 135 140 Ser Gin Ser Val Ser Ser Ser Phe Leu Ala Trp Tyr Gin Gin Lys Pro 145 150 155 160 Gly Gin Ala Pro Arg Leu Leu lie Tyr Tyr Ala Ser Ser Arg Ala Thr 165 170 175 Gly lie Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 180 185 190 Leu Thr lie Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys 195 200 205 Gln Gln Thr Gly Arg lie Pro Pro Thr Phe Gly Gin Gly Thr Lys Val 210 215 220 Glu lie Lys Ser Ser Ser Ser Gly Ser Ser Ser Ser Gly Ser Ser Ser 225 230 235 240 Ser Gly Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro 245 250 255 Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser 260 265 270 Ser Phe Ser Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu 275 280 285 Trp Val Ser Ser lie Ser Gly Ser Ser Gly Thr Thr Tyr Tyr Ala Asp 290 295 300 Ser Val Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr 305 310 315 320 Leu Tyr Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 325 330 335 Tyr Cys Ala Lys Pro Phe Pro Tyr Phe Asp Tyr Trp Gly Gin Gly Thr 340 345 350 Leu Val Thr Val Gly Gly Ser Gly Gly Glu lie Val Leu Thr Gin Ser 355 360 365 Pro Gly Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys 370 375 380 Arg Ala Ser Gin Ser Val Ser Ser Ser Phe Leu Ala Trp Tyr Gin Gin 385 390 395 400 Lys Pro Gly Gin Ala Pro Arg Leu Leu lie Tyr Tyr Ala Ser Ser Arg 405 410 415 Ala Thr Gly lie Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp 420 425 430 Phe Thr Leu Thr lie Ser Arg Leu Glu Pro Gin Asp Phe Ala Val Tyr 435 440 445 Tyr Cys Gin Gin Thr Gly Arg lie Pro Pro Thr Phe Gly Gin Gly Thr 450 455 460 Lys Val Glu lie Lys 465 <210> 2 <211> 477 <212> PRT <213> Artificial Sequence <220> <223> DiaF8 <400> 2 Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Leu Phe 20 25 30 Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Ser Thr His Leu Tyr Leu Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Gly Gly Ser Gly Gly Glu Ile Val Leu Thr 115 120 125 Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu 130 135 140 Ser Cys Arg Ala Ser Gln Ser Val Ser Met Pro Phe Leu Ala Trp Tyr 145 150 155 160 Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr Gly Ala Ser 165 170 175 Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly 180 185 190 Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu Asp Phe Ala 195 200 205 Val Tyr Tyr Cys Gln Gln Met Arg Gly Arg Pro Pro Thr Phe Gly Gln 210 215 220 Gly Thr Lys Val Glu Ile Lys Ser Ser Ser Ser Gly Ser Ser Ser Ser 225 230 235 240 Gly Ser Ser Ser Ser Gly Glu Val Gln Leu Leu Glu Ser Gly Gly Gly 245 250 255 Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 260 265 270 Phe Thr Phe Ser Leu Phe Thr Met Ser Trp Val Arg Gln Ala Pro Gly 275 280 285 Lys Gly Leu Glu Trp Val Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr 290 295 300 Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn 305 310 315 320 Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp 325 330 335 Thr Ala Val Tyr Tyr Cys Ala Lys Ser Thr His Leu Tyr Leu Phe Asp 340 345 350 Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Ser Gly 355 360 365 Gly Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro 370 375 380 Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Met 385 390 395 400 Pro Phe Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu 405 410 415 Leu Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe 420 425 430 Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu 435 440 445 Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Met Arg Gly Arg 450 455 460 Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 465 470 475 <210> 3 <211> 475 <212> PRT <213> Artificial Sequence <220> <223> DiaNHS76 <400> 3 Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Tyr Ser Ile Ser Ser Gly 20 25 30 Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 Ile Gly Ser Ile Tyr His Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu 50 55 60 Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser 65 70 75 80 Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Lys Trp Ser Lys Phe Asp Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser Gly Gly Ser Gly Gly Ser Ser Glu Leu Thr Gln 115 120 125 Asp Pro Ala Val Ser Val Ala Leu Gly Gln Thr Val Arg Ile Thr Cys 130 135 140 Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Ala Ser Trp Tyr Gln Gln Lys 145 150 155 160 Pro Gly Gln Ala Pro Val Leu Val Ile Tyr Gly Lys Asn Asn Arg Pro 165 170 175 Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Ser Ser Gly Asn Thr Ala 180 185 190 Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu Asp Glu Ala Asp Tyr Tyr 195 200 205 Cys Asn Ser Arg Asp Ser Ser Gly Asn His Val Val Phe Gly Gly Gly 210 215 220 Thr Lys Leu Thr Val Leu Ser Ser Ser Ser Gly Ser Ser Ser Ser Gly 225 230 235 240 Ser Ser Ser Ser Gly Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu 245 250 255 Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Tyr 260 265 270 Ser Ile Ser Ser Gly Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly 275 280 285 Lys Gly Leu Glu Trp Ile Gly Ser Ile Tyr His Ser Gly Ser Thr Tyr 290 295 300 Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser 305 310 315 320 Lys Asn Gln Phe Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr 325 330 335 Ala Val Tyr Tyr Cys Ala Arg Gly Lys Trp Ser Lys Phe Asp Tyr Trp 340 345 350 Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Ser Gly Gly Ser 355 360 365 Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln Thr 370 375 380 Val Arg Ile Thr Cys Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Ala Ser 385 390 395 400 Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr Gly 405 410 415 Lys Asn Asn Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Ser 420 425 430 Ser Gly Asn Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu Asp 435 440 445 Glu Ala Asp Tyr Tyr Cys Asn Ser Arg Asp Ser Ser Gly Asn His Val 450 455 460 Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 465 470 475 <210> 4 <211> 476 <212> PRT <213> Artificial Sequence <220> <223> DiaNHS76F8 <400> 4 Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Tyr Ser Ile Ser Ser Gly 20 25 30 Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 Ile Gly Ser Ile Tyr His Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu 50 55 60 Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser 65 70 75 80 Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Lys Trp Ser Lys Phe Asp Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser Gly Gly Ser Gly Gly Glu lie Val Leu Thr Gin 115 120 125 Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser 130 135 140 Cys Arg Ala Ser Gin Ser Val Ser Met Pro Phe Leu Ala Trp Tyr Gin 145 150 155 160 Gln Lys Pro Gly Gin Ala Pro Arg Leu Leu lie Tyr Gly Ala Ser Ser 165 170 175 Arg Ala Thr Gly lie Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr 180 185 190 Asp Phe Thr Leu Thr lie Ser Arg Leu Glu Pro Glu Asp Phe Ala Val 195 200 205 Tyr Tyr Cys Gin Gin Met Arg Gly Arg Pro Pro Thr Phe Gly Gin Gly 210 215 220 Thr Lys Val Glu lie Lys Ser Ser Ser Ser Gly Ser Ser Ser Ser Gly 225 230 235 240 Ser Ser Ser Ser Gly Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu 245 250 255 Val Gin Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 260 265 270 Thr Phe Ser Leu Phe Thr Met Ser Trp Val Arg Gin Ala Pro Gly Lys 275 280 285 Gly Leu Glu Trp Val Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr 290 295 300 Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser 305 310 315 320 Lys Asn Thr Leu Tyr Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr 325 330 335 Ala Val Tyr Tyr Cys Ala Lys Ser Thr His Leu Tyr Leu Phe Asp Tyr 340 345 350 Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser Gly Gly Ser Gly Gly 355 360 365 Ser Ser Glu Leu Thr Gin Asp Pro Ala Val Ser Val Ala Leu Gly Gin 370 375 380 Thr Val Arg lie Thr Cys Gin Gly Asp Ser Leu Arg Ser Tyr Tyr Ala 385 390 395 400 Ser Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Val Leu Val lie Tyr 405 410 415 Gly Lys Asn Asn Arg Pro Ser Gly lie Pro Asp Arg Phe Ser Gly Ser Gly Lys Asn Asn Arg Pro Ser Gly lie Pro Asp Arg Phe Ser Gly Ser420 425 430 Ser Ser Gly Asn Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu 435 440 445 Asp Glu Ala Asp Tyr Tyr Cys Asn Ser Arg Asp Ser Ser Gly Asn His 450 455 460 Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 465 470 475 <210> 5 <211> 472 <212> PRT <213> Artificial Sequence <220> <223> DiaNHS76L19 <400> 5 Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Tyr Ser Ile Ser Ser Gly 20 25 30 Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 Ile Gly Ser Ile Tyr His Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu 50 55 60 Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser 65 70 75 80 Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Lys Trp Ser Lys Phe Asp Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser Gly Gly Ser Gly Gly Glu Ile Val Leu Thr Gln 115 120 125 Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser 130 135 140 Cys Arg Ala Ser Gln Ser Val Ser Ser Ser Phe Leu Ala Trp Tyr Gln 145 150 155 160 Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr Tyr Ala Ser Ser 165 170 175 Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr 180 185 190 Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu Asp Phe Ala Val 195 200 205 Tyr Tyr Cys Gln Gln Thr Gly Arg Ile Pro Pro Thr Phe Gly Gln Gly 210 215 220 Thr Lys Val Glu Ile Lys Ser Ser Ser Ser Gly Ser Ser Ser Ser Gly 225 230 235 240 Ser Ser Ser Ser Gly Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu 245 250 255 Val Gin Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 260 265 270 Thr Phe Ser Ser Phe Ser Met Ser Trp Val Arg Gin Ala Pro Gly Lys 275 280 285 Gly Leu Glu Trp Val Ser Ser lie Ser Gly Ser Ser Gly Thr Thr Tyr 290 295 300 Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser 305 310 315 320 Lys Asn Thr Leu Tyr Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr 325 330 335 Ala Val Tyr Tyr Cys Ala Lys Pro Phe Pro Tyr Phe Asp Tyr Trp Gly 340 345 350 Gln Gly Thr Leu Val Thr Val Gly Gly Ser Gly Gly Ser Ser Glu Leu 355 360 365 Thr Gin Asp Pro Ala Val Ser Val Ala Leu Gly Gin Thr Val Arg lie 370 375 380 Thr Cys Gin Gly Asp Ser Leu Arg Ser Tyr Tyr Ala Ser Trp Tyr Gin 385 390 395 400 Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr Gly Lys Asn Asn 405 410 415 Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Ser Ser Gly Asn 420 425 430 Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu Asp Glu Ala Asp 435 440 445 Tyr Tyr Cys Asn Ser Arg Asp Ser Ser Gly Asn His Val Val Phe Gly 450 455 460 Gly Gly Thr Lys Leu Thr Val Leu 465 470 <210> 6 <211> 476 <212> PRT <213> Artificial Sequence <220> <223> DiaF8NHS76 <400> 6 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Leu Phe 20 25 30 Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Ser Thr His Leu Tyr Leu Phe Asp Tyr Trp Gly Gin Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Gly Gly Ser Gly Gly Ser Ser Glu Leu Thr 115 120 125 Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gin Thr Val Arg lie Thr 130 135 140 Cys Gin Gly Asp Ser Leu Arg Ser Tyr Tyr Ala Ser Trp Tyr Gin Gin 145 150 155 160 Lys Pro Gly Gin Ala Pro Val Leu Val lie Tyr Gly Lys Asn Asn Arg 165 170 175 Pro Ser Gly lie Pro Asp Arg Phe Ser Gly Ser Ser Ser Gly Asn Thr 180 185 190 Ala Ser Leu Thr lie Thr Gly Ala Gin Ala Glu Asp Glu Ala Asp Tyr 195 200 205 Tyr Cys Asn Ser Arg Asp Ser Ser Gly Asn His Val Val Phe Gly Gly 210 215 220 Gly Thr Lys Leu Thr Val Leu Ser Ser Ser Ser Gly Ser Ser Ser Ser 225 230 235 240 Gly Ser Ser Ser Ser Gly Gln Val Gln Leu Gln Glu Ser Gly Pro Gly 245 250 255 Leu Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Ala Val Ser Gly 260 265 270 Tyr Ser Ile Ser Ser Gly Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro 275 280 285 Gly Lys Gly Leu Glu Trp Ile Gly Ser Ile Tyr His Ser Gly Ser Thr 290 295 300 Tyr Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr 305 310 315 320 Ser Lys Asn Gln Phe Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp 325 330 335 Thr Ala Val Tyr Tyr Cys Ala Arg Gly Lys Trp Ser Lys Phe Asp Tyr 340 345 350 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Ser Gly Gly 355 360 365 Glu Ile Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 370 375 380 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Met Pro 385 390 395 400 Phe Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 405 410 415 Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 420 425 430 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu 435 440 445 Pro Gin Asp Phe Ala Val Tyr Tyr Cys Gin Gin Met Arg Gly Arg Pro 450 455 460 Pro Thr Phe Gly Gin Gly Thr Lys Val Glu Ile Lys 465 470 475 <210> 7 <211> 473 <212> PRT <213> Artificial Sequence <220> <223> DiaF8L19 <400> 7 Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Leu Phe 20 25 30 Thr Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala He Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Ser Thr His Leu Tyr Leu Phe Asp Tyr Trp Gly Gin Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Gly Gly Ser Gly Gly Glu He Val Leu Thr 115 120 125 Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu 130 135 140 Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Ser Phe Leu Ala Trp Tyr 145 150 155 160 Gln Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu He Tyr Tyr Ala Ser 165 170 175 Ser Arg Ala Thr Gly lie Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly 180 185 190 Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu Pro Glu Asp Phe Ala 195 200 205 Val Tyr Tyr Cys Gin Gin Thr Gly Arg lie Pro Pro Thr Phe Gly Gin 210 215 220 Gly Thr Lys Val Glu lie Lys Ser Ser Ser Ser Gly Ser Ser Ser Ser 225 230 235 240 Gly Ser Ser Ser Ser Gly Glu Val Gin Leu Leu Glu Ser Gly Gly Gly 245 250 255 Leu Val Gin Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 260 265 270 Phe Thr Phe Ser Ser Phe Ser Met Ser Trp Val Arg Gin Ala Pro Gly 275 280 285 Lys Gly Leu Glu Trp Val Ser Ser lie Ser Gly Ser Ser Gly Thr Thr 290 295 300 Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr lie Ser Arg Asp Asn 305 310 315 320 Ser Lys Asn Thr Leu Tyr Leu Gin Met Asn Ser Leu Arg Ala Glu Asp 325 330 335 Thr Ala Val Tyr Tyr Cys Ala Lys Pro Phe Pro Tyr Phe Asp Tyr Trp 340 345 350 Gly Gln Gly Thr Leu Val Thr Val Gly Gly Ser Gly Gly Glu Ile Val 355 360 365 Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala 370 375 380 Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Met Pro Phe Leu Ala 385 390 395 400 Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr Gly 405 410 415 Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser Gly Ser Gly 420 425 430 Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu Asp 435 440 445 Phe Ala Val Tyr Tyr Cys Gln Gln Met Arg Gly Arg Pro Pro Thr Phe 450 455 460 Gly Gln Gly Thr Lys Val Glu Ile Lys 465 470 <210> 8 <211> 472 <212> PRT <213> Artificial Sequence <220> <223> DiaL19NHS76 <400> 8 Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Phe 20 25 30 Ser Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser lie Ser Gly Ser Ser Gly Thr Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Pro Phe Pro Tyr Phe Asp Tyr Trp Gly Gin Gly Thr Leu Val 100 105 110 Thr Val Gly Gly Ser Gly Gly Ser Ser Glu Leu Thr Gin Asp Pro Ala 115 120 125 Val Ser Val Ala Leu Gly Gin Thr Val Arg lie Thr Cys Gin Gly Asp 130 135 140 Ser Leu Arg Ser Tyr Tyr Ala Ser Trp Tyr Gin Gin Lys Pro Gly Gin 145 150 155 160 Ala Pro Val Leu Val lie Tyr Gly Lys Asn Asn Arg Pro Ser Gly lie 165 170 175 Pro Asp Arg Phe Ser Gly Ser Ser Ser Gly Asn Thr Ala Ser Leu Thr 180 185 190 lie Thr Gly Ala Gin Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Asn Ser 195 200 205 Arg Asp Ser Ser Gly Asn His Val Val Phe Gly Gly Gly Thr Lys Leu 210 215 220 Thr Val Leu Ser Ser Ser Ser Gly Ser Ser Ser Ser Gly Ser Ser Ser 225 230 235 240 Ser Gly Gin Val Gin Leu Gin Glu Ser Gly Pro Gly Leu Val Lys Pro 245 250 255 Ser Glu Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Tyr Ser lie Ser 260 265 270 Ser Gly Tyr Tyr Trp Gly Trp lie Arg Gin Pro Pro Gly Lys Gly Leu 275 280 285 Glu Trp lie Gly Ser lie Tyr His Ser Gly Ser Thr Tyr Tyr Asn Pro 290 295 300 Ser Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln 305 310 315 320 Phe Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr 325 330 335 Tyr Cys Ala Arg Gly Lys Trp Ser Lys Phe Asp Tyr Trp Gly Gln Gly 340 345 350 Thr Leu Val Thr Val Ser Ser Gly Gly Ser Gly Gly Glu Ile Val Leu 355 360 365 Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr 370 375 380 Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser Phe Leu Ala Trp 385 390 395 400 Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr Tyr Ala 405 410 415 Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser Gly Ser Gly Ser 420 425 430 Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu Asp Phe 435 440 445 Ala Val Tyr Tyr Cys Gln Gln Thr Gly Arg Ile Pro Pro Thr Phe Gly 450 455 460 Gln Gly Thr Lys Val Glu Ile Lys 465 470 <210> 9 <211> 473 <212> PRT <213> Artificial Sequence <220> <223> DiaL19F8 <400> 9 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Phe 20 25 30 Ser Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Ser Gly Ser Ser Gly Thr Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Pro Phe Pro Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Gly Gly Ser Gly Gly Glu lie Val Leu Thr Gin Ser Pro Gly 115 120 125 Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala 130 135 140 Ser Gin Ser Val Ser Met Pro Phe Leu Ala Trp Tyr Gin Gin Lys Pro 145 150 155 160 Gly Gin Ala Pro Arg Leu Leu lie Tyr Gly Ala Ser Ser Arg Ala Thr 165 170 175 Gly lie Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 180 185 190 Leu Thr lie Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys 195 200 205 Gln Gin Met Arg Gly Arg Pro Pro Thr Phe Gly Gin Gly Thr Lys Val 210 215 220 Glu lie Lys Ser Ser Ser Ser Gly Ser Ser Ser Ser Gly Ser Ser Ser 225 230 235 240 Ser Gly Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro 245 250 255 Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser 260 265 270 Leu Phe Thr Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu 275 280 285 Trp Val Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp 290 295 300 Ser Val Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr 305 310 315 320 Leu Tyr Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 325 330 335 Tyr Cys Ala Lys Ser Thr His Leu Tyr Leu Phe Asp Tyr Trp Gly Gin 340 345 350 Gly Thr Leu Val Thr Val Ser Ser Gly Gly Ser Gly Gly Glu lie Val 355 360 365 Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala 370 375 380 Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Ser Phe Leu Ala 385 390 395 400 Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu lie Tyr Tyr 405 410 415 Ala Ser Ser Arg Ala Thr Gly lie Pro Asp Arg Phe Ser Gly Ser Gly 420 425 430 Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu Asp 435 440 445 Phe Ala Val Tyr Tyr Cys Gln Gln Thr Gly Arg Ile Pro Pro Thr Phe 450 455 460 Gly Gln Gly Thr Lys Val Glu Ile Lys 465 470 <210> 10 <211> 108 <212> PRT <213> Artificial Sequence <220> <223> L19 VL <400> 10 Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30 Phe Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Tyr Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Thr Gly Arg Ile Pro 85 90 95 Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 11 <211> 114 <212> PRT <213> Artificial Sequence <220> <223> L19VH <400> 11 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Phe 20 25 30 Ser Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Ser Gly Ser Ser Gly Thr Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Pro Phe Pro Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val <210> 12 <211> 108 <212> PRT <213> Artificial Sequence <220> <223> F8 VL <400> 12 Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Met Pro 20 25 30 Phe Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Tyr Gly Ala Ser Ser Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Met Arg Gly Arg Pro 85 90 95 Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 13 <211> 118 <212> PRT <213> Artificial Sequence <220> <223> F8VH <400> 13 Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Leu Phe 20 25 30 Thr Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Ser Thr His Leu Tyr Leu Phe Asp Tyr Trp Gly Gin Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210> 14 <211> 108 <212> PRT <213> Artificial Sequence <220> <223> NHS76VL <400> 14 Ser Ser Glu Leu Thr Gin Asp Pro Ala Val Ser Val Ala Leu Gly Gin 1 5 10 15 Thr Val Arg lie Thr Cys Gin Gly Asp Ser Leu Arg Ser Tyr Tyr Ala 20 25 30 Ser Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Val Leu Val lie Tyr 35 40 45 Gly Lys Asn Asn Arg Pro Ser Gly lie Pro Asp Arg Phe Ser Gly Ser 50 55 60 Ser Ser Gly Asn Thr Ala Ser Leu Thr lie Thr Gly Ala Gin Ala Glu 65 70 75 80 Asp Glu Ala Asp Tyr Tyr Cys Asn Ser Arg Asp Ser Ser Gly Asn His 85 90 95 Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 <210> 15 <211> 117 <212> PRT <213> Artificial Sequence <220> <223> NHS76VH? <400> 15 Gln Val Gin Leu Gin Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Tyr Ser Ile Ser Ser Gly 20 25 30 Tyr Tyr Trp Gly Trp Ile Arg Gin Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 Ile Gly Ser Ile Tyr His Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu 50 55 60 Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gin Phe Ser 65 70 75 80 Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Lys Trp Ser Lys Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 16 <211> 193 <212> PRT <213> Mus musculus <400> 16 Arg Val Ile Pro Val Ser Gly Pro Ala Arg Cys Leu Ser Gin Ser Arg 1 5 10 15 Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys Thr Ala Arg Glu Lys 20 25 30 Leu Lys His Tyr Ser Cys Thr Ala Glu Asp lie Asp His Glu Asp lie 35 40 45 Thr Arg Asp Gin Thr Ser Thr Leu Lys Thr Cys Leu Pro Leu Glu Leu 50 55 60 His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr Ser Ser Thr Thr 65 70 75 80 Arg Gly Ser Cys Leu Pro Pro Gin Lys Thr Ser Leu Met Met Thr Leu 85 90 95 Cys Leu Gly Ser lie Tyr Glu Asp Leu Lys Met Tyr Gin Thr Glu Phe 100 105 110 Gln Ala lie Asn Ala Ala Leu Gin Asn His Asn His Gin Gin lie lie 115 120 125 Leu Asp Lys Gly Met Leu Val Ala lie Asp Glu Leu Met Gin Ser Leu 130 135 140 Asn His Asn Gly Glu Thr Leu Arg Gin Lys Pro Pro Val Gly Glu Ala 145 150 155 160 Asp Pro Tyr Arg Val Lys Met Lys Leu Cys lie Leu Leu His Ala Phe 165 170 175 Ser Thr Arg Val Val Thr lie Asn Arg Val Met Gly Tyr Leu Ser Ser 180 185 190 Ala <210> 17 <211> 313 <212> PRT <213> Mus musculus <400> 17 Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr 1 5 10 15 Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg His Gly Val Ile Gly 35 40 45 Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu Phe Leu Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu Lys 85 90 95 Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser 100 105 110 Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu Lys 115 120 125 Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val Thr 130 135 140 Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln Arg 145 150 155 160 Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys Pro 165 170 175 Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala Leu Glu Ala Arg Gln 180 185 190 Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe Ile Arg Asp Ile 195 200 205 Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met Lys Pro Leu Lys Asn 210 215 220 Ser Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro 225 230 235 240 His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg Ile Gln Arg Lys Lys 245 250 255 Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gln Lys Gly Ala Phe 260 265 270 Leu Val Glu Lys Thr Ser Thr Glu Val Gln Cys Lys Gly Gly Asn Val 275 280 285 Cys Val Gln Ala Gln Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp 290 295 300 Ala Cys Val Pro Cys Arg Val Arg Ser 305 310 <210> 18 <211> 197 <212> PRT <213> Homo sapiens <400> 18 Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys Leu 1 5 10 15 His His Ser Gln Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gln Lys 20 25 30 Ala Arg Gln Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu Ile Asp 35 40 45 His Glu Asp Ile Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys Leu 50 55 60 Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu Thr 65 70 75 80 Ser Phe Ile Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser Phe 85 90 95 Met Met Ala Leu Cys Leu Ser Ser Ile Tyr Glu Asp Leu Lys Met Tyr 100 105 110 Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro Lys 115 120 125 Arg Gin He Phe Leu Asp Gin Asn Met Leu Ala Val He Asp Glu Leu 130 135 140 Met Gin Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gin Lys Ser Ser 145 150 155 160 Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys He Lys Leu Cys He Leu 165 170 175 Leu His Ala Phe Arg He Arg Ala Val Thr He Asp Arg Val Met Ser 180 185 190 Tyr Leu Asn Ala Ser 195 <210> 19 <211> 306 <212> PRT <213> Homo sapiens <400> 19 He Trp Glu Leu Lys Lys Asp Val Tyr Val Val Glu Leu Asp Trp Tyr 1 5 10 15 Pro Asp Ala Pro Gly Glu Met Val Val Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Gly He Thr Trp Thr Leu Asp Gin Ser Ser Glu Val Leu Gly 35 40 45 Ser Gly Lys Thr Leu Thr He Gin Val Lys Glu Phe Gly Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Val Leu Ser His Ser Leu Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asp Gly lie Trp Ser Thr Asp lie Leu Lys 85 90 95 Asp Gln Lys Glu Pro Lys Asn Lys Thr Phe Leu Arg Cys Glu Ala Lys 100 105 110 Asn Tyr Ser Gly Arg Phe Thr Cys Trp Trp Leu Thr Thr lie Ser Thr 115 120 125 Asp Leu Thr Phe Ser Val Lys Ser Ser Arg Gly Ser Ser Asp Pro Gln 130 135 140 Gly Val Thr Cys Gly Ala Ala Thr Leu Ser Ala Glu Arg Val Arg Gly 145 150 155 160 Asp Asn Lys Glu Tyr Glu Tyr Ser Val Glu Cys Gln Glu Asp Ser Ala 165 170 175 Cys Pro Ala Ala Glu Glu Ser Leu Pro lie Glu Val Met Val Asp Ala 180 185 190 Val His Lys Leu Lys Tyr Glu Asn Tyr Thr Ser Ser Phe Phe lie Arg 195 200 205 Asp lie lie Lys Pro Asp Pro Pro Lys Asn Leu Gln Leu Lys Pro Leu 210 215 220 Lys Asn Ser Arg Gin Val Glu Val Ser Trp Glu Tyr Pro Asp Thr Trp 225 230 235 240 Ser Thr Pro His Ser Tyr Phe Ser Leu Thr Phe Cys Val Gin Val Gin 245 250 255 Gly Lys Ser Lys Arg Glu Lys Lys Asp Arg Val Phe Thr Asp Lys Thr 260 265 270 Ser Ala Thr Val He Cys Arg Lys Asn Ala Ser He Ser Val Arg Ala 275 280 285 Gln Asp Arg Tyr Tyr Ser Ser Ser Trp Ser Glu Trp Ala Ser Val Pro 290 295 300 Cys Ser 305 <210> 20 <211> 149 <212> PRT <213> Mus musculus <400> 20 Ala Pro Thr Ser Ser Ser Thr Ser Ser Ser Thr Ala Glu Ala Gin Gin 1 5 10 15 Gln Gin Gin Gin Gin Gin Gin Gin Gin His Leu Glu Gin Leu Leu 20 25 30 Met Asp Leu Gin Glu Leu Leu Ser Arg Met Glu Asn Tyr Arg Asn Leu 35 40 45 Lys Leu Pro Arg Met Leu Thr Phe Lys Phe Tyr Leu Pro Lys Gin Ala 50 55 60 Thr Glu Leu Lys Asp Leu Gin Cys Leu Glu Asp Glu Leu Gly Pro Leu 65 70 75 80 Arg His Val Leu Asp Leu Thr Gin Ser Lys Ser Phe Gin Leu Glu Asp 85 90 95 Ala Glu Asn Phe He Ser Asn He Arg Val Thr Val Val Lys Leu Lys 100 105 110 Gly Ser Asp Asn Thr Phe Glu Cys Gin Phe Asp Asp Glu Ser Ala Thr 115 120 125 Val Val Asp Phe Leu Arg Arg Trp He Ala Phe Cys Gin Ser He He 130 135 140 Ser Thr Ser Pro Gin 145 <210> 21 <211> 133 <212> PRT <213> Homo sapiens <400> 21 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gin Leu Gin Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gin Met He Leu Asn Gly He Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 22 <211> 133 <212> PRT <213> Mus musculus <400> 22 Gly Ile His Val Phe Ile Leu Gly Cys Val Ser Val Gly Leu Pro Lys 1 5 10 15 Thr Glu Ala Asn Trp Ile Asp Val Arg Tyr Asp Leu Glu Lys Ile Glu 20 25 30 Ser Leu Ile Gin Ser Ile His Ile Asp Thr Thr Leu Tyr Thr Asp Ser 35 40 45 Asp Phe His Pro Ser Cys Lys Val Thr Ala Met Asn Cys Phe Leu Leu 50 55 60 Glu Leu Gin Val Ile Leu His Glu Tyr Ser Asn Met Thr Leu Asn Glu 65 70 75 80 Thr Val Arg Asn Val Leu Tyr Leu Ala Asn Ser Thr Leu Ser Ser Asn 85 90 95 Lys Asn Val Ala Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu 100 105 110 Lys Thr Phe Thr Glu Phe Leu Gin Ser Phe Ile Arg Ile Val Gin Met 115 120 125 Phe Ile Asn Thr Ser 130 <210> 23 <211> 133 <212> PRT <213> Homo sapiens <400> 23 Gly Ile His Val Phe Ile Leu Gly Cys Phe Ser Ala Gly Leu Pro Lys 1 5 10 15 Thr Glu Ala Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu 20 25 30 Asp Leu Ile Gin Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser 35 40 45 Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu 50 55 60 Glu Leu Gin Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp 65 70 75 80 Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn 85 90 95 Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu 100 105 110 Lys Asn Ile Lys Glu Phe Leu Gin Ser Phe Val His Ile Val Gin Met 115 120 125 Phe Ile Asn Thr Ser 130 <210> 24 <211> 129 <212> PRT <213> Mus musculus <400> 24 Glu Cys His Ile Lys Asp Lys Glu Gly Lys Ala Tyr Glu Ser Val Leu 1 5 10 15 Met Ile Ser Ile Asp Glu Leu Asp Lys Met Thr Gly Thr Asp Ser Asn 20 25 30 Cys Pro Asn Asn Glu Pro Asn Phe Phe Arg Lys His Val Cys Asp Asp 35 40 45 Thr Lys Glu Ala Ala Phe Leu Asn Arg Ala Ala Arg Lys Leu Lys Gin 50 55 60 Phe Leu Lys Met Asn Ile Ser Glu Glu Phe Asn Val His Leu Leu Thr 65 70 75 80 Val Ser Gin Gly Thr Gin Thr Leu Val Asn Cys Thr Ser Lys Glu Glu 85 90 95 Lys Asn Val Lys Glu Gin Lys Lys Asn Asp Ala Cys Phe Leu Lys Arg 100 105 110 Leu Leu Arg Glu Ile Lys Thr Cys Trp Asn Lys Ile Leu Lys Gly Ser 115 120 125 Ile <210> 25 <211> 152 <212> PRT <213> Homo sapiens <400> 25 Asp Cys Asp Ile Glu Gly Lys Asp Gly Lys Gin Tyr Glu Ser Val Leu 1 5 10 15 Met Val Ser Ile Asp Gin Leu Leu Asp Ser Met Lys Glu Ile Gly Ser 20 25 30 Asn Cys Leu Asn Asn Glu Phe Asn Phe Phe Lys Arg His Ile Cys Asp 35 40 45 Ala Asn Lys Glu Gly Met Phe Leu Phe Arg Ala Ala Arg Lys Leu Arg 50 55 60 Gln Phe Leu Lys Met Asn Ser Thr Gly Asp Phe Asp Leu His Leu Leu 65 70 75 80 Lys Val Ser Glu Gly Thr Thr Ile Leu Leu Asn Cys Thr Gly Gln Val 85 90 95 Lys Gly Arg Lys Pro Ala Ala Leu Gly Glu Ala Gln Pro Thr Lys Ser 100 105 110 Leu Glu Glu Asn Lys Ser Leu Lys Glu Gln Lys Lys Leu Asn Asp Leu 115 120 125 Cys Phe Leu Lys Arg Leu Leu Gln Glu Ile Lys Thr Cys Trp Asn Lys 130 135 140 Ile Leu Met Gly Thr Lys Glu His 145 150 <210> 26 <211> 129 <212> PRT <213> Mus musculus <400> 26 His Lys Ser Ser Pro Gln Gly Pro Asp Arg Leu Leu Ile Arg Leu Arg 1 5 10 15 His Leu Ile Asp Ile Val Glu Gin Leu Lys Ile Tyr Glu Asn Asp Leu 20 25 30 Asp Pro Glu Leu Leu Ser Ala Pro Gin Asp Val Lys Gly His Cys Glu 35 40 45 His Ala Ala Phe Ala Cys Phe Gin Lys Ala Lys Leu Lys Pro Ser Asn 50 55 60 Pro Gly Asn Asn Lys Thr Phe He He Asp Leu Val Ala Gin Leu Arg 65 70 75 80 Arg Arg Leu Pro Ala Arg Arg Gly Gly Lys Lys Gin Lys His He Ala 85 90 95 Lys Cys Pro Ser Cys Asp Ser Tyr Gin Lys Arg Thr Pro Lys Glu Phe 100 105 110 Leu Gin Arg Leu Lys Trp Leu Leu Gin Lys Met He His Gin His Leu 115 120 125 Ser <210> 27 <211> 133 <212> PRT <213> Homo sapiens <400> 27 Gln Gly Gin Asp Arg His Met He Arg Met Arg Gin Leu He Asp He 1 5 10 15 Val Asp Gin Leu Lys Asn Tyr Val Asn Asp Leu Val Pro Gin Phe Leu 20 25 30 Pro Ala Pro Glu Asp Val Glu Thr Asn Cys Glu Trp Ser Ala Phe Ser 35 40 45 Cys Phe Gin Lys Ala Gin Leu Lys Ser Ala Asn Thr Gly Asn Asn Glu 50 55 60 Arg He He Asn Val Ser He Lys Lys Leu Lys Arg Lys Pro Pro Ser 65 70 75 80 Thr Asn Ala Gly Arg Arg Gin Lys His Arg Leu Thr Cys Pro Ser Cys 85 90 95 Asp Ser Tyr Glu Lys Lys Pro Pro Lys Glu Phe Leu Glu Arg Phe Lys 100 105 110 Ser Leu Leu Gin Lys Met He His Gin His Leu Ser Ser Arg Thr His 115 120 125 Gly Ser Glu Asp Ser 130 <210> 28 <211> 206 <212> PRT <213> Mus musculus <400> 28 Gly Thr Pro Asp Cys Tyr Phe Ser His Ser Pro He Ser Ser Asn Phe 1 5 10 15 Lys Val Lys Phe Arg Glu Leu Thr Asp His Leu Leu Lys Asp Tyr Pro 20 25 30 Val Thr Val Ala Val Asn Leu Gin Asp Glu Lys His Cys Lys Ala Leu 35 40 45 Trp Ser Leu Phe Leu Ala Gin Arg Trp lie Glu Gin Leu Lys Thr Val 50 55 60 Ala Gly Ser Lys Met Gin Thr Leu Leu Glu Asp Val Asn Thr Glu lie 65 70 75 80 His Phe Val Thr Ser Cys Thr Phe Gin Pro Leu Pro Glu Cys Leu Arg 85 90 95 Phe Val Gin Thr Asn lie Ser His Leu Leu Lys Asp Thr Cys Thr Gin 100 105 110 Leu Leu Ala Leu Lys Pro Cys lie Gly Lys Ala Cys Gin Asn Phe Ser 115 120 125 Arg Cys Leu Glu Val Gin Cys Gin Pro Asp Ser Ser Thr Leu Leu Pro 130 135 140 Pro Arg Ser Pro lie Ala Leu Glu Ala Thr Glu Leu Pro Glu Pro Arg 145 150 155 160 Pro Arg Gin Leu Leu Leu Leu Leu Leu Leu Leu Leu Pro Leu Thr Leu 165 170 175 Val Leu Leu Ala Ala Ala Trp Gly Leu Arg Trp Gin Arg Ala Arg Arg 180 185 190 Arg Gly Glu Leu His Pro Gly Val Pro Leu Pro Ser His Pro 195 200 205 <210> 29 <211> 209 <212> PRT <213> Homo sapiens <400> 29 Thr Gin Asp Cys Ser Phe Gin His Ser Pro lie Ser Ser Asp Phe Ala 1 5 10 15 Val Lys lie Arg Glu Leu Ser Asp Tyr Leu Leu Gin Asp Tyr Pro Val 20 25 30 Thr Val Ala Ser Asn Leu Gin Asp Glu Glu Leu Cys Gly Gly Leu Trp 35 40 45 Arg Leu Val Leu Ala Gin Arg Trp Met Glu Arg Leu Lys Thr Val Ala 50 55 60 Gly Ser Lys Met Gin Gly Leu Leu Glu Arg Val Asn Thr Glu lie His 65 70 75 80 Phe Val Thr Lys Cys Ala Phe Gin Pro Pro Pro Ser Cys Leu Arg Phe 85 90 95 Val Gin Thr Asn lie Ser Arg Leu Leu Gin Glu Thr Ser Glu Gin Leu 100 105 110 Val Ala Leu Lys Pro Trp lie Thr Arg Gin Asn Phe Ser Arg Cys Leu 115 120 125 Glu Leu Gin Cys Gin Pro Asp Ser Ser Thr Leu Pro Pro Pro Trp Ser 130 135 140 Pro Arg Pro Leu Glu Ala Thr Ala Pro Thr Ala Pro Gln Pro Pro Leu 145 150 155 160 Leu Leu Leu Leu Leu Leu Pro Val Gly Leu Leu Leu Leu Leu Ala Ala Ala 165 170 175 Trp Cys Leu His Trp Gln Arg Thr Arg Arg Arg Thr Pro Arg Pro Gly 180 185 190 Glu Gln Val Pro Pro Val Pro Ser Pro Gln Asp Leu Leu Leu Val Glu 195 200 205 His <210> 30 <211> 124 <212> PRT <213> Mus musculus <400> 30 Ala Pro Thr Arg Ser Pro Ile Thr Val Thr Arg Pro Trp Lys His Val 1 5 10 15 Glu Ala Ile Lys Glu Ala Leu Asn Leu Leu Asp Asp Met Pro Val Thr 20 25 30 Leu Asn Glu Glu Val Glu Val Val Ser Asn Glu Phe Ser Phe Lys Lys 35 40 45 Leu Thr Cys Val Gln Thr Arg Leu Lys Ile Phe Glu Gln Gly Leu Arg 50 55 60 Gly Asn Phe Thr Lys Leu Lys Gly Ala Leu Asn Met Thr Ala Ser Tyr 65 70 75 80 Tyr Gin Thr Tyr Cys Pro Pro Thr Pro Glu Thr Asp Cys Glu Thr Gin 85 90 95 Val Thr Thr Tyr Ala Asp Phe He Asp Ser Leu Lys Thr Phe Leu Thr 100 105 110 Asp He Pro Phe Glu Cys Lys Lys Pro Gly Gin Lys 115 120 <210> 31 <211> 127 <212> PRT <213> Homo sapiens <400> 31 Ala Pro Ala Arg Ser Pro Ser Pro Ser Thr Gin Pro Trp Glu His Val 1 5 10 15 Asn Ala He Gin Glu Ala Arg Arg Leu Leu Asn Leu Ser Arg Asp Thr 20 25 30 Ala Ala Glu Met Asn Glu Thr Val Glu Val He Ser Glu Met Phe Asp 35 40 45 Leu Gin Glu Pro Thr Cys Leu Gin Thr Arg Leu Glu Leu Tyr Lys Gin 50 55 60 Gly Leu Arg Gly Ser Leu Thr Lys Leu Lys Gly Pro Leu Thr Met Met 65 70 75 80 Ala Ser His Tyr Lys Gin His Cys Pro Pro Thr Pro Glu Thr Ser Cys 85 90 95 Ala Thr Gin He He Thr Phe Glu Ser Phe Lys Glu Asn Leu Lys Asp 100 105 110 Phe Leu Leu Val He Pro Phe Asp Cys Trp Glu Pro Val Gin Glu 115 120 125 <210> 32 <211> 824 <212> PRT <213> Artificial Sequence <220> <223> mIL12b IL12a IL2 GMCSF <400> 32 Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr 1 5 10 15 Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Asp He Thr Trp Thr Ser Asp Gin Arg His Gly Val He Gly 35 40 45 Ser Gly Lys Thr Leu Thr He Thr Val Lys Glu Phe Leu Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asn Gly He Trp Ser Thr Glu He Leu Lys 85 90 95 Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser 100 105 110 Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu Lys 115 120 125 Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val Thr 130 135 140 Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln Arg 145 150 155 160 Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys Pro 165 170 175 Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala Leu Glu Ala Arg Gln 180 185 190 Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe Ile Arg Asp Ile 195 200 205 Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met Lys Pro Leu Lys Asn 210 215 220 Ser Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro 225 230 235 240 His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg Ile Gln Arg Lys Lys 245 250 255 Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gln Lys Gly Ala Phe 260 265 270 Leu Val Glu Lys Thr Ser Thr Glu Val Gln Cys Lys Gly Gly Asn Val 275 280 285 Cys Val Gln Ala Gln Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp 290 295 300 Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly Gly Gly Ser Gly Gly 305 310 315 320 Gly Gly Ser Gly Gly Gly Gly Ser Arg Val Ile Pro Val Ser Gly Pro 325 330 335 Ala Arg Cys Leu Ser Gln Ser Arg Asn Leu Leu Lys Thr Thr Asp Asp 340 345 350 Met Val Lys Thr Ala Arg Glu Lys Leu Lys His Tyr Ser Cys Thr Ala 355 360 365 Glu Asp Ile Asp His Glu Asp Ile Thr Arg Asp Gln Thr Ser Thr Leu 370 375 380 Lys Thr Cys Leu Pro Leu Glu Leu His Lys Asn Glu Ser Cys Leu Ala 385 390 395 400 Thr Arg Glu Thr Ser Ser Thr Thr Arg Gly Ser Cys Leu Pro Pro Gln 405 410 415 Lys Thr Ser Leu Met Met Thr Leu Cys Leu Gly Ser Ile Tyr Glu Asp 420 425 430 Leu Lys Met Tyr Gln Thr Glu Phe Gln Ala Ile Asn Ala Ala Leu Gln 435 440 445 Asn His Asn His Gln Gln Ile Ile Leu Asp Lys Gly Met Leu Val Ala 450 455 460 Ile Asp Glu Leu Met Gln Ser Leu Asn His Asn Gly Glu Thr Leu Arg 465 470 475 480 Gln Lys Pro Pro Val Gly Glu Ala Asp Pro Tyr Arg Val Lys Met Lys 485 490 495 Leu Cys Ile Leu Leu His Ala Phe Ser Thr Arg Val Val Thr Ile Asn 500 505 510 Arg Val Met Gly Tyr Leu Ser Ser Ala Gly Gly Gly Gly Ser Gly Gly 515 520 525 Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Ser 530 535 540 Ser Ser Thr Ala Glu Ala Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln 545 550 555 560 Gln Gln His Leu Glu Gln Leu Leu Met Asp Leu Gln Glu Leu Leu Ser 565 570 575 Arg Met Glu Asn Tyr Arg Asn Leu Lys Leu Pro Arg Met Leu Thr Phe 580 585 590 Lys Phe Tyr Leu Pro Lys Gln Ala Thr Glu Leu Lys Asp Leu Gln Cys 595 600 605 Leu Glu Asp Glu Leu Gly Pro Leu Arg His Val Leu Asp Leu Thr Gln 610 615 620 Ser Lys Ser Phe Gln Leu Glu Asp Ala Glu Asn Phe Ile Ser Asn Ile 625 630 635 640 Arg Val Thr Val Val Lys Leu Lys Gly Ser Asp Asn Thr Phe Glu Cys 645 650 655 Gln Phe Asp Asp Glu Ser Ala Thr Val Val Asp Phe Leu Arg Arg Trp 660 665 670 Ile Ala Phe Cys Gln Ser Ile Ile Ser Thr Ser Pro Gln Gly Gly Gly 675 680 685 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Arg 690 695 700 Ser Pro Ile Thr Val Thr Arg Pro Trp Lys His Val Glu Ala Ile Lys 705 710 715 720 Glu Ala Leu Asn Leu Leu Asp Asp Met Pro Val Thr Leu Asn Glu Glu 725 730 735 Val Glu Val Val Ser Asn Glu Phe Ser Phe Lys Lys Leu Thr Cys Val 740 745 750 Gln Thr Arg Leu Lys Ile Phe Glu Gin Gly Leu Arg Gly Asn Phe Thr 755 760 765 Lys Leu Lys Gly Ala Leu Asn Met Thr Ala Ser Tyr Tyr Gin Thr Tyr 770 775 780 Cys Pro Pro Thr Pro Glu Thr Asp Cys Glu Thr Gin Val Thr Thr Tyr 785 790 795 800 Ala Asp Phe He Asp Ser Leu Lys Thr Phe Leu Thr Asp He Pro Phe 805 810 815 Glu Cys Lys Lys Pro Gly Gin Lys 820 <210> 33 <211> 804 <212> PRT <213> Artificial Sequence <220> <223> mIL12b IL12a IL7 GMCSF <400> 33 Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr 1 5 10 15 Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg His Gly Val Ile Gly 35 40 45 Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu Phe Leu Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu Lys 85 90 95 Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser 100 105 110 Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu Lys 115 120 125 Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val Thr 130 135 140 Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln Arg 145 150 155 160 Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys Pro 165 170 175 Thr Ala Glu Glu Thr Leu Pro lie Glu Leu Ala Leu Glu Ala Arg Gin 180 185 190 Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe lie Arg Asp lie 195 200 205 Ile Lys Pro Asp Pro Pro Lys Asn Leu Gin Met Lys Pro Leu Lys Asn 210 215 220 Ser Gin Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro 225 230 235 240 His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg lie Gin Arg Lys Lys 245 250 255 Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gin Lys Gly Ala Phe 260 265 270 Leu Val Glu Lys Thr Ser Thr Glu Val Gin Cys Lys Gly Gly Asn Val 275 280 285 Cys Val Gin Ala Gin Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp 290 295 300 Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly Gly Gly Ser Gly Gly 305 310 315 320 Gly Gly Ser Gly Gly Gly Gly Ser Arg Val lie Pro Val Ser Gly Pro 325 330 335 Ala Arg Cys Leu Ser Gin Ser Arg Asn Leu Leu Lys Thr Thr Asp Asp 340 345 350 Met Val Lys Thr Ala Arg Gin Lys Leu Lys His Tyr Ser Cys Thr Ala 355 360 365 Glu Asp He Asp His Gin Asp He Thr Arg Asp Gin Thr Ser Thr Leu 370 375 380 Lys Thr Cys Leu Pro Leu Gin Leu His Lys Asn Gin Ser Cys Leu Ala 385 390 395 400 Thr Arg Gin Thr Ser Ser Thr Thr Arg Gly Ser Cys Leu Pro Pro Gin 405 410 415 Lys Thr Ser Leu Met Met Thr Leu Cys Leu Gin Ser He Tyr Gin Asp Gin 420 425 430 Leu Lys Met Tyr Gin Thr Gin Phe Gin Ala He Asn Ala Ala Leu Gin 435 440 445 Asn His Asn His Gin Gin He He Leu Asp Lys Gin Met Leu Val Ala 450 455 460 He Asp Gin Leu Met Gin Ser Leu Asn His Asn Gin Gin Thr Leu Arg 465 470 475 480 Gln Lys Pro Pro Val Gin Gin Ala Asp Pro Tyr Gin Val Gin Met Gin 485 490 495 Leu Cys Ile Leu Leu His Ala Phe Ser Thr Arg Val Val Thr Ile Asn 500 505 510 Arg Val Met Gly Tyr Leu Ser Ser Ala Gly Gly Gly Gly Ser Gly Gly 515 520 525 Gly Gly Ser Gly Gly Gly Gly Ser Glu Cys His Ile Lys Asp Lys Glu 530 535 540 Gly Lys Ala Tyr Glu Ser Val Leu Met Ile Ser Ile Asp Glu Leu Asp 545 550 555 560 Lys Met Thr Gly Thr Asp Ser Asn Cys Pro Asn Asn Glu Pro Asn Phe 565 570 575 Phe Arg Lys His Val Cys Asp Asp Thr Lys Glu Ala Ala Phe Leu Asn 580 585 590 Arg Ala Ala Arg Lys Leu Lys Gin Phe Leu Lys Met Asn Ile Ser Glu 595 600 605 Glu Phe Asn Val His Leu Leu Thr Val Ser Gin Gly Thr Gin Thr Leu 610 615 620 Val Asn Cys Thr Ser Lys Glu Glu Lys Asn Val Lys Glu Gin Lys Lys 625 630 635 640 Asn Asp Ala Cys Phe Leu Lys Arg Leu Leu Arg Glu Ile Lys Thr Cys 645 650 655 Trp Asn Lys Ile Leu Lys Gly Ser Ile Gly Gly Gly Gly Ser Gly Gly 660 665 670 Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Arg Ser Pro Ile Thr 675 680 685 Val Thr Arg Pro Trp Lys His Val Glu Ala Ile Lys Glu Ala Leu Asn 690 695 700 Leu Leu Asp Asp Met Pro Val Thr Leu Asn Glu Glu Val Glu Val Val 705 710 715 720 Ser Asn Glu Phe Ser Phe Lys Lys Leu Thr Cys Val Gln Thr Arg Leu 725 730 735 Lys Ile Phe Glu Gln Gly Leu Arg Gly Asn Phe Thr Lys Leu Lys Gly 740 745 750 Ala Leu Asn Met Thr Ala Ser Tyr Tyr Gln Thr Tyr Cys Pro Pro Thr 755 760 765 Pro Glu Thr Asp Cys Glu Thr Gln Val Thr Thr Tyr Ala Asp Phe Ile 770 775 780 Asp Ser Leu Lys Thr Phe Leu Thr Asp Ile Pro Phe Glu Cys Lys Lys 785 790 795 800 Pro Gly Gln Lys <210> 34 <211> 808 <212> PRT <213> Artificial Sequence <220> <223> mIL12bIL12aIL15GMCSF <400> 34 Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr 1 5 10 15 Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg His Gly Val Ile Gly 35 40 45 Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu Phe Leu Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu Lys 85 90 95 Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser 100 105 110 Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu Lys 115 120 125 Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val Thr 130 135 140 Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln Arg 145 150 155 160 Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys Pro 165 170 175 Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala Leu Glu Ala Arg Gln 180 185 190 Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe Ile Arg Asp Ile 195 200 205 Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met Lys Pro Leu Lys Asn 210 215 220 Ser Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro 225 230 235 240 His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg Ile Gln Arg Lys Lys 245 250 255 Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gln Lys Gly Ala Phe 260 265 270 Leu Val Glu Lys Thr Ser Thr Glu Val Gln Cys Lys Gly Gly Asn Val 275 280 285 Cys Val Gin Ala Gin Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp 290 295 300 Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly Gly Gly Ser Gly Gly 305 310 315 320 Gly Gly Ser Gly Gly Gly Gly Ser Arg Val He Pro Val Ser Gly Pro 325 330 335 Ala Arg Cys Leu Ser Gin Ser Arg Asn Leu Leu Lys Thr Thr Asp Asp 340 345 350 Met Val Lys Thr Ala Arg Glu Lys Leu Lys His Tyr Ser Cys Thr Ala 355 360 365 Glu Asp He Asp His Glu Asp He Thr Arg Asp Gin Thr Ser Thr Leu 370 375 380 Lys Thr Cys Leu Pro Leu Glu Leu His Lys Asn Glu Ser Cys Leu Ala 385 390 395 400 Thr Arg Glu Thr Ser Ser Thr Thr Arg Gly Ser Cys Leu Pro Pro Gin 405 410 415 Lys Thr Ser Leu Met Met Thr Leu Cys Leu Gly Ser He Tyr Glu Asp 420 425 430 Leu Lys Met Tyr Gin Thr Glu Phe Gin Ala He Asn Ala Ala Leu Gin 435 440 445 Asn His Asn His Gin Gin lie lie Leu Asp Lys Gly Met Leu Val Ala 450 455 460 Ile Asp Glu Leu Met Gin Ser Leu Asn His Asn Gly Glu Thr Leu Arg 465 470 475 480 Gln Lys Pro Pro Val Gly Glu Ala Asp Pro Tyr Arg Val Lys Met Lys 485 490 495 Leu Cys lie Leu Leu His Ala Phe Ser Thr Arg Val Val Thr lie Asn 500 505 510 Arg Val Met Gly Tyr Leu Ser Ser Ala Gly Gly Gly Gly Ser Gly Gly 515 520 525 Gly Gly Ser Gly Gly Gly Gly Ser Gly lie His Val Phe lie Leu Gly 530 535 540 Cys Val Ser Val Gly Leu Pro Lys Thr Glu Ala Asn Trp lie Asp Val 545 550 555 560 Arg Tyr Asp Leu Glu Lys lie Glu Ser Leu lie Gin Ser lie His lie 565 570 575 Asp Thr Thr Leu Tyr Thr Asp Ser Asp Phe His Pro Ser Cys Lys Val 580 585 590 Thr Ala Met Asn Cys Phe Leu Leu Glu Leu Gin Val lie Leu His Glu 595 600 605 Tyr Ser Asn Met Thr Leu Asn Glu Thr Val Arg Asn Val Leu Tyr Leu 610 615 620 Ala Asn Ser Thr Leu Ser Ser Asn Lys Asn Val Ala Glu Ser Gly Cys 625 630 635 640 Lys Glu Cys Glu Glu Leu Glu Glu Lys Thr Phe Thr Glu Phe Leu Gln 645 650 655 Ser Phe Ile Arg Ile Val Gln Met Phe Ile Asn Thr Ser Gly Gly Gly 660 665 670 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Arg 675 680 685 Ser Pro Ile Thr Val Thr Arg Pro Trp Lys His Val Glu Ala Ile Lys 690 695 700 Glu Ala Leu Asn Leu Leu Asp Asp Met Pro Val Thr Leu Asn Glu Glu 705 710 715 720 Val Glu Val Val Ser Asn Glu Phe Ser Phe Lys Lys Leu Thr Cys Val 725 730 735 Gln Thr Arg Leu Lys Ile Phe Glu Gln Gly Leu Arg Gly Asn Phe Thr 740 745 750 Lys Leu Lys Gly Ala Leu Asn Met Thr Ala Ser Tyr Tyr Gin Thr Tyr 755 760 765 Cys Pro Pro Thr Pro Glu Thr Asp Cys Glu Thr Gin Val Thr Thr Tyr 770 775 780 Ala Asp Phe He Asp Ser Leu Lys Thr Phe Leu Thr Asp He Pro Phe 785 790 795 800 Glu Cys Lys Lys Pro Gly Gin Lys 805 <210> 35 <211> 804 <212> PRT <213> Artificial Sequence <220> <223> mIL12b IL12a IL21 GMCSF <400> 35 Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr 1 5 10 15 Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Asp He Thr Trp Thr Ser Asp Gin Arg His Gly Val He Gly 35 40 45 Ser Gly Lys Thr Leu Thr He Thr Val Lys Glu Phe Leu Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu Lys 85 90 95 Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser 100 105 110 Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu Lys 115 120 125 Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val Thr 130 135 140 Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln Arg 145 150 155 160 Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys Pro 165 170 175 Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala Leu Glu Ala Arg Gln 180 185 190 Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe Ile Arg Asp Ile 195 200 205 Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met Lys Pro Leu Lys Asn 210 215 220 Ser Gin Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro 225 230 235 240 His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg Ile Gin Arg Lys Lys 245 250 255 Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gin Lys Gly Ala Phe 260 265 270 Leu Val Glu Lys Thr Ser Thr Glu Val Gin Cys Lys Gly Gly Asn Val 275 280 285 Cys Val Gin Ala Gin Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp 290 295 300 Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly Gly Gly Ser Gly Gly 305 310 315 320 Gly Gly Ser Gly Gly Gly Gly Ser Arg Val Ile Pro Val Ser Gly Pro 325 330 335 Ala Arg Cys Leu Ser Gin Ser Arg Asn Leu Leu Lys Thr Thr Asp Asp 340 345 350 Met Val Lys Thr Ala Arg Glu Lys Leu Lys His Tyr Ser Cys Thr Ala 355 360 365 Glu Asp Ile Asp His Glu Asp Ile Thr Arg Asp Gin Thr Ser Thr Leu 370 375 380 Lys Thr Cys Leu Pro Leu Glu Leu His Lys Asn Glu Ser Cys Leu Ala 385 390 395 400 Thr Arg Glu Thr Ser Ser Thr Thr Arg Gly Ser Cys Leu Pro Pro Gln 405 410 415 Lys Thr Ser Leu Met Met Thr Leu Cys Leu Gly Ser Ile Tyr Glu Asp 420 425 430 Leu Lys Met Tyr Gln Thr Glu Phe Gln Ala Ile Asn Ala Ala Leu Gln 435 440 445 Asn His Asn His Gln Gln Ile Ile Leu Asp Lys Gly Met Leu Val Ala 450 455 460 Ile Asp Glu Leu Met Gln Ser Leu Asn His Asn Gly Glu Thr Leu Arg 465 470 475 480 Gln Lys Pro Pro Val Gly Glu Ala Asp Pro Tyr Arg Val Lys Met Lys 485 490 495 Leu Cys Ile Leu Leu His Ala Phe Ser Thr Arg Val Val Thr Ile Asn 500 505 510 Arg Val Met Gly Tyr Leu Ser Ser Ala Gly Gly Gly Gly Ser Gly Gly 515 520 525 Gly Gly Ser Gly Gly Gly Gly Ser His Lys Ser Ser Pro Gln Gly Pro 530 535 540 Asp Arg Leu Leu Ile Arg Leu Arg His Leu Ile Asp Ile Val Glu Gin 545 550 555 560 Leu Lys Ile Tyr Glu Asn Asp Leu Asp Pro Glu Leu Leu Ser Ala Pro 565 570 575 Gln Asp Val Lys Gly His Cys Glu His Ala Ala Phe Ala Cys Phe Gin 580 585 590 Lys Ala Lys Leu Lys Pro Ser Asn Pro Gly Asn Asn Lys Thr Phe Ile 595 600 605 Ile Asp Leu Val Ala Gin Leu Arg Arg Arg Leu Pro Ala Arg Arg Gly 610 615 620 Gly Lys Lys Gin Lys His Ile Ala Lys Cys Pro Ser Cys Asp Ser Tyr 625 630 635 640 Glu Lys Arg Thr Pro Lys Glu Phe Leu Glu Arg Leu Lys Trp Leu Leu 645 650 655 Gln Lys Met Ile His Gin His Leu Ser Gly Gly Gly Gly Ser Gly Gly 660 665 670 Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Arg Ser Pro Ile Thr 675 680 685 Val Thr Arg Pro Trp Lys His Val Glu Ala Ile Lys Glu Ala Leu Asn 690 695 700 Leu Leu Asp Asp Met Pro Val Thr Leu Asn Glu Glu Val Glu Val Val 705 710 715 720 Ser Asn Glu Phe Ser Phe Lys Lys Leu Thr Cys Val Gln Thr Arg Leu 725 730 735 Lys Ile Phe Glu Gln Gly Leu Arg Gly Asn Phe Thr Lys Leu Lys Gly 740 745 750 Ala Leu Asn Met Thr Ala Ser Tyr Tyr Gln Thr Tyr Cys Pro Pro Thr 755 760 765 Pro Glu Thr Asp Cys Glu Thr Gln Val Thr Thr Tyr Ala Asp Phe Ile 770 775 780 Asp Ser Leu Lys Thr Phe Leu Thr Asp Ile Pro Phe Glu Cys Lys Lys 785 790 795 800 Pro Gly Gln Lys <210> 36 <211> 906 <212> PRT <213> Artificial Sequence <220> <223> mIL12bIL12aIL2 FLT3L <400> 36 Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr 1 5 10 15 Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg His Gly Val Ile Gly 35 40 45 Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu Phe Leu Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu Lys 85 90 95 Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser 100 105 110 Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu Lys 115 120 125 Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val Thr 130 135 140 Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln Arg 145 150 155 160 Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys Pro 165 170 175 Thr Ala Glu Glu Thr Leu Pro lie Glu Leu Ala Leu Glu Ala Arg Gin 180 185 190 Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe lie Arg Asp lie 195 200 205 Ile Lys Pro Asp Pro Pro Lys Asn Leu Gin Met Lys Pro Leu Lys Asn 210 215 220 Ser Gin Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro 225 230 235 240 His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg lie Gin Arg Lys Lys 245 250 255 Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gin Lys Gly Ala Phe 260 265 270 Leu Val Glu Lys Thr Ser Thr Glu Val Gin Cys Lys Gly Gly Asn Val 275 280 285 Cys Val Gin Ala Gin Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp 290 295 300 Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly Gly Gly Ser Gly Gly 305 310 315 320 Gly Gly Ser Gly Gly Gly Gly Ser Arg Val lie Pro Val Ser Gly Pro 325 330 335 Ala Arg Cys Leu Ser Gin Ser Arg Asn Leu Leu Lys Thr Thr Asp Asp 340 345 350 Met Val Lys Thr Ala Arg Glu Lys Leu Lys His Tyr Ser Cys Thr Ala 355 360 365 Glu Asp He Asp His Glu Asp He Thr Arg Asp Gin Thr Ser Thr Leu 370 375 380 Lys Thr Cys Leu Pro Leu Glu Leu His Lys Asn Glu Ser Cys Leu Ala 385 390 395 400 Thr Arg Glu Thr Ser Ser Thr Thr Arg Gly Ser Cys Leu Pro Pro Gin 405 410 415 Lys Thr Ser Leu Met Met Thr Leu Cys Leu Gly Ser He Tyr Glu Asp 420 425 430 Leu Lys Met Tyr Gin Thr Glu Phe Gin Ala He Asn Ala Ala Leu Gin 435 440 445 Asn His Asn His Gin Gin He He Leu Asp Lys Gly Met Leu Val Ala 450 455 460 He Asp Glu Leu Met Gin Ser Leu Asn His Asn Gly Glu Thr Leu Arg 465 470 475 480 Gln Lys Pro Pro Val Gly Glu Ala Asp Pro Tyr Arg Val Lys Met Lys 485 490 495 Leu Cys Ile Leu Leu His Ala Phe Ser Thr Arg Val Val Thr Ile Asn 500 505 510 Arg Val Met Gly Tyr Leu Ser Ser Ala Gly Gly Gly Gly Ser Gly Gly 515 520 525 Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Ser 530 535 540 Ser Ser Thr Ala Glu Ala Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln 545 550 555 560 Gln Gln His Leu Glu Gln Leu Leu Met Asp Leu Gln Glu Leu Leu Ser 565 570 575 Arg Met Glu Asn Tyr Arg Asn Leu Lys Leu Pro Arg Met Leu Thr Phe 580 585 590 Lys Phe Tyr Leu Pro Lys Gln Ala Thr Glu Leu Lys Asp Leu Gln Cys 595 600 605 Leu Glu Asp Glu Leu Gly Pro Leu Arg His Val Leu Asp Leu Thr Gln 610 615 620 Ser Lys Ser Phe Gln Leu Glu Asp Ala Glu Asn Phe Ile Ser Asn Ile 625 630 635 640 Arg Val Thr Val Val Lys Leu Lys Gly Ser Asp Asn Thr Phe Glu Cys 645 650 655 Gln Phe Asp Asp Glu Ser Ala Thr Val Val Asp Phe Leu Arg Arg Trp 660 665 670 Ile Ala Phe Cys Gln Ser Ile Ile Ser Thr Ser Pro Gln Gly Gly Gly 675 680 685 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Thr Pro Asp 690 695 700 Cys Tyr Phe Ser His Ser Pro Ile Ser Ser Asn Phe Lys Val Lys Phe 705 710 715 720 Arg Glu Leu Thr Asp His Leu Leu Lys Asp Tyr Pro Val Thr Val Ala 725 730 735 Val Asn Leu Gln Asp Glu Lys His Cys Lys Ala Leu Trp Ser Leu Phe 740 745 750 Leu Ala Gln Arg Trp Ile Glu Gln Leu Lys Thr Val Ala Gly Ser Lys 755 760 765 Met Gln Thr Leu Leu Glu Asp Val Asn Thr Glu Ile His Phe Val Thr 770 775 780 Ser Cys Thr Phe Gln Pro Leu Pro Glu Cys Leu Arg Phe Val Gln Thr 785 790 795 800 Asn Ile Ser His Leu Leu Lys Asp Thr Cys Thr Gin Leu Leu Ala Leu 805 810 815 Lys Pro Cys Ile Gly Lys Ala Cys Gin Asn Phe Ser Arg Cys Leu Glu 820 825 830 Val Gin Cys Gin Pro Asp Ser Ser Thr Leu Leu Pro Pro Arg Ser Pro 835 840 845 Ile Ala Leu Glu Ala Thr Glu Leu Pro Glu Pro Arg Pro Arg Gin Leu 850 855 860 Leu Leu Leu Leu Leu Leu Leu Leu Pro Leu Thr Leu Val Leu Leu Ala 865 870 875 880 Ala Ala Trp Gly Leu Arg Trp Gin Arg Ala Arg Arg Arg Gly Glu Leu 885 890 895 His Pro Gly Val Pro Leu Pro Ser His Pro 900 905 <210> 37 <211> 886 <212> PRT <213> Artificial Sequence <220> <223> mIL12bIL12aIL7 FLT3L <400> 37 Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr 1 5 10 15 Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg His Gly Val Ile Gly 35 40 45 Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu Phe Leu Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu Lys 85 90 95 Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser 100 105 110 Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu Lys 115 120 125 Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val Thr 130 135 140 Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln Arg 145 150 155 160 Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys Pro 165 170 175 Thr Ala Glu Glu Thr Leu Pro lie Glu Leu Ala Leu Glu Ala Arg Gin 180 185 190 Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe lie Arg Asp lie 195 200 205 Ile Lys Pro Asp Pro Pro Lys Asn Leu Gin Met Lys Pro Leu Lys Asn 210 215 220 Ser Gin Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro 225 230 235 240 His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg lie Gin Arg Lys Lys 245 250 255 Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gin Lys Gly Ala Phe 260 265 270 Leu Val Glu Lys Thr Ser Thr Glu Val Gin Cys Lys Gly Gly Asn Val 275 280 285 Cys Val Gin Ala Gin Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp 290 295 300 Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly Gly Gly Ser Gly Gly 305 310 315 320 Gly Gly Ser Gly Gly Gly Gly Ser Arg Val lie Pro Val Ser Gly Pro 325 330 335 Ala Arg Cys Leu Ser Gin Ser Arg Asn Leu Leu Lys Thr Thr Asp Asp 340 345 350 Met Val Lys Thr Ala Arg Glu Lys Leu Lys His Tyr Ser Cys Thr Ala 355 360 365 Glu Asp He Asp His Glu Asp He Thr Arg Asp Gin Thr Ser Thr Leu 370 375 380 Lys Thr Cys Leu Pro Leu Glu Leu His Lys Asn Glu Ser Cys Leu Ala 385 390 395 400 Thr Arg Glu Thr Ser Ser Thr Thr Arg Gly Ser Cys Leu Pro Pro Gin 405 410 415 Lys Thr Ser Leu Met Met Thr Leu Cys Leu Gly Ser He Tyr Glu Asp 420 425 430 Leu Lys Met Tyr Gin Thr Glu Phe Gin Ala He Asn Ala Ala Leu Gin 435 440 445 Asn His Asn His Gin Gin He He Leu Asp Lys Gly Met Leu Val Ala 450 455 460 He Asp Glu Leu Met Gin Ser Leu Asn His Asn Gly Glu Thr Leu Arg 465 470 475 480 Gln Lys Pro Pro Val Gly Glu Ala Asp Pro Tyr Arg Val Lys Met Lys 485 490 495 Leu Cys Ile Leu Leu His Ala Phe Ser Thr Arg Val Val Thr Ile Asn 500 505 510 Arg Val Met Gly Tyr Leu Ser Ser Ala Gly Gly Gly Gly Ser Gly Gly 515 520 525 Gly Gly Ser Gly Gly Gly Gly Ser Glu Cys His Ile Lys Asp Lys Glu 530 535 540 Gly Lys Ala Tyr Glu Ser Val Leu Met Ile Ser Ile Asp Glu Leu Asp 545 550 555 560 Lys Met Thr Gly Thr Asp Ser Asn Cys Pro Asn Asn Glu Pro Asn Phe 565 570 575 Phe Arg Lys His Val Cys Asp Asp Thr Lys Glu Ala Ala Phe Leu Asn 580 585 590 Arg Ala Ala Arg Lys Leu Lys Gin Phe Leu Lys Met Asn Ile Ser Glu 595 600 605 Glu Phe Asn Val His Leu Leu Thr Val Ser Gin Gly Thr Gin Thr Leu 610 615 620 Val Asn Cys Thr Ser Lys Glu Glu Lys Asn Val Lys Glu Gin Lys Lys 625 630 635 640 Asn Asp Ala Cys Phe Leu Lys Arg Leu Leu Arg Glu Ile Lys Thr Cys 645 650 655 Trp Asn Lys Ile Leu Lys Gly Ser Ile Gly Gly Gly Gly Ser Gly Gly 660 665 670 Gly Gly Ser Gly Gly Gly Gly Ser Gly Thr Pro Asp Cys Tyr Phe Ser 675 680 685 His Ser Pro Ile Ser Ser Asn Phe Lys Val Lys Phe Arg Glu Leu Thr 690 695 700 Asp His Leu Leu Lys Asp Tyr Pro Val Thr Val Ala Val Asn Leu Gln 705 710 715 720 Asp Glu Lys His Cys Lys Ala Leu Trp Ser Leu Phe Leu Ala Gln Arg 725 730 735 Trp Ile Glu Gln Leu Lys Thr Val Ala Gly Ser Lys Met Gln Thr Leu 740 745 750 Leu Glu Asp Val Asn Thr Glu Ile His Phe Val Thr Ser Cys Thr Phe 755 760 765 Gln Pro Leu Pro Glu Cys Leu Arg Phe Val Gln Thr Asn Ile Ser His 770 775 780 Leu Leu Lys Asp Thr Cys Thr Gln Leu Leu Ala Leu Lys Pro Cys Ile 785 790 795 800 Gly Lys Ala Cys Gln Asn Phe Ser Arg Cys Leu Glu Val Gln Cys Gln 805 810 815 Pro Asp Ser Ser Thr Leu Leu Pro Pro Arg Ser Pro Ile Ala Leu Glu 820 825 830 Ala Thr Glu Leu Pro Glu Pro Arg Pro Arg Gln Leu Leu Leu Leu Leu 835 840 845 Leu Leu Leu Leu Pro Leu Thr Leu Val Leu Leu Ala Ala Ala Trp Gly 850 855 860 Leu Arg Trp Gln Arg Ala Arg Arg Arg Gly Glu Leu His Pro Gly Val 865 870 875 880 Pro Leu Pro Ser His Pro 885 <210> 38 <211> 890 <212> PRT <213> Artificial Sequence <220> <223> mIL12bIL12aIL15FLT3L <400> 38 Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr 1 5 10 15 Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg His Gly Val Ile Gly 35 40 45 Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu Phe Leu Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu Lys 85 90 95 Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser 100 105 110 Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu Lys 115 120 125 Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val Thr 130 135 140 Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln Arg 145 150 155 160 Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys Pro 165 170 175 Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala Leu Glu Ala Arg Gln 180 185 190 Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe Ile Arg Asp Ile 195 200 205 Ile Lys Pro Asp Pro Pro Lys Asn Leu Gin Met Lys Pro Leu Lys Asn 210 215 220 Ser Gin Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro 225 230 235 240 His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg Ile Gin Arg Lys Lys 245 250 255 Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gin Lys Gly Ala Phe 260 265 270 Leu Val Glu Lys Thr Ser Thr Glu Val Gin Cys Lys Gly Gly Asn Val 275 280 285 Cys Val Gin Ala Gin Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp 290 295 300 Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly Gly Gly Ser Gly Gly 305 310 315 320 Gly Gly Ser Gly Gly Gly Gly Ser Arg Val Ile Pro Val Ser Gly Pro 325 330 335 Ala Arg Cys Leu Ser Gin Ser Arg Asn Leu Leu Lys Thr Thr Asp Asp 340 345 350 Met Val Lys Thr Ala Arg Glu Lys Leu Lys His Tyr Ser Cys Thr Ala 355 360 365 Glu Asp lie Asp His Glu Asp lie Thr Arg Asp Gin Thr Ser Thr Leu 370 375 380 Lys Thr Cys Leu Pro Leu Glu Leu His Lys Asn Glu Ser Cys Leu Ala 385 390 395 400 Thr Arg Glu Thr Ser Ser Thr Thr Arg Gly Ser Cys Leu Pro Pro Gin 405 410 415 Lys Thr Ser Leu Met Met Thr Leu Cys Leu Gly Ser lie Tyr Glu Asp 420 425 430 Leu Lys Met Tyr Gin Thr Glu Phe Gin Ala lie Asn Ala Ala Leu Gin 435 440 445 Asn His Asn His Gin Gin lie lie Leu Asp Lys Gly Met Leu Val Ala 450 455 460 lie Asp Glu Leu Met Gin Ser Leu Asn His Asn Gly Glu Thr Leu Arg 465 470 475 480 Gln Lys Pro Pro Val Gly Glu Ala Asp Pro Tyr Arg Val Lys Met Lys 485 490 495 Leu Cys lie Leu Leu His Ala Phe Ser Thr Arg Val Val Thr lie Asn 500 505 510 Arg Val Met Gly Tyr Leu Ser Ser Ala Gly Gly Gly Gly Ser Gly Gly 515 520 525 Gly Gly Ser Gly Gly Gly Gly Ser Gly Ile His Val Phe Ile Leu Gly 530 535 540 Cys Val Ser Val Gly Leu Pro Lys Thr Glu Ala Asn Trp Ile Asp Val 545 550 555 560 Arg Tyr Asp Leu Glu Lys Ile Glu Ser Leu Ile Gln Ser Ile His Ile 565 570 575 Asp Thr Thr Leu Tyr Thr Asp Ser Asp Phe His Pro Ser Cys Lys Val 580 585 590 Thr Ala Met Asn Cys Phe Leu Leu Glu Leu Gln Val Ile Leu His Glu 595 600 605 Tyr Ser Asn Met Thr Leu Asn Glu Thr Val Arg Asn Val Leu Tyr Leu 610 615 620 Ala Asn Ser Thr Leu Ser Ser Asn Lys Asn Val Ala Glu Ser Gly Cys 625 630 635 640 Lys Glu Cys Glu Glu Leu Glu Glu Lys Thr Phe Thr Glu Phe Leu Gln 645 650 655 Ser Phe Ile Arg Ile Val Gln Met Phe Ile Asn Thr Ser Gly Gly Gly 660 665 670 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Thr Pro Asp 675 680 685 Cys Tyr Phe Ser His Ser Pro Ile Ser Ser Asn Phe Lys Val Lys Phe 690 695 700 Arg Glu Leu Thr Asp His Leu Leu Lys Asp Tyr Pro Val Thr Val Ala 705 710 715 720 Val Asn Leu Gln Asp Glu Lys His Cys Lys Ala Leu Trp Ser Leu Phe 725 730 735 Leu Ala Gln Arg Trp Ile Glu Gln Leu Lys Thr Val Ala Gly Ser Lys 740 745 750 Met Gln Thr Leu Leu Glu Asp Val Asn Thr Glu Ile His Phe Val Thr 755 760 765 Ser Cys Thr Phe Gln Pro Leu Pro Glu Cys Leu Arg Phe Val Gln Thr 770 775 780 Asn Ile Ser His Leu Leu Lys Asp Thr Cys Thr Gln Leu Leu Ala Leu 785 790 795 800 Lys Pro Cys Ile Gly Lys Ala Cys Gln Asn Phe Ser Arg Cys Leu Glu 805 810 815 Val Gln Cys Gln Pro Asp Ser Ser Thr Leu Leu Pro Pro Arg Ser Pro 820 825 830 Ile Ala Leu Glu Ala Thr Glu Leu Pro Glu Pro Arg Pro Arg Gin Leu 835 840 845 Leu Leu Leu Leu Leu Leu Leu Leu Pro Leu Thr Leu Val Leu Leu Ala 850 855 860 Ala Ala Trp Gly Leu Arg Trp Gin Arg Ala Arg Arg Arg Gly Glu Leu 865 870 875 880 His Pro Gly Val Pro Leu Pro Ser His Pro 885 890 <210> 39 <211> 886 <212> PRT <213> Artificial Sequence <220> <223> mIL12bIL12aIL21 FLT3L <400> 39 Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr 1 5 10 15 Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Asp Ile Thr Trp Thr Ser Asp Gin Arg His Gly Val Ile Gly 35 40 45 Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu Phe Leu Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu Lys 85 90 95 Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser 100 105 110 Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu Lys 115 120 125 Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val Thr 130 135 140 Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln Arg 145 150 155 160 Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys Pro 165 170 175 Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala Leu Glu Ala Arg Gln 180 185 190 Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe Ile Arg Asp Ile 195 200 205 Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met Lys Pro Leu Lys Asn 210 215 220 Ser Gin Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro 225 230 235 240 His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg Ile Gin Arg Lys Lys 245 250 255 Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gin Lys Gly Ala Phe 260 265 270 Leu Val Glu Lys Thr Ser Thr Glu Val Gin Cys Lys Gly Gly Asn Val 275 280 285 Cys Val Gin Ala Gin Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp 290 295 300 Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly Gly Gly Ser Gly Gly 305 310 315 320 Gly Gly Ser Gly Gly Gly Gly Ser Arg Val Ile Pro Val Ser Gly Pro 325 330 335 Ala Arg Cys Leu Ser Gin Ser Arg Asn Leu Leu Lys Thr Thr Asp Asp 340 345 350 Met Val Lys Thr Ala Arg Glu Lys Leu Lys His Tyr Ser Cys Thr Ala 355 360 365 Glu Asp Ile Asp His Glu Asp Ile Thr Arg Asp Gin Thr Ser Thr Leu 370 375 380 Lys Thr Cys Leu Pro Leu Glu Leu His Lys Asn Glu Ser Cys Leu Ala 385 390 395 400 Thr Arg Glu Thr Ser Ser Thr Thr Arg Gly Ser Cys Leu Pro Pro Gln 405 410 415 Lys Thr Ser Leu Met Met Thr Leu Cys Leu Gly Ser Ile Tyr Glu Asp 420 425 430 Leu Lys Met Tyr Gln Thr Glu Phe Gln Ala Ile Asn Ala Ala Leu Gln 435 440 445 Asn His Asn His Gln Gln Ile Ile Leu Asp Lys Gly Met Leu Val Ala 450 455 460 Ile Asp Glu Leu Met Gln Ser Leu Asn His Asn Gly Glu Thr Leu Arg 465 470 475 480 Gln Lys Pro Pro Val Gly Glu Ala Asp Pro Tyr Arg Val Lys Met Lys 485 490 495 Leu Cys Ile Leu Leu His Ala Phe Ser Thr Arg Val Val Thr Ile Asn 500 505 510 Arg Val Met Gly Tyr Leu Ser Ser Ala Gly Gly Gly Gly Ser Gly Gly 515 520 525 Gly Gly Ser Gly Gly Gly Gly Ser His Lys Ser Ser Pro Gln Gly Pro 530 535 540 Asp Arg Leu Leu Ile Arg Leu Arg His Leu Ile Asp Ile Val Glu Gin 545 550 555 560 Leu Lys Ile Tyr Glu Asn Asp Leu Asp Pro Glu Leu Leu Ser Ala Pro 565 570 575 Gln Asp Val Lys Gly His Cys Glu His Ala Ala Phe Ala Cys Phe Gin 580 585 590 Lys Ala Lys Leu Lys Pro Ser Asn Pro Gly Asn Asn Lys Thr Phe Ile 595 600 605 Ile Asp Leu Val Ala Gin Leu Arg Arg Arg Leu Pro Ala Arg Arg Gly 610 615 620 Gly Lys Lys Gin Lys His Ile Ala Lys Cys Pro Ser Cys Asp Ser Tyr 625 630 635 640 Glu Lys Arg Thr Pro Lys Glu Phe Leu Glu Arg Leu Lys Trp Leu Leu 645 650 655 Gln Lys Met Ile His Gin His Leu Ser Gly Gly Gly Gly Ser Gly Gly 660 665 670 Gly Gly Ser Gly Gly Gly Gly Ser Gly Thr Pro Asp Cys Tyr Phe Ser 675 680 685 His Ser Pro Ile Ser Ser Asn Phe Lys Val Lys Phe Arg Glu Leu Thr 690 695 700 Asp His Leu Leu Lys Asp Tyr Pro Val Thr Val Ala Val Asn Leu Gln 705 710 715 720 Asp Glu Lys His Cys Lys Ala Leu Trp Ser Leu Phe Leu Ala Gln Arg 725 730 735 Trp Ile Glu Gln Leu Lys Thr Val Ala Gly Ser Lys Met Gln Thr Leu 740 745 750 Leu Glu Asp Val Asn Thr Glu Ile His Phe Val Thr Ser Cys Thr Phe 755 760 765 Gln Pro Leu Pro Glu Cys Leu Arg Phe Val Gln Thr Asn Ile Ser His 770 775 780 Leu Leu Lys Asp Thr Cys Thr Gln Leu Leu Ala Leu Lys Pro Cys Ile 785 790 795 800 Gly Lys Ala Cys Gln Asn Phe Ser Arg Cys Leu Glu Val Gln Cys Gln 805 810 815 Pro Asp Ser Ser Thr Leu Leu Pro Pro Arg Ser Pro Ile Ala Leu Glu 820 825 830 Ala Thr Glu Leu Pro Glu Pro Arg Pro Arg Gln Leu Leu Leu Leu Leu 835 840 845 Leu Leu Leu Leu Pro Leu Thr Leu Val Leu Leu Ala Ala Ala Trp Gly 850 855 860 Leu Arg Trp Gin Arg Ala Arg Arg Arg Gly Glu Leu His Pro Gly Val 865 870 875 880 Pro Leu Pro Ser His Pro 885 <210> 40 <211> 808 <212> PRT <213> Artificial Sequence <220> <223> hIL12 bIL12 aIL2 GMCSF <400> 40 Ile Trp Glu Leu Lys Lys Asp Val Tyr Val Val Glu Leu Asp Trp Tyr 1 5 10 15 Pro Asp Ala Pro Gly Glu Met Val Val Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Gly Ile Thr Trp Thr Leu Asp Gin Ser Ser Glu Val Leu Gly 35 40 45 Ser Gly Lys Thr Leu Thr Ile Gin Val Lys Glu Phe Gly Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Val Leu Ser His Ser Leu Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asp Gly Ile Trp Ser Thr Asp Ile Leu Lys 85 90 95 Asp Gln Lys Glu Pro Lys Asn Lys Thr Phe Leu Arg Cys Glu Ala Lys 100 105 110 Asn Tyr Ser Gly Arg Phe Thr Cys Trp Trp Leu Thr Thr Ile Ser Thr 115 120 125 Asp Leu Thr Phe Ser Val Lys Ser Ser Arg Gly Ser Ser Asp Pro Gln 130 135 140 Gly Val Thr Cys Gly Ala Ala Thr Leu Ser Ala Glu Arg Val Arg Gly 145 150 155 160 Asp Asn Lys Glu Tyr Glu Tyr Ser Val Glu Cys Gln Glu Asp Ser Ala 165 170 175 Cys Pro Ala Ala Glu Glu Ser Leu Pro Ile Glu Val Met Val Asp Ala 180 185 190 Val His Lys Leu Lys Tyr Glu Asn Tyr Thr Ser Ser Phe Phe Ile Arg 195 200 205 Asp Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Leu Lys Pro Leu 210 215 220 Lys Asn Ser Arg Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Thr Trp 225 230 235 240 Ser Thr Pro His Ser Tyr Phe Ser Leu Thr Phe Cys Val Gln Val Gln 245 250 255 Gly Lys Ser Lys Arg Glu Lys Lys Asp Arg Val Phe Thr Asp Lys Thr 260 265 270 Ser Ala Thr Val Ile Cys Arg Lys Asn Ala Ser Ile Ser Val Arg Ala 275 280 285 Gln Asp Arg Tyr Tyr Ser Ser Ser Trp Ser Glu Trp Ala Ser Val Pro 290 295 300 Cys Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 305 310 315 320 Ser Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys 325 330 335 Leu His His Ser Gln Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gln 340 345 350 Lys Ala Arg Gln Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu Ile 355 360 365 Asp His Glu Asp Ile Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys 370 375 380 Leu Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu 385 390 395 400 Thr Ser Phe Ile Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser 405 410 415 Phe Met Met Ala Leu Cys Leu Ser Ser Ile Tyr Glu Asp Leu Lys Met 420 425 430 Tyr Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro 435 440 445 Lys Arg Gln Ile Phe Leu Asp Gln Asn Met Leu Ala Val Ile Asp Glu 450 455 460 Leu Met Gln Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gln Lys Ser 465 470 475 480 Ser Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys Ile Lys Leu Cys Ile 485 490 495 Leu Leu His Ala Phe Arg Ile Arg Ala Val Thr Ile Asp Arg Val Met 500 505 510 Ser Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 515 520 525 Gly Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln 530 535 540 Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly 545 550 555 560 Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys 565 570 575 Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu 580 585 590 Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser 595 600 605 Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val 610 615 620 Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr 625 630 635 640 Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr 645 650 655 Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Gly Gly Gly Gly Ser Gly 660 665 670 Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Ala Arg Ser Pro Ser 675 680 685 Pro Ser Thr Gln Pro Trp Glu His Val Asn Ala Ile Gln Glu Ala Arg 690 695 700 Arg Leu Leu Asn Leu Ser Arg Asp Thr Ala Ala Glu Met Asn Glu Thr 705 710 715 720 Val Glu Val Ile Ser Glu Met Phe Asp Leu Gln Glu Pro Thr Cys Leu 725 730 735 Gln Thr Arg Leu Glu Leu Tyr Lys Gln Gly Leu Arg Gly Ser Leu Thr 740 745 750 Lys Leu Lys Gly Pro Leu Thr Met Met Ala Ser His Tyr Lys Gln His 755 760 765 Cys Pro Pro Thr Pro Glu Thr Ser Cys Ala Thr Gln Ile Ile Thr Phe 770 775 780 Glu Ser Phe Lys Glu Asn Leu Lys Asp Phe Leu Leu Val Ile Pro Phe 785 790 795 800 Asp Cys Trp Glu Pro Val Gln Glu 805 <210> 41 <211> 827 <212> PRT <213> Artificial Sequence <220> <223> hIL12bIL12aIL7 GMCSF <400> 41 Ile Trp Glu Leu Lys Lys Asp Val Tyr Val Val Glu Leu Asp Trp Tyr 1 5 10 15 Pro Asp Ala Pro Gly Glu Met Val Val Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Gly lie Thr Trp Thr Leu Asp Gin Ser Ser Glu Val Leu Gly 35 40 45 Ser Gly Lys Thr Leu Thr lie Gin Val Lys Glu Phe Gly Asp Ala Gly 50 55 60 Gln Tyr Thr Cy s His Lys Gly Gly Glu Val Leu Ser His Ser Leu Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asp Gly lie Trp Ser Thr Asp lie Leu Lys 85 90 95 Asp Gin Lys Glu Pro Lys Asn Lys Thr Phe Leu Arg Cys Glu Ala Lys 100 105 110 Asn Tyr Ser Gly Arg Phe Thr Cy s Trp Trp Leu Thr Thr lie Ser Thr 115 120 125 Asp Leu Thr Phe Ser Val Lys Ser Ser Arg Gly Ser Ser Asp Pro Gin 130 135 140 Gly Val Thr Cy s Gly Ala Ala Thr Leu Ser Ala Glu Arg Val Arg Gly 145 150 155 160 Asp Asn Lys Glu Tyr Glu Tyr Ser Val Glu Cys Gin Glu Asp Ser Ala 165 170 175 Cys Pro Ala Ala Glu Glu Ser Leu Pro lie Glu Val Met Val Asp Ala 180 185 190 Val His Lys Leu Lys Tyr Glu Asn Tyr Thr Ser Ser Phe Phe Ile Arg 195 200 205 Asp Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Leu Lys Pro Leu 210 215 220 Lys Asn Ser Arg Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Thr Trp 225 230 235 240 Ser Thr Pro His Ser Tyr Phe Ser Leu Thr Phe Cys Val Gln Val Gln 245 250 255 Gly Lys Ser Lys Arg Glu Lys Lys Asp Arg Val Phe Thr Asp Lys Thr 260 265 270 Ser Ala Thr Val Ile Cys Arg Lys Asn Ala Ser Ile Ser Val Arg Ala 275 280 285 Gln Asp Arg Tyr Tyr Ser Ser Ser Trp Ser Glu Trp Ala Ser Val Pro 290 295 300 Cys Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 305 310 315 320 Ser Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys 325 330 335 Leu His His Ser Gln Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gln 340 345 350 Lys Ala Arg Gin Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu lie 355 360 365 Asp His Glu Asp He Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys 370 375 380 Leu Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu 385 390 395 400 Thr Ser Phe He Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser 405 410 415 Phe Met Met Ala Leu Cys Leu Ser Ser He Tyr Glu Asp Leu Lys Met 420 425 430 Tyr Gin Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro 435 440 445 Lys Arg Gin He Phe Leu Asp Gin Asn Met Leu Ala Val He Asp Glu 450 455 460 Leu Met Gin Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gin Lys Ser 465 470 475 480 Ser Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys He Lys Leu Cys He 485 490 495 Leu Leu His Ala Phe Arg He Arg Ala Val Thr He Asp Arg Val Met 500 505 510 Ser Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 515 520 525 Gly Gly Gly Gly Ser Asp Cys Asp Ile Glu Gly Lys Asp Gly Lys Gln 530 535 540 Tyr Glu Ser Val Leu Met Val Ser Ile Asp Gln Leu Leu Asp Ser Met 545 550 555 560 Lys Glu Ile Gly Ser Asn Cys Leu Asn Asn Glu Phe Asn Phe Phe Lys 565 570 575 Arg His Ile Cys Asp Ala Asn Lys Glu Gly Met Phe Leu Phe Arg Ala 580 585 590 Ala Arg Lys Leu Arg Gln Phe Leu Lys Met Asn Ser Thr Gly Asp Phe 595 600 605 Asp Leu His Leu Leu Lys Val Ser Glu Gly Thr Thr Ile Leu Leu Asn 610 615 620 Cys Thr Gly Gln Val Lys Gly Arg Lys Pro Ala Ala Leu Gly Glu Ala 625 630 635 640 Gln Pro Thr Lys Ser Leu Glu Glu Asn Lys Ser Leu Lys Glu Gln Lys 645 650 655 Lys Leu Asn Asp Leu Cys Phe Leu Lys Arg Leu Leu Gln Glu Ile Lys 660 665 670 Thr Cys Trp Asn Lys Ile Leu Met Gly Thr Lys Glu His Gly Gly Gly 675 680 685 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Ala Arg 690 695 700 Ser Pro Ser Pro Ser Thr Gln Pro Trp Glu His Val Asn Ala Ile Gln 705 710 715 720 Glu Ala Arg Arg Leu Leu Asn Leu Ser Arg Asp Thr Ala Ala Glu Met 725 730 735 Asn Glu Thr Val Glu Val Ile Ser Glu Met Phe Asp Leu Gln Glu Pro 740 745 750 Thr Cys Leu Gln Thr Arg Leu Glu Leu Tyr Lys Gln Gly Leu Arg Gly 755 760 765 Ser Leu Thr Lys Leu Lys Gly Pro Leu Thr Met Met Ala Ser His Tyr 770 775 780 Lys Gln His Cys Pro Pro Thr Pro Glu Thr Ser Cys Ala Thr Gln Ile 785 790 795 800 Ile Thr Phe Glu Ser Phe Lys Glu Asn Leu Lys Asp Phe Leu Leu Val 805 810 815 Ile Pro Phe Asp Cys Trp Glu Pro Val Gln Glu 820 825 <210> 42 <211> 808 <212> PRT <213> Artificial Sequence <220> <223> hIL12 bIL12 aIL15 GMCSF <400> 42 Ile Trp Glu Leu Lys Lys Asp Val Tyr Val Val Glu Leu Asp Trp Tyr 1 5 10 15 Pro Asp Ala Pro Gly Glu Met Val Val Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Gly Ile Thr Trp Thr Leu Asp Gln Ser Ser Glu Val Leu Gly 35 40 45 Ser Gly Lys Thr Leu Thr Ile Gln Val Lys Glu Phe Gly Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Val Leu Ser His Ser Leu Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asp Gly Ile Trp Ser Thr Asp Ile Leu Lys 85 90 95 Asp Gln Lys Glu Pro Lys Asn Lys Thr Phe Leu Arg Cys Glu Ala Lys 100 105 110 Asn Tyr Ser Gly Arg Phe Thr Cys Trp Trp Leu Thr Thr Ile Ser Thr 115 120 125 Asp Leu Thr Phe Ser Val Lys Ser Ser Arg Gly Ser Ser Asp Pro Gin 130 135 140 Gly Val Thr Cys Gly Ala Ala Thr Leu Ser Ala Glu Arg Val Arg Gly 145 150 155 160 Asp Asn Lys Glu Tyr Glu Tyr Ser Val Glu Cys Gin Glu Asp Ser Ala 165 170 175 Cys Pro Ala Ala Glu Glu Ser Leu Pro Ile Glu Val Met Val Asp Ala 180 185 190 Val His Lys Leu Lys Tyr Glu Asn Tyr Thr Ser Ser Phe Phe Ile Arg 195 200 205 Asp Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gin Leu Lys Pro Leu 210 215 220 Lys Asn Ser Arg Gin Val Glu Val Ser Trp Glu Tyr Pro Asp Thr Trp 225 230 235 240 Ser Thr Pro His Ser Tyr Phe Ser Leu Thr Phe Cys Val Gin Val Gin 245 250 255 Gly Lys Ser Lys Arg Glu Lys Lys Asp Arg Val Phe Thr Asp Lys Thr 260 265 270 Ser Ala Thr Val Ile Cys Arg Lys Asn Ala Ser Ile Ser Val Arg Ala 275 280 285 Gln Asp Arg Tyr Tyr Ser Ser Ser Trp Ser Glu Trp Ala Ser Val Pro 290 295 300 Cys Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 305 310 315 320 Ser Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys 325 330 335 Leu His His Ser Gln Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gln 340 345 350 Lys Ala Arg Gln Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu Ile 355 360 365 Asp His Glu Asp Ile Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys 370 375 380 Leu Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu 385 390 395 400 Thr Ser Phe Ile Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser 405 410 415 Phe Met Met Ala Leu Cys Leu Ser Ser Ile Tyr Glu Asp Leu Lys Met 420 425 430 Tyr Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro 435 440 445 Lys Arg Gin He Phe Leu Asp Gin Asn Met Leu Ala Val He Asp Glu 450 455 460 Leu Met Gin Ala Leu Asn Phe Asn Ser Gin Thr Val Pro Gin Lys Ser 465 470 475 480 Ser Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys He Lys Leu Cys He 485 490 495 Leu Leu His Ala Phe Arg He Arg Ala Val Thr He Asp Arg Val Met 500 505 510 Ser Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 515 520 525 Gly Gly Gly Gly Ser Gly He His Val Phe He Leu Gly Cys Phe Ser 530 535 540 Ala Gly Leu Pro Lys Thr Gin Ala Asn Trp Val Asn Val He Ser Asp 545 550 555 560 Leu Lys Lys He Gin Asp Leu He Gin Ser Met His He Asp Ala Thr 565 570 575 Leu Tyr Thr Gin Ser Gin Val His Pro Ser Cys Lys Val Thr Ala Met 580 585 590 Lys Cys Phe Leu Leu Glu Leu Gin Val He Ser Leu Glu Ser Gly Asp 595 600 605 Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn 610 615 620 Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys 625 630 635 640 Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val 645 650 655 His Ile Val Gln Met Phe Ile Asn Thr Ser Gly Gly Gly Gly Ser Gly 660 665 670 Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Ala Arg Ser Pro Ser 675 680 685 Pro Ser Thr Gln Pro Trp Glu His Val Asn Ala Ile Gln Glu Ala Arg 690 695 700 Arg Leu Leu Asn Leu Ser Arg Asp Thr Ala Ala Glu Met Asn Glu Thr 705 710 715 720 Val Glu Val Ile Ser Glu Met Phe Asp Leu Gln Glu Pro Thr Cys Leu 725 730 735 Gln Thr Arg Leu Glu Leu Tyr Lys Gln Gly Leu Arg Gly Ser Leu Thr 740 745 750 Lys Leu Lys Gly Pro Leu Thr Met Met Ala Ser His Tyr Lys Gin His 755 760 765 Cys Pro Pro Thr Pro Gin Thr Ser Cys Ala Thr Gin He He Thr Phe 770 775 780 Glu Ser Phe Lys Gin Asn Leu Lys Asp Phe Leu Leu Val He Pro Phe 785 790 795 800 Asp Cys Trp Gin Pro Val Gin Gin 805 <210> 43 <211> 808 <212> PRT <213> Artificial Sequence <220> <223> hIL12bIL12aIL21 GMCSF <400> 43 Ile Trp Gin Leu Lys Lys Asp Val Tyr Val Val Gin Leu Asp Trp Tyr 1 5 10 15 Pro Asp Ala Pro Gin Gin Met Val Val Leu Thr Cys Asp Thr Pro Gin 20 25 30 Glu Asp Gin He Thr Trp Thr Leu Asp Gin Ser Ser Gin Val Leu Gin 35 40 45 Ser Gin Lys Thr Leu Thr He Gin Val Gin Gin Phe Gin Asp Ala Gin 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Val Leu Ser His Ser Leu Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asp Gly lie Trp Ser Thr Asp lie Leu Lys 85 90 95 Asp Gln Lys Glu Pro Lys Asn Lys Thr Phe Leu Arg Cys Glu Ala Lys 100 105 110 Asn Tyr Ser Gly Arg Phe Thr Cys Trp Trp Leu Thr Thr lie Ser Thr 115 120 125 Asp Leu Thr Phe Ser Val Lys Ser Ser Arg Gly Ser Ser Asp Pro Gln 130 135 140 Gly Val Thr Cys Gly Ala Ala Thr Leu Ser Ala Glu Arg Val Arg Gly 145 150 155 160 Asp Asn Lys Glu Tyr Glu Tyr Ser Val Glu Cys Gln Glu Asp Ser Ala 165 170 175 Cys Pro Ala Ala Glu Glu Ser Leu Pro lie Glu Val Met Val Asp Ala 180 185 190 Val His Lys Leu Lys Tyr Glu Asn Tyr Thr Ser Ser Phe Phe lie Arg 195 200 205 Asp lie lie Lys Pro Asp Pro Pro Lys Asn Leu Gln Leu Lys Pro Leu 210 215 220 Lys Asn Ser Arg Gin Val Glu Val Ser Trp Glu Tyr Pro Asp Thr Trp 225 230 235 240 Ser Thr Pro His Ser Tyr Phe Ser Leu Thr Phe Cys Val Gin Val Gin 245 250 255 Gly Lys Ser Lys Arg Glu Lys Lys Asp Arg Val Phe Thr Asp Lys Thr 260 265 270 Ser Ala Thr Val He Cys Arg Lys Asn Ala Ser He Ser Val Arg Ala 275 280 285 Gln Asp Arg Tyr Tyr Ser Ser Ser Trp Ser Glu Trp Ala Ser Val Pro 290 295 300 Cys Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 305 310 315 320 Ser Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys 325 330 335 Leu His His Ser Gin Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gin 340 345 350 Lys Ala Arg Gin Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu He 355 360 365 Asp His Glu Asp He Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys 370 375 380 Leu Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu 385 390 395 400 Thr Ser Phe Ile Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser 405 410 415 Phe Met Met Ala Leu Cys Leu Ser Ser Ile Tyr Glu Asp Leu Lys Met 420 425 430 Tyr Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro 435 440 445 Lys Arg Gln Ile Phe Leu Asp Gln Asn Met Leu Ala Val Ile Asp Glu 450 455 460 Leu Met Gln Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gln Lys Ser 465 470 475 480 Ser Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys Ile Lys Leu Cys Ile 485 490 495 Leu Leu His Ala Phe Arg Ile Arg Ala Val Thr Ile Asp Arg Val Met 500 505 510 Ser Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 515 520 525 Gly Gly Gly Gly Ser Gln Gly Gln Asp Arg His Met Ile Arg Met Arg 530 535 540 Gln Leu Ile Asp Ile Val Asp Gln Leu Lys Asn Tyr Val Asn Asp Leu 545 550 555 560 Val Pro Glu Phe Leu Pro Ala Pro Glu Asp Val Glu Thr Asn Cys Glu 565 570 575 Trp Ser Ala Phe Ser Cys Phe Gln Lys Ala Gln Leu Lys Ser Ala Asn 580 585 590 Thr Gly Asn Asn Glu Arg Ile Ile Asn Val Ser Ile Lys Lys Leu Lys 595 600 605 Arg Lys Pro Pro Ser Thr Asn Ala Gly Arg Arg Gln Lys His Arg Leu 610 615 620 Thr Cys Pro Ser Cys Asp Ser Tyr Glu Lys Lys Pro Pro Lys Glu Phe 625 630 635 640 Leu Glu Arg Phe Lys Ser Leu Leu Gln Lys Met Ile His Gln His Leu 645 650 655 Ser Ser Arg Thr His Gly Ser Glu Asp Ser Gly Gly Gly Gly Ser Gly 660 665 670 Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Ala Arg Ser Pro Ser 675 680 685 Pro Ser Thr Gln Pro Trp Glu His Val Asn Ala Ile Gln Glu Ala Arg 690 695 700 Arg Leu Leu Asn Leu Ser Arg Asp Thr Ala Ala Glu Met Asn Glu Thr 705 710 715 720 Val Glu Val Ile Ser Glu Met Phe Asp Leu Gln Glu Pro Thr Cys Leu 725 730 735 Gln Thr Arg Leu Glu Leu Tyr Lys Gln Gly Leu Arg Gly Ser Leu Thr 740 745 750 Lys Leu Lys Gly Pro Leu Thr Met Met Ala Ser His Tyr Lys Gln His 755 760 765 Cys Pro Pro Thr Pro Glu Thr Ser Cys Ala Thr Gln Ile Ile Thr Phe 770 775 780 Glu Ser Phe Lys Glu Asn Leu Lys Asp Phe Leu Leu Val Ile Pro Phe 785 790 795 800 Asp Cys Trp Glu Pro Val Gln Glu 805 <210> 44 <211> 890 <212> PRT <213> Artificial Sequence <220> <223> hIL12bIL12aIL2 FLT3L <400> 44 Ile Trp Glu Leu Lys Lys Asp Val Tyr Val Val Glu Leu Asp Trp Tyr 1 5 10 15 Pro Asp Ala Pro Gly Glu Met Val Val Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Gly Ile Thr Trp Thr Leu Asp Gln Ser Ser Glu Val Leu Gly 35 40 45 Ser Gly Lys Thr Leu Thr Ile Gln Val Lys Glu Phe Gly Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Val Leu Ser His Ser Leu Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asp Gly Ile Trp Ser Thr Asp Ile Leu Lys 85 90 95 Asp Gln Lys Glu Pro Lys Asn Lys Thr Phe Leu Arg Cys Glu Ala Lys 100 105 110 Asn Tyr Ser Gly Arg Phe Thr Cys Trp Trp Leu Thr Thr Ile Ser Thr 115 120 125 Asp Leu Thr Phe Ser Val Lys Ser Ser Arg Gly Ser Ser Asp Pro Gln 130 135 140 Gly Val Thr Cys Gly Ala Ala Thr Leu Ser Ala Glu Arg Val Arg Gly 145 150 155 160 Asp Asn Lys Glu Tyr Glu Tyr Ser Val Glu Cys Gln Glu Asp Ser Ala 165 170 175 Cys Pro Ala Ala Glu Glu Ser Leu Pro lie Glu Val Met Val Asp Ala 180 185 190 Val His Lys Leu Lys Tyr Glu Asn Tyr Thr Ser Ser Phe Phe lie Arg 195 200 205 Asp lie lie Lys Pro Asp Pro Pro Lys Asn Leu Gin Leu Lys Pro Leu 210 215 220 Lys Asn Ser Arg Gin Val Glu Val Ser Trp Glu Tyr Pro Asp Thr Trp 225 230 235 240 Ser Thr Pro His Ser Tyr Phe Ser Leu Thr Phe Cys Val Gin Val Gin 245 250 255 Gly Lys Ser Lys Arg Glu Lys Lys Asp Arg Val Phe Thr Asp Lys Thr 260 265 270 Ser Ala Thr Val lie Cys Arg Lys Asn Ala Ser lie Ser Val Arg Ala 275 280 285 Gln Asp Arg Tyr Tyr Ser Ser Ser Trp Ser Glu Trp Ala Ser Val Pro 290 295 300 Cys Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 305 310 315 320 Ser Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys 325 330 335 Leu His His Ser Gin Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gin 340 345 350 Lys Ala Arg Gin Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu He 355 360 365 Asp His Glu Asp He Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys 370 375 380 Leu Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu 385 390 395 400 Thr Ser Phe He Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser 405 410 415 Phe Met Met Ala Leu Cys Leu Ser Ser He Tyr Glu Asp Leu Lys Met 420 425 430 Tyr Gin Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro 435 440 445 Lys Arg Gin He Phe Leu Asp Gin Asn Met Leu Ala Val He Asp Glu 450 455 460 Leu Met Gin Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gin Lys Ser 465 470 475 480 Ser Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys He Lys Leu Cys He 485 490 495 Leu Leu His Ala Phe Arg Ile Arg Ala Val Thr Ile Asp Arg Val Met 500 505 510 Ser Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 515 520 525 Gly Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln 530 535 540 Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly 545 550 555 560 Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys 565 570 575 Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu 580 585 590 Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser 595 600 605 Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val 610 615 620 Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr 625 630 635 640 Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr 645 650 655 Phe Cys Gln Ser lie lie Ser Thr Leu Thr Gly Gly Gly Gly Ser Gly 660 665 670 Gly Gly Gly Ser Gly Gly Gly Gly Ser Thr Gin Asp Cys Ser Phe Gin 675 680 685 His Ser Pro lie Ser Ser Asp Phe Ala Val Lys lie Arg Glu Leu Ser 690 695 700 Asp Tyr Leu Leu Gin Asp Tyr Pro Val Thr Val Ala Ser Asn Leu Gin 705 710 715 720 Asp Glu Glu Leu Cys Gly Gly Leu Trp Arg Leu Val Leu Ala Gin Arg 725 730 735 Trp Met Glu Arg Leu Lys Thr Val Ala Gly Ser Lys Met Gin Gly Leu 740 745 750 Leu Glu Arg Val Asn Thr Glu lie His Phe Val Thr Lys Cys Ala Phe 755 760 765 Gln Pro Pro Pro Ser Cys Leu Arg Phe Val Gin Thr Asn lie Ser Arg 770 775 780 Leu Leu Gin Glu Thr Ser Glu Gin Leu Val Ala Leu Lys Pro Trp lie 785 790 795 800 Thr Arg Gin Asn Phe Ser Arg Cys Leu Glu Leu Gin Cys Gin Pro Asp 805 810 815 Ser Ser Thr Leu Pro Pro Pro Trp Ser Pro Arg Pro Leu Glu Ala Thr 820 825 830 Ala Pro Thr Ala Pro Gin Pro Pro Leu Leu Leu Leu Leu Leu Leu Pro 835 840 845 Val Gly Leu Leu Leu Leu Ala Ala Ala Trp Cys Leu His Trp Gin Arg 850 855 860 Thr Arg Arg Arg Thr Pro Arg Pro Gly Glu Gin Val Pro Pro Val Pro 865 870 875 880 Ser Pro Gin Asp Leu Leu Leu Val Glu His 885 890 <210> 45 <211> 909 <212> PRT <213> Artificial Sequence <220> <223> hIL12b IL12a IL7 FLT3L <400> 45 Ile Trp Glu Leu Lys Lys Asp Val Tyr Val Val Glu Leu Asp Trp Tyr 1 5 10 15 Pro Asp Ala Pro Gly Glu Met Val Val Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Gly lie Thr Trp Thr Leu Asp Gin Ser Ser Glu Val Leu Gly 35 40 45 Ser Gly Lys Thr Leu Thr lie Gin Val Lys Glu Phe Gly Asp Ala Gly 50 55 60 Gln Tyr Thr Cy s His Lys Gly Gly Glu Val Leu Ser His Ser Leu Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asp Gly lie Trp Ser Thr Asp lie Leu Lys 85 90 95 Asp Gin Lys Glu Pro Lys Asn Lys Thr Phe Leu Arg Cys Glu Ala Lys 100 105 110 Asn Tyr Ser Gly Arg Phe Thr Cy s Trp Trp Leu Thr Thr lie Ser Thr 115 120 125 Asp Leu Thr Phe Ser Val Lys Ser Ser Arg Gly Ser Ser Asp Pro Gin 130 135 140 Gly Val Thr Cy s Gly Ala Ala Thr Leu Ser Ala Glu Arg Val Arg Gly 145 150 155 160 Asp Asn Lys Glu Tyr Glu Tyr Ser Val Glu Cys Gin Glu Asp Ser Ala 165 170 175 Cys Pro Ala Ala Glu Glu Ser Leu Pro lie Glu Val Met Val Asp Ala 180 185 190 Val His Lys Leu Lys Tyr Glu Asn Tyr Thr Ser Ser Phe Phe Ile Arg 195 200 205 Asp Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Leu Lys Pro Leu 210 215 220 Lys Asn Ser Arg Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Thr Trp 225 230 235 240 Ser Thr Pro His Ser Tyr Phe Ser Leu Thr Phe Cys Val Gln Val Gln 245 250 255 Gly Lys Ser Lys Arg Glu Lys Lys Asp Arg Val Phe Thr Asp Lys Thr 260 265 270 Ser Ala Thr Val Ile Cys Arg Lys Asn Ala Ser Ile Ser Val Arg Ala 275 280 285 Gln Asp Arg Tyr Tyr Ser Ser Ser Trp Ser Glu Trp Ala Ser Val Pro 290 295 300 Cys Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 305 310 315 320 Ser Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys 325 330 335 Leu His His Ser Gln Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gln 340 345 350 Lys Ala Arg Gin Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu lie 355 360 365 Asp His Glu Asp He Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys 370 375 380 Leu Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu 385 390 395 400 Thr Ser Phe He Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser 405 410 415 Phe Met Met Ala Leu Cys Leu Ser Ser He Tyr Glu Asp Leu Lys Met 420 425 430 Tyr Gin Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro 435 440 445 Lys Arg Gin He Phe Leu Asp Gin Asn Met Leu Ala Val He Asp Glu 450 455 460 Leu Met Gin Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gin Lys Ser 465 470 475 480 Ser Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys He Lys Leu Cys He 485 490 495 Leu Leu His Ala Phe Arg He Arg Ala Val Thr He Asp Arg Val Met 500 505 510 Ser Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 515 520 525 Gly Gly Gly Gly Ser Asp Cys Asp Ile Glu Gly Lys Asp Gly Lys Gln 530 535 540 Tyr Glu Ser Val Leu Met Val Ser Ile Asp Gln Leu Leu Asp Ser Met 545 550 555 560 Lys Glu Ile Gly Ser Asn Cys Leu Asn Asn Glu Phe Asn Phe Phe Lys 565 570 575 Arg His Ile Cys Asp Ala Asn Lys Glu Gly Met Phe Leu Phe Arg Ala 580 585 590 Ala Arg Lys Leu Arg Gln Phe Leu Lys Met Asn Ser Thr Gly Asp Phe 595 600 605 Asp Leu His Leu Leu Lys Val Ser Glu Gly Thr Thr Ile Leu Leu Asn 610 615 620 Cys Thr Gly Gln Val Lys Gly Arg Lys Pro Ala Ala Leu Gly Glu Ala 625 630 635 640 Gln Pro Thr Lys Ser Leu Glu Glu Asn Lys Ser Leu Lys Glu Gln Lys 645 650 655 Lys Leu Asn Asp Leu Cys Phe Leu Lys Arg Leu Leu Gln Glu Ile Lys 660 665 670 Thr Cys Trp Asn Lys Ile Leu Met Gly Thr Lys Glu His Gly Gly Gly 675 680 685 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Thr Gln Asp Cys 690 695 700 Ser Phe Gln His Ser Pro Ile Ser Ser Asp Phe Ala Val Lys Ile Arg 705 710 715 720 Glu Leu Ser Asp Tyr Leu Leu Gln Asp Tyr Pro Val Thr Val Ala Ser 725 730 735 Asn Leu Gln Asp Glu Glu Leu Cys Gly Gly Leu Trp Arg Leu Val Leu 740 745 750 Ala Gln Arg Trp Met Glu Arg Leu Lys Thr Val Ala Gly Ser Lys Met 755 760 765 Gln Gly Leu Leu Glu Arg Val Asn Thr Glu Ile His Phe Val Thr Lys 770 775 780 Cys Ala Phe Gln Pro Pro Pro Ser Cys Leu Arg Phe Val Gln Thr Asn 785 790 795 800 Ile Ser Arg Leu Leu Gln Glu Thr Ser Glu Gln Leu Val Ala Leu Lys 805 810 815 Pro Trp lie Thr Arg Gin Asn Phe Ser Arg Cys Leu Glu Leu Gin Cys 820 825 830 Gln Pro Asp Ser Ser Thr Leu Pro Pro Pro Trp Ser Pro Arg Pro Leu 835 840 845 Glu Ala Thr Ala Pro Thr Ala Pro Gin Pro Pro Leu Leu Leu Leu Leu 850 855 860 Leu Leu Pro Val Gly Leu Leu Leu Leu Ala Ala Ala Trp Cys Leu His 865 870 875 880 Trp Gin Arg Thr Arg Arg Arg Thr Pro Arg Pro Gly Glu Gin Val Pro 885 890 895 Pro Val Pro Ser Pro Gin Asp Leu Leu Leu Val Glu His 900 905 <210> 46 <211> 890 <212> PRT <213> Artificial Sequence <220> <223> hIL12bIL12aIL15 FLT3L <400> 46 Ile Trp Glu Leu Lys Lys Asp Val Tyr Val Val Glu Leu Asp Trp Tyr 1 5 10 15 Pro Asp Ala Pro Gly Glu Met Val Val Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Gly lie Thr Trp Thr Leu Asp Gin Ser Ser Glu Val Leu Gly 35 40 45 Ser Gly Lys Thr Leu Thr lie Gin Val Lys Glu Phe Gly Asp Ala Gly 50 55 60 Gln Tyr Thr Cy s His Lys Gly Gly Glu Val Leu Ser His Ser Leu Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asp Gly lie Trp Ser Thr Asp lie Leu Lys 85 90 95 Asp Gin Lys Glu Pro Lys Asn Lys Thr Phe Leu Arg Cys Glu Ala Lys 100 105 110 Asn Tyr Ser Gly Arg Phe Thr Cy s Trp Trp Leu Thr Thr lie Ser Thr 115 120 125 Asp Leu Thr Phe Ser Val Lys Ser Ser Arg Gly Ser Ser Asp Pro Gin 130 135 140 Gly Val Thr Cy s Gly Ala Ala Thr Leu Ser Ala Glu Arg Val Arg Gly 145 150 155 160 Asp Asn Lys Glu Tyr Glu Tyr Ser Val Glu Cys Gin Glu Asp Ser Ala 165 170 175 Cys Pro Ala Ala Glu Glu Ser Leu Pro lie Glu Val Met Val Asp Ala 180 185 190 Val His Lys Leu Lys Tyr Glu Asn Tyr Thr Ser Ser Phe Phe Ile Arg 195 200 205 Asp Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Leu Lys Pro Leu 210 215 220 Lys Asn Ser Arg Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Thr Trp 225 230 235 240 Ser Thr Pro His Ser Tyr Phe Ser Leu Thr Phe Cys Val Gln Val Gln 245 250 255 Gly Lys Ser Lys Arg Glu Lys Lys Asp Arg Val Phe Thr Asp Lys Thr 260 265 270 Ser Ala Thr Val Ile Cys Arg Lys Asn Ala Ser Ile Ser Val Arg Ala 275 280 285 Gln Asp Arg Tyr Tyr Ser Ser Ser Trp Ser Glu Trp Ala Ser Val Pro 290 295 300 Cys Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 305 310 315 320 Ser Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys 325 330 335 Leu His His Ser Gln Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gln 340 345 350 Lys Ala Arg Gin Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu lie 355 360 365 Asp His Glu Asp He Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys 370 375 380 Leu Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu 385 390 395 400 Thr Ser Phe He Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser 405 410 415 Phe Met Met Ala Leu Cys Leu Ser Ser He Tyr Glu Asp Leu Lys Met 420 425 430 Tyr Gin Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro 435 440 445 Lys Arg Gin He Phe Leu Asp Gin Asn Met Leu Ala Val He Asp Glu 450 455 460 Leu Met Gin Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gin Lys Ser 465 470 475 480 Ser Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys He Lys Leu Cys He 485 490 495 Leu Leu His Ala Phe Arg He Arg Ala Val Thr He Asp Arg Val Met 500 505 510 Ser Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 515 520 525 Gly Gly Gly Gly Ser Gly Ile His Val Phe Ile Leu Gly Cys Phe Ser 530 535 540 Ala Gly Leu Pro Lys Thr Glu Ala Asn Trp Val Asn Val Ile Ser Asp 545 550 555 560 Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr 565 570 575 Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met 580 585 590 Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp 595 600 605 Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn 610 615 620 Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys 625 630 635 640 Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val 645 650 655 His Ile Val Gln Met Phe Ile Asn Thr Ser Gly Gly Gly Gly Ser Gly 660 665 670 Gly Gly Gly Ser Gly Gly Gly Gly Ser Thr Gin Asp Cys Ser Phe Gin 675 680 685 His Ser Pro Ile Ser Ser Asp Phe Ala Val Lys Ile Arg Glu Leu Ser 690 695 700 Asp Tyr Leu Leu Gin Asp Tyr Pro Val Thr Val Ala Ser Asn Leu Gin 705 710 715 720 Asp Glu Glu Leu Cys Gly Gly Leu Trp Arg Leu Val Leu Ala Gin Arg 725 730 735 Trp Met Glu Arg Leu Lys Thr Val Ala Gly Ser Lys Met Gin Gly Leu 740 745 750 Leu Glu Arg Val Asn Thr Glu Ile His Phe Val Thr Lys Cys Ala Phe 755 760 765 Gln Pro Pro Pro Ser Cys Leu Arg Phe Val Gin Thr Asn Ile Ser Arg 770 775 780 Leu Leu Gin Glu Thr Ser Glu Gin Leu Val Ala Leu Lys Pro Trp Ile 785 790 795 800 Thr Arg Gin Asn Phe Ser Arg Cys Leu Glu Leu Gin Cys Gin Pro Asp 805 810 815 Ser Ser Thr Leu Pro Pro Pro Trp Ser Pro Arg Pro Leu Glu Ala Thr 820 825 830 Ala Pro Thr Ala Pro Gln Pro Pro Leu Leu Leu Leu Leu Leu Leu Pro 835 840 845 Val Gly Leu Leu Leu Leu Ala Ala Ala Trp Cys Leu His Trp Gln Arg 850 855 860 Thr Arg Arg Arg Thr Pro Arg Pro Gly Glu Gln Val Pro Pro Val Pro 865 870 875 880 Ser Pro Gln Asp Leu Leu Leu Val Glu His 885 890 <210> 47 <211> 890 <212> PRT <213> Artificial Sequence <220> <223> hIL12bIL12aIL21 FLT3L <400> 47 Ile Trp Glu Leu Lys Lys Asp Val Tyr Val Val Glu Leu Asp Trp Tyr 1 5 10 15 Pro Asp Ala Pro Gly Glu Met Val Val Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Gly Ile Thr Trp Thr Leu Asp Gln Ser Ser Glu Val Leu Gly 35 40 45 Ser Gly Lys Thr Leu Thr lie Gin Val Lys Glu Phe Gly Asp Ala Gly 50 55 60 Gln Tyr Thr Cy s His Lys Gly Gly Glu Val Leu Ser His Ser Leu Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asp Gly lie Trp Ser Thr Asp lie Leu Lys 85 90 95 Asp Gin Lys Glu Pro Lys Asn Lys Thr Phe Leu Arg Cy s Glu Ala Lys 100 105 110 Asn Tyr Ser Gly Arg Phe Thr Cy s Trp Trp Leu Thr Thr lie Ser Thr 115 120 125 Asp Leu Thr Phe Ser Val Lys Ser Ser Arg Gly Ser Ser Asp Pro Gin 130 135 140 Gly Val Thr Cy s Gly Ala Ala Thr Leu Ser Ala Glu Arg Val Arg Gly 145 150 155 160 Asp Asn Lys Glu Tyr Glu Tyr Ser Val Glu Cy s Gin Glu Asp Ser Ala 165 170 175 Cys Pro Ala Ala Glu Glu Ser Leu Pro lie Glu Val Met Val Asp Ala 180 185 190 Val His Lys Leu Lys Tyr Glu Asn Tyr Thr Ser Ser Phe Phe lie Arg 195 200 205 Asp Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gin Leu Lys Pro Leu 210 215 220 Lys Asn Ser Arg Gin Val Glu Val Ser Trp Gin Tyr Pro Asp Thr Trp 225 230 235 240 Ser Thr Pro His Ser Tyr Phe Ser Leu Thr Phe Cys Val Gin Val Gin 245 250 255 Gly Lys Ser Lys Arg Gin Lys Lys Asp Arg Val Phe Thr Asp Lys Thr 260 265 270 Ser Ala Thr Val Ile Cys Arg Lys Asn Ala Ser Ile Ser Val Arg Ala 275 280 285 Gln Asp Arg Tyr Tyr Ser Ser Ser Trp Ser Gin Trp Ala Ser Val Pro 290 295 300 Cys Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 305 310 315 320 Ser Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys 325 330 335 Leu His His Ser Gin Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gin 340 345 350 Lys Ala Arg Gin Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu Ile 355 360 365 Asp His Glu Asp lie Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys 370 375 380 Leu Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu 385 390 395 400 Thr Ser Phe lie Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser 405 410 415 Phe Met Met Ala Leu Cys Leu Ser Ser lie Tyr Glu Asp Leu Lys Met 420 425 430 Tyr Gin Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro 435 440 445 Lys Arg Gin lie Phe Leu Asp Gin Asn Met Leu Ala Val lie Asp Glu 450 455 460 Leu Met Gin Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gin Lys Ser 465 470 475 480 Ser Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys lie Lys Leu Cys lie 485 490 495 Leu Leu His Ala Phe Arg lie Arg Ala Val Thr lie Asp Arg Val Met 500 505 510 Ser Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 515 520 525 Gly Gly Gly Gly Ser Gin Gin Asp Arg His Met lie Arg Met Arg 530 535 540 Gln Leu lie Asp lie Val Asp Gin Leu Lys Asn Tyr Val Asn Asp Leu 545 550 555 560 Val Pro Glu Phe Leu Pro Ala Pro Glu Asp Val Glu Thr Asn Cys Glu 565 570 575 Trp Ser Ala Phe Ser Cys Phe Gin Lys Ala Gin Leu Lys Ser Ala Asn 580 585 590 Thr Gly Asn Asn Glu Arg lie lie Asn Val Ser lie Lys Lys Leu Lys 595 600 605 Arg Lys Pro Pro Ser Thr Asn Ala Gly Arg Arg Gin Lys His Arg Leu 610 615 620 Thr Cys Pro Ser Cys Asp Ser Tyr Glu Lys Lys Pro Pro Lys Glu Phe 625 630 635 640 Leu Glu Arg Phe Lys Ser Leu Leu Gin Lys Met lie His Gin His Leu 645 650 655 Ser Ser Arg Thr His Gly Ser Glu Asp Ser Gly Gly Gly Gly Ser Gly 660 665 670 Gly Gly Gly Ser Gly Gly Gly Gly Ser Thr Gin Asp Cys Ser Phe Gin 675 680 685 His Ser Pro lie Ser Ser Asp Phe Ala Val Lys lie Arg Glu Leu Ser 690 695 700 Asp Tyr Leu Leu Gin Asp Tyr Pro Val Thr Val Ala Ser Asn Leu Gin 705 710 715 720 Asp Glu Glu Leu Cys Gly Gly Leu Trp Arg Leu Val Leu Ala Gin Arg 725 730 735 Trp Met Glu Arg Leu Lys Thr Val Ala Gly Ser Lys Met Gin Gly Leu 740 745 750 Leu Glu Arg Val Asn Thr Glu lie His Phe Val Thr Lys Cys Ala Phe 755 760 765 Gln Pro Pro Pro Ser Cys Leu Arg Phe Val Gin Thr Asn lie Ser Arg 770 775 780 Leu Leu Gin Glu Thr Ser Glu Gin Leu Val Ala Leu Lys Pro Trp lie 785 790 795 800 Thr Arg Gin Asn Phe Ser Arg Cys Leu Glu Leu Gin Cys Gin Pro Asp 805 810 815 Ser Ser Thr Leu Pro Pro Pro Trp Ser Pro Arg Pro Leu Glu Ala Thr 820 825 830 Ala Pro Thr Ala Pro Gln Pro Pro Leu Leu Leu Leu Leu Leu Leu Pro 835 840 845 Val Gly Leu Leu Leu Leu Ala Ala Ala Trp Cys Leu His Trp Gln Arg 850 855 860 Thr Arg Arg Arg Thr Pro Arg Pro Gly Glu Gln Val Pro Pro Val Pro 865 870 875 880 Ser Pro Gln Asp Leu Leu Leu Val Glu His 885 890 <210> 48 <211> 1316 <212> PRT <213> Artificial Sequence <220> <223> mIL12bIL12aIL2DiaNHS76F8GMCSF-Thr <400> 48 Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr 1 5 10 15 Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg His Gly Val Ile Gly 35 40 45 Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu Phe Leu Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu Lys 85 90 95 Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser 100 105 110 Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu Lys 115 120 125 Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val Thr 130 135 140 Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln Arg 145 150 155 160 Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys Pro 165 170 175 Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala Leu Glu Ala Arg Gln 180 185 190 Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe Ile Arg Asp Ile 195 200 205 Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met Lys Pro Leu Lys Asn 210 215 220 Ser Gin Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro 225 230 235 240 His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg Ile Gin Arg Lys Lys 245 250 255 Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gin Lys Gly Ala Phe 260 265 270 Leu Val Glu Lys Thr Ser Thr Glu Val Gin Cys Lys Gly Gly Asn Val 275 280 285 Cys Val Gin Ala Gin Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp 290 295 300 Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly Gly Gly Ser Gly Gly 305 310 315 320 Gly Gly Ser Gly Gly Gly Gly Ser Arg Val Ile Pro Val Ser Gly Pro 325 330 335 Ala Arg Cys Leu Ser Gin Ser Arg Asn Leu Leu Lys Thr Thr Asp Asp 340 345 350 Met Val Lys Thr Ala Arg Glu Lys Leu Lys His Tyr Ser Cys Thr Ala 355 360 365 Glu Asp Ile Asp His Glu Asp Ile Thr Arg Asp Gin Thr Ser Thr Leu 370 375 380 Lys Thr Cys Leu Pro Leu Glu Leu His Lys Asn Glu Ser Cys Leu Ala 385 390 395 400 Thr Arg Glu Thr Ser Ser Thr Thr Arg Gly Ser Cys Leu Pro Pro Gln 405 410 415 Lys Thr Ser Leu Met Met Thr Leu Cys Leu Gly Ser Ile Tyr Glu Asp 420 425 430 Leu Lys Met Tyr Gln Thr Glu Phe Gln Ala Ile Asn Ala Ala Leu Gln 435 440 445 Asn His Asn His Gln Gln Ile Ile Leu Asp Lys Gly Met Leu Val Ala 450 455 460 Ile Asp Glu Leu Met Gln Ser Leu Asn His Asn Gly Glu Thr Leu Arg 465 470 475 480 Gln Lys Pro Pro Val Gly Glu Ala Asp Pro Tyr Arg Val Lys Met Lys 485 490 495 Leu Cys Ile Leu Leu His Ala Phe Ser Thr Arg Val Val Thr Ile Asn 500 505 510 Arg Val Met Gly Tyr Leu Ser Ser Ala Gly Gly Gly Gly Ser Gly Gly 515 520 525 Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Ser 530 535 540 Ser Ser Thr Ala Glu Ala Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln 545 550 555 560 Gln Gln His Leu Glu Gln Leu Leu Met Asp Leu Gln Glu Leu Leu Ser 565 570 575 Arg Met Glu Asn Tyr Arg Asn Leu Lys Leu Pro Arg Met Leu Thr Phe 580 585 590 Lys Phe Tyr Leu Pro Lys Gln Ala Thr Glu Leu Lys Asp Leu Gln Cys 595 600 605 Leu Glu Asp Glu Leu Gly Pro Leu Arg His Val Leu Asp Leu Thr Gln 610 615 620 Ser Lys Ser Phe Gln Leu Glu Asp Ala Glu Asn Phe Ile Ser Asn Ile 625 630 635 640 Arg Val Thr Val Val Lys Leu Lys Gly Ser Asp Asn Thr Phe Glu Cys 645 650 655 Gln Phe Asp Asp Glu Ser Ala Thr Val Val Asp Phe Leu Arg Arg Trp 660 665 670 Ile Ala Phe Cys Gln Ser Ile Ile Ser Thr Ser Pro Gln Gly Gly Gly 675 680 685 Gly Ser Leu Val Pro Arg Gly Ser Gly Gly Gly Gly Ser Gln Val Gln 690 695 700 Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu Thr Leu Ser 705 710 715 720 Leu Thr Cys Ala Val Ser Gly Tyr Ser Ile Ser Ser Gly Tyr Tyr Trp 725 730 735 Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile Gly Ser 740 745 750 Ile Tyr His Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Lys Ser Arg 755 760 765 Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu Lys Leu 770 775 780 Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Arg Gly 785 790 795 800 Lys Trp Ser Lys Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val 805 810 815 Ser Ser Gly Gly Ser Gly Gly Glu Ile Val Leu Thr Gln Ser Pro Gly 820 825 830 Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala 835 840 845 Ser Gin Ser Val Ser Met Pro Phe Leu Ala Trp Tyr Gin Gin Lys Pro 850 855 860 Gly Gin Ala Pro Arg Leu Leu lie Tyr Gly Ala Ser Ser Arg Ala Thr 865 870 875 880 Gly lie Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 885 890 895 Leu Thr lie Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys 900 905 910 Gln Gin Met Arg Gly Arg Pro Pro Thr Phe Gly Gin Gly Thr Lys Val 915 920 925 Glu lie Lys Ser Ser Ser Ser Gly Ser Ser Ser Ser Gly Ser Ser Ser 930 935 940 Ser Gly Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro 945 950 955 960 Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser 965 970 975 Leu Phe Thr Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu 980 985 990 Trp Val Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp 995 1000 1005 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn 1010 1015 1020 Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala 1025 1030 1035 Val Tyr Tyr Cys Ala Lys Ser Thr His Leu Tyr Leu Phe Asp Tyr 1040 1045 1050 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Ser Gly 1055 1060 1065 Gly Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu 1070 1075 1080 Gly Gln Thr Val Arg Ile Thr Cys Gln Gly Asp Ser Leu Arg Ser 1085 1090 1095 Tyr Tyr Ala Ser Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val 1100 1105 1110 Leu Val Ile Tyr Gly Lys Asn Asn Arg Pro Ser Gly Ile Pro Asp 1115 1120 1125 Arg Phe Ser Gly Ser Ser Ser Gly Asn Thr Ala Ser Leu Thr Ile 1130 1135 1140 Thr Gly Ala Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Asn Ser 1145 1150 1155 Arg Asp Ser Ser Gly Asn His Val Val Phe Gly Gly Gly Thr Lys 1160 1165 1170 Leu Thr Val Leu Ser Ser Ser Ser Gly Leu Val Pro Arg Gly Ser 1175 1180 1185 Ser Ser Ser Gly Ala Pro Thr Arg Ser Pro Ile Thr Val Thr Arg 1190 1195 1200 Pro Trp Lys His Val Glu Ala Ile Lys Glu Ala Leu Asn Leu Leu 1205 1210 1215 Asp Asp Met Pro Val Thr Leu Asn Glu Glu Val Glu Val Val Ser 1220 1225 1230 Asn Glu Phe Ser Phe Lys Lys Leu Thr Cys Val Gln Thr Arg Leu 1235 1240 1245 Lys Ile Phe Glu Gln Gly Leu Arg Gly Asn Phe Thr Lys Leu Lys 1250 1255 1260 Gly Ala Leu Asn Met Thr Ala Ser Tyr Tyr Gln Thr Tyr Cys Pro 1265 1270 1275 Pro Thr Pro Glu Thr Asp Cys Glu Thr Gln Val Thr Thr Tyr Ala 1280 1285 1290 Asp Phe Ile Asp Ser Leu Lys Thr Phe Leu Thr Asp Ile Pro Phe 1295 1300 1305 ​Glu Cys Lys Lys Pro Gly Gin Lys 1310 1315 <210> 49 <211> 1300 <212> PRT <213> Artificial Sequence <220> <223> hIL12 bIL12 aIL2DiaNHS76F8 GMCSF-Thr <400> 49 Ile Trp Glu Leu Lys Lys Asp Val Tyr Val Val Glu Leu Asp Trp Tyr 1 5 10 15 Pro Asp Ala Pro Gly Glu Met Val Val Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Gly Ile Thr Trp Thr Leu Asp Gin Ser Ser Glu Val Leu Gly 35 40 45 Ser Gly Lys Thr Leu Thr Ile Gin Val Lys Glu Phe Gly Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Val Leu Ser His Ser Leu Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asp Gly Ile Trp Ser Thr Asp Ile Leu Lys 85 90 95 Asp Gin Lys Glu Pro Lys Asn Lys Thr Phe Leu Arg Cys Glu Ala Lys 100 105 110 Asn Tyr Ser Gly Arg Phe Thr Cys Trp Trp Leu Thr Thr Ile Ser Thr 115 120 125 Asp Leu Thr Phe Ser Val Lys Ser Ser Arg Gly Ser Ser Asp Pro Gln 130 135 140 Gly Val Thr Cys Gly Ala Ala Thr Leu Ser Ala Glu Arg Val Arg Gly 145 150 155 160 Asp Asn Lys Glu Tyr Glu Tyr Ser Val Glu Cys Gln Glu Asp Ser Ala 165 170 175 Cys Pro Ala Ala Glu Glu Ser Leu Pro Ile Glu Val Met Val Asp Ala 180 185 190 Val His Lys Leu Lys Tyr Glu Asn Tyr Thr Ser Ser Phe Phe Ile Arg 195 200 205 Asp Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Leu Lys Pro Leu 210 215 220 Lys Asn Ser Arg Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Thr Trp 225 230 235 240 Ser Thr Pro His Ser Tyr Phe Ser Leu Thr Phe Cys Val Gln Val Gln 245 250 255 Gly Lys Ser Lys Arg Glu Lys Lys Asp Arg Val Phe Thr Asp Lys Thr 260 265 270 Ser Ala Thr Val lie Cys Arg Lys Asn Ala Ser lie Ser Val Arg Ala 275 280 285 Gln Asp Arg Tyr Tyr Ser Ser Ser Trp Ser Glu Trp Ala Ser Val Pro 290 295 300 Cys Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 305 310 315 320 Ser Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys 325 330 335 Leu His His Ser Gin Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gin 340 345 350 Lys Ala Arg Gin Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu lie 355 360 365 Asp His Glu Asp lie Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys 370 375 380 Leu Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu 385 390 395 400 Thr Ser Phe lie Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser 405 410 415 Phe Met Met Ala Leu Cys Leu Ser Ser lie Tyr Glu Asp Leu Lys Met 420 425 430 Tyr Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro 435 440 445 Lys Arg Gln Ile Phe Leu Asp Gln Asn Met Leu Ala Val Ile Asp Glu 450 455 460 Leu Met Gln Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gln Lys Ser 465 470 475 480 Ser Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys Ile Lys Leu Cys Ile 485 490 495 Leu Leu His Ala Phe Arg Ile Arg Ala Val Thr Ile Asp Arg Val Met 500 505 510 Ser Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 515 520 525 Gly Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln 530 535 540 Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly 545 550 555 560 Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys 565 570 575 Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu 580 585 590 Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser 595 600 605 Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val 610 615 620 Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr 625 630 635 640 Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr 645 650 655 Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Gly Gly Gly Gly Ser Leu 660 665 670 Val Pro Arg Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu 675 680 685 Ser Gly Pro Gly Leu Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys 690 695 700 Ala Val Ser Gly Tyr Ser Ile Ser Ser Gly Tyr Tyr Trp Gly Trp Ile 705 710 715 720 Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile Gly Ser Ile Tyr His 725 730 735 Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr lie 740 745 750 Ser Val Asp Thr Ser Lys Asn Gin Phe Ser Leu Lys Leu Ser Ser Val 755 760 765 Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Arg Gly Lys Trp Ser 770 775 780 Lys Phe Asp Tyr Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser Gly 785 790 795 800 Gly Ser Gly Gly Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser 805 810 815 Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser 820 825 830 Val Ser Met Pro Phe Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala 835 840 845 Pro Arg Leu Leu lie Tyr Gly Ala Ser Ser Arg Ala Thr Gly lie Pro 850 855 860 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie 865 870 875 880 Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Met 885 890 895 Arg Gly Arg Pro Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 900 905 910 Ser Ser Ser Ser Gly Ser Ser Ser Ser Gly Glu 915 920 925 Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly Ser 930 935 940 Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Leu Phe Thr 945 950 955 960 Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val Ser 965 970 975 Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys 980 985 990 Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 995 1000 1005 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 1010 1015 1020 Ala Lys Ser Thr His Leu Tyr Leu Phe Asp Tyr Trp Gly Gin Gly 1025 1030 1035 Thr Leu Val Thr Val Ser Ser Gly Gly Ser Gly Gly Ser Ser Glu 1040 1045 1050 Leu Thr Gin Asp Pro Ala Val Ser Val Ala Leu Gly Gin Thr Val 1055 1060 1065 Arg He Thr Cy s Gin Gly Asp Ser Leu Arg Ser Tyr Tyr Ala Ser 1070 1075 1080 Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Val Leu Val He Tyr 1085 1090 1095 Gly Lys Asn Asn Arg Pro Ser Gly He Pro Asp Arg Phe Ser Gly 1100 1105 1110 Ser Ser Ser Gly Asn Thr Ala Ser Leu Thr He Thr Gly Ala Gin 1115 1120 1125 Ala Glu Asp Glu Ala Asp Tyr Tyr Cy s Asn Ser Arg Asp Ser Ser 1130 1135 1140 Gly Asn His Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 1145 1150 1155 Ser Ser Ser Ser Gly Leu Val Pro Arg Gly Ser Ser Ser Ser Gly 1160 1165 1170 Ala Pro Ala Arg Ser Pro Ser Pro Ser Thr Gin Pro Trp Glu His 1175 1180 1185 Val Asn Ala He Gin Glu Ala Arg Arg Leu Leu Asn Leu Ser Arg 1190 1195 1200 Asp Thr Ala Ala Glu Met Asn Glu Thr Val Glu Val Ile Ser Glu 1205 1210 1215 Met Phe Asp Leu Gln Glu Pro Thr Cys Leu Gln Thr Arg Leu Glu 1220 1225 1230 Leu Tyr Lys Gln Gly Leu Arg Gly Ser Leu Thr Lys Leu Lys Gly 1235 1240 1245 Pro Leu Thr Met Met Ala Ser His Tyr Lys Gln His Cys Pro Pro 1250 1255 1260 Thr Pro Glu Thr Ser Cys Ala Thr Gln Ile Ile Thr Phe Glu Ser 1265 1270 1275 Phe Lys Glu Asn Leu Lys Asp Phe Leu Leu Val Ile Pro Phe Asp 1280 1285 1290 Cys Trp Glu Pro Val Gln Glu 1295 1300 <210> 50 <211> 1316 <212> PRT <213> Artificial Sequence <220> <223> mIL12bIL12aIL2DiaF8GMCSF <400> 50 Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr 1 5 10 15 Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg His Gly Val Ile Gly 35 40 45 Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu Phe Leu Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu Lys 85 90 95 Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser 100 105 110 Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu Lys 115 120 125 Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val Thr 130 135 140 Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln Arg 145 150 155 160 Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys Pro 165 170 175 Thr Ala Glu Glu Thr Leu Pro lie Glu Leu Ala Leu Glu Ala Arg Gin 180 185 190 Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe lie Arg Asp lie 195 200 205 Ile Lys Pro Asp Pro Pro Lys Asn Leu Gin Met Lys Pro Leu Lys Asn 210 215 220 Ser Gin Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro 225 230 235 240 His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg lie Gin Arg Lys Lys 245 250 255 Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gin Lys Gly Ala Phe 260 265 270 Leu Val Glu Lys Thr Ser Thr Glu Val Gin Cys Lys Gly Gly Asn Val 275 280 285 Cys Val Gin Ala Gin Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp 290 295 300 Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly Gly Gly Ser Gly Gly 305 310 315 320 Gly Gly Ser Gly Gly Gly Gly Ser Arg Val lie Pro Val Ser Gly Pro 325 330 335 Ala Arg Cys Leu Ser Gin Ser Arg Asn Leu Leu Lys Thr Thr Asp Asp 340 345 350 Met Val Lys Thr Ala Arg Glu Lys Leu Lys His Tyr Ser Cys Thr Ala 355 360 365 Glu Asp He Asp His Glu Asp He Thr Arg Asp Gin Thr Ser Thr Leu 370 375 380 Lys Thr Cys Leu Pro Leu Glu Leu His Lys Asn Glu Ser Cys Leu Ala 385 390 395 400 Thr Arg Glu Thr Ser Ser Thr Thr Arg Gly Ser Cys Leu Pro Pro Gin 405 410 415 Lys Thr Ser Leu Met Met Thr Leu Cys Leu Gly Ser He Tyr Glu Asp 420 425 430 Leu Lys Met Tyr Gin Thr Glu Phe Gin Ala He Asn Ala Ala Leu Gin 435 440 445 Asn His Asn His Gin Gin He He Leu Asp Lys Gly Met Leu Val Ala 450 455 460 He Asp Glu Leu Met Gin Ser Leu Asn His Asn Gly Glu Thr Leu Arg 465 470 475 480 Gln Lys Pro Pro Val Gly Glu Ala Asp Pro Tyr Arg Val Lys Met Lys 485 490 495 Leu Cys Ile Leu Leu His Ala Phe Ser Thr Arg Val Val Thr Ile Asn 500 505 510 Arg Val Met Gly Tyr Leu Ser Ser Ala Gly Gly Gly Gly Ser Gly Gly 515 520 525 Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Ser 530 535 540 Ser Ser Thr Ala Glu Ala Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln 545 550 555 560 Gln Gln His Leu Glu Gln Leu Leu Met Asp Leu Gln Glu Leu Leu Ser 565 570 575 Arg Met Glu Asn Tyr Arg Asn Leu Lys Leu Pro Arg Met Leu Thr Phe 580 585 590 Lys Phe Tyr Leu Pro Lys Gln Ala Thr Glu Leu Lys Asp Leu Gln Cys 595 600 605 Leu Glu Asp Glu Leu Gly Pro Leu Arg His Val Leu Asp Leu Thr Gln 610 615 620 Ser Lys Ser Phe Gln Leu Glu Asp Ala Glu Asn Phe Ile Ser Asn Ile 625 630 635 640 Arg Val Thr Val Val Lys Leu Lys Gly Ser Asp Asn Thr Phe Glu Cys 645 650 655 Gln Phe Asp Asp Glu Ser Ala Thr Val Val Asp Phe Leu Arg Arg Trp 660 665 670 Ile Ala Phe Cys Gln Ser Ile Ile Ser Thr Ser Pro Gln Gly Gly Gly 675 680 685 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu 690 695 700 Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu 705 710 715 720 Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Leu Phe Thr Met Ser Trp 725 730 735 Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ala Ile Ser 740 745 750 Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe 755 760 765 Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn 770 775 780 Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Lys Ser Thr 785 790 795 800 His Leu Tyr Leu Phe Asp Tyr Trp Gly Gin Gly Thr Leu Val Thr Val 805 810 815 Ser Ser Gly Gly Ser Gly Gly Glu lie Val Leu Thr Gin Ser Pro Gly 820 825 830 Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala 835 840 845 Ser Gin Ser Val Ser Met Pro Phe Leu Ala Trp Tyr Gin Gin Lys Pro 850 855 860 Gly Gin Ala Pro Arg Leu Leu lie Tyr Gly Ala Ser Ser Arg Ala Thr 865 870 875 880 Gly lie Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 885 890 895 Leu Thr lie Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys 900 905 910 Gln Gin Met Arg Gly Arg Pro Pro Thr Phe Gly Gin Gly Thr Lys Val 915 920 925 Glu lie Lys Ser Ser Ser Ser Gly Ser Ser Ser Ser Gly Ser Ser Ser 930 935 940 Ser Gly Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro 945 950 955 960 Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser 965 970 975 Leu Phe Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu 980 985 990 Trp Val Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp 995 1000 1005 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn 1010 1015 1020 Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala 1025 1030 1035 Val Tyr Tyr Cys Ala Lys Ser Thr His Leu Tyr Leu Phe Asp Tyr 1040 1045 1050 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Ser Gly 1055 1060 1065 Gly Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser 1070 1075 1080 Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val 1085 1090 1095 Ser Met Pro Phe Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala 1100 1105 1110 Pro Arg Leu Leu Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile 1115 1120 1125 Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu 1130 1135 1140 Thr Ile Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys 1145 1150 1155 Gln Gln Met Arg Gly Arg Pro Pro Thr Phe Gly Gln Gly Thr Lys 1160 1165 1170 Val Glu Ile Lys Ser Ser Ser Ser Gly Ser Ser Ser Ser Gly Ser 1175 1180 1185 Ser Ser Ser Gly Ala Pro Thr Arg Ser Pro Ile Thr Val Thr Arg 1190 1195 1200 Pro Trp Lys His Val Glu Ala Ile Lys Glu Ala Leu Asn Leu Leu 1205 1210 1215 Asp Asp Met Pro Val Thr Leu Asn Glu Glu Val Glu Val Val Ser 1220 1225 1230 Asn Glu Phe Ser Phe Lys Lys Leu Thr Cys Val Gln Thr Arg Leu 1235 1240 1245 Lys Ile Phe Glu Gln Gly Leu Arg Gly Asn Phe Thr Lys Leu Lys 1250 1255 1260 Gly Ala Leu Asn Met Thr Ala Ser Tyr Tyr Gin Thr Tyr Cys Pro 1265 1270 1275 Pro Thr Pro Glu Thr Asp Cys Glu Thr Gin Val Thr Thr Tyr Ala 1280 1285 1290 Asp Phe He Asp Ser Leu Lys Thr Phe Leu Thr Asp He Pro Phe 1295 1300 1305 Glu Cys Lys Lys Pro Gly Gin Lys 1310 1315 <210> 51 <211> 1300 <212> PRT <213> Artificial Sequence <220> <223> hIL12 bIL12 aIL2DiaF8 GMCSF <400> 51 Ile Trp Glu Leu Lys Lys Asp Val Tyr Val Val Glu Leu Asp Trp Tyr 1 5 10 15 Pro Asp Ala Pro Gly Glu Met Val Val Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Gly He Thr Trp Thr Leu Asp Gin Ser Ser Glu Val Leu Gly 35 40 45 Ser Gly Lys Thr Leu Thr He Gin Val Lys Glu Phe Gly Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Val Leu Ser His Ser Leu Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asp Gly lie Trp Ser Thr Asp lie Leu Lys 85 90 95 Asp Gln Lys Glu Pro Lys Asn Lys Thr Phe Leu Arg Cys Glu Ala Lys 100 105 110 Asn Tyr Ser Gly Arg Phe Thr Cys Trp Trp Leu Thr Thr lie Ser Thr 115 120 125 Asp Leu Thr Phe Ser Val Lys Ser Ser Arg Gly Ser Ser Asp Pro Gln 130 135 140 Gly Val Thr Cys Gly Ala Ala Thr Leu Ser Ala Glu Arg Val Arg Gly 145 150 155 160 Asp Asn Lys Glu Tyr Glu Tyr Ser Val Glu Cys Gln Glu Asp Ser Ala 165 170 175 Cys Pro Ala Ala Glu Glu Ser Leu Pro lie Glu Val Met Val Asp Ala 180 185 190 Val His Lys Leu Lys Tyr Glu Asn Tyr Thr Ser Ser Phe Phe lie Arg 195 200 205 Asp lie lie Lys Pro Asp Pro Pro Lys Asn Leu Gln Leu Lys Pro Leu 210 215 220 Lys Asn Ser Arg Gin Val Glu Val Ser Trp Glu Tyr Pro Asp Thr Trp 225 230 235 240 Ser Thr Pro His Ser Tyr Phe Ser Leu Thr Phe Cys Val Gin Val Gin 245 250 255 Gly Lys Ser Lys Arg Glu Lys Lys Asp Arg Val Phe Thr Asp Lys Thr 260 265 270 Ser Ala Thr Val He Cys Arg Lys Asn Ala Ser He Ser Val Arg Ala 275 280 285 Gln Asp Arg Tyr Tyr Ser Ser Ser Trp Ser Glu Trp Ala Ser Val Pro 290 295 300 Cys Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 305 310 315 320 Ser Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys 325 330 335 Leu His His Ser Gin Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gin 340 345 350 Lys Ala Arg Gin Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu He 355 360 365 Asp His Glu Asp He Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys 370 375 380 Leu Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu 385 390 395 400 Thr Ser Phe Ile Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser 405 410 415 Phe Met Met Ala Leu Cys Leu Ser Ser Ile Tyr Glu Asp Leu Lys Met 420 425 430 Tyr Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro 435 440 445 Lys Arg Gln Ile Phe Leu Asp Gln Asn Met Leu Ala Val Ile Asp Glu 450 455 460 Leu Met Gln Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gln Lys Ser 465 470 475 480 Ser Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys Ile Lys Leu Cys Ile 485 490 495 Leu Leu His Ala Phe Arg Ile Arg Ala Val Thr Ile Asp Arg Val Met 500 505 510 Ser Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 515 520 525 Gly Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln 530 535 540 Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly 545 550 555 560 Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys 565 570 575 Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu 580 585 590 Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser 595 600 605 Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val 610 615 620 Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr 625 630 635 640 Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr 645 650 655 Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Gly Gly Gly Gly Ser Gly 660 665 670 Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Leu Glu Ser 675 680 685 Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala 690 695 700 Ala Ser Gly Phe Thr Phe Ser Leu Phe Thr Met Ser Trp Val Arg Gln 705 710 715 720 Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ala Ile Ser Gly Ser Gly 725 730 735 Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser 740 745 750 Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg 755 760 765 Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Lys Ser Thr His Leu Tyr 770 775 780 Leu Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly 785 790 795 800 Gly Ser Gly Gly Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser 805 810 815 Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser 820 825 830 Val Ser Met Pro Phe Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala 835 840 845 Pro Arg Leu Leu lie Tyr Gly Ala Ser Ser Arg Ala Thr Gly lie Pro 850 855 860 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie 865 870 875 880 Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Met 885 890 895 Arg Gly Arg Pro Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 900 905 910 Ser Ser Ser Ser Gly Ser Ser Ser Ser Gly Ser Ser Ser Ser Gly Glu 915 920 925 Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly Ser 930 935 940 Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Leu Phe Thr 945 950 955 960 Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val Ser 965 970 975 Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys 980 985 990 Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 995 1000 1005 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 1010 1015 1020 Ala Lys Ser Thr His Leu Tyr Leu Phe Asp Tyr Trp Gly Gln Gly 1025 1030 1035 Thr Leu Val Thr Val Ser Ser Gly Gly Ser Gly Gly Glu Ile Val 1040 1045 1050 Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly Glu Arg 1055 1060 1065 Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Met Pro Phe 1070 1075 1080 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 1085 1090 1095 Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe 1100 1105 1110 Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg 1115 1120 1125 Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Met Arg 1130 1135 1140 Gly Arg Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 1145 1150 1155 Ser Ser Ser Ser Gly Ser Ser Ser Ser Gly Ser Ser Ser Ser Gly 1160 1165 1170 Ala Pro Ala Arg Ser Pro Ser Pro Ser Thr Gin Pro Trp Glu His 1175 1180 1185 Val Asn Ala lie Gin Glu Ala Arg Arg Leu Leu Asn Leu Ser Arg 1190 1195 1200 Asp Thr Ala Ala Glu Met Asn Glu Thr Val Glu Val lie Ser Glu 1205 1210 1215 Met Phe Asp Leu Gin Glu Pro Thr Cys Leu Gin Thr Arg Leu Glu 1220 1225 1230 Leu Tyr Lys Gin Gly Leu Arg Gly Ser Leu Thr Lys Leu Lys Gly 1235 1240 1245 Pro Leu Thr Met Met Ala Ser His Tyr Lys Gin His Cys Pro Pro 1250 1255 1260 Thr Pro Glu Thr Ser Cys Ala Thr Gin lie lie Thr Phe Glu Ser 1265 1270 1275 Phe Lys Glu Asn Leu Lys Asp Phe Leu Leu Val lie Pro Phe Asp 1280 1285 1290 Cys Trp Glu Pro Val Gin Glu 1295 1300 <210> 52 <211> 559 <212> PRT <213> Artificial Sequence <220> <223> Second polypeptide chain of mIL12bscF8-scF8IL12aIL2GMCSF (murine mIL12bscF8 chain) <400> 52 Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr 1 5 10 15 Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg His Gly Val Ile Gly 35 40 45 Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu Phe Leu Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu Lys 85 90 95 Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser 100 105 110 Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu Lys 115 120 125 Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val Thr 130 135 140 Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln Arg 145 150 155 160 Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys Pro 165 170 175 Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala Leu Glu Ala Arg Gln 180 185 190 Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe Ile Arg Asp Ile 195 200 205 Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met Lys Pro Leu Lys Asn 210 215 220 Ser Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro 225 230 235 240 His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg Ile Gln Arg Lys Lys 245 250 255 Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gln Lys Gly Ala Phe 260 265 270 Leu Val Glu Lys Thr Ser Thr Glu Val Gln Cys Lys Gly Gly Asn Val 275 280 285 Cys Val Gin Ala Gin Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp 290 295 300 Ala Cys Val Pro Cys Arg Val Arg Ser Gly Ser Ala Asp Gly Gly Glu 305 310 315 320 Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly Ser 325 330 335 Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Leu Phe Thr 340 345 350 Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val Ser 355 360 365 Ala He Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys 370 375 380 Gly Arg Phe Thr He Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 385 390 395 400 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 405 410 415 Lys Ser Thr His Leu Tyr Leu Phe Asp Tyr Trp Gly Gin Gly Thr Leu 420 425 430 Val Thr Val Ser Ser Ser Ser Gly Asp Gly Ser Ser Gly Gly Ser Gly 435 440 445 Gly Ala Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu 450 455 460 Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val 465 470 475 480 Ser Met Pro Phe Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro 485 490 495 Arg Leu Leu Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp 500 505 510 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 515 520 525 Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Met Arg 530 535 540 Gly Arg Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 545 550 555 <210> 53 <211> 742 <212> PRT <213> Artificial Sequence <220> <223> First polypeptide chain of mIL12bscF8-scF8IL12aIL2GMCSF (murine mscF8IL12aIL2GMCSF chain) <400> 53 Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Leu Phe 20 25 30 Thr Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala He Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Ser Thr His Leu Tyr Leu Phe Asp Tyr Trp Gly Gin Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ser Ser Gly Asp Gly Ser Ser Gly Gly Ser 115 120 125 Gly Gly Ala Ser Glu He Val Leu Thr Gin Ser Pro Gly Thr Leu Ser 130 135 140 Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser 145 150 155 160 Val Ser Met Pro Phe Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala 165 170 175 Pro Arg Leu Leu lie Tyr Gly Ala Ser Ser Arg Ala Thr Gly lie Pro 180 185 190 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie 195 200 205 Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Met 210 215 220 Arg Gly Arg Pro Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 225 230 235 240 Gly Ser Ala Asp Gly Gly Arg Val lie Pro Val Ser Gly Pro Ala Arg 245 250 255 Cys Leu Ser Gin Ser Arg Asn Leu Leu Lys Thr Thr Asp Asp Met Val 260 265 270 Lys Thr Ala Arg Glu Lys Leu Lys His Tyr Ser Cys Thr Ala Glu Asp 275 280 285 Ile Asp His Glu Asp lie Thr Arg Asp Gin Thr Ser Thr Leu Lys Thr 290 295 300 Cys Leu Pro Leu Glu Leu His Lys Asn Glu Ser Cys Leu Ala Thr Arg 305 310 315 320 Glu Thr Ser Ser Thr Thr Arg Gly Ser Cys Leu Pro Pro Gln Lys Thr 325 330 335 Ser Leu Met Met Thr Leu Cys Leu Gly Ser Ile Tyr Glu Asp Leu Lys 340 345 350 Met Tyr Gln Thr Glu Phe Gln Ala Ile Asn Ala Ala Leu Gln Asn His 355 360 365 Asn His Gln Gln Ile Ile Leu Asp Lys Gly Met Leu Val Ala Ile Asp 370 375 380 Glu Leu Met Gln Ser Leu Asn His Asn Gly Glu Thr Leu Arg Gln Lys 385 390 395 400 Pro Pro Val Gly Glu Ala Asp Pro Tyr Arg Val Lys Met Lys Leu Cys 405 410 415 Ile Leu Leu His Ala Phe Ser Thr Arg Val Val Thr Ile Asn Arg Val 420 425 430 Met Gly Tyr Leu Ser Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly 435 440 445 Ser Gly Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Ser Ser Ser 450 455 460 Thr Ala Glu Ala Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln 465 470 475 480 His Leu Glu Gin Leu Leu Met Asp Leu Gin Glu Leu Leu Ser Arg Met 485 490 495 Glu Asn Tyr Arg Asn Leu Lys Leu Pro Arg Met Leu Thr Phe Lys Phe 500 505 510 Tyr Leu Pro Lys Gin Ala Thr Glu Leu Lys Asp Leu Gin Cys Leu Glu 515 520 525 Asp Glu Leu Gly Pro Leu Arg His Val Leu Asp Leu Thr Gin Ser Lys 530 535 540 Ser Phe Gin Leu Glu Asp Ala Glu Asn Phe He Ser Asn He Arg Val 545 550 555 560 Thr Val Val Lys Leu Lys Gly Ser Asp Asn Thr Phe Glu Cys Gin Phe 565 570 575 Asp Asp Glu Ser Ala Thr Val Val Asp Phe Leu Arg Arg Trp He Ala 580 585 590 Phe Cys Gin Ser He He Ser Thr Ser Pro Gin Gly Gly Gly Gly Ser 595 600 605 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Arg Ser Pro 610 615 620 He Thr Val Thr Arg Pro Trp Lys His Val Glu Ala He Lys Glu Ala 625 630 635 640 Leu Asn Leu Leu Asp Asp Met Pro Val Thr Leu Asn Glu Glu Val Glu 645 650 655 Val Val Ser Asn Glu Phe Ser Phe Lys Lys Leu Thr Cys Val Gln Thr 660 665 670 Arg Leu Lys Ile Phe Glu Gin Gly Leu Arg Gly Asn Phe Thr Lys Leu 675 680 685 Lys Gly Ala Leu Asn Met Thr Ala Ser Tyr Tyr Gin Thr Tyr Cys Pro 690 695 700 Pro Thr Pro Glu Thr Asp Cys Glu Thr Gin Val Thr Thr Tyr Ala Asp 705 710 715 720 Phe He Asp Ser Leu Lys Thr Phe Leu Thr Asp He Pro Phe Glu Cys 725 730 735 Lys Lys Pro Gly Gin Lys 740 <210> 54 <211> 551 <212> PRT <213> Artificial Sequence <220> <223> Second Polypeptide Chain of hIL12bscNHS76-scNHS76IL12aIL2GMCSF (human hIL12bscNHS76 chain ) <400> 54 Ile Trp Glu Leu Lys Lys Asp Val Tyr Val Val Glu Leu Asp Trp Tyr 1 5 10 15 Pro Asp Ala Pro Gly Glu Met Val Val Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Gly Ile Thr Trp Thr Leu Asp Gln Ser Ser Glu Val Leu Gly 35 40 45 Ser Gly Lys Thr Leu Thr Ile Gln Val Lys Glu Phe Gly Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Val Leu Ser His Ser Leu Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asp Gly Ile Trp Ser Thr Asp Ile Leu Lys 85 90 95 Asp Gln Lys Glu Pro Lys Asn Lys Thr Phe Leu Arg Cys Glu Ala Lys 100 105 110 Asn Tyr Ser Gly Arg Phe Thr Cys Trp Trp Leu Thr Thr Ile Ser Thr 115 120 125 Asp Leu Thr Phe Ser Val Lys Ser Ser Arg Gly Ser Ser Asp Pro Gln 130 135 140 Gly Val Thr Cys Gly Ala Ala Thr Leu Ser Ala Glu Arg Val Arg Gly 145 150 155 160 Asp Asn Lys Glu Tyr Glu Tyr Ser Val Glu Cys Gln Glu Asp Ser Ala 165 170 175 Cys Pro Ala Ala Glu Glu Ser Leu Pro Ile Glu Val Met Val Asp Ala 180 185 190 Val His Lys Leu Lys Tyr Glu Asn Tyr Thr Ser Ser Phe Phe Ile Arg 195 200 205 Asp Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Leu Lys Pro Leu 210 215 220 Lys Asn Ser Arg Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Thr Trp 225 230 235 240 Ser Thr Pro His Ser Tyr Phe Ser Leu Thr Phe Cys Val Gln Val Gln 245 250 255 Gly Lys Ser Lys Arg Glu Lys Lys Asp Arg Val Phe Thr Asp Lys Thr 260 265 270 Ser Ala Thr Val Ile Cys Arg Lys Asn Ala Ser Ile Ser Val Arg Ala 275 280 285 Gln Asp Arg Tyr Tyr Ser Ser Ser Trp Ser Glu Trp Ala Ser Val Pro 290 295 300 Cys Ser Gly Ser Ala Asp Gly Gly Gln Val Gln Leu Gln Glu Ser Gly 305 310 315 320 Pro Gly Leu Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Ala Val 325 330 335 Ser Gly Tyr Ser lie Ser Ser Gly Tyr Tyr Trp Gly Trp lie Arg Gin 340 345 350 Pro Pro Gly Lys Gly Leu Glu Trp lie Gly Ser lie Tyr His Ser Gly 355 360 365 Ser Thr Tyr Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr lie Ser Val 370 375 380 Asp Thr Ser Lys Asn Gin Phe Ser Leu Lys Leu Ser Ser Val Thr Ala 385 390 395 400 Ala Asp Thr Ala Val Tyr Tyr Cys Ala Arg Gly Lys Trp Ser Lys Phe 405 410 415 Asp Tyr Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser Ser Ser Gly 420 425 430 Asp Gly Ser Ser Gly Gly Ser Gly Gly Ala Ser Ser Ser Glu Leu Thr 435 440 445 Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gin Thr Val Arg lie Thr 450 455 460 Cys Gin Gly Asp Ser Leu Arg Ser Tyr Tyr Ala Ser Trp Tyr Gin Gin 465 470 475 480 Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr Gly Lys Asn Asn Arg 485 490 495 Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Ser Ser Gly Asn Thr 500 505 510 Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu Asp Glu Ala Asp Tyr 515 520 525 Tyr Cys Asn Ser Arg Asp Ser Ser Gly Asn His Val Val Phe Gly Gly 530 535 540 Gly Thr Lys Leu Thr Val Leu 545 550 <210> 55 <211> 732 <212> PRT <213> Artificial Sequence (Artificial Sequence) <220> <223> First polypeptide chain of hIL12bscNHS76-scNHS76IL12aIL2GMCSF (human hscNHS76IL12aIL 2GMCSF chain) <400> 55 Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Tyr Ser Ile Ser Ser Gly 20 25 30 Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 Ile Gly Ser lie Tyr His Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu 50 55 60 Lys Ser Arg Val Thr lie Ser Val Asp Thr Ser Lys Asn Gin Phe Ser 65 70 75 80 Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Lys Trp Ser Lys Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110 Val Thr Val Ser Ser Ser Ser Gly Asp Gly Ser Ser Gly Gly Ser Gly 115 120 125 Gly Ala Ser Ser Ser Glu Leu Thr Gin Asp Pro Ala Val Ser Val Ala 130 135 140 Leu Gly Gin Thr Val Arg lie Thr Cys Gin Gly Asp Ser Leu Arg Ser 145 150 155 160 Tyr Tyr Ala Ser Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Val Leu 165 170 175 Val lie Tyr Gly Lys Asn Asn Arg Pro Ser Gly lie Pro Asp Arg Phe 180 185 190 Ser Gly Ser Ser Ser Gly Asn Thr Ala Ser Leu Thr lie Thr Gly Ala 195 200 205 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Asn Ser Arg Asp Ser Ser 210 215 220 Gly Asn His Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 225 230 235 240 Ser Ala Asp Gly Gly Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly 245 250 255 Met Phe Pro Cys Leu His His Ser Gln Asn Leu Leu Arg Ala Val Ser 260 265 270 Asn Met Leu Gln Lys Ala Arg Gln Thr Leu Glu Phe Tyr Pro Cys Thr 275 280 285 Ser Glu Glu Ile Asp His Glu Asp Ile Thr Lys Asp Lys Thr Ser Thr 290 295 300 Val Glu Ala Cys Leu Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu 305 310 315 320 Asn Ser Arg Glu Thr Ser Phe Ile Thr Asn Gly Ser Cys Leu Ala Ser 325 330 335 Arg Lys Thr Ser Phe Met Met Ala Leu Cys Leu Ser Ser Ile Tyr Glu 340 345 350 Asp Leu Lys Met Tyr Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu 355 360 365 Leu Met Asp Pro Lys Arg Gin He Phe Leu Asp Gin Asn Met Leu Ala 370 375 380 Val He Asp Glu Leu Met Gin Ala Leu Asn Phe Asn Ser Glu Thr Val 385 390 395 400 Pro Gin Lys Ser Ser Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys He 405 410 415 Lys Leu Cys He Leu Leu His Ala Phe Arg He Arg Ala Val Thr He 420 425 430 Asp Arg Val Met Ser Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly 435 440 445 Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr 450 455 460 Lys Lys Thr Gin Leu Gin Leu Glu His Leu Leu Leu Asp Leu Gin Met 465 470 475 480 He Leu Asn Gly He Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met 485 490 495 Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His 500 505 510 Leu Gin Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn 515 520 525 Leu Ala Gin Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu lie Ser 530 535 540 Asn lie Asn Val lie Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe 545 550 555 560 Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr lie Val Glu Phe Leu Asn 565 570 575 Arg Trp lie Thr Phe Cys Gin Ser lie lie Ser Thr Leu Thr Gly Gly 580 585 590 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Ala 595 600 605 Arg Ser Pro Ser Pro Ser Thr Gin Pro Trp Glu His Val Asn Ala lie 610 615 620 Gln Glu Ala Arg Arg Leu Leu Asn Leu Ser Arg Asp Thr Ala Ala Glu 625 630 635 640 Met Asn Glu Thr Val Glu Val lie Ser Glu Met Phe Asp Leu Gin Glu 645 650 655 Pro Thr Cys Leu Gin Thr Arg Leu Glu Leu Tyr Lys Gin Gly Leu Arg 660 665 670 Gly Ser Leu Thr Lys Leu Lys Gly Pro Leu Thr Met Met Ala Ser His 675 680 685 Tyr Lys Gln His Cys Pro Pro Thr Pro Glu Thr Ser Cys Ala Thr Gln 690 695 700 Ile Ile Thr Phe Glu Ser Phe Lys Glu Asn Leu Lys Asp Phe Leu Leu 705 710 715 720 Val Ile Pro Phe Asp Cys Trp Glu Pro Val Gln Glu 725 730 <210> 56 <211> 357 <212> PRT <213> Artificial Sequence <220> <223> First polypeptide chain of mIL12aIL2IL12bGMCSF <400> 56 Arg Val Ile Pro Val Ser Gly Pro Ala Arg Cys Leu Ser Gln Ser Arg 1 5 10 15 Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys Thr Ala Arg Glu Lys 20 25 30 Leu Lys His Tyr Ser Cys Thr Ala Glu Asp Ile Asp His Glu Asp Ile 35 40 45 Thr Arg Asp Gln Thr Ser Thr Leu Lys Thr Cys Leu Pro Leu Glu Leu 50 55 60 His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr Ser Ser Thr Thr 65 70 75 80 Arg Gly Ser Cys Leu Pro Pro Gln Lys Thr Ser Leu Met Met Thr Leu 85 90 95 Cys Leu Gly Ser Ile Tyr Glu Asp Leu Lys Met Tyr Gln Thr Glu Phe 100 105 110 Gln Ala Ile Asn Ala Ala Leu Gln Asn His Asn His Gln Gln Ile Ile 115 120 125 Leu Asp Lys Gly Met Leu Val Ala Ile Asp Glu Leu Met Gln Ser Leu 130 135 140 Asn His Asn Gly Glu Thr Leu Arg Gln Lys Pro Pro Val Gly Glu Ala 145 150 155 160 Asp Pro Tyr Arg Val Lys Met Lys Leu Cys Ile Leu Leu His Ala Phe 165 170 175 Ser Thr Arg Val Val Thr Ile Asn Arg Val Met Gly Tyr Leu Ser Ser 180 185 190 Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 195 200 205 Ala Pro Thr Ser Ser Ser Thr Ser Ser Ser Thr Ala Glu Ala Gln Gln 210 215 220 Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln His Leu Glu Gln Leu Leu 225 230 235 240 Met Asp Leu Gin Glu Leu Leu Ser Arg Met Glu Asn Tyr Arg Asn Leu 245 250 255 Lys Leu Pro Arg Met Leu Thr Phe Lys Phe Tyr Leu Pro Lys Gin Ala 260 265 270 Thr Glu Leu Lys Asp Leu Gin Cys Leu Glu Asp Glu Leu Gly Pro Leu 275 280 285 Arg His Val Leu Asp Leu Thr Gin Ser Lys Ser Phe Gin Leu Glu Asp 290 295 300 Ala Glu Asn Phe He Ser Asn He Arg Val Thr Val Val Lys Leu Lys 305 310 315 320 Gly Ser Asp Asn Thr Phe Glu Cys Gin Phe Asp Asp Glu Ser Ala Thr 325 330 335 Val Val Asp Phe Leu Arg Arg Trp He Ala Phe Cys Gin Ser He He 340 345 350 Ser Thr Ser Pro Gin 355 <210> 57 <211> 452 <212> PRT <213> Artificial Sequence <220> <223> Second Polypeptide Chain of mIL12aIL2IL12bGMCSF <400> 57 Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr 1 5 10 15 Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg His Gly Val Ile Gly 35 40 45 Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu Phe Leu Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu Lys 85 90 95 Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser 100 105 110 Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu Lys 115 120 125 Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val Thr 130 135 140 Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln Arg 145 150 155 160 Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gin Glu Asp Val Thr Cys Pro 165 170 175 Thr Ala Glu Glu Thr Leu Pro lie Glu Leu Ala Leu Glu Ala Arg Gin 180 185 190 Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe lie Arg Asp lie 195 200 205 lie Lys Pro Asp Pro Pro Lys Asn Leu Gin Met Lys Pro Leu Lys Asn 210 215 220 Ser Gin Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro 225 230 235 240 His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg lie Gin Arg Lys Lys 245 250 255 Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gin Lys Gly Ala Phe 260 265 270 Leu Val Glu Lys Thr Ser Thr Glu Val Gin Cys Lys Gly Gly Asn Val 275 280 285 Cys Val Gin Ala Gin Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp 290 295 300 Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly Gly Gly Ser Gly Gly 305 310 315 320 Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Arg Ser Pro Ile Thr 325 330 335 Val Thr Arg Pro Trp Lys His Val Glu Ala Ile Lys Glu Ala Leu Asn 340 345 350 Leu Leu Asp Asp Met Pro Val Thr Leu Asn Glu Glu Val Glu Val Val 355 360 365 Ser Asn Glu Phe Ser Phe Lys Lys Leu Thr Cys Val Gln Thr Arg Leu 370 375 380 Lys Ile Phe Glu Gln Gly Leu Arg Gly Asn Phe Thr Lys Leu Lys Gly 385 390 395 400 Ala Leu Asn Met Thr Ala Ser Tyr Tyr Gln Thr Tyr Cys Pro Pro Thr 405 410 415 Pro Glu Thr Asp Cys Glu Thr Gln Val Thr Thr Tyr Ala Asp Phe Ile 420 425 430 Asp Ser Leu Lys Thr Phe Leu Thr Asp Ile Pro Phe Glu Cys Lys Lys 435 440 445 Pro Gly Gln Lys 450 <210> 58 <211> 337 <212> PRT <213> Artificial Sequence <220> <223> First polypeptide chain of mIL12aIL7IL12bGMCSF <400> 58 Arg Val lie Pro Val Ser Gly Pro Ala Arg Cys Leu Ser Gin Ser Arg 1 5 10 15 Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys Thr Ala Arg Glu Lys 20 25 30 Leu Lys His Tyr Ser Cys Thr Ala Glu Asp lie Asp His Glu Asp lie 35 40 45 Thr Arg Asp Gin Thr Ser Thr Leu Lys Thr Cys Leu Pro Leu Glu Leu 50 55 60 His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr Ser Ser Thr Thr 65 70 75 80 Arg Gly Ser Cys Leu Pro Pro Gin Lys Thr Ser Leu Met Met Thr Leu 85 90 95 Cys Leu Gly Ser lie Tyr Glu Asp Leu Lys Met Tyr Gin Thr Glu Phe 100 105 110 Gln Ala lie Asn Ala Ala Leu Gin Asn His Asn His Gin Gin lie lie 115 120 125 Leu Asp Lys Gly Met Leu Val Ala lie Asp Glu Leu Met Gin Ser Leu 130 135 140 Asn His Asn Gly Glu Thr Leu Arg Gin Lys Pro Pro Val Gly Glu Ala 145 150 155 160 Asp Pro Tyr Arg Val Lys Met Lys Leu Cys Ile Leu Leu His Ala Phe 165 170 175 Ser Thr Arg Val Val Thr Ile Asn Arg Val Met Gly Tyr Leu Ser Ser 180 185 190 Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 195 200 205 Glu Cys His Ile Lys Asp Lys Glu Gly Lys Ala Tyr Glu Ser Val Leu 210 215 220 Met Ile Ser Ile Asp Glu Leu Asp Lys Met Thr Gly Thr Asp Ser Asn 225 230 235 240 Cys Pro Asn Asn Glu Pro Asn Phe Phe Arg Lys His Val Cys Asp Asp 245 250 255 Thr Lys Glu Ala Ala Phe Leu Asn Arg Ala Ala Arg Lys Leu Lys Gin 260 265 270 Phe Leu Lys Met Asn Ile Ser Glu Glu Phe Asn Val His Leu Leu Thr 275 280 285 Val Ser Gin Gly Thr Gin Thr Leu Val Asn Cys Thr Ser Lys Glu Glu 290 295 300 Lys Asn Val Lys Glu Gin Lys Lys Asn Asp Ala Cys Phe Leu Lys Arg 305 310 315 320 Leu Leu Arg Glu He Lys Thr Cys Trp Asn Lys He Leu Lys Gly Ser 325 330 335 He <210> 59 <211> 452 <212> PRT <213> Artificial Sequence <220> <223> Second Polypeptide Chain of mIL12aIL7IL12bGMCSF <400> 59 Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr 1 5 10 15 Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Asp He Thr Trp Thr Ser Asp Gin Arg His Gly Val He Gly 35 40 45 Ser Gly Lys Thr Leu Thr He Thr Val Lys Glu Phe Leu Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asn Gly He Trp Ser Thr Glu He Leu Lys 85 90 95 Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser 100 105 110 Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu Lys 115 120 125 Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val Thr 130 135 140 Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln Arg 145 150 155 160 Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys Pro 165 170 175 Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala Leu Glu Ala Arg Gln 180 185 190 Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe Ile Arg Asp Ile 195 200 205 Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met Lys Pro Leu Lys Asn 210 215 220 Ser Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro 225 230 235 240 His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg Ile Gln Arg Lys Lys 245 250 255 Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gln Lys Gly Ala Phe 260 265 270 Leu Val Glu Lys Thr Ser Thr Glu Val Gln Cys Lys Gly Gly Asn Val 275 280 285 Cys Val Gln Ala Gln Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp 290 295 300 Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly Gly Gly Ser Gly Gly 305 310 315 320 Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Arg Ser Pro Ile Thr 325 330 335 Val Thr Arg Pro Trp Lys His Val Glu Ala Ile Lys Glu Ala Leu Asn 340 345 350 Leu Leu Asp Asp Met Pro Val Thr Leu Asn Glu Glu Val Glu Val Val 355 360 365 Ser Asn Glu Phe Ser Phe Lys Lys Leu Thr Cys Val Gln Thr Arg Leu 370 375 380 Lys Ile Phe Glu Gln Gly Leu Arg Gly Asn Phe Thr Lys Leu Lys Gly 385 390 395 400 Ala Leu Asn Met Thr Ala Ser Tyr Tyr Gln Thr Tyr Cys Pro Pro Thr 405 410 415 Pro Glu Thr Asp Cys Glu Thr Gln Val Thr Thr Tyr Ala Asp Phe Ile 420 425 430 Asp Ser Leu Lys Thr Phe Leu Thr Asp Ile Pro Phe Glu Cys Lys Lys 435 440 445 Pro Gly Gln Lys 450 <210> 60 <211> 337 <212> PRT <213> Artificial Sequence <220> <223> First polypeptide chain of mIL12aIL21IL12bGMCSF <400> 60 Arg Val Ile Pro Val Ser Gly Pro Ala Arg Cys Leu Ser Gln Ser Arg 1 5 10 15 Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys Thr Ala Arg Glu Lys 20 25 30 Leu Lys His Tyr Ser Cys Thr Ala Glu Asp Ile Asp His Glu Asp Ile 35 40 45 Thr Arg Asp Gln Thr Ser Thr Leu Lys Thr Cys Leu Pro Leu Glu Leu 50 55 60 His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr Ser Ser Thr Thr 65 70 75 80 Arg Gly Ser Cys Leu Pro Pro Gin Lys Thr Ser Leu Met Met Thr Leu 85 90 95 Cys Leu Gly Ser Ile Tyr Gin Asp Leu Lys Met Tyr Gin Thr Gin Phe 100 105 110 Gln Ala Ile Asn Ala Ala Leu Gin Asn His Asn His Gin Gin Ile Ile 115 120 125 Leu Asp Lys Gly Met Leu Val Ala Ile Asp Gin Leu Met Gin Ser Leu 130 135 140 Asn His Asn Gly Gin Thr Leu Arg Gin Lys Pro Pro Val Gin Gin Ala 145 150 155 160 Asp Pro Tyr Arg Val Lys Met Lys Leu Cys Ile Leu Leu His Ala Phe 165 170 175 Ser Thr Arg Val Val Thr Ile Asn Arg Val Met Gin Tyr Leu Ser Ser 180 185 190 Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 195 200 205 His Lys Ser Ser Pro Gin Gly Pro Asp Arg Leu Leu Ile Arg Leu Arg 210 215 220 His Leu Ile Asp Ile Val Gin Gin Leu Lys Ile Tyr Gin Asn Asp Leu 225 230 235 240 Asp Pro Glu Leu Leu Ser Ala Pro Gin Asp Val Lys Gly His Cys Glu 245 250 255 His Ala Ala Phe Ala Cys Phe Gin Lys Ala Lys Leu Lys Pro Ser Asn 260 265 270 Pro Gly Asn Asn Lys Thr Phe He He Asp Leu Val Ala Gin Leu Arg 275 280 285 Arg Arg Leu Pro Ala Arg Arg Gly Gly Lys Lys Gin Lys His He Ala 290 295 300 Lys Cys Pro Ser Cys Asp Ser Tyr Gin Lys Arg Thr Pro Lys Glu Phe 305 310 315 320 Leu Gin Arg Leu Lys Trp Leu Leu Gin Lys Met He His Gin His Leu 325 330 335 Ser <210> 61 <211> 452 <212> PRT <213> Artificial Sequence <220> <223> Second Polypeptide Chain of mIL12aIL21IL12bGMCSF <400> 61 Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr 1 5 10 15 Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Asp lie Thr Trp Thr Ser Asp Gin Arg His Gly Val lie Gly 35 40 45 Ser Gly Lys Thr Leu Thr lie Thr Val Lys Glu Phe Leu Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asn Gly lie Trp Ser Thr Glu lie Leu Lys 85 90 95 Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser 100 105 110 Gly Arg Phe Thr Cys Ser Trp Leu Val Gin Arg Asn Met Asp Leu Lys 115 120 125 Phe Asn lie Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val Thr 130 135 140 Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gin Arg 145 150 155 160 Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gin Glu Asp Val Thr Cys Pro 165 170 175 Thr Ala Glu Glu Thr Leu Pro lie Glu Leu Ala Leu Glu Ala Arg Gin 180 185 190 Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe Ile Arg Asp Ile 195 200 205 Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met Lys Pro Leu Lys Asn 210 215 220 Ser Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro 225 230 235 240 His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg Ile Gln Arg Lys Lys 245 250 255 Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gln Lys Gly Ala Phe 260 265 270 Leu Val Glu Lys Thr Ser Thr Glu Val Gln Cys Lys Gly Gly Asn Val 275 280 285 Cys Val Gln Ala Gln Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp 290 295 300 Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly Gly Gly Ser Gly Gly 305 310 315 320 Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Arg Ser Pro Ile Thr 325 330 335 Val Thr Arg Pro Trp Lys His Val Glu Ala Ile Lys Glu Ala Leu Asn 340 345 350 Leu Leu Asp Asp Met Pro Val Thr Leu Asn Glu Glu Val Glu Val Val 355 360 365 Ser Asn Glu Phe Ser Phe Lys Lys Leu Thr Cys Val Gln Thr Arg Leu 370 375 380 Lys Ile Phe Glu Gln Gly Leu Arg Gly Asn Phe Thr Lys Leu Lys Gly 385 390 395 400 Ala Leu Asn Met Thr Ala Ser Tyr Tyr Gln Thr Tyr Cys Pro Pro Thr 405 410 415 Pro Glu Thr Asp Cys Glu Thr Gln Val Thr Thr Tyr Ala Asp Phe Ile 420 425 430 Asp Ser Leu Lys Thr Phe Leu Thr Asp Ile Pro Phe Glu Cys Lys Lys 435 440 445 Pro Gly Gln Lys 450 <210> 62 <211> 345 <212> PRT <213> Artificial Sequence <220> <223> First polypeptide chain of hIL12aIL2IL12bGMCSF <400> 62 Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys Leu 1 5 10 15 His His Ser Gin Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gin Lys 20 25 30 Ala Arg Gin Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu He Asp 35 40 45 His Glu Asp He Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys Leu 50 55 60 Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu Thr 65 70 75 80 Ser Phe He Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser Phe 85 90 95 Met Met Ala Leu Cys Leu Ser Ser He Tyr Glu Asp Leu Lys Met Tyr 100 105 110 Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro Lys 115 120 125 Arg Gin He Phe Leu Asp Gin Asn Met Leu Ala Val He Asp Glu Leu 130 135 140 Met Gin Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gin Lys Ser Ser 145 150 155 160 Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys He Lys Leu Cys He Leu 165 170 175 Leu His Ala Phe Arg Ile Arg Ala Val Thr Ile Asp Arg Val Met Ser 180 185 190 Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 195 200 205 Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu 210 215 220 Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile 225 230 235 240 Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe 245 250 255 Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu 260 265 270 Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys 275 280 285 Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile 290 295 300 Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala 305 310 315 320 Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe 325 330 335 Cys Gln Ser lie lie Ser Thr Leu Thr 340 345 <210> 63 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> Second polypeptide chain of hIL12aIL2IL12bGMCSF <400> 63 Ile Trp Glu Leu Lys Lys Asp Val Tyr Val Val Glu Leu Asp Trp Tyr 1 5 10 15 Pro Asp Ala Pro Gly Glu Met Val Val Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Gly lie Thr Trp Thr Leu Asp Gln Ser Ser Glu Val Leu Gly 35 40 45 Ser Gly Lys Thr Leu Thr lie Gin Val Lys Glu Phe Gly Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Val Leu Ser His Ser Leu Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asp Gly lie Trp Ser Thr Asp lie Leu Lys 85 90 95 Asp Gin Lys Glu Pro Lys Asn Lys Thr Phe Leu Arg Cys Glu Ala Lys 100 105 110 Asn Tyr Ser Gly Arg Phe Thr Cys Trp Trp Leu Thr Thr Ile Ser Thr 115 120 125 Asp Leu Thr Phe Ser Val Lys Ser Ser Arg Gly Ser Ser Asp Pro Gln 130 135 140 Gly Val Thr Cys Gly Ala Ala Thr Leu Ser Ala Glu Arg Val Arg Gly 145 150 155 160 Asp Asn Lys Glu Tyr Glu Tyr Ser Val Glu Cys Gln Glu Asp Ser Ala 165 170 175 Cys Pro Ala Ala Glu Glu Ser Leu Pro Ile Glu Val Met Val Asp Ala 180 185 190 Val His Lys Leu Lys Tyr Glu Asn Tyr Thr Ser Ser Phe Phe Ile Arg 195 200 205 Asp Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Leu Lys Pro Leu 210 215 220 Lys Asn Ser Arg Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Thr Trp 225 230 235 240 Ser Thr Pro His Ser Tyr Phe Ser Leu Thr Phe Cys Val Gln Val Gln 245 250 255 Gly Lys Ser Lys Arg Glu Lys Lys Asp Arg Val Phe Thr Asp Lys Thr 260 265 270 Ser Ala Thr Val lie Cys Arg Lys Asn Ala Ser lie Ser Val Arg Ala 275 280 285 Gln Asp Arg Tyr Tyr Ser Ser Ser Trp Ser Glu Trp Ala Ser Val Pro 290 295 300 Cys Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 305 310 315 320 Ser Ala Pro Ala Arg Ser Pro Ser Pro Ser Thr Gin Pro Trp Glu His 325 330 335 Val Asn Ala lie Gin Glu Ala Arg Arg Leu Leu Asn Leu Ser Arg Asp 340 345 350 Thr Ala Ala Glu Met Asn Glu Thr Val Glu Val lie Ser Glu Met Phe 355 360 365 Asp Leu Gin Glu Pro Thr Cys Leu Gin Thr Arg Leu Glu Leu Tyr Lys 370 375 380 Gln Gly Leu Arg Gly Ser Leu Thr Lys Leu Lys Gly Pro Leu Thr Met 385 390 395 400 Met Ala Ser His Tyr Lys Gin His Cys Pro Pro Thr Pro Glu Thr Ser 405 410 415 Cys Ala Thr Gin lie lie Thr Phe Glu Ser Phe Lys Glu Asn Leu Lys 420 425 430 Asp Phe Leu Leu Val Ile Pro Phe Asp Cys Trp Glu Pro Val Gln Glu 435 440 445 <210> 64 <211> 364 <212> PRT <213> Artificial Sequence <220> <223> First polypeptide chain of hIL12aIL7IL12bGMCSF <400> 64 Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys Leu 1 5 10 15 His His Ser Gln Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gln Lys 20 25 30 Ala Arg Gln Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu Ile Asp 35 40 45 His Glu Asp Ile Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys Leu 50 55 60 Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu Thr 65 70 75 80 Ser Phe Ile Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser Phe 85 90 95 Met Met Ala Leu Cys Leu Ser Ser Ile Tyr Glu Asp Leu Lys Met Tyr 100 105 110 Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro Lys 115 120 125 Arg Gln Ile Phe Leu Asp Gln Asn Met Leu Ala Val Ile Asp Glu Leu 130 135 140 Met Gln Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gln Lys Ser Ser 145 150 155 160 Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys Ile Lys Leu Cys Ile Leu 165 170 175 Leu His Ala Phe Arg Ile Arg Ala Val Thr Ile Asp Arg Val Met Ser 180 185 190 Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 195 200 205 Gly Gly Gly Ser Asp Cys Asp Ile Glu Gly Lys Asp Gly Lys Gln Tyr 210 215 220 Glu Ser Val Leu Met Val Ser Ile Asp Gln Leu Leu Asp Ser Met Lys 225 230 235 240 Glu Ile Gly Ser Asn Cys Leu Asn Asn Glu Phe Asn Phe Phe Lys Arg 245 250 255 His Ile Cys Asp Ala Asn Lys Glu Gly Met Phe Leu Phe Arg Ala Ala 260 265 270 Arg Lys Leu Arg Gin Phe Leu Lys Met Asn Ser Thr Gly Asp Phe Asp 275 280 285 Leu His Leu Leu Lys Val Ser Glu Gly Thr Thr Ile Leu Leu Asn Cys 290 295 300 Thr Gly Gin Val Lys Gly Arg Lys Pro Ala Ala Leu Gly Glu Ala Gin 305 310 315 320 Pro Thr Lys Ser Leu Glu Glu Asn Lys Ser Leu Lys Glu Gin Lys Lys 325 330 335 Leu Asn Asp Leu Cys Phe Leu Lys Arg Leu Leu Gin Glu Ile Lys Thr 340 345 350 Cys Trp Asn Lys Ile Leu Met Gly Thr Lys Glu His 355 360 <210> 65 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> Second Polypeptide Chain of hIL12aIL7IL12bGMCSF <400> 65 Ile Trp Glu Leu Lys Lys Asp Val Tyr Val Val Glu Leu Asp Trp Tyr 1 5 10 15 Pro Asp Ala Pro Gly Glu Met Val Val Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Gly lie Thr Trp Thr Leu Asp Gin Ser Ser Glu Val Leu Gly 35 40 45 Ser Gly Lys Thr Leu Thr lie Gin Val Lys Glu Phe Gly Asp Ala Gly 50 55 60 Gln Tyr Thr Cy s His Lys Gly Gly Glu Val Leu Ser His Ser Leu Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asp Gly lie Trp Ser Thr Asp lie Leu Lys 85 90 95 Asp Gin Lys Glu Pro Lys Asn Lys Thr Phe Leu Arg Cys Glu Ala Lys 100 105 110 Asn Tyr Ser Gly Arg Phe Thr Cy s Trp Trp Leu Thr Thr lie Ser Thr 115 120 125 Asp Leu Thr Phe Ser Val Lys Ser Ser Arg Gly Ser Ser Asp Pro Gin 130 135 140 Gly Val Thr Cy s Gly Ala Ala Thr Leu Ser Ala Glu Arg Val Arg Gly 145 150 155 160 Asp Asn Lys Glu Tyr Glu Tyr Ser Val Glu Cys Gin Glu Asp Ser Ala 165 170 175 Cys Pro Ala Ala Glu Glu Ser Leu Pro lie Glu Val Met Val Asp Ala 180 185 190 Val His Lys Leu Lys Tyr Glu Asn Tyr Thr Ser Ser Phe Phe Ile Arg 195 200 205 Asp Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Leu Lys Pro Leu 210 215 220 Lys Asn Ser Arg Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Thr Trp 225 230 235 240 Ser Thr Pro His Ser Tyr Phe Ser Leu Thr Phe Cys Val Gln Val Gln 245 250 255 Gly Lys Ser Lys Arg Glu Lys Lys Asp Arg Val Phe Thr Asp Lys Thr 260 265 270 Ser Ala Thr Val Ile Cys Arg Lys Asn Ala Ser Ile Ser Val Arg Ala 275 280 285 Gln Asp Arg Tyr Tyr Ser Ser Ser Trp Ser Glu Trp Ala Ser Val Pro 290 295 300 Cys Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 305 310 315 320 Ser Ala Pro Ala Arg Ser Pro Ser Pro Ser Thr Gln Pro Trp Glu His 325 330 335 Val Asn Ala Ile Gln Glu Ala Arg Arg Leu Leu Asn Leu Ser Arg Asp 340 345 350 Thr Ala Ala Glu Met Asn Glu Thr Val Glu Val Ile Ser Glu Met Phe 355 360 365 Asp Leu Gln Glu Pro Thr Cys Leu Gln Thr Arg Leu Glu Leu Tyr Lys 370 375 380 Gln Gly Leu Arg Gly Ser Leu Thr Lys Leu Lys Gly Pro Leu Thr Met 385 390 395 400 Met Ala Ser His Tyr Lys Gln His Cys Pro Pro Thr Pro Glu Thr Ser 405 410 415 Cys Ala Thr Gln Ile Ile Thr Phe Glu Ser Phe Lys Glu Asn Leu Lys 420 425 430 Asp Phe Leu Leu Val Ile Pro Phe Asp Cys Trp Glu Pro Val Gln Glu 435 440 445 <210> 66 <211> 345 <212> PRT <213> Artificial Sequence <220> <223> First polypeptide chain of hIL12aIL21IL12bGMCSF <400> 66 Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys Leu 1 5 10 15 His His Ser Gln Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gln Lys 20 25 30 Ala Arg Gin Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu He Asp 35 40 45 His Glu Asp He Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys Leu 50 55 60 Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu Thr 65 70 75 80 Ser Phe He Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser Phe 85 90 95 Met Met Ala Leu Cys Leu Ser Ser He Tyr Glu Asp Leu Lys Met Tyr 100 105 110 Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro Lys 115 120 125 Arg Gin He Phe Leu Asp Gin Asn Met Leu Ala Val He Asp Glu Leu 130 135 140 Met Gin Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gin Lys Ser Ser 145 150 155 160 Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys He Lys Leu Cys He Leu 165 170 175 Leu His Ala Phe Arg He Arg Ala Val Thr He Asp Arg Val Met Ser 180 185 190 Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 195 200 205 Gly Gly Gly Ser Gln Gly Gln Asp Arg His Met Ile Arg Met Arg Gln 210 215 220 Leu Ile Asp Ile Val Asp Gln Leu Lys Asn Tyr Val Asn Asp Leu Val 225 230 235 240 Pro Glu Phe Leu Pro Ala Pro Glu Asp Val Glu Thr Asn Cys Glu Trp 245 250 255 Ser Ala Phe Ser Cys Phe Gln Lys Ala Gln Leu Lys Ser Ala Asn Thr 260 265 270 Gly Asn Asn Glu Arg Ile Ile Asn Val Ser Ile Lys Lys Leu Lys Arg 275 280 285 Lys Pro Pro Ser Thr Asn Ala Gly Arg Arg Gln Lys His Arg Leu Thr 290 295 300 Cys Pro Ser Cys Asp Ser Tyr Glu Lys Lys Pro Pro Lys Glu Phe Leu 305 310 315 320 Glu Arg Phe Lys Ser Leu Leu Gln Lys Met Ile His Gln His Leu Ser 325 330 335 Ser Arg Thr His Gly Ser Glu Asp Ser 340 345 <210> 67 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> Second polypeptide chain of hIL12aIL21IL12bGMCSF <400> 67 Ile Trp Glu Leu Lys Lys Asp Val Tyr Val Val Glu Leu Asp Trp Tyr 1 5 10 15 Pro Asp Ala Pro Gly Glu Met Val Val Leu Thr Cys Asp Thr Pro Glu 20 25 30 Glu Asp Gly Ile Thr Trp Thr Leu Asp Gln Ser Ser Glu Val Leu Gly 35 40 45 Ser Gly Lys Thr Leu Thr Ile Gln Val Lys Glu Phe Gly Asp Ala Gly 50 55 60 Gln Tyr Thr Cys His Lys Gly Gly Glu Val Leu Ser His Ser Leu Leu 65 70 75 80 Leu Leu His Lys Lys Glu Asp Gly Ile Trp Ser Thr Asp Ile Leu Lys 85 90 95 Asp Gln Lys Glu Pro Lys Asn Lys Thr Phe Leu Arg Cys Glu Ala Lys 100 105 110 Asn Tyr Ser Gly Arg Phe Thr Cys Trp Trp Leu Thr Thr Ile Ser Thr 115 120 125 Asp Leu Thr Phe Ser Val Lys Ser Ser Arg Gly Ser Ser Asp Pro Gin 130 135 140 Gly Val Thr Cys Gly Ala Ala Thr Leu Ser Ala Glu Arg Val Arg Gly 145 150 155 160 Asp Asn Lys Glu Tyr Glu Tyr Ser Val Glu Cys Gin Glu Asp Ser Ala 165 170 175 Cys Pro Ala Ala Glu Glu Ser Leu Pro Ile Glu Val Met Val Asp Ala 180 185 190 Val His Lys Leu Lys Tyr Glu Asn Tyr Thr Ser Ser Phe Phe Ile Arg 195 200 205 Asp Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gin Leu Lys Pro Leu 210 215 220 Lys Asn Ser Arg Gin Val Glu Val Ser Trp Glu Tyr Pro Asp Thr Trp 225 230 235 240 Ser Thr Pro His Ser Tyr Phe Ser Leu Thr Phe Cys Val Gin Val Gin 245 250 255 Gly Lys Ser Lys Arg Glu Lys Lys Asp Arg Val Phe Thr Asp Lys Thr 260 265 270 Ser Ala Thr Val Ile Cys Arg Lys Asn Ala Ser Ile Ser Val Arg Ala 275 280 285​​​​​​​​​​​​​​​​​​​ Gln Asp Arg Tyr Tyr Ser Ser Ser Trp Ser Glu Trp Ala Ser Val Pro 290 295 300 Cys Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 305 310 315 320 Ser Ala Pro Ala Arg Ser Pro Ser Pro Ser Thr Gln Pro Trp Glu His 325 330 335 Val Asn Ala Ile Gln Glu Ala Arg Arg Leu Leu Asn Leu Ser Arg Asp 340 345 350 Thr Ala Ala Glu Met Asn Glu Thr Val Glu Val Ile Ser Glu Met Phe 355 360 365 Asp Leu Gln Glu Pro Thr Cys Leu Gln Thr Arg Leu Glu Leu Tyr Lys 370 375 380 Gln Gly Leu Arg Gly Ser Leu Thr Lys Leu Lys Gly Pro Leu Thr Met 385 390 395 400 Met Ala Ser His Tyr Lys Gln His Cys Pro Pro Thr Pro Glu Thr Ser 405 410 415 Cys Ala Thr Gln Ile Ile Thr Phe Glu Ser Phe Lys Glu Asn Leu Lys 420 425 430 Asp Phe Leu Leu Val Ile Pro Phe Asp Cys Trp Glu Pro Val Gln Glu 435 440 445 <210> 68 <211> 2472 <212> DNA <213> Artificial Sequence <220> <223> Nucleic acid encoding mIL12bIL12aIL2 GMCSF <400> 68 atgtgggagc tggagaaaga cgtttatgtt gtagaggtgg actggactcc cgatgcccct 60 ggagaaacag tgaacctcac ctgtgacacg cctgaagaag atgacatcac ctggacctca 120 gaccagagac atggagtcat aggctctgga aagaccctga ccatcactgt caaagagttt 180 ctagatgctg gccagtacac ctgccacaaa ggaggcgaga ctctgagcca ctcacatctg 240 ctgctccaca agaaggaaaa tggaatttgg tccactgaaa ttttaaaaaa tttcaaaaac 300 aagactttcc tgaagtgtga agcaccaaat tactccggac ggttcacgtg ctcatggctg 360 gtgcaaagaa acatggactt gaagttcaac atcaagagca gtagcagttc ccctgactct 420 cgggcagtga catgtggaat ggcgtctctg tctgcagaga aggtcacact ggaccaaagg 480 gactatgaga agtattcagt gtcctgccag gaggatgtca cctgcccaac tgccgaggag 540 gactatgaga agtattcagt gtcctgccag gaggatgtca cctgcccaac tgccgaggag 540accctgccca ttgaactggc gttggaagca cggcagcaga ataaatatga gaactacagc 600 accagcttct tcatcaggga catcatcaaa ccagacccgc ccaagaactt gcagatgaag 660 cctttgaaga actcacaggt ggaggtcagc tgggagtacc ctgactcctg gagcactccc 720 cattcctact tctccctcaa gttctttgtt cgaatccagc gcaagaaaga aaagatgaag 780 gagacagagg aggggtgtaa ccagaaaggt gcgttcctcg tagagaagac atctaccgaa 840 gtccaatgca aaggcgggaa tgtctgcgtg caagctcagg atcgctatta caattcctcg 900 tgcagcaagt gggcatgtgt tccctgcagg gtccgatccg gaggcggtgg aagtggcggt 960 ggaggctctg gaggtggcgg aagcagggtc attccagtct ctggacctgc caggtgtctt 1020 agccagtccc gaaacctgct gaagaccaca gatgacatgg tgaagacggc cagagaaaaa 1080 ctgaaacatt attcctgcac tgctgaagac atcgatcatg aagacatcac acgggaccaa 1140 accagcacat tgaagacctg tttaccactg gaactacaca agaacgagag ttgcctggct 1200 actagagaga cttcttccac aacaagaggg agctgcctgc ccccacagaa gacgtctttg 1260 atgatgaccc tgtgccttgg tagcatctat gaggacttga agatgtacca gacagagttc 1320 caggccatca acgcagcact tcagaatcac aaccatcagc agatcattct agacaagggc 1380 atgctggtgg ccatcgatga gctgatgcag tctctgaatc ataatggcga gactctgcgc 1440 cagaaacctc ctgtgggaga agcagaccct tacagagtga aaatgaagct ctgcatcctg 1500 cttcacgcct tcagcacccg cgtcgtgacc atcaacaggg tgatgggcta tctgagctcc 1560 gccggtggag gaggttctgg aggcggtgga agtggtggcg gaggtagcgc acccacttca 1620 agctccactt caagctctac agcggaagca cagcagcagc agcagcagca gcagcagcag 1680 cagcagcacc tggagcagct gttgatggac ctacaggagc tcctgagcag gatggagaat 1740 tacaggaacc tgaaactccc caggatgctc accttcaaat tttacttgcc caagcaggcc 1800 acagaattga aagatcttca gtgcctagaa gatgaacttg gacctctgcg gcatgttctg 1860 gatttgactc aaagcaaaag ctttcaattg gaagatgctg agaatttcat cagcaatatc 1920 agagtaactg ttgtaaaact aaagggctct gacaacacat ttgagtgcca attcgatgat 1980 gagtcagcaa ctgtggtgga ctttctgagg agatggatag ccttctgtca aagcatcatc 2040 tcaacaagcc ctcaaggcgg agggggttca ggtgggggcg ggagcggagg tggcggatcg 2100 gcacccaccc gctcacccat cactgtcacc cggccttgga agcatgtaga ggccatcaaa 2160 gaagccctga acctcctgga tgacatgcct gtcacgttga atgaagaggt agaagtcgtc 2220<000545~6>tctaacgagt tctccttcaa gaagctaaca tgtgtgcaga cccgcctgaa gatattcgag 2280 cagggtctac ggggcaattt caccaaactc aagggcgcct tgaacatgac agccagctac 2340 taccagacat actgcccccc aactccggaa acggactgtg aaacacaagt taccacctat 2400 gcggatttca tagacagcct taaaaccttt ctgactgata tcccctttga atgcaaaaaa 2460 [[ID=~6]]ccaggccaaa aa 2472 <210> 69 <211> 3948 <212> DNA <213> Artificial Sequence <220><~005466><223> Nucleic acid encoding mIL12bIL12aIL2DiaNHS76F8GMCSF-Thr <400> 69<~005468>atgtgggagc tggagaaaga cgtttatgtt gtagaggtgg actggactcc cgatgcccct 60 It should be noted that there seems to be some inconsistent or unclear formatting in the original text (such as the "~" in some tags), and this translation attempts to handle it as accurately as possible while maintaining the original structure.ggagaaacag tgaacctcac ctgtgacacg cctgaagaag atgacatcac ctggacctca 120 gaccagagac atggagtcat aggctctgga aagaccctga ccatcactgt caaagagttt 180 ctagatgctg gccagtacac ctgccacaaa ggaggcgaga ctctgagcca ctcacatctg 240 ctgctccaca agaaggaaaa tggaatttgg tccactgaaa ttttaaaaaa tttcaaaaac 300 aagactttcc tgaagtgtga agcaccaaat tactccggac ggttcacgtg ctcatggctg 360 gtgcaaagaa acatggactt gaagttcaac atcaagagca gtagcagttc ccctgactct 420 cgggcagtga catgtggaat ggcgtctctg tctgcagaga aggtcacact ggaccaaagg 480 gactatgaga agtattcagt gtcctgccag gaggatgtca cctgcccaac tgccgaggag 540 accctgccca ttgaactggc gttggaagca cggcagcaga ataaatatga gaactacagc 600 accagcttct tcatcaggga catcatcaaa ccagacccgc ccaagaactt gcagatgaag 660 cctttgaaga actcacaggt ggaggtcagc tgggagtacc ctgactcctg gagcactccc 720 cattcctact tctccctcaa gttctttgtt cgaatccagc gcaagaaaga aaagatgaag 780 GAGACAGAGG AGGAGGAGGA GAGGAGGAGA GAGGAGGAGA GAGGAGGAGA 60 GTCCTGCTGA GAGCTGCCTG GAGAAGAGCC TGCCTGAGCC TGCCTGAGCC 60 TGCAGCAAGT GGGCATGTGT TCCCTGCAGG GTCGATCCGG AGGCGGTGGA AGTGGCGGT 60 GGAGGCTCTG GAGGTGGCGG AAGCAGGGTC ATTCCTAGTC TCTGGACCTG CCAGGTGTCTT 60 AGCCAGTCCC GAAACCTGCT GAAGACCACA GATGACATGG TGAAGACGGC CAGAGAAAAA 60 CTGAAACATT ATTCCCGCAC TGCTGAAGAC ATCGATCATG AAGACATCAC ACGGGACCAA 60 ACCAGCACAT TGAAGACCTG TTTACCACCG AACTACACAA GAACGAGAGT GCCTGGCT 60 ACTAGAGAGA CTTCTTCCAC AACAAGAGGG AGCTGCCTGC CCCCACAGAA GACGTCTTTG 60 ATGATGACCC TGUGCCTTGG TAGCATCTAT GAGGACTTGA AGATGTACCA GACAGAGTTC 60 CAGGCCATCA ACGCAGCACT TCAGAATCAC AACCATCAGC AGATCATTCT AGACAAGGGC 60 ATGCTGGTGG CCATCGATGA GCTGATGCAG TCTCTGAAAT CATAATGGCG AGACTCTGCGC 60 CAGAAACCTC CTGTGGGAGA AGCAGACCCG ATAGAGTGAA AATGAAGCT CTGCTCCTG 60 cttcacgcct tcagcacccg cgtcgtgacc atcaacaggg tgatgggcta tctgagctcc 1560 gccggtggag gaggttctgg aggcggtgga agtggtggcg gaggtagcgc acccacttca 1620 agctccactt caagctctac agcggaagca cagcagcagc agcagcagca gcagcagcag 1680 cagcagcacc tggagcagct gttgatggac ctacaggagc tcctgagcag gatggagaat 1740 tacaggaacc tgaaactccc caggatgctc accttcaaat tttacttgcc caagcaggcc 1800 acagaattga aagatcttca gtgcctagaa gatgaacttg gacctctgcg gcatgttctg 1860 gatttgactc aaagcaaaag ctttcaattg gaagatgctg agaatttcat cagcaatatc 1920 agagtaactg ttgtaaaact aaagggctct gacaacacat ttgagtgcca attcgatgat 1980 gagtcagcaa ctgtggtgga ctttctgagg agatggatag ccttctgtca aagcatcatc 2040 tcaacaagcc ctcaaggcgg agggggttca ctcgtcccaa gaggaagtgg aggtggcgga 2100 tcgcaggtgc agctgcagga gagcggcccc ggcctggtga agcccagcga gaccctgagc 2160 ctgacctgcg ccgtgagcgg ctacagcatc agcagcggct actactgggg ctggatcagg 2220 CAGCCCCCCC GCAAGGGCCT GGAGTGGATC GGCAGCATCT ACCACAGCGG CAGCACCTAC 2280 TACAACCCCC A GCCTGAAGAG CAGGGTGACC ATCAGCGTGG ACACCAGCAA GAACCAGTTC 2340 AGCCTGAAGC TGAGCAGCGT GACC GCCGCC GACACC GCCGT GTACTACTG CGCCAGGGGC 2400 AAGTGGAGCA AGTT CGACTACTGGGGCCAGGGCAC CCTGGTGACCGTGAGCAGCGGCGGT 2460 AGCGGAGGGG AAATTGTGTT GACGCAGTCT CCAGGCACCCT GTCTTTGTC TCCAGGGGAA 2520 AGAGCCACCCC TCTCCTGCAG GGCCAGTCAG AGTGTTAGCA TGCCTTTTTT AGCCTGGTAC 2580 CAGCAGAAAC CTGGCCAGGC TCCCAGGCTC CTCATCTATG GTGATCCAGC AGGGCCACT 2640 GGCATCCCAG ACAGGTTCAA TGGCAGTGGG TCTGGGACAG ACTTC ACTCT CACCATCAGC 2700 AGACTGGAGC CTGAAGATTT TGCAGTGTA TTACTGTCAG CAGATGCGTG GTGGCCGCCG 2760 ACGTTTGGCC AAGGGACCAA GGTGGAAATC AAATCTTCAA GTAGCGGTTT AAGTT CCTCT 2820 GGATCGTCAA GCTCTGGTGA GGTGCAGCTG TTGGAGTCTG GGGGAGGCTT GGTACAGCCT 2880 GGGGGGTCCC TGAGACTCTC CTGTGCAGCC TCTGGATTC ACCTTTAGCC TGT TTACGATG 2940 agctgggtcc gccaggctcc agggaagggg ctggagtggg tctcagctat tagtggtagt 3000 ggtggtagca catactacgc agactccgtg aagggccggt tcaccatctc cagagacaat 3060 tccaagaaca cgctgtatct gcaaatgaac agcctgagag ccgaggacac ggccgtatat 3120 tactgtgcga aaagtactca tttgtatctt tttgactact ggggccaggg aaccctggtc 3180 accgtctcca gtggcggtag cggagggagc agcgagctga cccaggaccc cgccgtgagc 3240 gtggccctgg gccagaccgt gaggatcacc tgccagggcg acagcctgag gagctactac 3300 gccagctggt accagcagaa gcccggccag gcccccgtgc tggtgatcta cggcaagaac 3360 aacaggccca gcggcatccc cgacaggttc agcggcagca gcagcggcaa caccgccagc 3420 ctgaccatca ccggcgccca ggccgaggac gaggccgact actactgcaa cagcagggac 3480 agcagcggca accacgtggt gttcggcggc ggcaccaagc tgaccgtgct gtcttcaagt 3540 agcggtctcg tcccaagagg aagttcaagc tctggtgcac ccacccgctc acccatcact 3600 gtcacccggc cttggaagca tgtagaggcc atcaaagaag ccctgaacct cctggatgac 3660 atgcctgtca cgttgaatga agaggtagaa gtcgtctcta acgagttctc cttcaagaag 3720 ctaacatgtg tgcagacccg cctgaagata ttcgagcagg gtctacgggg caatttcacc 3780 aaactcaagg gcgccttgaa catgacagcc agctactacc agacatactg ccccccaact 3840 ccggaaacgg actgtgaaac acaagttacc acctatgcgg atttcataga cagccttaaa 3900 acctttctga ctgatatccc ctttgaatgc aaaaaaccag gccaaaaa 3948 <210> 70 <211> 2412 <212> DNA <213> Artificial Sequence <220> <223> Nucleic acid encoding mIL12bIL12aIL7 GMCSF <400> 70 atgtgggagc tggagaaaga cgtttatgtt gtagaggtgg actggactcc cgatgcccct 60 ggagaaacag tgaacctcac ctgtgacacg cctgaagaag atgacatcac ctggacctca 120 gaccagagac atggagtcat aggctctgga aagaccctga ccatcactgt caaagagttt 180 ctagatgctg gccagtacac ctgccacaaa ggaggcgaga ctctgagcca ctcacatctg 240 ctgctccaca agaaggaaaa tggaatttgg tccactgaaa ttttaaaaaa tttcaaaaac 300 aagactttcc tgaagtgtga agcaccaaat tactccggac ggttcacgtg ctcatggctg 360 gtgcaaagaa acatggactt gaagttcaac atcaagagca gtagcagttc ccctgactct 420 cgggcagtga catgtggaat ggcgtctctg tctgcagaga aggtcacact ggaccaaagg 480 gactatgaga agtattcagt gtcctgccag gaggatgtca cctgcccaac tgccgaggag 540 accctgccca ttgaactggc gttggaagca cggcagcaga ataaatatga gaactacagc 600 accagcttct tcatcaggga catcatcaaa ccagacccgc ccaagaactt gcagatgaag 660 cctttgaaga actcacaggt ggaggtcagc tgggagtacc ctgactcctg gagcactccc 720 cattcctact tctccctcaa gttctttgtt cgaatccagc gcaagaaaga aaagatgaag 780 gagacagagg aggggtgtaa ccagaaaggt gcgttcctcg tagagaagac atctaccgaa 840 gtccaatgca aaggcgggaa tgtctgcgtg caagctcagg atcgctatta caattcctcg 900 tgcagcaagt gggcatgtgt tccctgcagg gtccgatccg gaggcggtgg aagtggcggt 960 ggaggctctg gaggtggcgg aagcagggtc attccagtct ctggacctgc caggtgtctt 1020 agccagtccc gaaacctgct gaagaccaca gatgacatgg tgaagacggc cagagaaaaa 1080 ctgaaacatt attcctgcac tgctgaagac atcgatcatg aagacatcac acgggaccaa 1140 accagcacat tgaagacctg tttaccactg gaactacaca agaacgagag ttgcctggct 1200 actagagaga cttcttccac aacaagaggg agctgcctgc ccccacagaa gacgtctttg 1260 atgatgaccc tgtgccttgg tagcatctat gaggacttga agatgtacca gacagagttc 1320 caggccatca acgcagcact tcagaatcac aaccatcagc agatcattct agacaagggc 1380 atgctggtgg ccatcgatga gctgatgcag tctctgaatc ataatggcga gactctgcgc 1440 cagaaacctc ctgtgggaga agcagaccct tacagagtga aaatgaagct ctgcatcctg 1500 cttcacgcct tcagcacccg cgtcgtgacc atcaacaggg tgatgggcta tctgagctcc 1560 gccggtggag gaggttctgg aggcggtgga agtggtggcg gaggtagcga gtgccacatt 1620 aaagacaaag aaggtaaagc atatgagagt gtactgatga tcagcatcga tgaattggac 1680 aaaatgacag gaactgatag taattgcccg aataatgaac caaacttttt tagaaaacat 1740 gtatgtgatg atacaaagga agctgctttt ctaaatcgtg ctgctcgcaa gttgaagcaa 1800 tttcttaaaa tgaatatcag tgaagaattc aatgtccact tactaacagt atcacaaggc 1860 acacaacac tggtgaactg cacaagtaag gaaaaaaa acgtaagga acagaaaaag 1920 aatgatgcat gtttcctaaa gagactactg agaaataa aaacttgttg gaataaaatt 1980 ttgaagggca gtataggcgg aggggttca ggtggggggcg ggagcggagg tggcggatcg 2040 gcacccacc gctcacccat cactgtcacc cggccttgga agcatgtaga ggccatcaaa 2100 gaagccctga acctcctgga tgacatgcct gtcacgttga atgaagaggt agaagtcgtc 2160 2220 cagggtctac ggggcaattt caccaaactc aagggcgcct tgaacatgac agccagctac 2280 taccagacat actgcccccc aactccggaa acggactgtg aaacacaagt taccacctat 2340 gcggatttca tagacagcct taaaaccttt ctgactgata tcccctttga atgcaaaaaa 2400 ccaggccaaa aa 2412 <210> 71 <211> 2412 <212> DNA <213> Artificial Sequence <220> <223> Nucleic acid encoding mIL12bIL12aIL21 GMCSF <400> 71 atgtgggagc tggagaaaga cgtttatgtt gtagaggtgg actggactcc cgatgcccct 60 ggagaaacag tgaacctcac ctgtgacacg cctgaagaag atgacatcac ctggacctca 120 gaccagagac atggagtcat aggctctgga aagaccctga ccatcactgt caaagagttt 180 ctagatgctg gccagtacac ctgccacaaa ggaggcgaga ctctgagcca ctcacatctg 240 ctgctccaca agaaggaaaa tggaatttgg tccactgaaa ttttaaaaaa tttcaaaaac 300 aagactttcc tgaagtgtga agcaccaaat tactccggac ggttcacgtg ctcatggctg 360 gtgcaaagaa acatggactt gaagttcaac atcaagagca gtagcagttc ccctgactct 420 cgggcagtga catgtggaat ggcgtctctg tctgcagaga aggtcacact ggaccaaagg 480 gactatgaga agtattcagt gtcctgccag gaggatgtca cctgcccaac tgccgaggag 540 accctgccca ttgaactggc gttggaagca cggcagcaga ataaatatga gaactacagc 600 accagcttct tcatcaggga catcatcaaa ccagacccgc ccaagaactt gcagatgaag 660 cctttgaaga actcacaggt ggaggtcagc tgggagtacc ctgactcctg gagcactccc 720 cattcctact tctccctcaa gttctttgtt cgaatccagc gcaagaaaga aaagatgaag 780 gagacagagg aggggtgtaa ccagaaaggt gcgttcctcg tagagaagac atctaccgaa 840 gtccaatgca aaggcgggaa tgtctgcgtg caagctcagg atcgctatta caattcctcg 900 tgcagcaagt gggcatgtgt tccctgcagg gtccgatccg gaggcggtgg aagtggcggt 960 ggaggctctg gaggtggcgg aagcagggtc attccagtct ctggacctgc caggtgtctt 1020 agccagtccc gaaacctgct gaagaccaca gatgacatgg tgaagacggc cagagaaaaa 1080 ctgaaacatt attcctgcac tgctgaagac atcgatcatg aagacatcac acgggaccaa 1140 accagcacat tgaagacctg tttaccactg gaactacaca agaacgagag ttgcctggct 1200 actagagaga cttcttccac aacaagaggg agctgcctgc ccccacagaa gacgtctttg 1260 atgatgaccc tgtgccttgg tagcatctat gaggacttga agatgtacca gacagagttc 1320 caggccatca acgcagcact tcagaatcac aaccatcagc agatcattct agacaagggc 1380 atgctggtgg ccatcgatga gctgatgcag tctctgaatc ataatggcga gactctgcgc 1440 cagaaacctc ctgtgggaga agcagaccct tacagagtga aaatgaagct ctgcatcctg 1500 cttcacgcct tcagcacccg cgtcgtgacc atcaacaggg tgatgggcta tctgagctcc 1560 gccggtggag gaggttctgg aggcggtgga agtggtggcg gaggtagcca taaatcaagc 1620 ccccaagggc cagatcgcct cctgattaga cttcgtcacc ttattgacat tgttgaacag 1680 ctgaaaatct atgaaaatga cttggatcct gaacttctat cagctccaca agatgtaaag 1740 gggcactgtg agcatgcagc ttttgcctgt tttcagaagg ccaaactcaa gccatcaaac 1800 cctggaaaca ataagacatt catcattgac ctcgtggccc agctcaggag gaggctgcct 1860 gccaggaggg gaggaaagaa acagaagcac atagctaaat gcccttcctg tgattcgtat 1920 gagaaaagga cacccaaaga attcctagaa agactaaaat ggctccttca aaagatgatt 1980 catcagcatc tctccggcgg agggggttca ggtgggggcg ggagcggagg tggcggatcg 2040 gcacccaccc gctcacccat cactgtcacc cggccttgga agcatgtaga ggccatcaaa 2100 gaagccctga acctcctgga tgacatgcct gtcacgttga atgaagaggt agaagtcgtc 2160 tctaacgagt tctccttcaa gaagctaaca tgtgtgcaga cccgcctgaa gatattcgag 2220 cagggtctac ggggcaattt caccaaactc aagggcgcct tgaacatgac agccagctac 2280 taccagacat actgcccccc aactccggaa acggactgtg aaacacaagt taccacctat 2340 gcggatttca tagacagcct taaaaccttt ctgactgata tcccctttga atgcaaaaaa 2400 ccaggccaaa aa 2412 <210> 72 <211> 2613 <212> DNA <213> Artificial Sequence <220> <223> Nucleic acid encoding mIL12aIL2IL12bGMCSF <400> 72 atgtgtcaat cacgctacct cctctttttg gccacccttg ccctcctaaa ccacctcagt 60 ttggccaggg tcattccagt ctctggacct gccaggtgtc ttagccagtc ccgaaacctg 120 ctgaagacca cagatgacat ggtgaagacg gccagagaaa aactgaaaca ttattcctgc 180 actgctgaag acatcgatca tgaagacatc acacgggacc aaaccagcac attgaagacc 300. tgtttaccac tggactaca caagaacgag agttgcctgg ctactagaga gacttcttcc acaacaagag ggagctgcct gcccccacag aagacgtctt tgatgatgac cctgtgcctt ggtagcatct atgaggactt gaagatgtac cagacagagt tccaggccat caacgcagca cttcagaatc acaaccatca gcagatcatt ctagacaagg gcatgctggt ggccatcgat gagctgatgc agtctctga tcataatggc gagactctgc gccagaacc tcctgtggga 540 gaagcagacc cttacagagt gaaaatgaag ctctgcatcc tgcttcacgc cttcagcacc cgcgtcgtga ccatcaacag ggtgatgggc tatctgagct ccgccggtgg aggaggttct ggaggcggtg gaagtggtgg cggaggtagc gcacccactt caagctccac ttcaagctct 720 acagcggaag cacagcagca gcagcagcagc cctggagcag 780. cctggagcag ctgttgatgg acctacagga gctcctgagc aggatggaga attack cctgaaactc 840 cccaggatgc tcaccttcaa attttacttg cccaagcagg ccacagaatt gaaagatctt cagtgcctag aagatgaact tggacctctg cggcatgttc tggatttgac tcaaagcaaa 960 agctttcaat tggaagatgc tgagaatttc atcagcaata tcagagtaac tgttgtaaaa 1020 ctaaagggct ctgacaacac atttgagtgc caattcgatg atgagtcagc aactgtggtg 1080 gactttctga ggagatggat agccttctgt caaagcatca tctcaacaag ccctcaagag 1140 ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgtgtcct 1200 cagaagctaa ccatctcctg gtttgccatc gttttgctgg tgtctccact catggccatg 1260 tgggagctgg agaaagacgt ttatgttgta gaggtggact ggactcccga tgcccctgga 1320 gaaacagtga acctcacctg tgacacgcct gaagaagatg acatcacctg gacctcagac 1380 cagagacatg gagtcatagg ctctggaaag accctgacca tcactgtcaa agagtttcta 1440 gatgctggcc agtacacctg ccacaaagga ggcgagactc tgagccactc acatctgctg 1500 ctccacaaga aggaaaatgg aatttggtcc actgaaattt taaaaaattt caaaaacaag 1560 actttcctga agtgtgaagc accaaattac tccggacggt tcacgtgctc atggctggtg 1620 caaagaaaca tggacttgaa gttcaacatc aagagcagta gcagttcccc tgactctcgg 1680 gcagtgacat gtggaatggc gtctctgtct gcagagaagg tcacactgga ccaaagggac 1740 tatgagaagt attcagtgtc ctgccaggag gatgtcacct gcccaactgc cgaggagacc 1800 ctgcccattg aactggcgtt ggaagcacgg cagcagaata aatatgagaa ctacagcacc 1860 agcttcttca tcagggacat catcaaacca gacccgccca agaacttgca gatgaagcct 1920 ttgaagaact cacaggtgga ggtcagctgg gagtaccctg actcctggag cactccccat 1980 tcctacttct ccctcaagtt ctttgttcga atccagcgca agaaagaaaa gatgaaggag 2040 acagaggagg ggtgtaacca gaaaggtgcg ttcctcgtag agaagacatc taccgaagtc 2100 caatgcaaag gcgggaatgt ctgcgtgcaa gctcaggatc gctattacaa ttcctcgtgc 2160 agcaagtggg catgtgttcc ctgcagggtc cgatccggag gcggtggaag tggcggtgga 2220 ggctctggag gtggcggaag cgcacccacc cgctcaccca tcactgtcac ccggccttgg 2280 aagcatgtag aggccatcaa agaagccctg aacctcctgg atgacatgcc tgtcacgttg 2340 aatgaagagg tagaagtcgt ctctaacgag ttctccttca agaagctaac atgtgtgcag 2400 acccgcctga agatattcga gcagggtcta cggggcaatt tcaccaaact caagggcgcc 2460 ttgaacatga cagccagcta ctaccagaca tactgccccc caactccgga aacggactgt 2520 gaaacacaag ttaccaccta tgcggatttc atagacagcc ttaaaacctt tctgactgat 2580 atcccctttg aatgcaaaaa accaggccaa aaa 2613 <210> 73 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Linker 1 – Used for connections between cytokines and between cytokines and their targets. <400> 73 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 1 5 10 15 <210> 74 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Linker 2 – The link between cytokines and targets <400> 74 Gly Gly Gly Gly Ser Leu Val Pro Arg Gly Ser Gly Gly Gly Gly Ser 1 5 10 15 <210> 75 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Linker 3 - for linking between cytokines, between cytokine and targeting <400> 75 Ser Ser Ser Ser Gly Ser Ser Ser Ser Gly Ser Ser Ser Ser Gly 1 5 10 15 <210> 76 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Linker 4 - for linking between cytokine and targeting <400> 76 Ser Ser Ser Ser Gly Leu Val Pro Arg Gly Ser Ser Ser Ser Gly 1 5 10 15

Claims

1. A liquid preparation for treating lung cancer, comprising glycerol and a fusion protein, the mass fraction of the glycerol being 70% or more, and the mass fraction of the fusion protein being 1%-20%; the fusion protein is mIL12bIL12aIL2GMCSF, mIL12bIL12aIL2DiaNHS76F8GMCSF-Thr, mIL12bIL12aIL7GMCSF, mIL12bIL12aIL21GMCSF or mIL12aIL2IL12bGMCSF, wherein, The amino acid sequence of the mIL12bIL12aIL2GMCSF is shown as SEQ ID NO. 32, the amino acid sequence of the mIL12bIL12aIL2DiaNHS76F8GMCSF-Thr is shown as SEQ ID NO. 48, the amino acid sequence of the mIL12bIL12aIL7GMCSF is shown as SEQ ID NO. 33, the amino acid sequence of the mIL12bIL12aIL21GMCSF is shown as SEQ ID NO. 35, and the mIL12aIL2IL12bGMCSF is a heterodimer consisting of a first polypeptide chain mIL12aIL2 and a second polypeptide chain mIL12bGMCSF, the amino acid sequence of the first polypeptide chain is shown as SEQ ID NO. 56, and the amino acid sequence of the second polypeptide chain is shown as SEQ ID NO.

57.

2. The liquid formulation according to claim 1, wherein, The nucleotide sequence encoding the mIL12bIL12aIL2GMCSF is shown as SEQ ID NO.

68.

3. The liquid formulation according to claim 1, wherein, The nucleotide sequence encoding the mIL12bIL12aIL2DiaNHS76F8GMCSF-Thr is shown as SEQ ID NO.

69.

4. The liquid formulation according to claim 1, wherein, The nucleotide sequence encoding the mIL12bIL12aIL7GMCSF is shown as SEQ ID NO.

70.

5. The liquid formulation of claim 1, wherein, The nucleotide sequence encoding the mIL12bIL12aIL21GMCSF is shown as SEQ ID NO.

71.

6. The liquid formulation of claim 1, wherein, The nucleotide sequence encoding the mIL12aIL2IL12bGMCSF is shown as SEQ ID NO.

72.

7. A medicament comprising the liquid formulation of any one of claims 1-6.

8. The medicament of claim 7, wherein the liquid formulation is formulated as an injection.

9. The medicament of claim 7 or 8, further comprising a diluent, wherein the diluent and the liquid formulation do not mix with each other.

10. Use of the liquid formulation of any one of claims 1-6 in the manufacture of a medicament for treating lung cancer.

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