RU2816646C2 - Multivalent pd-l1-binding compounds for treating malignant neoplasms - Google Patents

Abstract

FIELD: biotechnology; medicine.

SUBSTANCE: present invention relates to multivalent compounds which bind programmed death ligand 1 (PD-L1), which are administered to a mammal with a malignant growth for treating a mammal. In some cases, the multivalent PD-L1-binding compound can include two or more polypeptides of programmed death protein 1 (PD-1) (and/or fragments thereof, capable of binding to PD-L1).

EFFECT: invention makes it possible to increase efficiency of PD-1 for neutralization of PD-L1 present on PD-L1-positive malignant cells in therapy of malignant neoplasms.

12 cl, 20 dwg, 7 ex

Description

BACKGROUND OF THE ART

Field of technology

[0002] The present invention relates to methods and materials for the treatment of malignant neoplasms. The invention relates to methods and materials for the production of multivalent PD-L1 binding compounds. The present invention also provides methods and materials for expressing PD-1 polypeptides in cells of a mammal having a cancer. In particular, the invention relates to compositions containing PD-1 polypeptides, the PD-1 polypeptides may be in the form of one or more multivalent programmed death protein ligand 1 (PD-L1) binding compounds. In some cases, a multivalent PD-L1 binding compound may comprise two or more programmed death protein 1 (PD-1) polypeptides (and/or fragments thereof having the ability to bind PD-L1). Compositions containing PD-L1 binding compounds are administered to a mammal having a cancer in an amount effective to treat the cancer, and are optionally administered in combination with one or more cancer therapies.

[0003] PD-L1, also called B7-H1, is an immune checkpoint protein that regulates the immune system through binding to the PD-1 receptor. In the tumor microenvironment, overexpression of PD-L1 on tumor cells promotes suppression of antitumor immunity (Dong et al. , Nat Med . 8:793-800, (2002); Hamanishi et al. , Int. J. Clin. Oncol . 21:462- 473 (2016); Dong et al. , Nat. Med . 5:1365-1369 (1999); Chen et al. , J. Clin. Invest . 125:3384-3391 (2015); He et al. , Sci. Rep . 5:13110 (2015); Chen et al. , Clin. Cancer Res . 18:6580-6587 (2012); Ohaegbulam et al. , Trends Mol. Med . 21:24-33 (2015); and Postow et al. , J. Clin. Oncol . 33:1974-1982 (2015)).

[0004] Antigen presenting cells (APCs) take up antigens released by malignant cells and present them to T cells. Malignant cells can also present antigens to activated T cells in the context of the major histocompatibility complex. Following T cell activation, PD-1 receptors are expressed on T cells and inhibit immune responses through recruitment of PD-L1 and PD-L2 ligands on APCs and PD-L1 on malignant cells. Thus, monoclonal antibody (mAb)-mediated specific blockade of the PD-1/PD-L1/PD-L2 pathway may enhance antitumor immunity. In addition to T cells and APCs, PD-1 and PD-L1 can be induced on other immune cells.

[0005] In a malignant disease state, the interaction of PD-L1 present on a PD-L1-positive malignant cell with PD-1 present on a T cell can reduce functional T cell signaling, preventing the immune system from attacking the PD-L1-positive cell. positive malignant cell. Soluble PD-1 (sPD-1) can act as a decoy by binding to PD-L1 present on PD-L1-positive malignant cells. For example, if sPD-1 is bound to PD-L1 present on a PD-L1 positive cancer cell, PD-L1 cannot interact with PD-1 present on the T cell, thus allowing the T cell to function by attacking the PD -L1-positive malignant cell.

SUMMARY OF THE INVENTION

[0006] The present invention relates to methods and materials for producing one or more multivalent PD-L1 binding compounds. The present invention relates to compositions containing PD-L1 binding compounds containing PD-1 polypeptides, the PD-1 polypeptides may be in the form of one or more multivalent PD-L1 binding compounds. PD-L1-binding compounds can act as a decoy by binding to PD-L1 present on PD-L1-positive cancer cells.

[0007] The present invention relates to methods and materials containing PD-1 polypeptides, the PD-1 polypeptides may be in the form of one or more multivalent PD-L1 binding compounds administered to a mammal having a cancer for treating the mammal. In some cases, a multivalent PD-L1 binding compound includes two or more amino acid segments that can bind PD-L1 (eg, PD-1 polypeptides and/or fragments thereof). A multivalent PD-L1 binding compound comprises two or more PD-1 polypeptides (or fragments thereof having the ability to bind PD-L1) such that the compound can bind two or more PD-L1 polypeptides. The present invention also relates to methods for producing PD-L1 binding compounds and compositions containing recombinant Ad containing at least one amino acid segment containing a PD-1 polypeptide forming a PD-L1 binding compound or a multivalent PD-L1 binding compound, presented in this description.

[0008] In an embodiment, mouse PD-1 polypeptides and/or human PD-1 polypeptides fused to a backbone polypeptide can form a polypeptide conjugate (e.g., a polypeptide conjugate comprising multiple (e.g., two or more) associated amino acid chains) thus that if a framework polypeptide fused to one or more PD-1 polypeptides is a polypeptide conjugate, the polypeptide conjugate can form a multivalent PD-L1 binding compound comprising two or more PD-1 polypeptides.

[0009] In an embodiment, the polypeptide conjugate may comprise three amino acid chains, wherein the backbone polypeptide may be a sigma-1 polypeptide derived from mammalian orthoreovirus 3, and wherein the polypeptide conjugate may comprise three PD-1 polypeptides.

[00010] In an embodiment, the polypeptide conjugate may comprise four amino acid chains, wherein the framework polypeptide may be a streptavidin polypeptide, and where the polypeptide conjugate may comprise four PD-1 polypeptides.

[00011] The present invention also provides compositions containing recombinant Ad containing nucleic acids encoding PD-1 polypeptides that bind to PD-L1. In an embodiment, a mouse PD-1 polypeptide and/or a human PD-1 polypeptide is fused to the vitamin K-dependent gamma-carboxyglutamine domain of a single-chain anti-factor X antibody polypeptide (GLA domain or GLA-EGF FX polypeptide) and is present on adenoviral hexon polypeptides thereby that if a polypeptide is present on two or more viral hexon polypeptides present on the capsid of a viral particle, the viral particle can form a multivalent PD-L1 binding compound comprising from about 240 to about 720 PD-1 polypeptides.

[00012] The present invention relates to methods and materials for treating cancer by administering one or more recombinant adenoviruses (Ad) expressing PD-1 protein and/or amino acid segments that bind to PD-L1 (e.g., PD-1 polypeptides and/or or fragments thereof). Recombinant Ad expressing PD-1 can be administered in combination with antitumor immunotherapy in a therapeutic regimen.

[00013] By using multivalent PD-L1 binding compounds (eg, compared to monomeric PD-1 polypeptides), the effectiveness of PD-1 to neutralize PD-L1 present on PD-L1-positive cancer cells can be increased, PD escape can be prevented -L1-positive malignant immune surveillance cells and/or can allow anti-tumor agents (eg, anti-tumor immunotherapies) to more effectively target PD-L1-positive malignant cells. In some cases, adenovirus presentation of PD-1 polypeptides can be used to retarget therapeutic Ads to PD-L1 on a cell for nucleic acid delivery or to kill PD-L1-expressing cells via oncolytic cell death.

[00014] In an embodiment, the PD-L1 binding compound may also include a targeting molecule. The targeting molecule and cell fusion molecule may be a viral polypeptide, such as measles virus (MV) hemagglutinin (H) polypeptide, MV fusion (F) polypeptide, or vesicular stomatitis virus (VSV) glycoprotein (G) polypeptide.

[00015] The PD-L1 binding compound may also include one or more therapeutic polypeptides. The therapeutic polypeptide is selected from a 4-1BB ligand polypeptide (4-1BBL), an OX40 ligand polypeptide (OX40L), a CD40 ligand polypeptide (CD40L), or a granulocyte-macrophage colony-stimulating factor (GM-CSF) polypeptide. The therapeutic polypeptide may also be a polypeptide that activates glucocorticoid-inducible tumor necrosis factor receptor (TNFR) protein signaling (GITR).

[00016] The PD-L1 binding compound may also include a detectable polypeptide. The detected polypeptide may be green fluorescent protein (GFP) or luciferase polypeptide.

[00017] The present invention relates to recombinant vectors for the expression of PD-L1 binding compounds containing nucleic acids encoding PD-1 polypeptides, the PD-1 polypeptides can be expressed with one or more targeting polypeptides and/or one or more therapeutic polypeptides and/ or proteins (eg collagen, elastin, laminin and fibrinogen). PD-1 polypeptides can be expressed as fusion proteins with heterologous polypeptides. Nucleic acids encoding PD-1 polypeptides, as well as nucleic acids encoding targeting polypeptides, therapeutic polypeptides and proteins, can be contained in expression vectors, optionally containing expression cassettes allowing expression of the polypeptides in prokaryotic or eukaryotic cells. The vector may be a viral vector. The viral vector may be Ad, adeno-associated virus (AAV), or lentivirus. The viral vector may be Ad selected from Ad657, Ad6/57/6, and variants thereof. Ad may be a conditionally replicative Ad (CRAd). The viral vector may be an oncolytic viral vector.

[00018] In another aspect, the present invention provides viral vectors for the expression of PD-L1 binding compounds containing nucleic acids encoding an amino acid chain, including a PD-1 polypeptide and a framework polypeptide as described herein (e.g., a polypeptide conjugate that may comprise a plurality of amino acid chains, wherein each amino acid chain includes a PD-1 polypeptide and a framework polypeptide; and wherein the plurality of amino acid chains may form a polypeptide conjugate).

[00019] In another aspect, the present invention relates to methods of treating a mammal having a cancer. The methods may include or consist essentially of administering a composition containing a PD-L1 binding compound to a mammal having a cancer.

[00020] The method of the invention includes administering an effective amount of compositions containing PD-L1 binding compounds provided herein. In an embodiment, PD-L1 binding compounds are administered to a mammal in combination with one or more anticancer therapeutic agents, thereby reducing the number of malignant cells present in the mammal. The one or more antitumor therapeutic agents may include an immunotherapy agent targeting PD-1 or PD-L1. The immunotherapeutic agent may be selected from nivolumab, pembrolizumab, atezolizumab, avelumab, cemiplimab and durvalumab.

[00021] In another aspect, the invention relates to methods of treating a mammal having a cancer, comprising or consisting essentially of administering a composition containing a multivalent PD-L1 binding compound to the mammal having a cancer. The multivalent PD-L1 binding compound may comprise a polypeptide conjugate as provided herein (eg, a polypeptide conjugate comprising multiple amino acid chains, wherein each amino acid chain includes a PD-1 polypeptide and a framework polypeptide). The malignancy may be prostate cancer, breast cancer, ovarian cancer, lung cancer (eg, non-small cell lung cancer), hepatocellular carcinoma, pancreatic cancer, kidney cancer, melanoma, brain malignancy, colon cancer, lymphoma, myeloma, lymphocytic leukemia or myelogenous leukemia. Administration may include systemic or local administration (eg, intravenous, intratumoral, intramuscular, intraorgan, and lymph node administration).

[00022] One aspect of the invention relates to multivalent programmed death protein 1 (PD-L1) ligand binding compounds comprising multiple amino acid chains, where each amino acid chain contains at least one programmed death protein 1 (PD-1) polypeptide.

[00023] An additional aspect of the invention relates to such a multivalent PD-L1 binding compound, wherein the multivalent PD-L1 binding compound is a polypeptide conjugate containing a framework polypeptide selected from Ig polypeptides, sigma-1 polypeptides and streptavidin polypeptides.

[00024] An additional aspect of the invention relates to such a multivalent PD-L1 binding compound, wherein the multivalent PD-L1 binding compound is a polypeptide conjugate containing more than one framework polypeptide.

[00025] An additional aspect of the invention relates to such a multivalent PD-L1 binding compound, wherein the PD-1 polypeptide is human PD-1 or mouse PD-1.

[00026] An additional aspect of the invention relates to such a multivalent PD-L1 binding compound, wherein the plurality of amino acid chains comprise a therapeutic polypeptide, a targeting polypeptide, or an antigenic polypeptide.

[00027] An additional aspect of the invention relates to such a multivalent PD-L1 binding compound, wherein the targeting polypeptide is selected from a measles virus hemagglutinin (MVH) polypeptide, a measles virus fusion polypeptide (MVF), and a vesicular stomatitis virus glycoprotein (VSVG) polypeptide.

[00028] A further aspect of the invention relates to such a multivalent PD-L1 binding compound, wherein the therapeutic polypeptide is selected from a 4-1BB ligand polypeptide (4-1BBL), an OX40 ligand polypeptide (OX40L), a CD40 ligand polypeptide (CD40L), a granulocyte ligand polypeptide macrophage colony-stimulating factor (GM-CSF) and GITR agonist.

[00029] An additional aspect of the invention relates to such a multivalent PD-L1 binding compound, wherein each amino acid chain comprising at least one programmed death protein 1 (PD-1) polypeptide is associated with a recombinant adenovirus (Ad), and the plurality of amino acid chains present on the shell polypeptide of recombinant Ad.

[00030] A further aspect of the invention provides such a multivalent PD-L1 binding compound, wherein the recombinant Ad comprises Ad capsid hexon polypeptides of strain Ad6 and at least one Ad capsid hexon polypeptide of strain Ad57.

[00031] An additional aspect of the invention relates to such a multivalent PD-L1 binding compound wherein the Ad capsid hexon polypeptides of strain Ad6 contain HVR 1-7 polypeptides from Ad strain Ad57.

[00032] An additional aspect of the invention relates to such a multivalent PD-L1 binding compound wherein the Ad capsid hexon polypeptides of strain Ad6 contain HVR 2-6 polypeptides from Ad strain Ad57.

[00033] An additional aspect of the invention relates to such a multivalent PD-L1 binding compound, wherein the programmed death protein 1 (PD-1) polypeptide is human PD-1.

[00034] An additional aspect of the invention relates to such a multivalent PD-L1 binding compound wherein the PD-1 polypeptide is fused to the vitamin K-dependent gamma-carboxyglutamine domain of a single chain anti-factor X antibody polypeptide (GLA domain of the FX polypeptide).

[00035] An additional aspect of the invention relates to such a multivalent PD-L1 binding compound, wherein each amino acid chain contains a targeting molecule selected from a measles virus hemagglutinin (MVH) polypeptide, a measles virus fusion polypeptide (MVF), and a vesicular stomatitis virus glycoprotein (VSVG) polypeptide ).

[00036] An additional aspect of the invention relates to such a multivalent PD-L1 binding compound, wherein each amino acid chain contains one or more therapeutic polypeptides selected from a 4-1BB ligand polypeptide (4-1BBL), an OX40 ligand polypeptide (OX40L), a ligand polypeptide CD40 (CD40L) and granulocyte-macrophage colony-stimulating factor polypeptide (GM-CSF).

[00037] A further aspect of the invention relates to such a pharmaceutical composition comprising a multivalent PD-L1 binding compound and a pharmaceutically acceptable carrier.

[00038] A further aspect of the invention relates to a method of treating cancer in an individual in need thereof, comprising administering a multivalent PD-L1 binding compound.

[00039] An additional aspect of the invention relates to such a method, further comprising administering to a mammal one or more antitumor therapeutic agents.

[00040] An additional aspect of the invention relates to such a method, wherein the antitumor therapeutic agent is an immunotherapeutic agent targeting PD-1.

[00041] A further aspect of the invention relates to such a method, wherein the immunotherapeutic agent is selected from the group consisting of nivolumab, pembrolizumab, atezolizumab, avelumab, cemiplimab and durvalumab.

[00042] Unless otherwise specified, all technical and scientific terms used in this description have the meaning commonly understood by one skilled in the art to which the invention belongs. Although methods and materials similar or equivalent to those presented herein may be used in the practice of the present invention, suitable methods and materials are described below. All publications, patent applications, patents and other references mentioned in this description are incorporated herein by reference in their entirety. In the event of a conflict, this description, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and are not intended to be limiting.

[00043] Details of one or more embodiments of the invention are set forth in the accompanying drawings and description below. Other features, objects and advantages of the invention will be apparent from the description and drawings, as well as the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

[00044] Figure 1A shows a schematic diagram of examples of engineered multivalent PD-L1 binding compounds. A multivalent PD-L1 binding compound containing a PD-1 polypeptide fused to an Ig polypeptide, which is a dimeric polypeptide conjugate, includes two amino acid chains, each of which includes one PD-1 polypeptide. The multivalent PD-L1 binding compound, which is a trimeric polypeptide conjugate, contains three amino acid chains, each of which includes one PD-1 polypeptide fused to a sigma polypeptide. The multivalent PD-L1 binding compound, which is a tetrameric polypeptide conjugate, contains four amino acid chains, each of which includes one PD-1 polypeptide fused to a streptavidin polypeptide.

[00045] The amino acid chain includes one PD-1 polypeptide fused to the GLA domain of an FX polypeptide. Fusion proteins can be designed with any substance at the N-terminus or C-terminus. The fusion protein bound to the surface of the adenoviral particle forms a multivalent PD-L1 binding compound.

[00046] Figure 1B shows engineered PD-L1 binding compounds resulting from the incorporation of PD-1 motifs into natural biopolymers like collagen, laminin, fibronectin or elastin. Fusion of PD-1 with protein monomers and expression in cells allows their assembly into multivalent polymers displaying multiple PD-L1-binding motifs.

[00047] Figure 2A shows nucleic acid vectors that can encode amino acid chains that can be used to produce multivalent PD-L1 binding compounds. Figure 2A shows a diagram of a nucleic acid vector that can encode an amino acid chain comprising one mouse PD-1 (mPD-1) polypeptide fused to an IgG polypeptide (top left) of a nucleic acid vector that can encode an amino acid chain comprising one polypeptide Human PD-1 (hPD-1) fused to an IgG polypeptide (top right), and an example cloning strategy (bottom) to replace an IgG-encoding nucleic acid with a nucleic acid encoding the GLA domain of an FX polypeptide to produce a nucleic acid vector that can encode an amino acid chain comprising a PD-1 polypeptide fused to the GLA domain of an FX polypeptide, or to replace a nucleic acid encoding an IgG with a nucleic acid encoding a sigma-1 polypeptide to produce a nucleic acid vector that can encode an amino acid chain comprising a PD polypeptide -1 fused to sigma-1 polypeptide.

[00048] Figure 2B includes images of gels showing the expression of the amino acid chain containing the PD-1 polypeptide. Lane 1: Molecular mass marker (base pairs; bp); Lane 2: PD-1 polypeptide fused to the GLA domain of the FX polypeptide; lane 3: molecular weight marker (bp); Lane 4: amino acid chain including a PD-1 polypeptide fused to a sigma-1 polypeptide.

[00049] Figure 3 shows a diagram of a nucleic acid vector that can encode an amino acid chain comprising an hPD-1 polypeptide fused to the GLA domain of an FX polypeptide (top left), a nucleic acid vector that can encode an amino acid chain comprising an hPD-1 polypeptide fused to a sigma-1 polypeptide (top right), and an example of a cloning strategy (bottom) to replace a nucleic acid encoding an hPD-1 polypeptide with a nucleic acid encoding an mPD-1 polypeptide.

[00050] Figure 4 shows a diagram of examples of nucleic acid vectors that can encode amino acid chains that can be used to produce multivalent PD-L1 binding compounds.

[00051] Figure 5 shows a diagram of examples of adenoviral (Ad) vectors that can encode amino acid chains that can be used to produce multivalent PD-L1 binding compounds.

[00052] Figure 6 shows a diagram of the Ad genome containing a nucleic acid encoding an immune checkpoint polypeptide that can activate T cells (4-1BBL), a nucleic acid encoding one or more PD-1 polypeptides (PD-1-X where X is a dimeric, trimeric, tetrameric or polymeric binding framework), and a nucleic acid encoding an antigen, such as a cancer antigen or an infectious disease antigen (an HPV epitope). A map of Ad genome restriction sites is also shown.

[00053] Figure 7 shows an alignment of Ad5, 6 and 57, showing variations in the hexon and E3 regions. (A) Pustell DNA alignment of the Ad6 and Ad57 genomes. The boxes indicate the hexon and E3 regions where variability is greatest between the two viruses. (B) ClustalW alignment of hypervariable region amino acids in hexon proteins from Ad5, Ad6, and Ad57.

[00054] Figure 8 shows the design of Ad657 resulting from the replacement of HVR Ad6 with HVR Ad57. Abbreviation: HVR, hypervariable regions.

[00055] Figure 9 is a diagram of replication-competent Ad (RC-Ad) where E1 expression is controlled by the native E1 promoter; CRAd-probasine-E1A (Ad-PB) variant, where E1 expression is controlled by the prostate-specific probasine promoter; CRAd-dl1101, where the binding of the p300 pathway, which is susceptible to the IFN pathway in normal cells, is ablated; CRAd-dl1107, where ablation of pRB binding allows the virus to kill malignant cells with RB pathway disruptions, but it is repressed in RB ⁺ normal cells; CRAd-dl1101/07, where the p300 pathway susceptible to the IFN pathway is ablated, ablation of RB binding allows the virus to kill malignant cells, ablation of pRB binding allows the virus to kill RB, but it is repressed in RB ⁺ normal cells.

[00056] Figure 10 shows the amino acid sequences of the N-terminal portion of the wild-type E1A polypeptide and the N-terminus of the E1A variants CRAd, dl1101, dl1107 and dl1101/1107.

[00057] Figure 11 shows a diagram of different E3 immune surveillance genes in Ad species C of Ad6 and Ad species D of Ad26. Both Ads express variants with different sizes and sequences of the E3 genes 12.5K, 6.7K, 19K, 10.4K (RIDα), 14.5K (RIDβ), and 14.7K, as well as depictions of the functions of these encoded E3 proteins.

[00058] Figure 12 shows regions on HVR Ad that can be modified, for example, by PEGylation or "bapylation" with biotin acceptor peptides (BAPs).

[00059] Figure 13 is a diagram showing Ad therapeutic cycles. A) Ad serotype switching pattern. B) Schematic of an example therapeutic cycle where Ad6 and Ad657 can be used for multiple rounds of treatment through serotype switching in combination with covalent polymer conjugation.

[00060] Figure 14 shows the conjugation of polyethylene glycol (PEG) to Ad657-HVR1-C. A) PAGE electrophoresis with SDS of Ad proteins with and without PEGylation. Arrows indicate increases in size due to chemical attachment of PEG to hexon. B) Effects of targeted PEGylation with maleimide-PEG and non-targeting NHS-PEG on virus infection.

[00061] Figure 15 shows chimeric HVR constructs that combine different HVRs from different Ad serotypes to modulate natural interactions with cells and blood factors that improve pharmacology, in combination with the insertion of cell-binding and cell-non-targeting peptides into different HVRs for changes in cell entry and cell avoidance. In this example, a single amino acid cysteine is incorporated into HVR1 and HVR5 Ad657 to modulate the pharmacology and target conjugation of polymers like polyethylene glycol or other imaging agents like fluorophores.

[00062] Figure 16 shows plasmid maps for typical CRAds and peptide combinations. Hexons as well as insertions of cell-targeting peptides into individual HVRs are shown.

[00063] Figure 17 shows plasmid maps for typical CRAds and peptide combinations. Hexons as well as insertions of cell-targeting peptides into individual HVRs are shown.

[00064] Figure 18 shows plasmid maps for typical CRAds and peptide combinations. Hexons as well as insertions of cell-targeting peptides into individual HVRs are shown.

[00065] Figure 19. PD-1/PD-L1 blockade assay - Fusion protein purified from cells infected with pAd6/57/6-dl1107-DE3-RSV-hPD-1-HA-Ig-I on a protein A column This protein was titrated in parallel with a positive control, anti-PD-1 monoclonal antibody from Promega (Madison, WI, USA).

[00066] Figure 20. Tumor growth assessment - B16-CAR melanoma cells were injected subcutaneously into C57BL/6 mice and tumors were injected with 3e11 viral particles of the indicated vectors. The CRAd+PD-L1 trap is pAd6/57/6-dl1107-ΔE3-RSV-hPD-1-HA-Ig-I. The CRAd+PD-L1+immunostimulator trap corresponds to tumors co-injected with pAd6/57/6-dl1107-ΔE3-RSV-hPD-1-HA-Ig-I and a second adenovirus expressing 4-1BBL.

DETAILED DESCRIPTION OF THE INVENTION

[00067] The present invention relates to methods and compositions for the treatment of malignant neoplasms containing PD-1 polypeptides. PD-1 polypeptides may be in the form of multivalent PD-L1 binding compounds.

[00068] In some cases, a multivalent PD-L1 binding compound contains two or more amino acid segments that can bind PD-L1 (eg, PD-1 polypeptides and/or fragments thereof). For example, a multivalent PD-L1 binding compound comprising two or more PD-1 polypeptides (and/or fragments thereof having the ability to bind PD-L1) can bind to two or more PD-L1 polypeptides. In some cases, a multivalent PD-L1 binding compound may comprise a single amino acid chain comprising two or more amino acid segments that can bind PD-L1.

[00069] In some cases, a multivalent PD-L1 binding compound may include a polypeptide conjugate comprising multiple (eg, two or more) amino acid chains, each of which includes one or more amino acid segments that can bind to PD-L1. For example, an amino acid chain comprising one or more amino acid segments that can bind to PD-L1 may be a fusion polypeptide comprising one or more PD-1 polypeptides fused to a framework polypeptide that may form a polypeptide conjugate with one or more other polypeptides comprising one or more amino acid segments that can bind to PD-L1 (eg, a polypeptide conjugate comprising multiple linked amino acid chains). If a framework polypeptide fused to one or more PD-1 polypeptides is in a polypeptide conjugate, the polypeptide conjugate may include two or more PD-1 polypeptides.

[00070] In an embodiment, the viral particle containing the PD-1 polypeptide may be in the form of a multivalent PD-L1 binding compound, where two or more viral envelope polypeptides are modified to include one or more amino acid chains comprising one or more amino acids segments that can bind to PD-L1.

[00071] In an embodiment, the PD-L1 binding compound comprises a recombinant virus genetically modified to express a PD-1 polypeptide, wherein the PD-1 polypeptide is displayed on the viral envelope. In a further embodiment, the recombinant virus is genetically modified to express a PD-1 polypeptide in combination with one or more polypeptides heterologous to the recombinant virus backbone, such as therapeutic polypeptides, targeting molecules/polypeptides, and antigenic polypeptides.

[00072] In an embodiment, the PD-L1 binding compound comprises recombinant Ad (i.e., Ad657, Ad6/57/6, and variants thereof) modified in one or more hypervariable regions (HVRs) of the viral capsid protein hexon, containing at least at least one PD-1 polypeptide. In an embodiment, the recombinant Ad is further modified to contain heterologous polypeptides, such as targeting polypeptides, therapeutic polypeptides and/or antigens. PD-1 polypeptides, as well as heterologous polypeptides, are expressed following viral replication. Once the virus is assembled, the expressed polypeptides can be displayed on the surface of the virus as a component of the virus capsid structure.

[00073] In a further embodiment, the PD-L1 binding compound comprises recombinant Ad genetically modified to express a PD-1 polypeptide as a fusion protein with one or more heterologous polypeptides, wherein the fusion protein is displayed on the capsid of the viral particle. Heterologous polypeptides may include the GLA domain of an FX polypeptide, CAR polypeptides, CD46 polypeptides, desmoglein polypeptides, integrin polypeptides, single chain antibody polypeptides, Camelid antibody polypeptides, capsid polypeptides, and envelope binding polypeptides.

[00074] In some cases, the amino acid chain comprising one or more PD-1 polypeptides and a polypeptide that can bind to a viral envelope polypeptide may be a fusion polypeptide comprising one or more PD-1 polypeptides and a polypeptide that can bind to a viral polypeptide shells. In an embodiment, a fusion protein of PD-1 and a heterologous polypeptide can be expressed in a host cell, purified, and mixed with Ad such that the fusion protein is associated with the surface of Ad by covalently attaching the heterologous polypeptide to Ad. If a first fusion polypeptide comprising one or more PD-1 polypeptides fused to a polypeptide that can bind to a viral envelope polypeptide is associated with a first viral envelope polypeptide on a viral particle, and a second fusion polypeptide comprising one or more PD-1 polypeptides, fused to a polypeptide that can bind to a viral envelope polypeptide is linked to a second viral envelope polypeptide on the viral particle, the viral particle will include two or more PD-1 polypeptides.

[00075] In some cases, the PD-L1 binding compound may comprise a viral particle where one or more viral envelope polypeptides are modified to include one or more amino acid chains including one or more amino acid segments that can bind PD-L1 ( for example, PD-1 polypeptides and/or fragments thereof). PD-1 polypeptides can be expressed during viral replication and are assembled and displayed on the viral envelope. In an embodiment, Ad (ie, Ad657, Ad6/57/6, and variants thereof) is modified in the HVR region to express a PD-1 polypeptide in combination with one or more heterologous polypeptides selected from therapeutic polypeptides, targeting polypeptides, and antigenic polypeptides. The PD-1 polypeptide can be expressed as a fusion protein with one or more heterologous polypeptides. For example, a viral particle is genetically modified to express two or more amino acid chains, each of which includes at least one PD-1 polypeptide and a heterologous polypeptide, where the amino acid chains are exposed on the surface of the viral particle as a component of the viral capsid, thereby forming a multivalent PD -L1-binding compound. In some cases, the viral particle is genetically modified to express two or more amino acid chains, each of which includes a PD-1 polypeptide, and is modified to express one or more amino acid chains comprising a heterologous polypeptide, wherein the amino acid chains are exposed on the surface of the viral particle as a component of the capsid virus, thereby forming a multivalent PD-L1 binding compound. In some cases, the amino acid chain includes two or more PD-1 polypeptides and a heterologous polypeptide exposed on the surface of the viral particle.

[00076] In an embodiment, the amino acid chain containing one or more PD-1 polypeptides may be a fusion protein containing one or more PD-1 polypeptides fused to a heterologous polypeptide. The heterologous polypeptide can bind to the surface of the Ad enveloped virus particle. In an embodiment, the fusion proteins are expressed in a host cell, purified, and mixed with Ad such that the fusion protein coats the surface of the Ad viral particle, thereby forming a multivalent PD-L1 binding compound.

[00077] In some cases, a multivalent PD-L1 binding compound provided herein may have increased affinity for PD-L1 (eg, compared to a monomeric PD-1 polypeptide). In some cases, a multivalent PD-L1 binding compound provided herein may have increased avidity for PD-L1 (eg, compared to a monomeric PD-1 polypeptide).

[00078] The present invention relates to methods and materials for producing compositions containing PD-1 polypeptides and multivalent PD-L1 binding compounds, as well as methods and materials for producing recombinant viruses containing nucleic acid molecules encoding an amino acid chain that can be used for the production of multivalent PD-L1 binding compounds presented herein.

[00079] Multivalent PD-L1 binding compounds as provided herein (e.g., a fusion polypeptide comprising one or two or more PD-1 polypeptides, or a polypeptide conjugate comprising two or more amino acid chains each comprising one or more PD-1 polypeptides, or a viral particle, wherein the viral envelope proteins contain one or more amino acid chains comprising one or more PD-1 polypeptides, or a viral particle, wherein one or more PD-1 polypeptides are associated with the surface of the viral particle), may include any suitable number of amino acid segments that can bind to PD-L1 (eg, PD-1 polypeptides and/or fragments thereof that can bind to PD-L1).

[00080] In some cases, a multivalent PD-L1 binding compound provided herein may include two or more (e.g., 2, 3, 4, 5, 6, 240, 720, or more) amino acid segments that can bind with PD-L1. In some cases, a multivalent PD-L1 binding compound provided herein may include from about two to about 720 amino acid segments that can bind to PD-L1. For example, if the multivalent PD-L1 binding compound provided herein is a single amino acid chain comprising two or more amino acid segments that can bind to PD-L1, the multivalent PD-L1 binding compound may comprise about 2, 3, 4, 5, 6, 7 or 8 PD-1 polypeptides (or PD-L1 binding fragments thereof). For example, if the multivalent PD-L1 binding compound provided herein is a polypeptide conjugate comprising a plurality of amino acid chains, each of which includes one or more amino acid segments that can bind PD-L1, the multivalent PD-L1 binding compound may include approximately 2, 3, 4, 5, 6, 7 or 8 PD-1 polypeptides (or PD-L1 binding fragments thereof).

[00081] For example, if the multivalent PD-L1 binding compound provided herein is a viral particle, where two or more viral envelope polypeptides (e.g., capsid polypeptides) are modified to include one or more amino acid chains comprising one or more amino acid segments that can bind to PD-L1, a multivalent PD-L1 binding compound may include from about 240 PD-1 polypeptides to about 720 PD-1 polypeptides (or PD-L1 binding fragments thereof). In cases where the multivalent PD-L1 binding compound provided herein includes two amino acid segments that can bind to PD-L1 (or a viral particle where one or more viral envelope polypeptides are modified to include one or more amino acid chains comprising two or more amino acid segments that can bind to PD-L1), a multivalent PD-L1 binding compound may be referred to as a dimeric or bivalent PD-L1 binding compound. In cases where the multivalent PD-L1 binding compound provided herein includes three amino acid segments that can bind to PD-L1, the multivalent PD-L1 binding compound may be referred to as a trimeric or trivalent PD-L1 binding compound. In cases where the multivalent PD-L1 binding compound provided herein includes four amino acid segments that can bind to PD-L1, the multivalent PD-L1 binding compound may be referred to as a tetrameric or tetravalent PD-L1 binding compound.

[00082] Multivalent PD-L1 binding compounds provided herein may include any suitable amino acid segments that can bind PD-L1.

[00083] In an embodiment, the amino acid segment that can bind to PD-L1 may include a PD-1 polypeptide. In some cases, the amino acid segment that can bind PD-L1 may include any fragment of a PD-1 polypeptide, provided that the fragment retains the ability to bind PD-L1. In some cases, the PD-1 polypeptide may be an sPD-1 polypeptide. The PD-1 polypeptide can be obtained from any suitable animal. In some cases, the PD-1 polypeptide can be obtained from a mammal. For example, the PD-1 polypeptide may be an hPD-1 polypeptide. For example, the PD-1 polypeptide may be an mPD-1 polypeptide. The PD-1 polypeptide may include any suitable PD-1 polypeptide sequence. In some cases, the PD-1 polypeptide can be modified (eg, to interact with PD-L1 with higher affinity). Examples of PD-1 polypeptide sequences (and nucleic acids encoding such polypeptides) may be as listed in the National Center for Biotechnology Information (NCBI) databases, for example, accession number AI928135 (version AI928135.1), accession number AY238517 ( version AY238517.1) and registration number CR988122 (version CR988122.1), registration number U64863 (version U64863.1), registration number NM_008798 (version NM_008798.2) and registration number NP_032824 (version NP_032824.1). In some cases, the PD-1 polypeptide contains the amino acid sequence set forth in SEQ ID NO: 5. In some cases, the PD-1 polypeptide contains the amino acid sequence set forth in SEQ ID NO: 6.

[00084] In some cases, the amino acid segment that can bind to PD-L1 (e.g., PD-1 polypeptides and/or fragments thereof) may have a different sequence from the wild-type PD-1 polypeptide (e.g., PD-1 polypeptide wild type having the amino acid sequence set forth in SEQ ID NO: 5 or SEQ ID NO: 6), sometimes referred to as a sequence variant. For example, the sequence of a PD-1 polypeptide (and/or a fragment thereof having the ability to bind to PD-L1) may have at least 80% sequence identity to SEQ ID NO: 5 or SEQ ID NO: 6. In some embodiments, the amino acid the segment that can bind PD-L1 may have at least 85% sequence identity, 90% sequence identity, 95% sequence identity, or at least 99% sequence identity to SEQ ID NO: 5 or SEQ ID NO: 6 Percentage sequence identity is calculated by determining the number of matching positions in aligned polypeptide sequences, dividing the number of matching positions by the total number of aligned amino acids accordingly and multiplying by 100. The term "matching position" refers to a position at which identical amino acids occur in the same position in aligned sequences. The term “total number of aligned amino acids” refers to the minimum number of amino acids in a PD-1 polypeptide required to align a second sequence, and does not include alignments (eg, forced alignments) with non-PD-1 sequences, such as sequences fused to a PD-1 polypeptide. 1 (eg, amino acids from an IgG polypeptide, streptavidin polypeptide, sigma-1, or the GLA domain of an FX polypeptide fused to a PD-1 polypeptide). The total number of aligned amino acids may correspond to the entire PD-1 sequence or may correspond to fragments of the full-length PD-1 sequence. Sequences can be aligned using the algorithm described by Altschul et al. ( Nucleic Acids Res. , 25:3389-3402 (1997)) and included in the BLAST (Basic Local Alignment Search Tool) programs available at ncbi.nlm.nih.gov on the Internet. BLAST searches or alignments can be performed to determine the percent sequence identity between a PD-1 polypeptide and any other sequence or portion thereof using the algorithm of Altschul et al . BLASTN is a program used for alignment and identity comparison between nucleic acid sequences, while BLASTP is a program used for alignment and identity comparison between amino acid sequences. When using BLAST programs to calculate the percent identity between a PD-1 sequence and another sequence, the default parameters of the corresponding programs are used.

[00085] In some cases, a multivalent PD-L1 binding compound may include a polypeptide conjugate comprising multiple amino acid chains, each of which includes one or more amino acid segments that can bind PD-L1 (e.g., PD-1 polypeptides and/or their fragments). As used herein, the term "multiple amino acid chains", each of which includes one or more amino acid segments that can bind PD-L1, refers to two or more (for example, 2, 3, 4, 5, 6, 7, 8 or more) amino acid chains, each of which includes one or more amino acid segments that can bind to PD-L1. For example, an amino acid chain comprising one or more PD-1 polypeptides may be a fusion polypeptide comprising one or more PD-1 polypeptides fused to a framework polypeptide, which may form a polypeptide conjugate with one or more other polypeptides comprising one or more amino acids segments that can bind to PD-L1 (eg, a polypeptide conjugate comprising multiple linked amino acid chains). If a framework polypeptide fused to one or more PD-1 polypeptides is in a polypeptide conjugate, the polypeptide conjugate may include two or more PD-1 polypeptides. In some cases, a polypeptide conjugate comprising multiple amino acid chains, each of which includes one or more PD-1 polypeptides, may be a homomeric PD-1 polypeptide (e.g., may include two or more identical amino acid chains, each of which includes one or more polypeptides PD-1). In some cases, a polypeptide conjugate comprising multiple amino acid chains, each of which includes one or more PD-1 polypeptides, may be a heteromeric PD-1 polypeptide (e.g., may include two or more different amino acid chains, each of which includes one or more polypeptides PD-1).

[00086] An amino acid chain comprising one or more amino acid segments that can bind to PD-L1 (e.g., PD-1 polypeptides and/or fragments thereof) present in a polypeptide conjugate provided herein (e.g., a polypeptide conjugate comprising a plurality of amino acid chains, each of which includes one or more PD-1 polypeptides) may include one or more amino acid segments that can bind to PD-L1 fused to any suitable framework polypeptide. For example, an amino acid chain comprising one or more PD-1 polypeptides present in a polypeptide conjugate provided herein may be a fusion polypeptide comprising one or more PD-1 polypeptides fused to any suitable framework polypeptide. In some cases, the framework polypeptide may have the ability to form a polypeptide conjugate (eg, a polypeptide conjugate comprising multiple linked amino acid chains). For example, two or more amino acid chains comprising one or more PD-1 polypeptides fused to a backbone polypeptide can form a polypeptide conjugate comprising two or more PD-1 polypeptides. If two or more amino acid chains comprising one or more PD-1 polypeptides fused to a backbone polypeptide form a polypeptide conjugate, the two or more amino acid chains may be linked (eg, through a covalent bond or ionic bond). The framework polypeptide can be any suitable polypeptide that can form a polypeptide conjugate with another polypeptide. The framework polypeptide may naturally form a polypeptide conjugate, or it may be engineered to form a polypeptide conjugate. Non-limiting examples of framework polypeptides that can form a polypeptide conjugate may include Ig polypeptides (eg, IgG polypeptides, such as IgG2 polypeptides), sigma-1 polypeptides, and streptavidin polypeptides.

[00087] An amino acid chain comprising one or more amino acid segments that can bind to PD-L1 (e.g., PD-1 polypeptides and/or fragments thereof), which may be present in a polypeptide conjugate provided herein (e.g., a polypeptide conjugate comprising a plurality of amino acid chains, each of which includes one or more PD-1 polypeptides), may include any suitable amino acid sequence. For example, an amino acid chain comprising one or more PD-1 polypeptides fused to a framework polypeptide may comprise any suitable amino acid sequence. Non-limiting examples of amino acid sequences of amino acid chains comprising one or more PD-1 polypeptides and a framework polypeptide include the amino acid sequences set forth in SEQ ID NO: 7, 10, 11, 12, 13, 16, 17, 18, 19, 20, 21 , 24, 25, 26 or 27. In some cases, the amino acid sequence of the amino acid chain comprising one or more PD-1 polypeptides and the framework polypeptide may have a sequence different from one of the amino acid sequences set forth in SEQ ID NO: 7, 10, 11, 12, 13, 16, 17, 18, 19, 20, 21, 24, 25, 26, or 27, sometimes referred to as a sequence variant. For example, the amino acid sequence of a fusion polypeptide comprising one or more PD-1 polypeptides and a framework polypeptide may have at least 80% sequence identity to any of the amino acid sequences set forth in SEQ ID NO: 7, 10, 11, 12, 13 , 16, 17, 18, 19, 20, 21, 24, 25, 26, or 27. In some embodiments, the amino acid sequence of a fusion polypeptide comprising one or more PD-1 polypeptides and a framework polypeptide may have at least 85% identity sequence identity, 90% sequence identity, 95% sequence identity, or at least 99% sequence identity with respect to any of the amino acid sequences shown in SEQ ID NO: 7, 10, 11, 12, 13, 16, 17, 18, 19, 20, 21, 24, 25, 26 or 27. Percent sequence identity is calculated by determining the number of matching positions in the aligned polypeptide sequences, dividing the number of matching positions by the total number of aligned amino acids as appropriate and multiplying by 100. The term "matching position" refers to the position , in which identical amino acids occur at the same position in aligned sequences. Sequences can be aligned using the algorithm described by Altschul et al. ( Nucleic Acids Res. , 25:3389-3402 (1997)) and included in the BLAST (Basic Local Alignment Search Tool) programs available at ncbi.nlm.nih.gov on the Internet. BLAST searches or alignments can be performed to determine the percent sequence identity between a polypeptide and any other sequence or portion thereof using the algorithm of Altschul et al . BLASTN is a program used for alignment and identity comparison between nucleic acid sequences, while BLASTP is a program used for alignment and identity comparison between amino acid sequences. When using BLAST programs to calculate the percent identity between a polypeptide sequence and another sequence, the default parameters of the corresponding programs are used. In some cases, an amino acid chain comprising one or more PD-1 polypeptides, which may be present in a polypeptide conjugate provided herein (e.g., a polypeptide conjugate comprising a plurality of amino acid chains, each of which includes one or more PD-1 polypeptides), may be as indicated in example 7.

[00088] In some cases, an amino acid chain comprising one or more amino acid segments that can bind to PD-L1 (e.g., PD-1 polypeptides and/or fragments thereof) can be fused to a framework polypeptide, which can form a polypeptide conjugate, comprising two linked amino acid chains (eg, an IgG polypeptide, such as an IgG2 polypeptide). For example, two amino acid chains, each comprising one PD-1 polypeptide fused to an IgG2 polypeptide, can form a polypeptide conjugate comprising two PD-1 polypeptides. Non-limiting examples of amino acid sequences of amino acid chains comprising a single PD-1 polypeptide fused to an IgG2 polypeptide include the amino acid sequences set forth in SEQ ID NO: 7, SEQ ID NO: 13 or SEQ ID NO: 21.

[00089] In some cases, an amino acid chain comprising one or more amino acid segments that can bind to PD-L1 (e.g., PD-1 polypeptides and/or fragments thereof) can be fused to a framework polypeptide, which can form a polypeptide conjugate, comprising three linked amino acid chains (for example, sigma-1 polypeptide). For example, three amino acid chains, each comprising one PD-1 polypeptide fused to a sigma-1 polypeptide, can form a polypeptide conjugate comprising three PD-1 polypeptides. Non-limiting examples of amino acid sequences of amino acid chains comprising one PD-1 polypeptide fused to a sigma-1 polypeptide include the amino acid sequences set forth in SEQ ID NO: 10, SEQ ID NO: 16, or SEQ ID NO: 24.

[00090] In some cases, an amino acid chain comprising one or more amino acid segments that can bind to PD-L1 (e.g., PD-1 polypeptides and/or fragments thereof) can be fused to a backbone polypeptide, which can form a polypeptide conjugate, comprising four linked amino acid chains (for example, streptavidin polypeptide). For example, four amino acid chains, each comprising one PD-1 polypeptide fused to a streptavidin polypeptide, can form a polypeptide conjugate comprising four PD-1 polypeptides. Non-limiting examples of amino acid sequences of amino acid chains comprising a single PD-1 polypeptide fused to a streptavidin polypeptide include the amino acid sequences set forth in SEQ ID NO: 11 or SEQ ID NO: 17.

[00091] In some cases, the multivalent PD-L1 binding compound may comprise a viral particle where two or more viral envelope polypeptides are modified to include one or more amino acid chains comprising one or more amino acid segments that can bind PD-L1 (eg, PD-1 polypeptides and/or fragments thereof). For example, a viral particle can be modified by coating the viral particle with two or more amino acid chains, each of which includes one or more PD-1 polypeptides and a polypeptide that can bind to the viral envelope polypeptide. An amino acid chain comprising one or more amino acid segments that can bind PD-L1 (e.g., PD-1 polypeptides and/or fragments thereof) that can be used to coat a viral particle to produce a multivalent PD-L1 binding compound (e.g., viral particle, where two or more viral envelope polypeptides are modified to include one or more amino acid chains comprising one or more PD-1 polypeptides) may include one or more amino acid segments that can bind to PD-L1 fused to any a suitable polypeptide that can bind to the viral envelope polypeptide. In some cases, a polypeptide that can bind to a viral envelope polypeptide may be covalently linked to one or more viral envelope polypeptides. In some cases, a polypeptide that can bind to a viral envelope polypeptide may bind to the viral envelope polypeptide with high affinity (eg, from about 0.25 μM to about 100 μM). The polypeptide that can bind to the viral envelope polypeptide may naturally bind to the viral envelope polypeptide or is engineered to bind to the viral envelope polypeptide. The polypeptide that can bind to the viral envelope polypeptide can bind to any suitable viral envelope polypeptide.

[00092] Non-limiting examples of viral envelope polypeptides include capsid polypeptides (eg, hexon polypeptides, fiber polypeptides, penton polypeptides) and adenovirus IX polypeptides. The polypeptide that can bind to the viral envelope polypeptide can bind to the viral envelope polypeptide present on any suitable type of viral particle. The virus particle may be a replication-competent (RC) virus particle (eg, helper-dependent (HD) virus particles and single-cycle (SC) virus particles), the virus particle may be a replication-defective (RD) virus particle, or the virus may be conditionally -replicative viral particle (CRAd). In some cases, the viral particle may be an oncolytic viral particle.

[00093] In some cases, the viral particle may be a viral vector. Non-limiting examples of viral particles that can be coated with one or more amino acid chains including one or more PD-1 polypeptides and a polypeptide that can bind to a viral envelope polypeptide include Ad, adeno-associated viruses (AAV), lentiviruses, enteroviruses, reoviruses, poxviruses, measles virus and herpes viruses.

[00094] Non-limiting examples of polypeptides that may be present in a viral envelope polypeptide include a GLA domain polypeptide (eg, the GLA domain of an FX polypeptide), CAR polypeptides, CD46 polypeptides, desmoglein polypeptides, integrin polypeptides, single chain antibody polypeptides, Camelid antibody polypeptides, polypeptides capsid and envelope-binding polypeptides.

[00095] An amino acid chain comprising one or more amino acid segments that can bind PD-L1 (e.g., PD-1 polypeptides and/or fragments thereof) that can be used to coat a viral particle to produce a multivalent PD-L1 binding compound (eg, a viral particle where one or two or more viral envelope polypeptides are modified to include one or more amino acid chains comprising one or more PD-1 polypeptides) may include any suitable amino acid sequence. For example, a viral particle can be contacted with a composition comprising two or more amino acid chains, each of which includes one or more PD-1 polypeptides and a polypeptide that can covalently bind to a viral envelope polypeptide such that the amino acid chains coat the viral particle . In some cases, the amino acid chain comprising one or more PD-1 polypeptides and a polypeptide that can bind to a viral envelope polypeptide may be a fusion polypeptide comprising one or more PD-1 polypeptides and a polypeptide that can bind to a viral envelope polypeptide. The amino acid chain comprising one or more PD-1 polypeptides and a polypeptide that can bind to a viral envelope polypeptide may include any suitable amino acid sequence. Non-limiting examples of amino acid sequences of amino acid chains comprising one or more PD-1 polypeptides and a polypeptide that can bind to a viral envelope polypeptide include the amino acid sequences set forth in SEQ ID NO: 8, 9, 14, 15, 22 or 23. In some In cases where the amino acid sequence of the amino acid chain comprising one or more PD-1 polypeptides and a polypeptide that can bind to the viral envelope polypeptide may have a sequence different from one of the amino acid sequences set forth in SEQ ID NO: 8, 9, 14, 15, 22 or 23, sometimes referred to as a sequence variant. For example, the amino acid sequence of a fusion polypeptide comprising one or more PD-1 polypeptides and a polypeptide that can bind to a viral envelope polypeptide may have at least 80% sequence identity to any of the amino acid sequences set forth in SEQ ID NO: 8. 9, 14, 15, 22, or 23. In some embodiments, the amino acid sequence of a fusion polypeptide comprising one or more PD-1 polypeptides and a polypeptide that can bind to a viral envelope polypeptide may have at least 85% sequence identity, 90% sequence identity, 95% sequence identity, or at least 99% sequence identity with respect to any of the amino acid sequences shown in SEQ ID NO: 8, 9, 14, 15, 22 or 23. The percentage of sequence identity is calculated by determining the number of matching positions in aligned polypeptide sequences by dividing the number of matching positions by the total number of aligned amino acids accordingly and multiplying by 100. The term “matching position” refers to a position in which identical amino acids occur at the same position in the aligned sequences. Sequences can be aligned using the algorithm described by Altschul et al. ( Nucleic Acids Res. , 25:3389-3402 (1997)) and included in the BLAST (Basic Local Alignment Search Tool) programs. In some cases, the amino acid chain comprising one or more PD-1 polypeptides and a polypeptide that can bind to a viral envelope polypeptide may be as set forth in the examples.

[00096] In some cases, an amino acid chain comprising one or more amino acid segments that can bind to PD-L1 (e.g., PD-1 polypeptides and/or fragments thereof) fused to a polypeptide that can bind to a viral envelope polypeptide (e.g. , GLA domain of an FX polypeptide) can be used to coat (eg, modify one or more viral envelope polypeptides) a viral particle to produce a coated viral particle having two or more amino acid chains including one or more amino acid segments that can bind to PD -L1 associated with one or more viral envelope polypeptides present on the surface of the viral particle.

[00097] For example, one or more amino acid chains, each comprising one PD-1 polypeptide fused to the GLA domain of an FX polypeptide, may be linked to two or more viral hexon polypeptides and/or other viral envelope proteins to coat the viral particle with two or more amino acid chains, each of which includes one PD-1 polypeptide fused to the GLA domain of an FX polypeptide. If the viral particle is Ad, 240 hexon polypeptide homotrimers (e.g., polypeptide conjugates comprising three hexon polypeptides) may be present on the viral particle (see, e.g., Chen et al. , Human gene therapy , 21:739-749 (2010)) . In some cases, a fusion polypeptide comprising one PD-1 polypeptide and the GLA domain of an FX polypeptide can be used to coat an Ad particle to produce a multivalent PD-L1 binding compound comprising approximately 240 PD-1 polypeptides. For example, if a fusion polypeptide comprising one PD-1 polypeptide fused to the GLA domain of an FX polypeptide is associated with each hexon polypeptide trimer on an Ad virus particle, the virus particle will include 240 PD-1 polypeptides. In some cases, a fusion polypeptide comprising one PD-1 polypeptide and the GLA domain of an FX polypeptide can be used to coat an Ad particle to produce a multivalent PD-L1 binding compound that may include from about 240 PD-1 polypeptides to about 720 PD-1 polypeptides. 1. For example, if a fusion polypeptide comprising one PD-1 polypeptide fused to the GLA domain of an FX polypeptide is linked to each hexon polypeptide on an Ad virus particle, the virus particle will include 720 PD-1 polypeptides.

[00098] In some cases, multivalent PD-L1 binding compounds provided herein (e.g., a fusion polypeptide comprising two or more PD-1 polypeptides, or a polypeptide conjugate comprising one or two or more amino acid chains, each which comprises one or more PD-1 polypeptides, or a viral particle, wherein two or more viral envelope polypeptides comprise one or more amino acid chains comprising one or more PD-1 polypeptides, or a viral particle, wherein one or more viral envelope polypeptides comprise one or more amino acid chains comprising two or more PD-1 polypeptides) may also include one or more additional molecules/polypeptides.

[00099] Non-limiting examples of molecules that can be included in the multivalent PD-L1 binding compound provided herein include targeting molecules (eg, targeting polypeptides and targeting nucleic acid sequences), antigens, therapeutic molecules, and detectable polypeptides. In some cases, if the multivalent PD-L1 binding compound is a polypeptide conjugate comprising a plurality of amino acid chains, each of which includes one or more PD-1 polypeptides and/or PD-1 polypeptide fragments having the ability to bind PD-L1, and the polypeptide conjugate includes one or more additional molecules, the one or more additional molecules may be included in at least one (e.g., 1, 2, 3, 4 or more) amino acid chain comprising one or more PD-1 polypeptides and/or polypeptide fragments PD-1, having the ability to bind to PD-L1, present in the polypeptide conjugate. For example, where the multivalent PD-L1 binding compound is a polypeptide conjugate comprising a plurality of amino acid chains, each of which includes one or more PD-1 polypeptides and/or PD-1 polypeptide fragments having the ability to bind PD-L1, and the polypeptide conjugate includes one or more additional molecules, one or more additional molecules may be included in each amino acid chain comprising one or more PD-1 polypeptides and/or PD-1 polypeptide fragments having the ability to bind to PD-L1 present in the polypeptide conjugate. In some cases, if the multivalent PD-L1 binding compound is a viral particle, where two or more viral envelope polypeptides are modified to include one or more amino acid chains comprising one or more PD-1 polypeptides and/or PD-1 polypeptide fragments having the ability to bind to PD-L1 (or a viral particle, where one or more viral envelope polypeptides comprise one or more amino acid chains comprising two or more amino acid segments comprising one or more PD-1 polypeptides and/or PD-1 polypeptide fragments having the ability to bind PD-L1), one or more additional molecules may be included in at least one (e.g., 1, 2, 3, 4 or more) amino acid chain comprising one or more PD-1 polypeptides and/or polypeptide PD-1 fragments used to modify the viral envelope polypeptide on the viral particle. For example, if the multivalent PD-L1 binding compound is a viral particle, where two or more viral envelope polypeptides are modified to include one or more amino acid chains comprising one or more PD-1 polypeptides and/or PD-1 polypeptide fragments having ability to bind to PD-L1 (or a viral particle where one or more viral envelope polypeptides are modified to include one or more amino acid chains comprising two or more amino acid segments comprising one or more PD-1 polypeptides and/or PD polypeptide fragments -1 having the ability to bind to PD-L1), one or more additional molecules can be included in each amino acid chain comprising one or more PD-1 polypeptides and/or PD-1 polypeptide fragments used to modify the viral envelope polypeptide on the viral particle .

[000100] If the PD-L1 binding compounds provided herein also include one or more targeting molecules (eg, targeting polypeptides and targeting nucleic acid sequences), the targeting molecule can be any suitable targeting molecule. In some cases, the targeting molecule may be a polypeptide (eg, a targeting polypeptide). In some cases, the targeting molecule may be a nucleic acid sequence (eg, a targeting nucleic acid sequence). In some cases, a targeting molecule can attract a T cell to a PD-L1 positive cell (eg, a PD-L1 positive cancer cell). For example, if a multivalent PD-L1 binding compound including a targeting molecule is bound to PD-L1 on a PD-L1 positive cell, the targeting molecule on the multivalent PD-L1 binding compound can attract T cells to the PD-L1 positive cell . For example, if the multivalent PD-L1 binding compound includes a targeting molecule, the targeting molecule on the multivalent PD-L1 binding compound can direct the multivalent PD-L1 binding compound to a target location (eg, a target tissue) in a mammal. Examples of targeting molecules that can be included in the multivalent PD-L1 binding compound provided herein include viral polypeptides, antigens (eg, peptide antigens), miRNA targeting sequences (eg, tissue-specific miRNA targeting sequences), antibodies, growth factors, and carbohydrates. If the targeting polypeptide is a viral polypeptide, the viral polypeptide can be derived from any suitable virus (e.g., measles virus (MV), canine rabies virus (CDV), vesicular stomatitis virus (VSV), gibbon leukemia virus (GALV), Zika virus, immunodeficiency virus human (HIV) and human endogenous retrovirus (HERV)). If the targeting polypeptide is a viral polypeptide, the viral polypeptide may be any suitable viral polypeptide (e.g., a hemagglutinin (H) polypeptide such as a non-targeting H polypeptide, a fusion polypeptide (F), a viral glycoprotein polypeptide (G), an envelope polypeptide (env; e.g. env HERV), syncytin-1 and syncytin-2). In some cases, the viral polypeptide may be an MV H polypeptide. In some cases, the viral polypeptide may be an MV F polypeptide. In some cases, the viral polypeptide may be a VSV G polypeptide.

[000101] If the PD-L1 binding compounds provided herein also include one or more therapeutic molecules, the therapeutic molecule may be any suitable therapeutic molecule. In some cases, the therapeutic molecule may be a therapeutic polypeptide. In some cases, a therapeutic molecule may reduce or eliminate PD-L1 inactivation of one or more T cells. In some cases, a therapeutic molecule may activate and/or lead to the accumulation of one or more pharmaceutical agents (eg, anticancer drugs). Examples of therapeutic molecules that can be included in the multivalent PD-L1 binding compound provided herein include 4-1BB ligand polypeptide (4-1BBL), OX40 ligand polypeptide (OX40L), granulocyte-macrophage colony-stimulating factor (GM-CSF) polypeptide ), anti-PD-1 antibodies, nucleic acid encoding anti-PD-1 antibodies, thymidine kinase, cytosine deaminase and iodine symporter. In some cases, the therapeutic molecule may be a therapeutic antibody. The therapeutic antibody may be an agonistic antibody targeting GITR. Signaling through GITR enhances T cell proliferation and effector functions and protects T cells from activation-induced cell death, which in turn increases the proportion of memory T cells. Combining GITR agonists with the PD-L1-binding compounds of the invention may exhibit significant synergistic effects.

[000102] If the PD-L1 binding compounds provided herein also include one or more detectable polypeptides, the detectable polypeptide may be any suitable detectable polypeptide. In some cases, a detectable polypeptide can be detected (eg, a clinician can detect) to determine the location of one or more PD-L1 binding compounds provided herein. In some cases, a detectable polypeptide can be detected (eg, a clinician can detect) to monitor the persistence of one or more multivalent PD-L1 binding compounds provided herein. Examples of detectable polypeptides that can be included in the PD-L1 binding compound provided herein are selected from fluorescent polypeptides (eg, green fluorescent protein (GFP)), luciferase polypeptides, peptide tags, and sodium iodine symporter.

[000103] Also provided herein are nucleic acids (e.g., nucleic acid vectors) that may encode the amino acid chains described herein (e.g., an amino acid chain comprising one or more PD-1 polypeptides fused to a framework polypeptide, or an amino acid chain chain comprising one or more PD-1 polypeptides fused to a polypeptide that can bind to a viral envelope polypeptide) that can be used to produce the multivalent PD-L1 binding compounds provided herein (e.g., a fusion polypeptide comprising two or more than a PD-1 polypeptide, or a polypeptide conjugate comprising two or more amino acid chains, each of which includes one or more PD-1 polypeptides, or a viral particle, wherein one or two or more viral envelope polypeptides are modified to include one or more amino acid chains comprising one or more PD-1 polypeptides, or a viral particle, wherein the one or more viral envelope polypeptides are modified to include one or more amino acid chains comprising two or more PD-1 polypeptides). In some cases, the nucleic acid may encode an amino acid chain that can be used to produce a polypeptide conjugate comprising two or more amino acid chains, each of which includes one or more PD-1 polypeptides. For example, the nucleic acid may encode an amino acid chain including one or more PD-1 polypeptides and a framework polypeptide. In some cases, the nucleic acid may encode an amino acid chain that can be used to produce a viral particle, wherein one or more viral envelope polypeptides comprise one or more amino acid chains comprising one or more PD-1 polypeptides. For example, the nucleic acid may encode an amino acid chain including one or more PD-1 polypeptides and a polypeptide that can bind to a viral envelope polypeptide.

[000104] Nucleic acid (e.g., nucleic acid vectors) encoding the amino acid chains described herein (e.g., an amino acid chain comprising one or more PD-1 polypeptides fused to a framework polypeptide, or an amino acid chain comprising one or more polypeptides PD-1 fused to a polypeptide that can bind to a viral envelope polypeptide) that can be used to produce the multivalent PD-L1 binding compounds provided herein may be a nucleic acid selected from DNA (e.g., DNA constructs), RNA (eg, mRNA) or combinations thereof. In some cases, the nucleic acid encoding an amino acid chain provided herein may be a vector (eg, an expression vector or a viral vector). The vector may be an RC vector (eg, HD vectors and SC vectors), or the vector may be an RD vector. If the vector is a viral vector, the viral vector can be derived from any suitable type of virus. In some cases, the viral vector can be derived from an oncolytic virus. Non-limiting examples of viruses from which viral vectors can be derived include Ad, adeno-associated viruses (AAV), lentiviruses, enteroviruses, reoviruses, poxviruses, measles virus, and herpes viruses.

[000105] In some cases, the nucleic acid encoding an amino acid chain provided herein may also include one or more regulatory elements (eg, to regulate expression of the amino acid chain). Non-limiting examples of regulatory elements that can be included in a nucleic acid encoding an amino acid chain provided herein include promoters (e.g., constitutive promoters, tissue/cell-specific promoters, and inducible promoters, such as chemically activated promoters and light-activated promoters) , enhancers, nucleic acid sequences (for example, “suicide” genes), the expression of which can cause apoptosis, necrosis and other forms of cell death.

[000106] In some cases, nucleic acids (e.g., nucleic acid vectors) encoding amino acid chains provided herein (e.g., an amino acid chain including one or more PD-1 polypeptides fused to a framework polypeptide, or an amino acid chain including one or more PD-1 polypeptides fused to a polypeptide that can bind to a viral envelope polypeptide) that can be used to produce the multivalent PD-L1 binding compounds provided herein (e.g., a fusion polypeptide comprising two or more PD-L1 polypeptides). 1, or a polypeptide conjugate comprising two or more amino acid chains, each of which includes one or more PD-1 polypeptides, or a viral particle, wherein two or more viral envelope polypeptides are modified to include one or more amino acid chains comprising one or more than PD-1 polypeptides, or a viral particle where one or more viral envelope polypeptides are modified to include one or more amino acid chains comprising two or more PD-1 polypeptides can result in the production of two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) amino acid chains presented herein.

[000107] In some cases, nucleic acids (e.g., nucleic acid vectors) encoding the amino acid chains provided herein (e.g., an amino acid chain including one or more PD-1 polypeptides fused to a framework polypeptide, or an amino acid chain including one or more PD-1 polypeptides fused to a polypeptide that can bind to a viral envelope polypeptide) that can be used to produce the multivalent PD-L1 binding compounds provided herein (e.g., a fusion polypeptide comprising two or more PD-L1 polypeptides). 1, or a polypeptide conjugate comprising two or more amino acid chains, each of which includes one or more PD-1 polypeptides, or a viral particle, wherein two or more viral envelope polypeptides are modified to include one or more amino acid chains comprising one or more PD-1 polypeptides, or a viral particle where one or more viral envelope polypeptides are modified to include one or more amino acid chains comprising two or more PD-1 polypeptides), can result in the continuous production of one or more (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) amino acid chains presented in the present description.

[000108] In some cases, nucleic acids (e.g., nucleic acid vectors) encoding amino acid chains provided herein (e.g., an amino acid chain including one or more PD-1 polypeptides fused to a framework polypeptide, or an amino acid chain including one or more PD-1 polypeptides fused to a polypeptide that can bind to a viral envelope polypeptide) that can be used to produce the multivalent PD-L1 binding compounds provided herein (e.g., a fusion polypeptide comprising two or more PD-L1 polypeptides). 1, or a polypeptide conjugate comprising two or more amino acid chains, each of which includes one or more PD-1 polypeptides, or a viral particle, wherein two or more viral envelope polypeptides are modified to include one or more amino acid chains comprising one or more than PD-1 polypeptides, or a viral particle where one or more viral envelope polypeptides are modified to include one or more amino acid chains comprising two or more PD-1 polypeptides) can result in the production of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) amino acid chains provided herein while the nucleic acid persists (eg, until the nucleic acid is degraded).

[000109] In some cases, the amino acid chains encoded by the nucleic acid presented herein can be used to produce the multivalent PD-L1 binding compounds presented herein. For example, two or more amino acid chains comprising one or more PD-1 polypeptides fused to a backbone polypeptide may assemble (eg, self-assemble) into a polypeptide conjugate comprising two or more amino acid chains each comprising one or more PD-1 polypeptides. 1 to obtain the polypeptide conjugate presented herein. If two or more amino acid chains comprising one or more PD-1 polypeptides fused to a backbone polypeptide are assembled into a polypeptide conjugate comprising two or more amino acid chains each comprising one or more PD-1 polypeptides to produce a polypeptide conjugate, amino acid chains can be assembled in vivo or in vitro . For example, one or more amino acid chains comprising one or more PD-1 polypeptides fused to a polypeptide that can bind to a viral envelope polypeptide can bind to two or more viral envelope polypeptides present on the surface of a viral particle to coat it to produce coated viral particle as described herein. If one or more amino acid chains comprising one or more PD-1 polypeptides fused to a polypeptide that can bind to a viral envelope polypeptide is coated on a viral particle to produce a coated viral particle, the viral particle can be coated in vivo or in vitro .

[000110] In some cases, multivalent PD-L1 binding compounds presented herein can be purified. The term "purified" polypeptide, protein or nucleic acid refers to a polypeptide or nucleic acid that is a major component in a mixture of components, for example, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more , 80% or more, 90% or more, 95% or more, or 99% or more by weight. For example, the purified multivalent PD-L1 binding compound may constitute approximately 30% or more by weight of a composition containing one or more multivalent PD-L1 binding compounds. Polypeptides can be purified by methods including, but not limited to, affinity chromatography and immunosorbent affinity columns. For example, a purified nucleic acid encoding an amino acid chain that can be used to produce a multivalent PD-L1 binding compound provided herein may constitute approximately 30% or more by weight of a composition containing one or more amino acid chains that can be used to preparing a multivalent PD-L1 binding compound as provided herein. Nucleic acid can be purified by methods including, but not limited to, phenol-chloroform extraction and column purification (eg, minicolumn purification).

[000111] Alternatively, the invention also provides multivalent PD-1 binding compounds comprising two or more amino acid segments that can bind PD-1 (e.g., PD-L1 polypeptides and/or fragments thereof), as well as methods for producing and the use of multivalent PD-1 binding compounds comprising two or more amino acid segments that can bind to PD-1 (eg, PD-L1 polypeptides and/or fragments thereof). In some cases, a multivalent PD-1 binding compound may include two or more amino acid segments that can bind to PD-1 (eg, PD-L1 polypeptides and/or fragments thereof). For example, a multivalent PD-1 binding compound comprising two or more PD-L1 polypeptides (and/or fragments thereof having the ability to bind PD-1) can bind to two or more PD-1 polypeptides. In some cases, a multivalent PD-1 binding compound may comprise a single amino acid chain comprising two or more amino acid segments that can bind to PD-1.

[000112] The present invention also relates to methods and materials for the treatment of cancer, for example, the administration of a composition containing one or more multivalent PD-L1 binding compounds presented herein. One or more multivalent PD-L1 binding compounds can be administered to an individual in combination with therapeutic polypeptides. In an embodiment, the multivalent PD-L1 binding compound comprises a therapeutic polypeptide administered to an individual for the treatment of cancer. In another embodiment, a multivalent PD-L1 binding compound is administered to an individual together with an Ad expressing a therapeutic polypeptide for the treatment of cancer.

[000113] The present invention relates to viral particles where two or more viral envelope polypeptides contain one or more amino acid chains comprising one or more PD-1 polypeptides. The viral particle may be a recombinant viral particle containing PD-1 polypeptides, including PD-1 fusion proteins, in the capsid protein of the viral particle. The viral particle may be a viral particle coated with PD-1 polypeptides, such as PD-1 fusion proteins.

[000114] In the embodiment where the PD-L1 binding compound contains a recombinant virus, the virus is a recombinant Ad, for example, Ad strain Ad657 or Ad strain Ad6/57/6 and variants thereof. Recombinant Ad can be modified to be conditionally replicative Ad (CRAd). Such recombinant Ad are described in US Patent Application No. 16/690,733, the disclosure of which is incorporated herein by reference in its entirety.

[000115] In an embodiment, the present invention provides a recombinant, conditionally replicative Ad viral particle comprising one or more amino acid chains, each of which includes one or two or more PD-1 polypeptides, or a nucleic acid encoding amino acid chains, which can be used to produce multivalent PD-L1 binding compounds.

[000116] In some cases, recombinant Ad provided herein (eg, recombinant Ad having oncolytic antitumor activity, such as recombinant Ad657) may include an Ad genome containing one or more nucleic acid deletions. The nucleic acid deletion may be a complete deletion (eg, a deletion of a nucleic acid encoding a polypeptide) or a partial deletion (eg, a deletion of one or more nucleotides in a nucleic acid encoding a polypeptide). Deletion of a nucleic acid may reduce or eliminate transcription and translation of the polypeptide encoded by the deleted nucleic acid. In some cases, the nucleic acid encoding a polypeptide associated with the production of infectious progeny may be deleted. Examples of nucleic acids that can be deleted and/or modified in the recombinant Ad provided herein may encode E1 (eg, E1A and E1B), E2, E3, E4, pIIIA, fiber, E1B, and include viral enhancers and promoters. For example, recombinant Ad provided herein (eg, recombinant Ad having oncolytic antitumor activity, such as recombinant Ad657) may comprise an Ad genome containing a deletion of one or more nucleotides in the nucleic acid encoding an E1 polypeptide. In some cases, the recombinant Ad provided herein may include one or more substitutions in the nucleic acid encoding the E1 polypeptide.

[0001117] In specific embodiments, the recombinant Ad provided herein is modified to contain a probasine promoter containing, for example, the nucleic acid SEQ ID NO: 41, modified to contain a dl1101 deletion in the nucleic acid encoding the E1 polypeptide; the recombinant Ad provided herein is modified to contain a dl1107 deletion in the nucleic acid encoding the E1 polypeptide; The recombinant Ad provided herein is modified to contain a dl1101 deletion and a dl1107 deletion. The dl1101 modification in E1a prevents binding to p300 and renders the virus susceptible to interferon (IFN) repression. The dl1107 mutation in E1a prevents it from binding to pRB to block viral replication in cells with intact pRB pathways. See N-terminal amino acid sequences of the E1A polypeptide, e.g., wild-type E1A Ad, and variants CRAd-657-dl1101, CRAd-657-dl1107, and CRAd-657-dl1101/1107 in Figure 10.

[000118] The pattern of mutations in Ad6, Ad657 and variants thereof, including mutations in the E1 protein to convert the virus to conditionally replicative Ad (CRAd), is shown in Figure 9 and Figure 10. These include dl1101 and/or dl1107, blocking binding to p300 and pRB, respectively.

[000119] Figure 10 shows the N-terminal amino acid sequences of E1A in wild-type Ad, as well as the Ad variants E1Adl1101, E1A dl1107 and E1A dl1101/1107.

[000120] Replacement of the Ad E1 promoter with the prostate-specific probasin-E1 promoter DNA sequence (SEQ ID NO: 41) to produce CRAd, Ad-PB is also shown (Figure 9). The probasine promoter is androgen-dependent, so it will work in androgen-sensitive tumors like LNCaP, but not androgen-resistant tumors like DU145.

[000121] Replication-competent Ads, as well as CRAds, of the invention are genetically modified by methods known to those skilled in the art to express one or more therapeutic polypeptides. The therapeutic polypeptides may be immunostimulatory polypeptides, for example, CD40L, 4-1BBL, OX40, GM-CSF, and combinations thereof.

[000122] Additionally, in an embodiment, the replication-competent Ad as well as the CRAd of the invention are genetically modified by methods known to those skilled in the art to express both PD-1 polypeptides and one or more therapeutic polypeptides as fusion proteins.

[000123] Replication-competent Ads as well as CRAds of the invention can be genetically modified by methods known to those skilled in the art to contain modified capsid fiber/protrusion proteins. The Ad fiber protein is a complex of three apparently identical subunits that mediates the initial attachment stage. Native fiber protein Ad6 (SEQ ID NO: 35) binds to CAR.

[000124] In an additional aspect of the invention, recombinant fiber-modified Ads are produced containing different fiber proteins or modifications in fiber/knob proteins that are not native to the parent Ad. Recombinant Ads were produced, including CRAds containing capsid proteins of different Ad strains and heterologous fiber/knob proteins, for example, recombinant Ads containing heterologous Ad35 fiber polypeptide or chimpanzee C68 fiber polypeptide, +/- K7 peptide.

[000125] Chimeric Ad, the AdF35 fiber chimera, has the amino acid sequence of SEQ ID NO: 36, is shorter than the Ad5 and Ad6 fiber proteins and retargets the virus to CD46.

[000126] The recombinant fiber-modified Ad contains a K7 fiber having the sequence SEQ ID NO: 37, targeting the virus to heparin sulfate proteoglycans and negative charges on cells.

[000127] Recombinant chimeric Ad, fiber 6/FC68, containing the sequence SEQ ID NO: 38, is a chimeric Ad containing fiber protein from chimpanzee adenovirus C68. The fiber protein is shorter than the Ad5 or Ad6 fiber proteins and binds to CAR.

[000128] Recombinant chimeric Ad fiber 6/FC68-K7 containing the sequence SEQ ID NO: 39 is a chimeric Ad containing fiber protein from chimpanzee adenovirus C68. The fiber protein is shorter than the Ad5 or Ad6 fiber proteins. Fiber 6/FC68-K7 binds to CAR and retargets heparin sulfate and negative charges.

[000129] Recombinant chimeric Ad fiber 6/FC68-HI-K7 containing the sequence SEQ ID NO: 40 is a chimeric Ad containing fiber protein from chimpanzee adenovirus C68. The fiber protein is shorter than the Ad5 or Ad6 fiber proteins. Fiber 6/FC68-HI-K7 binds to CAR and retargets heparin sulfate and negative charges.

[000130] In some cases, recombinant Ad provided herein (eg, recombinant Ad having oncolytic antitumor activity, such as recombinant Ad657) may include an Ad genome containing one or more nucleic acid insertions. For example, the nucleic acid insertion may include a nucleic acid encoding a polypeptide, such as a therapeutic polypeptide. The nucleic acid can be inserted into any suitable location in the genome of the recombinant Ad provided herein. In some cases, a nucleic acid encoding a polypeptide can be inserted into an HVR (eg, HVR loop 5) of the recombinant Ad genome provided herein. For example, if a nucleic acid encoding a polypeptide is inserted into the HVR of a recombinant Ad genome as provided herein, the nucleic acid encoding the polypeptide may express one or more polypeptides, and the expressed polypeptides may be inserted into the capsid of the recombinant Ad. In cases where a nucleic acid encoding a polypeptide is inserted into the HVR region of the genome of a recombinant Ad as provided herein, the recombinant Ad can present from about 1 to about 720 polypeptides encoded by the inserted nucleic acid on its surface. The nucleic acid insertion may be a nucleic acid encoding any suitable polypeptide. In some cases, the nucleic acid insertion may encode a therapeutic polypeptide, polypeptide antigen, or cytokine.

[000131] In general, Ad can be modified to include the CRAd modifications presented herein.

[000132] In an embodiment, the recombinant Ad is a conditionally replicative Ad with a dl1101 modification in E1a to prevent binding to p300 and rendering the virus susceptible to IFN repression, and a dl1107 mutation in E1a to prevent its binding to pRB to block viral replication in cells with intact pRB pathways, and E3A deletion. CRAd expresses human PD-1 fused to human immunoglobulin.

[000133] In an embodiment, the recombinant Ad (eg, Ad657) is a conditionally replicative Ad with a dl1101 modification in E1a preventing binding to p300 and rendering the virus susceptible to IFN repression, and a deletion of E3A. CRAd expresses human PD-1 fused to human factor X (GLA) on the surface of the viral particle, where CRAd is retargeted to cells expressing PD-L1 on their surface.

[000134] In an embodiment, the recombinant Ad is a conditionally replicative Ad (e.g., Ad657) with the dl1101 modification in E1a preventing binding to p300 and making the virus susceptible to IFN repression, and the dl1107 mutation in E1a preventing it from binding to pRB to block replication virus in cells with intact pRB pathways and E3A deletion. CRAd expresses human PD-1 fused to human factor X (GLA) on the surface of the viral particle, where CRAd is retargeted to cells expressing PD-L1 on their surface.

[000135] Viral particles containing PD-1 polypeptides bound on the surface of the viral particle can be used to treat a mammal having a cancer (eg, a cancer comprising one or more PD-L1 positive cancer cells). The viral particles can be administered to an individual in an amount effective to treat the cancer, and, optionally, they can be administered in combination with antineoplastic therapeutic agents, including immunotherapies and chemotherapeutic agents.

[000136] In one aspect of the invention, a composition comprising one or more multivalent PD-L1 binding compounds or a nucleic acid encoding an amino acid chain that can be used to prepare the multivalent PD-L1 binding compounds described herein and, optionally, one or more antitumor therapeutic agents may be administered to a mammal having a cancer to treat the mammal. The multivalent PD-L1 binding compound provided herein can bind to PD-L1 on PD-L1 positive cancer cells. Binding of PD-1 polypeptides may neutralize PD-L1 function and thus may prevent PD-L1-positive cells (eg, PD-L1-positive malignant cells) from evading immune surveillance and/or may allow antitumor agents (eg, antitumor immunotherapies) to means) to more effectively target PD-L1-positive cells. In cases where the nucleic acid encoding a multivalent PD-L1 binding compound as provided herein is an oncolytic Ad vector encoding a multivalent PD-L1 binding compound as provided herein, the oncolytic Ad can infect a cell and can regulate T- cellular responses directed towards the infected cell. Oncolytic Ad can infect any suitable cell type. In some cases, oncolytic Ad can infect a PD-L1-positive cell (eg, a PD-L1-positive malignant cell). In some cases, oncolytic Ad can infect nondividing cells (eg, kidney cells).

[000137] In some cases, a composition containing or consisting essentially of one or more PD-L1 binding compounds provided herein can be used to treat a mammal having a cancer (e.g., a cancer comprising one or more PD-L1-positive malignant cells). For example, a composition containing one or more multivalent PD-L1 binding compounds and, optionally, one or more antitumor therapeutic agents can be administered to a mammal having a cancer to treat the mammal.

[000138] Alternatively, the methods and materials presented herein can be used to treat other diseases or disorders associated with PD-L1 positive cells. In some cases, a composition containing one or more PD-L1 binding compounds provided herein can be used to treat a mammal having an infectious disease (e.g., an infectious disease involving one or more PD-L1 positive macrophages, such as infiltrating tumor macrophages). Non-limiting examples of infectious diseases that may involve one or more PD-L1 positive cells include HIV, hepatitis and malaria. In some cases, a composition containing one or more PD-L1 binding compounds provided herein can be used to treat a mammal having an autoimmune disease (e.g., an autoimmune disease involving one or more PD-L1 positive macrophages, such as infiltrating tumor macrophages).

[000139] Any suitable mammal having a malignancy can be treated as described herein. For example, humans and other primates, such as marmosets, having cancer can be treated with a composition containing one or more PD-L1 binding compounds provided herein, and optionally in combination with one or more methods of treating cancer. neoplasms to reduce the severity of a malignant neoplasm, reduce the symptom of a malignant neoplasm and/or reduce the number of malignant cells present in the body of a mammal, human or other primate. In some cases, dogs, cats, horses, cows, pigs, sheep, mice and rats with cancer can be treated with a composition containing one or more multivalent PD-L1 binding compounds presented herein, viruses containing PD -L1 binding compounds, or a nucleic acid encoding an amino acid chain that can be used to produce the multivalent PD-L1 binding compounds provided herein and, optionally, by one or more methods of treating cancers provided herein .

[000140] When treating a mammal (eg, human) having a cancer as provided herein, the cancer may be any suitable cancer. The malignancy may include one or more PD-L1-positive malignant cells. The malignancy may include one or more solid tumors. A malignant neoplasm can be a hemoblastosis. Non-limiting examples of cancers that may be treated as provided herein include breast cancer, colorectal cancer, kidney cancer, lung cancer (eg, non-small cell lung cancer), ovarian cancer, melanoma, brain cancer, sarcoma, prostate cancer glands, pancreatic cancer, head and neck cancer, liver cancer, retinoblastoma, lymphoma and leukemia.

[000141] In some cases, a mammal can be identified as having a cancer (eg, a cancer comprising one or more PD-L1 positive cancer cells). Any suitable method may be used to identify a mammal having a malignant neoplasm. For example, imaging techniques and biopsy techniques can be used to identify mammals (eg, humans) with cancer.

[000142] Once a mammal has been identified as having a cancer, it can be administered a composition containing one or more PD-L1 binding compounds provided herein and, optionally, can be treated with one or more antineoplastic therapeutic agents and/or agents for treatment of malignant neoplasms. The one or more anticancer therapeutic agents may include any suitable agent for the treatment of cancer. In some cases, treatment of malignant neoplasms may include surgery. In some cases, treatment of malignant neoplasms may include radiation therapy. In some cases, treatment of malignancies may include pharmacotherapy such as chemotherapy, hormonal therapy, targeted therapy and/or cytotoxic therapy. Non-limiting examples of treatment of malignancies include the administration of one or more receptor tyrosine kinase inhibitors (eg, erlotinib), the administration of one or more PD1/PD-L1 inhibitors (eg, nivolumab, pembrolizumab, atezolizumab, avelumab, cemiplimab and durvalumab), the administration of one or more immunotherapies agents, such as immunotherapies that can target PD1/PD-L1 (eg, nivolumab, pembrolizumab, atezolizumab, avelumab, cemiplimab, durvalumab, alemtuzumab, ipilimumab, ofatumumab and rituximab), administration of one or more anti-GITR antibodies, administration one or more platinum compounds (eg, cisplatin or carboplatin), administration of one or more taxanes (eg, paclitaxel, docetaxel or albumin-bound paclitaxel such as nab-paclitaxel), administration of altretamine, administration of capecitabine, administration of cyclophosphamide, administration of etoposide (vp -16), gemcitabine administration, ifosfamide administration, irinotecan administration (cpt-11), liposomal doxorubicin administration, melphalan administration, pemetrexed administration, topotecan administration, vinorelbine administration, administration of one or more luteinizing hormone releasing hormone (LHRH) agonists (such as goserelin and leuprolide), administration of one or more anti-estrogen therapeutics (such as tamoxifen), administration of one or more aromatase inhibitors (such as letrozole, anastrozole and exemestane), administration of one or more angiogenesis inhibitors (such as bevacizumab), administration of one or more poly(ADP)-ribose polymerase (PARP) inhibitors (such as olaparib, rucaparib and niraparib), external beam radiotherapy, brachytherapy, radiophosphorus and any combination thereof. In cases where a mammal having a cancer is treated with a composition containing one or more multivalent PD-L1 binding compounds or a nucleic acid encoding an amino acid chain that can be used to prepare the multivalent PD-L1 binding compounds presented herein, and treated with one or more cancer treatment agents, a composition containing one or more cancer treatment agents may be administered simultaneously or independently. For example, a composition containing one or more multivalent PD-L1 binding compounds or a nucleic acid encoding an amino acid chain that can be used to prepare the multivalent PD-L1 binding compounds provided herein can be administered first, and one or more agents for the treatment of malignant neoplasms can be administered second, or vice versa.

[000143] In some cases, a composition containing one or more PD-L1 binding compounds provided herein can be formulated in a pharmaceutically acceptable composition for administration to a mammal having a cancer. For example, a therapeutically effective amount of one or more multivalent PD-L1 binding compounds or a nucleic acid encoding an amino acid chain that can be used to prepare the multivalent PD-L1 binding compounds provided herein can be formulated together with one or more pharmaceutically acceptable carriers (additives) and/or diluents. The pharmaceutical composition may further comprise a therapeutic polypeptide and/or a recombinant virus genetically modified to express the therapeutic polypeptide. A pharmaceutical composition containing or consisting essentially of a multivalent PD-L1 binding compound and, optionally, a therapeutic polypeptide may be formulated for administration in solid or liquid form, including, but not limited to, sterile solutions, suspensions, sustained release formulations. release, tablets, capsules, pills, powders, nanoparticles and granules. Pharmaceutically acceptable carriers and excipients that may be used in the pharmaceutical composition provided herein include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffering agents such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, sodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silicon dioxide, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene block copolymers, polyethylene glycol and lanolin.

[000144] A pharmaceutical composition containing or consisting essentially of one or more multivalent PD-L1 binding compounds as provided herein, and optionally a therapeutic polypeptide, is formulated for oral or parenteral (including subcutaneous, intramuscular, intravenous , intratumoral and intradermal) administration.

[000145] When administered orally, a pharmaceutical composition containing one or more PD-L1 binding compounds or a nucleic acid encoding an amino acid chain that can be used to prepare the multivalent PD-L1 binding compounds provided herein may be in the form of a pill , tablets or capsules.

[000146] Compositions suitable for parenteral administration include aqueous and non-aqueous, sterile, injectable solutions that may contain antioxidants, buffers, bacteriostatic agents and solutes that render the composition isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions, which may include suspending agents and thickening agents. The formulations may be in single-dose containers or multi-dose containers, such as sealed ampoules and vials, and may be stored in a lyophilized state, requiring only the addition of a sterile liquid carrier, such as water for injection, immediately prior to use. Extemporaneous injection solutions and suspensions can be prepared from sterile powders, granules and tablets. Such injectable solutions may be in the form of, for example, a sterile injectable aqueous or oily suspension. This suspension can be formulated using, for example, suitable dispersing agents or humectants (such as, for example, Tween 80) and suspending agents. The sterile injectable preparation may be a sterile injectable solution or suspension in a non-toxic, parenterally acceptable diluent or solvent, for example, a solution in 1,3-butanediol. Non-limiting examples of acceptable carriers and solvents that can be used include mannitol, water, Ringer's solution and isotonic sodium chloride solution. Additionally, sterile fatty oils can be used as a solvent or suspending medium. In some cases, a light fatty oil such as synthetic mono- or di-glycerides may be used. Fatty acids such as oleic acid and its glyceride derivatives can be used to formulate injectables, as can natural pharmaceutically acceptable oils such as olive oil or castor oil, including the oils in their polyoxyethylated versions. In some cases, these oil solutions or suspensions may contain a long-chain alcohol diluent or dispersant.

[000147] Specific embodiments presented herein may be further limited in the claims by using the expressions “consisting of” or “consisting essentially of.” When used in claims filed or added as amendments, the transitional term "consisting of" excludes any element, step or ingredient not defined in the claims. The transitional term "consisting essentially of" limits the scope of the claim to certain materials or steps, as well as materials or steps that do not significantly affect the basic and novel characteristics.

[000148] In some cases, a pharmaceutically acceptable composition containing one or more PD-L1 binding compounds provided herein can be administered locally or systemically. For example, a composition containing a compound that inhibits the function of the PD-L1 intracellular domain can be administered locally by injection into or near a malignancy (eg, tumor) in a mammal (eg, human). For example, a composition containing a compound that inhibits PD-L1 intracellular domain function may be administered systemically by oral administration or by injection (eg, subcutaneous, intramuscular, intravenous, intratumoral, and intradermal injection) to a mammal (eg, human).

[000149] Effective dosages may vary depending on the severity of the malignancy, the route of administration, the age and general health of the individual, the use of excipients, the possibility of concomitant use with other therapeutic agents, such as the use of other agents, and the judgment of the attending physician.

[000150] An effective amount of a composition containing one or more PD-L1 binding compounds provided herein can be any amount that can reduce the severity of a cancer, reduce a symptom of a cancer, and/or reduce the number of cancer cells present in the body mammal without causing significant toxicity to the mammal. The effective amount may remain constant, or it may be adjusted as a sliding scale or variable dose depending on the mammal's response to treatment. Various factors may influence the exact effective amount used for a particular indication. For example, the frequency of administration, duration of treatment, use of multiple drugs, route of administration, and severity of the condition (eg, cancer) may require that the precise effective amount administered be increased or decreased.

[000151] The frequency of administration may be any frequency that results in sensitization of cancer cells of a mammal to one or more cancer treatment agents (eg, one or more anticancer immunotherapies) without causing significant toxicity to the mammal. For example, the frequency of administration may be from about once a week to about three times a day, or from about twice a month to about six times a day, or from about twice a week to about once a day. The frequency of administration may remain constant or variable during treatment. A course of treatment using a composition containing one or more PD-L1 binding compounds provided herein may include periods of rest. For example, a composition containing one or more multivalent PD-L1 binding compounds or a nucleic acid encoding an amino acid chain that can be used to produce the multivalent PD-L1 binding compounds provided herein can be administered daily for a two-week period followed by a two-week rest period, and this treatment regimen can be repeated many times. As with the effective amount, various factors may influence the exact dosage frequency used for a particular indication. For example, the effective amount, duration of treatment, use of multiple drugs, route of administration, and severity of the condition (eg, cancer) may require increased or decreased frequency of administration.

[000152] An effective duration of administration of a composition containing one or more PD-L1 binding compounds provided herein may be any duration that can result in a decrease in the severity of the cancer, a decrease in the symptom of the cancer, and/or a decrease in the number of cancer cells, present in the body of a mammal without causing significant toxicity to the mammal. Thus, the effective duration may vary from a few days to several weeks, months or years. In general, the effective duration of treatment for malignancy can range from six months to one year. Many factors may influence the exact effective duration of a particular treatment. For example, the effective duration may vary depending on the frequency of administration, the effective amount, the use of multiple drugs, the route of administration and the severity of the condition being treated.

[000153] In some cases, the course of treatment and the severity of one or more symptoms associated with the condition being treated (eg, cancer) can be monitored. Any suitable method can be used to determine whether cancer cells in a mammal are sensitized to one or more cancer treatment agents (eg, one or more anticancer immunotherapies). For example, cancer response (eg, based on size and/or number of tumors) can be assessed using Office imaging techniques at different points in time.

[000154] The present invention will be further described in the following examples, without limiting the scope of the invention described in the claims.

EXAMPLES

Example 1: Construction of recombinant adenoviruses: insertion of individual HVRs from different Ad serotypes with insertion of cell-targeting/non-targeting peptides or new amino acids.

[000155] Recombinant adenoviruses have been constructed using recombinant DNA technology by methods known to those skilled in the art. Recombinant Ad (Ad657) is derived from the first Ad strain Ad6 (eg, may include the genome of the first Ad strain), and it may include the hexon capsid HVR from the second Ad strain, Ad57, see Figure 8. These embodiments are typically used in context of Ad, in which different HVRs of the capsid hexon from different Ads are combined (i.e., HVRs are rearranged). Figure 7 shows the alignment of Ad5, Ad6 and Ad57, showing changes in the hexon regions. For example, HVR1 Ad of strain Ad6 with HVR 2-7 Ad of strain Ad57 or HVR1 and 7 Ad of strain Ad6 with HVR 2-6 Ad of strain Ad57. In a further embodiment, the Ad6/56/6 virus contains HVR 1 and 7 from Ad strain Ad6 and HVR 2-6 from Ad strain Ad657.

[000156] In one aspect of the invention, chimeric Ads containing HVR1 Ad6 and HVR 2-7 Ad57 were prepared, the chimera referred to as the HVR Ad6/57 chimera containing a hexon having the amino acid sequence of SEQ ID NO: 28.

[000157] In yet another aspect of the invention, chimeric Ads containing HVR1 and 7 Ad6 and HVR 2-6 Ad57 were prepared, the chimera referred to as the HVR Ad6/57/6 chimera containing a hexon having the amino acid sequence of SEQ ID NO: 29.

[000158] To obtain the recombinant Ad strain Ad657, the nucleic acid encoding Ad57 HVR 1-7 was synthesized and inserted into the Ad6 hexon in a plasmid with an FRT-Zeocin®-FRT cassette between pVI and the hexon. It was recombined into different pAd6 plasmids to produce Ad657 and its variants. The amino acid sequence of the Ad657 hexon is shown in SEQ ID NO: 30.

[000159] In the context of Ad657 variants, the Ad57 HVR sequence was synthesized with HVR1 modified with cysteine, flexibility amino acids, and restriction sites to allow the insertion of other peptides. It was inserted into the Ad6 hexon in a plasmid with an FRT-Zeocin®-FRT cassette between pVI and the hexon. It was recombined into different pAd6 plasmids to produce Ad657 variants with a cysteine in HVR1, the variant designated Ad657-HVR1-XXA containing a hexon having the amino acid sequence SEQ ID NO: 42.

[000160] In the context of Ad657 variants, the Ad57 HVR sequence was synthesized with HVR5 modified with cysteine, flexibility amino acids, and restriction sites to allow the insertion of other peptides. It was inserted into the Ad6 hexon in a plasmid with an FRT-Zeocin®-FRT cassette between pVI and the hexon. It was recombined into different pAd6 plasmids to produce Ad657 variants with a cysteine in HVR5, the variant designated Ad657-HVR5-XXA containing a hexon having the amino acid sequence SEQ ID NO: 43.

[000161] In the context of Ad657 variants, the Ad57 HVR sequence was synthesized with HVR1 modified with cysteine, flexibility amino acids, and restriction sites to allow the insertion of other peptides. It was inserted into the Ad6 hexon in a plasmid with an FRT-Zeocin®-FRT cassette between pVI and the hexon. It was recombined into various pAd6 plasmids to produce Ad657 variants without the cysteine in HVR1 but with restriction sites allowing peptide insertions into HVR1, the variant designated Ad657-HVR1-XA containing a hexon having the amino acid sequence SEQ ID NO: 44.

[000162] In the context of Ad657 variants, the Ad57 HVR sequence was synthesized with HVR5 modified with cysteine, flexibility amino acids, and restriction sites to allow the insertion of other peptides. It was inserted into the Ad6 hexon in a plasmid with an FRT-Zeocin®-FRT cassette between pVI and the hexon. It was recombined into various pAd6 plasmids to produce Ad657 variants without the cysteine in HVR5 but with restriction sites allowing peptide insertions into HVR5, the variant designated Ad657-HVR5-XA containing a hexon having the amino acid sequence of SEQ ID NO: 45.

[000163] In the context of Ad657 variants, the Ad657 HVR1-XA sequence was modified by insertion of a biotin acceptor peptide into HVR1. It was recombined into different pAd6 plasmids to produce Ad657 variants with BAP in HVR1, the variant designated Ad657-HVR1-PSTCD, containing a hexon having the amino acid sequence SEQ ID NO: 46.

[000164] Insertion of a biotin acceptor peptide results in nontargeting of virus variants from the liver, allowing virus retargeting with avidin or streptavidin and biotinylated ligands and virus purification on monomeric avidin or streptavidin columns.

[000165] In the context of Ad657 variants, the Ad657 HVR1-XA sequence was modified by insertion of a biotin acceptor peptide (BAP) into HVR1. It was recombined into different pAd6 plasmids to produce Ad657 variants with BAP in HVR1, the variant designated Ad657-HVR5-PSTCD, containing a hexon having the amino acid sequence SEQ ID NO: 47.

[000166] In the context of Ad657 variants, the Ad657 HVR5-XA sequence was modified by insertion of a synthetic V1/V2 loop from the HIV envelope into HVR5, the variant designated Ad657-HVR5-V1/V2 containing a hexon having the amino acid sequence of SEQ ID NO: 48.

[000167] Insertion of a synthetic V1/V2 loop from the HIV envelope allows the antigen to be exposed to serve as a vaccine and also allows retargeting through binding to HIV envelope interacting proteins.

[000168] In the context of Ad657 variants, the Ad657 HVR5-XA sequence has been modified by inserting synthetic peptides from human papillomavirus (HPV) into HVR5, the variant designated Ad657-HVR5-HPV contains a hexon having the amino acid sequence of SEQ ID NO: 49.

[000169] Insertion of synthetic peptides from human papillomavirus allows HPV peptides to be exposed as antigens for vaccination purposes, as well as for retargeting through binding to HPV peptide-interacting proteins.

[000170] In another aspect of the invention, chimeric Ads containing HVR1 Ad6 and HVR 2-7 Ad57 were prepared, the chimera referred to as the HVR Ad6/57 chimera containing a hexon having the amino acid sequence of SEQ ID NO: 50.

[000171] In another aspect of the invention, chimeric Ads containing HVR1 and 7 Ad6 and HVR 2-6 Ad57 were prepared, the chimera referred to as the HVR Ad6/57/6 chimera containing a hexon having the amino acid sequence of SEQ ID NO: 51.

[000172] The plasmid maps in Figures 16-18 show a combination of insertions of individual HVRs from different Ad serotypes with the insertion of cell-targeting/non-targeting peptides, therapeutic polypeptides, or novel amino acids such as cysteine into the hexon for targeted chemical modification and shielding.

[000173] In some embodiments, cell binding peptides are incorporated into HVR 1 or HVR 5, with embodiments serving as examples of incorporating these and other peptides into any of HVR Ad.

Example 2. Targeted chemical conjugation of cysteine-modified, hexon-modified Ad657-HVR5C.

[000174] Figure 13 is a depiction of Ad variants showing the combination of insertion of individual HVRs from different Ad serotypes with insertion of new amino acids such as cysteine into hexon for targeted chemical modification and shielding.

[000175] This example demonstrates the ability to target polymer and other chemical modifications to cysteines embedded in the Ad hexon (Figure 15). Non-targeting PEG inactivates viral infection, while cysteine-targeted PEGylation results in preservation of viral function.

[000176] One aspect of the invention provides the use of polymers or embedded peptides/proteins for non-targeting, retargeting and shielding from antibodies, proteins, cells. Figures 12 and 14 show regions of HVR Ad that can be modified, for example, by PEGylation or "bapylation". Another example shows the insertion of a biotin acceptor peptide (BAP) into these HVRs, allowing vector retargeting with avidin or streptavidin and biotinylated ligands or with avidin or streptavidin fusion proteins. The BAP insertion also allows viruses to be purified on monomeric avidin or streptavidin columns to obtain a vector. Likewise, Ad57-HVR1-XXA and XA provide an example of the insertion of a cysteine into this site for targeted chemical modification by maleimide or other cysteine-reactive agents.

[000177] In an embodiment, the different Ad serotypes and/or variants include polymer shielding to allow multiple administrations of the Ad6 and Ad657 variants. An example of a therapeutic cycle is shown where Ad6 and Ad657 can be used for multiple rounds of treatment through serotype switching in combination with covalent polymer conjugation (Figure 13).

[000178] Ad657-HVR1C expressing GFP-luciferase was produced by cells and purified using CsCl gradients. The virus was covalently modified with 5 kDa polyethylene glycol (PEG). The virus was treated with NHS-PEG, which reacts randomly with amines/lysines on viral proteins, or maleimide-PEG, which specifically reacts with cysteine, which was inserted into HVR1 using the XXA shuttle plasmid. These unmodified or modified viruses were then purified by a final CsCl centrifugation and then desalted. The virus was separated on SDS PAGE gels, stained with SyproRuby and visualized (Figure 14). This suggests that NHS PEGylation results in random modification of many viral proteins, as evidenced by the increase in apparent protein mass (indicated by arrows). In contrast, the targeted reaction of maleimide-PEG with cysteine in HVR1 results in modification of only the hexon and does not damage other viral capsomer proteins. The effects of PEGylation on viral function are assessed.

Example 3. Conditional replicative Ad (CRAd).

[000179] The pattern of mutations in Ad6, Ad657 and variants thereof, including mutations in the E1 protein to convert the virus to conditionally replicative Ad (CRAd), is shown in Figure 9, Figure 10 and Figure 11. These include dl1101 and/or dl1107, blocking binding to p300 and pRB, respectively.

[000180] Figure 10 shows the N-terminal amino acid sequences of E1A in wild-type Ad, as well as the Ad variants E1Adl1101, E1A dl1107, and E1A dl1101/1107.

[000181] Also shown is replacement of the E1 Ad promoter with the prostate-specific probasine promoter and E1 DNA sequence SEQ ID NO: 31 to produce CRAd, Ad-PB (Figure 9). The probasine promoter is androgen-dependent, so it will act in androgen-sensitive tumors like LNCaP, but not in androgen-resistant tumors like DU145.

[000182] In an embodiment, a conditionally replicative Ad657 virus having the dl1101 modification in E1, and the dl1107 mutation in E1a, and the E3A deletion, is constructed to express human PD-1, fused to human immunoglobulin as a decoy protein for PD-L1. The virus also expresses GFP-luciferase from the CMV promoter. This conditionally replicative variant of the Ad657 virus is designated CrAd6d1101/1107DE3ADP-hPD-1-Ig-GL.

Example 4. Retargeted and non-targeted recombinant adenovirus.

[000183] Ad in vitro binds and enters cells through the combined interactions of its fiber and penton base proteins with cell surface receptors. The trimeric fiber binds to the coxsackie adenovirus receptor (CAR), and cells lacking CAR are relatively resistant to infection unless they also express α _v integrins, which can be bound by a penton-based RGD motif.

[000184] The Ad fiber protein is a complex of three apparently identical subunits that mediate the initial attachment step. The native Ad6 fiber protein contains the amino acid sequence set forth in SEQ ID NO: 35 and binds to CAR.

[000185] In an additional aspect of the invention, recombinant fiber-modified Ads are produced containing different fiber proteins not native to the parent Ad. Recombinant Ad, including CRAd, containing capsid proteins from different Ad strains were produced, for example, recombinant Ad containing heterologous Ad35 fiber polypeptide or chimpanzee C68 fiber polypeptide, +/- K7 peptide.

[000186] Chimeric Ad, the AdF35 fiber chimera, has the amino acid sequence of SEQ ID NO: 36, is shorter than the Ad5 and Ad6 fiber proteins and retargets the virus to CD46.

[000187] Recombinant fiber-modified Ad contains a K7 fiber having the sequence SEQ ID NO: 37, targeting the virus to heparin sulfate proteoglycans and negative charges on cells.

[000188] Recombinant chimeric Ad fiber 6/FC68 containing the sequence SEQ ID NO: 38 is a chimeric Ad containing fiber protein from chimpanzee adenovirus C68. The fiber protein is shorter than the Ad5 or Ad6 fiber proteins and is bound by CAR.

[000189] Recombinant chimeric Ad fiber 6/FC68-K7 containing the sequence SEQ ID NO: 39 is a chimeric Ad containing fiber protein from chimpanzee adenovirus C68. The fiber protein is shorter than the Ad5 or Ad6 fiber proteins. Fiber 6/FC68-K7 binds to CAR and retargets heparin sulfate and negative charges.

[000190] Recombinant chimeric Ad fiber 6/FC68-HI-K7 containing the sequence SEQ ID NO: 40 is a chimeric Ad containing fiber protein from chimpanzee adenovirus C68. The fiber protein is shorter than the Ad5 or Ad6 fiber proteins. Fiber 6/FC68-HI-K7 binds to CAR and retargets heparin sulfate and negative charges.

[000191] Coagulation factor X (FX) binds with nanomolar affinity to adenovirus species C hexons and thus allows adenovirus species C to efficiently transduce hepatocytes following IV injection.

[000192] Following the construction of CRAd viruses expressing human factor X (GLA) fused to human PD-1. Wild-type mouse and human PD-1 domains and mouse and human high-affinity PD-1 domains were fused to the N- or C-termini of the human GLA domain of FX-EGF and their expression was driven by the RSV or CMV promoter. In some cases, these fusion protein expression cassettes were inserted into the E3 domain of these viruses. In other cases, they were inserted between the fiber and E4 genes of the viruses.

[000193] Conditional replicative Ad657 had a dl1101 modification in E1a preventing it from binding to p300 and making it susceptible to IFN, an E3A deletion, an RSV expression cassette inserted into the E3 deletion. This virus expresses human factor X (GLA) fused to human PD-1 and an adenovirus retargeting protein to PD-L1 ⁺ cells (i.e., CrAd657d1101DE3ADP*hPD1-GLA-GL-L).

[000194] Conditional replicative Ad657 has a dl1101 modification in E1a that prevents it from binding to p300 and renders the virus susceptible to IFN repression, and a dl1107 mutation in E1a that prevents it from binding to pRB to block viral replication in cells with intact pRB pathways, E3 deletion and contains an RSV expression cassette inserted into the E3 deletion. This virus expresses human factor X (GLA) fused to human PD-1 (i.e., CrAd657*1101/1107DE3BSTRSVhPD1-GLA*IgIZI). During viral replication, PD-1 will be expressed in the cell as a monomer capable of binding to PD-L1 upon release from the cell. PD-1 can also be bound to the surface of Ad to retarget Ad to PD-L1-expressing cells.

[000195] A conditionally replicative Ad6/57/6 with a dl1107 mutation in E1a and an E3A deletion with an integrated RSV expression cassette inserted into the E3 deletion is engineered to express human PD-1 fused to human immunoglobulin as a PD-trap protein L1 (i.e. pAd/57/6-dl1107-ΔE3-RSV-hPD-1-HA-Ig-I). See figure 16.

[000196] The fusion protein was purified from cells infected with pAd6/57/6-dl1107-DE3-RSV-hPD-1-HA-Ig-I on a protein A column. This protein was titrated in parallel with a positive control anti-PD-1 monoclonal antibody in the PD-1/PD-L1 blockade assay from Promega (Madison, WI, USA). High-affinity human hPD-1-HA-Ig fusion protein inhibits PD-L1. See figure 19.

Example 5: Construction of recombinant Ad for expression of PD-L1 binding compounds in combination with heterologous polypeptides.

[000197] A conditionally replicative Ad virus (CRAd657 or CRAd6/57/6) is modified to express PD-1 polypeptides in combination with one or more heterologous polypeptides (eg, therapeutic polypeptides and/or targeting polypeptides). In one embodiment, an RSV-PD-1 fusion cassette is inserted into the E3 deletion of the virus and therapeutic transgenes are inserted as CMV, or MCMV, or RSV expression cassettes between the fiber and E4 genes by methods known to those skilled in the art. See figure 17 and figure 18.

Example 6: Evaluation of Tumor Growth in CRAd Expressing High Affinity Human hPD-1-HA-Ig Fusion Protein Alone or in Combination with an Immunotherapeutic Load.

[000198] B16-CAR melanoma cells were injected subcutaneously into C57BL/6 mice and the resulting tumors were injected with 3e11 viral particles of the multivalent PD-L1 binding compounds of the invention. The CRAd+PD-L1 trap is pAd6/57/6-dl1107-ΔE3-RSV-hPD-1-HA-Ig-I. The CRAd+PD-L1+immunostimulant decoy was co-injected into tumors using pAd6/57/6-dl1107-ΔE3-RSV-hPD-1-HA-Ig-I and a second adenovirus expressing 4-1BBL. CRAd expressing a high-affinity human hPD-1-HA-Ig fusion protein delays tumor growth alone or in combination with an immunotherapeutic challenge. See figure 20.

[000199] In an embodiment, CRAd657 or CRAd6/57/6 is modified to express PD-1 polypeptides in combination with one or more therapeutic polypeptides. The therapeutic polypeptide can be selected from GM-CSF, 4-1BBL, CD40L, OX40, and combinations thereof. In one aspect, the PD-1 polypeptide is expressed with a therapeutic polypeptide as a fusion protein. In an additional aspect, an RSV-PD-1 fusion cassette is inserted into the E3 deletion of the virus and therapeutic transgenes are inserted as CMV or MCMV or RSV expression cassettes between the fiber and E4 genes of the virus. See figures 17 and 18.

Example 7: Construction of Multivalent PD-L1 Binding Compounds.

[000200] Fusion proteins comprising a PD-1 polypeptide and either a framework polypeptide or the GLA domain of an FX polypeptide are prepared from genes that can be cloned by PCR using mutations and fusions from wild-type genes, mutant genes or synthetic genes by methods known in the art. specialists in this field. Fusion proteins are collected by cloning into expression vectors, culturing under conditions allowing expression and assembly of the proteins, and optionally purifying the proteins from cellular media.

[000201] Fusion proteins can be produced by transfecting plasmids or infecting a cell with vectors expressing PD-1 polypeptides and/or fragments thereof fused to a polypeptide that can bind to a viral envelope polypeptide, followed by purification with PD-1 or a partner by fusion on antibody columns, or PD-L1 columns, or biotin columns.

[000202] In an embodiment, Ad can be brought into contact with purified PD-1 fusion proteins under conditions that allow the surface of Ad to be coated with the fusion protein. For example, the GLA domain of an FX polypeptide binds to the surface of Ad such that PD-1 is exposed on the surface of Ad to bind to PD-L1 on the cell surface. An adenovirus in which the fusion partner is bound to the surface of Ad and PD-1 can bind to PD-L1 on the cell is a multimeric PD-L1-binding compound.

[000203] In an embodiment, nucleic acids encoding fusion proteins are inserted into plasmids or viral vectors or inserted into mRNA production vectors and transfected or infected in vitro cell cultures or infecting animals in vivo . Samples are analyzed for protein expression using antibodies against PD-1 and/or fusion protein partners.

[000204] Nucleic acid (e.g., DNA, mRNA, and viral vectors) encoding fusion proteins are administered to normal or tumor-bearing animals and changes in peripheral blood cells, lymph node cells, spleen cells, and tumor cell populations are analyzed by staining for CD3, CD4, CD8, PD-1, PD-L1 and T cell activation markers.

[000205] Nucleic acid (e.g., DNA, mRNA) or viral vectors encoding fusion proteins are administered to tumor-bearing animals and effects on tumor size, metastasis, dissemination, and survival are assessed.

Other embodiments.

[000206] It should be understood that although the invention has been described in combination with the detailed description, the above description is provided for illustrative purposes and not to limit the scope of the invention as defined by the scope of the claims. Other aspects, advantages and modifications are included within the scope of the claims.

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LIST OF SEQUENCES

<110> MAYO FOUNDATION FOR MEDICAL EDUCATION AND RESEARCH

<120> MULTIVALENT PD-L1 BINDING COMPOUNDS FOR TREATMENT

MALIGNANT NEOPLASMS

<130> ADZE 2 PCT SEQ

<150>US 62/839,916

<151> 2019-04-29

<160> 51

<170> Patent In version 3.5

<210> 1

<211> 465

<212> PROTEIN

<213> Human adenovirus 6

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Ala Tyr Asn Ala Leu Ala Pro Lys Gly Ala Pro Asn Ser Cys Glu Trp

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Glu Gln Asn Glu Thr Ala Gln Val Asp Ala Gln Glu Leu Asp Glu Glu

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Glu Asn Glu Ala Asn Glu Ala Gln Ala Arg Glu Gln Glu Gln Ala Lys

35 40 45

Lys Thr His Val Tyr Ala Gln Ala Pro Leu Ser Gly Ile Lys Ile Thr

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Lys Glu Gly Leu Gln Ile Gly Thr Ala Asp Ala Thr Val Ala Gly Ala

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Gly Lys Glu Ile Phe Ala Asp Lys Thr Phe Gln Pro Glu Pro Gln Val

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Gly Glu Ser Gln Trp Asn Glu Ala Asp Ala Thr Ala Ala Gly Gly Arg

100 105 110

Val Leu Lys Lys Thr Thr Pro Met Lys Pro Cys Tyr Gly Ser Tyr Ala

115 120 125

Arg Pro Thr Asn Ser Asn Gly Gly Gln Gly Val Leu Trp Leu Thr Asn

130 135 140

Gly Lys Leu Glu Ser Gln Val Glu Met Gln Phe Phe Ser Thr Ser Thr

145 150 155 160

Asn Ala Thr Asn Glu Val Asn Asn Ile Gln Pro Thr Val Val Leu Tyr

165 170 175

Ser Glu Asp Val Asn Met Glu Thr Pro Asp Thr His Leu Ser Tyr Lys

180 185 190

Pro Lys Met Gly Asp Lys Asn Ala Lys Val Ser Leu Gly Gln Gln Ala

195 200 205

Met Pro Asn Arg Pro Asn Tyr Ile Ala Phe Arg Asp Asn Phe Ile Gly

210 215 220

Leu Met Tyr Tyr Asn Ser Thr Gly Asn Met Gly Val Leu Ala Gly Gln

225 230 235 240

Ala Ser Gln Leu Asn Ala Val Val Asp Leu Gln Asp Arg Asn Thr Glu

245 250 255

Leu Ser Tyr Gln Leu Leu Leu Asp Ser Ile Gly Asp Arg Thr Arg Tyr

260 265 270

Phe Ser Met Trp Asn Gln Ala Val Asp Ser Tyr Asp Pro Asp Val Arg

275 280 285

Ile Ile Glu Asn His Gly Thr Glu Asp Glu Leu Pro Asn Tyr Cys Phe

290 295 300

Pro Leu Gly Gly Ile Gly Ile Thr Asp Thr Phe Gln Ala Val Lys Thr

305 310 315 320

Thr Ala Ala Asn Gly Asp Gln Gly Asn Thr Thr Trp Gln Lys Asp Ser

325 330 335

Thr Phe Ala Glu Arg Asn Glu Ile Gly Val Gly Asn Asn Phe Ala Met

340 345 350

Glu Ile Asn Leu Asn Ala Asn Leu Trp Arg Asn Phe Leu Tyr Ser Asn

355 360 365

Ile Ala Leu Tyr Leu Pro Asp Lys Leu Lys Tyr Asn Pro Thr Asn Val

370 375 380

Asp Ile Ser Asp Asn Pro Asn Thr Tyr Asp His Met Lys Arg Val Val

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Ala Pro Gly Leu Val Asp Cys Tyr Ile Asn Leu Gly Ala Arg Trp Ser

405 410 415

Leu Asp Tyr Met Asp Asn Val Asn Pro Phe Asn His His Arg Asn Ala

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Gly Leu Arg Tyr Arg Ser Met Leu Leu Gly Asn Gly Arg Tyr Val Pro

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Phe His Ile Gln Val Pro Gln Lys Phe Phe Ala Ile Lys Asn Leu Leu

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Met Ala Thr Pro Ser Met Met Pro Gln Trp Ser Tyr Met His Ile Ser

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Gly Gln Asp Ala Ser Glu Tyr Leu Ser Pro Gly Leu Val Gln Phe Ala

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Arg Ala Thr Glu Thr Tyr Phe Ser Leu Asn Asn Lys Phe Arg Asn Pro

35 40 45

Thr Val Ala Pro Thr His Asp Val Thr Thr Asp Arg Ser Gln Arg Leu

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Thr Leu Arg Phe Ile Pro Val Asp Arg Glu Asp Thr Ala Tyr Ser Tyr

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Lys Ala Arg Phe Thr Leu Ala Val Gly Asp Asn Arg Val Leu Asp Met

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Ala Ser Thr Tyr Phe Asp Ile Arg Gly Val Leu Asp Arg Gly Pro Thr

100 105 110

Phe Lys Pro Tyr Ser Gly Thr Ala Tyr Asn Ala Leu Ala Pro Lys Gly

115 120 125

Ala Pro Asn Ser Cys Glu Trp Asp Glu Asp Asp Thr Gln Val Gln Val

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Ala Ala Glu Asp Asp Gln Asp Asp Asp Glu Glu Glu Glu Gln Leu Pro

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Gln Gln Arg Asn Gly Lys Lys Thr His Val Tyr Ala Gln Ala Pro Phe

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Ala Gly Glu Ala Ile Asn Lys Asn Gly Leu Gln Ile Gly Thr Asn Gly

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Ala Ala Thr Glu Gly Asn Lys Glu Ile Tyr Ala Asp Lys Thr Tyr Gln

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Pro Glu Pro Gln Ile Gly Glu Ser Gln Trp Asn Glu Ala Glu Ser Ser

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Val Ala Gly Gly Arg Val Leu Lys Lys Thr Thr Pro Met Lys Pro Cys

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Tyr Gly Ser Tyr Ala Arg Pro Thr Asn Ser Asn Gly Gly Gln Gly Val

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Met Val Glu Gln Asn Gly Lys Leu Glu Ser Gln Val Glu Met Gln Phe

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Phe Ser Thr Ser Val Asn Ala Met Asn Glu Ala Asn Ala Ile Gln Pro

275 280 285

Lys Leu Val Leu Tyr Ser Glu Asp Val Asn Met Glu Thr Pro Asp Thr

290 295 300

His Leu Ser Tyr Lys Pro Gly Lys Ser Asp Asp Asn Ser Lys Ala Met

305 310 315 320

Leu Gly Gln Gln Ser Met Pro Asn Arg Pro Asn Tyr Ile Ala Phe Arg

325 330 335

Asp Asn Phe Ile Gly Leu Met Tyr Tyr Asn Ser Thr Gly Asn Met Gly

340 345 350

Val Leu Ala Gly Gln Ala Ser Gln Leu Asn Ala Val Val Asp Leu Gln

355 360 365

Asp Arg Asn Thr Glu Leu Ser Tyr Gln Leu Leu Leu Asp Ser Ile Gly

370 375 380

Asp Arg Thr Arg Tyr Phe Ser Met Trp Asn Gln Ala Val Asp Ser Tyr

385 390 395 400

Asp Pro Asp Val Arg Ile Ile Glu Asn His Gly Thr Glu Asp Glu Leu

405 410 415

Pro Asn Tyr Cys Phe Pro Leu Gly Gly Ile Gly Val Thr Asp Thr Tyr

420 425 430

Gln Ala Ile Lys Ala Thr Asn Gly Asn Gly Gly Ala Thr Thr Trp Ala

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Gln Asp Asn Thr Phe Ala Glu Arg Asn Glu Ile Gly Val Gly Asn Asn

450 455 460

Phe Ala Met Glu Ile Asn Leu Asn Ala Asn Leu Trp Arg Asn Phe Leu

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Tyr Ser Asn Ile Ala Leu Tyr Leu Pro Asp Lys Leu Lys Tyr Asn Pro

485 490 495

Thr Asn Val Glu Ile Ser Asp Asn Pro Asn Thr Tyr Asp Tyr Met Asn

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Lys Arg Val Val Ala Pro Gly Leu Val Asp Cys Tyr Ile Asn Leu Gly

515 520 525

Ala Arg Trp Ser Leu Asp Tyr Met Asp Asn Val Asn Pro Phe Asn His

530 535 540

His Arg Asn Ala Gly Leu Arg Tyr Arg Ser Met Leu Leu Gly Asn Gly

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Arg Tyr Val Pro Phe His Ile Gln Val Pro Gln Lys Phe Phe Ala Ile

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Lys Asn Leu Leu Leu Leu Pro Gly Ser Tyr Thr Tyr Glu Trp Asn Phe

580 585 590

Arg Lys Asp Val Asn Met Val Leu Gln Ser Ser Leu Gly Asn Asp Leu

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Arg Val Asp Gly Ala Ser Ile Lys Phe Asp Ser Ile Cys Leu Tyr Ala

610 615 620

Thr Phe Phe Pro Met Ala His Asn Thr Ala Ser Thr Leu Glu Ala Met

625 630 635 640

Leu Arg Asn Asp Thr Asn Asp Gln Ser Phe Asn Asp Tyr Leu Ser Ala

645 650 655

Ala Asn Met Leu Tyr Pro Ile Pro Ala Asn Ala Thr Asn Val Pro Ile

660 665 670

Ser Ile Pro Ser Arg Asn Trp Ala Ala Phe Arg Gly Trp Ala Phe Thr

675 680 685

Arg Leu Lys Thr Lys Glu Thr Pro Ser Leu Gly Ser Gly Tyr Asp Pro

690 695 700

Tyr Tyr Thr Tyr Ser Gly Ser Ile Pro Tyr Leu Asp Gly Thr Phe Tyr

705 710 715 720

Leu Asn His Thr Phe Lys Lys Val Ala Ile Thr Phe Asp Ser Ser Val

725 730 735

Ser Trp Pro Gly Asn Asp Arg Leu Leu Thr Pro Asn Glu Phe Glu Ile

740 745 750

Lys Arg Ser Val Asp Gly Glu Gly Tyr Asn Val Ala Gln Cys Asn Met

755 760 765

Thr Lys Asp Trp Phe Leu Val Gln Met Leu Ala Asn Tyr Asn Ile Gly

770 775 780

Tyr Gln Gly Phe Tyr Ile Pro Glu Ser Tyr Lys Asp Arg Met Tyr Ser

785 790 795 800

Phe Phe Arg Asn Phe Gln Pro Met Ser Arg Gln Val Val Asp Asp Thr

805 810 815

Lys Tyr Lys Asp Tyr Gln Gln Val Gly Ile Ile His Gln His Asn Asn

820 825 830

Ser Gly Phe Val Gly Tyr Leu Ala Pro Thr Met Arg Glu Gly Gln Ala

835 840 845

Tyr Pro Ala Asn Val Pro Tyr Pro Leu Ile Gly Lys Thr Ala Val Asp

850 855 860

Ser Ile Thr Gln Lys Lys Phe Leu Cys Asp Arg Thr Leu Trp Arg Ile

865 870 875 880

Pro Phe Ser Ser Asn Phe Met Ser Met Gly Ala Leu Thr Asp Leu Gly

885 890 895

Gln Asn Leu Leu Tyr Ala Asn Ser Ala His Ala Leu Asp Met Thr Phe

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Glu Val Asp Pro Met Asp Glu Pro Thr Leu Leu Tyr Val Leu Phe Glu

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Val Phe Asp Val Val Arg Val His Gln Pro His Arg Gly Val Ile Glu

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Thr Val Tyr Leu Arg Thr Pro Phe Ser Ala Gly Asn Ala Thr Thr

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Met Ala Thr Pro Ser Met Met Pro Gln Trp Ser Tyr Met His Ile Ser

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Gly Gln Asp Ala Ser Glu Tyr Leu Ser Pro Gly Leu Val Gln Phe Ala

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Arg Ala Thr Glu Thr Tyr Phe Ser Leu Asn Asn Lys Phe Arg Asn Pro

35 40 45

Thr Val Ala Pro Thr His Asp Val Thr Thr Asp Arg Ser Gln Arg Leu

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Thr Leu Arg Phe Ile Pro Val Asp Arg Glu Asp Thr Ala Tyr Ser Tyr

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Lys Ala Arg Phe Thr Leu Ala Val Gly Asp Asn Arg Val Leu Asp Met

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Ala Ser Thr Tyr Phe Asp Ile Arg Gly Val Leu Asp Arg Gly Pro Thr

100 105 110

Phe Lys Pro Tyr Ser Gly Thr Ala Tyr Asn Ala Leu Ala Pro Lys Gly

115 120 125

Ala Pro Asn Ser Cys Glu Trp Asp Glu Asp Asp Thr Gln Val Gln Val

130 135 140

Ala Ala Glu Asp Asp Gln Asp Asp Asp Glu Glu Glu Glu Gln Leu Pro

145 150 155 160

Gln Gln Arg Asn Gly Lys Lys Thr His Val Tyr Ala Gln Ala Pro Phe

165 170 175

Ala Gly Glu Ala Ile Asn Lys Asn Gly Leu Gln Ile Gly Thr Asn Gly

180 185 190

Ala Ala Thr Glu Gly Asn Lys Glu Ile Tyr Ala Asp Lys Thr Tyr Gln

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Pro Glu Pro Gln Ile Gly Glu Ser Gln Trp Asn Glu Ala Glu Ser Ser

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Val Ala Gly Gly Arg Val Leu Lys Lys Thr Thr Pro Met Lys Pro Cys

225 230 235 240

Tyr Gly Ser Tyr Ala Arg Pro Thr Asn Ser Asn Gly Gly Gln Gly Val

245 250 255

Met Val Glu Gln Asn Gly Lys Leu Glu Ser Gln Val Glu Met Gln Phe

260 265 270

Phe Ser Thr Ser Val Asn Ala Met Asn Glu Ala Asn Ala Ile Gln Pro

275 280 285

Lys Leu Val Leu Tyr Ser Glu Asp Val Asn Met Glu Thr Pro Asp Thr

290 295 300

His Leu Ser Tyr Lys Pro Gly Lys Ser Asp Asp Asn Ser Lys Ala Met

305 310 315 320

Leu Gly Gln Gln Ser Met Pro Asn Arg Pro Asn Tyr Ile Ala Phe Arg

325 330 335

Asp Asn Phe Ile Gly Leu Met Tyr Tyr Asn Ser Thr Gly Asn Met Gly

340 345 350

Val Leu Ala Gly Gln Ala Ser Gln Leu Asn Ala Val Val Asp Leu Gln

355 360 365

Asp Arg Asn Thr Glu Leu Ser Tyr Gln Leu Leu Leu Asp Ser Ile Gly

370 375 380

Asp Arg Thr Arg Tyr Phe Ser Met Trp Asn Gln Ala Val Asp Ser Tyr

385 390 395 400

Asp Pro Asp Val Arg Ile Ile Glu Asn His Gly Thr Glu Asp Glu Leu

405 410 415

Pro Asn Tyr Cys Phe Pro Leu Gly Gly Ile Gly Val Thr Asp Thr Tyr

420 425 430

Gln Ala Ile Lys Ala Thr Asn Gly Asn Gly Gly Ala Thr Thr Trp Ala

435 440 445

Gln Asp Asn Thr Phe Ala Glu Arg Asn Glu Ile Gly Val Gly Asn Asn

450 455 460

Phe Ala Met Glu Ile Asn Leu Asn Ala Asn Leu Trp Arg Asn Phe Leu

465 470 475 480

Tyr Ser Asn Ile Ala Leu Tyr Leu Pro Asp Lys Leu Lys Tyr Asn Pro

485 490 495

Thr Asn Val Glu Ile Ser Asp Asn Pro Asn Thr Tyr Asp Tyr Met Asn

500 505 510

Lys Arg Val Val Ala Pro Gly Leu Val Asp Cys Tyr Ile Asn Leu Gly

515 520 525

Ala Arg Trp Ser Leu Asp Tyr Met Asp Asn Val Asn Pro Phe Asn His

530 535 540

His Arg Asn Ala Gly Leu Arg Tyr Arg Ser Met Leu Leu Gly Asn Gly

545 550 555 560

Arg Tyr Val Pro Phe His Ile Gln Val Pro Gln Lys Phe Phe Ala Ile

565 570 575

Lys Asn Leu Leu Leu Leu Pro Gly Ser Tyr Thr Tyr Glu Trp Asn Phe

580 585 590

Arg Lys Asp Val Asn Met Val Leu Gln Ser Ser Leu Gly Asn Asp Leu

595 600 605

Arg Val Asp Gly Ala Ser Ile Lys Phe Asp Ser Ile Cys Leu Tyr Ala

610 615 620

Thr Phe Phe Pro Met Ala His Asn Thr Ala Ser Thr Leu Glu Ala Met

625 630 635 640

Leu Arg Asn Asp Thr Asn Asp Gln Ser Phe Asn Asp Tyr Leu Ser Ala

645 650 655

Ala Asn Met Leu Tyr Pro Ile Pro Ala Asn Ala Thr Asn Val Pro Ile

660 665 670

Ser Ile Pro Ser Arg Asn Trp Ala Ala Phe Arg Gly Trp Ala Phe Thr

675 680 685

Arg Leu Lys Thr Lys Glu Thr Pro Ser Leu Gly Ser Gly Tyr Asp Pro

690 695 700

Tyr Tyr Thr Tyr Ser Gly Ser Ile Pro Tyr Leu Asp Gly Thr Phe Tyr

705 710 715 720

Leu Asn His Thr Phe Lys Lys Val Ala Ile Thr Phe Asp Ser Ser Val

725 730 735

Ser Trp Pro Gly Asn Asp Arg Leu Leu Thr Pro Asn Glu Phe Glu Ile

740 745 750

Lys Arg Ser Val Asp Gly Glu Gly Tyr Asn Val Ala Gln Cys Asn Met

755 760 765

Thr Lys Asp Trp Phe Leu Val Gln Met Leu Ala Asn Tyr Asn Ile Gly

770 775 780

Tyr Gln Gly Phe Tyr Ile Pro Glu Ser Tyr Lys Asp Arg Met Tyr Ser

785 790 795 800

Phe Phe Arg Asn Phe Gln Pro Met Ser Arg Gln Val Val Asp Asp Thr

805 810 815

Lys Tyr Lys Asp Tyr Gln Gln Val Gly Ile Ile His Gln His Asn Asn

820 825 830

Ser Gly Phe Val Gly Tyr Leu Ala Pro Thr Met Arg Glu Gly Gln Ala

835 840 845

Tyr Pro Ala Asn Val Pro Tyr Pro Leu Ile Gly Lys Thr Ala Val Asp

850 855 860

Ser Ile Thr Gln Lys Lys Phe Leu Cys Asp Arg Thr Leu Trp Arg Ile

865 870 875 880

Pro Phe Ser Ser Asn Phe Met Ser Met Gly Ala Leu Thr Asp Leu Gly

885 890 895

Gln Asn Leu Leu Tyr Ala Asn Ser Ala His Ala Leu Asp Met Thr Phe

900 905 910

Glu Val Asp Pro Met Asp Glu Pro Thr Leu Leu Tyr Val Leu Phe Glu

915 920 925

Val Phe Asp Val Val Arg Val His Gln Pro His Arg Gly Val Ile Glu

930 935 940

Thr Val Tyr Leu Arg Thr Pro Phe Ser Ala Gly Asn Ala Thr Thr

945 950 955

<210> 4

<211> 962

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Hexon polypeptide

<400> 4

Met Ala Thr Pro Ser Met Met Pro Gln Trp Ser Tyr Met His Ile Ser

1 5 10 15

Gly Gln Asp Ala Ser Glu Tyr Leu Ser Pro Gly Leu Val Gln Phe Ala

20 25 30

Arg Ala Thr Glu Thr Tyr Phe Ser Leu Asn Asn Lys Phe Arg Asn Pro

35 40 45

Thr Val Ala Pro Thr His Asp Val Thr Thr Asp Arg Ser Gln Arg Leu

50 55 60

Thr Leu Arg Phe Ile Pro Val Asp Arg Glu Asp Thr Ala Tyr Ser Tyr

65 70 75 80

Lys Ala Arg Phe Thr Leu Ala Val Gly Asp Asn Arg Val Leu Asp Met

85 90 95

Ala Ser Thr Tyr Phe Asp Ile Arg Gly Val Leu Asp Arg Gly Pro Thr

100 105 110

Phe Lys Pro Tyr Ser Gly Thr Ala Tyr Asn Ala Leu Ala Pro Lys Gly

115 120 125

Ala Pro Asn Ser Cys Glu Trp Glu Gln Asn Glu Thr Ala Gln Val Asp

130 135 140

Ala Gln Glu Leu Asp Glu Glu Glu Asn Glu Ala Asn Glu Ala Gln Ala

145 150 155 160

Arg Glu Gln Glu Gln Ala Lys Lys Thr His Val Tyr Ala Gln Ala Pro

165 170 175

Leu Ser Gly Glu Ala Ile Asn Lys Asn Gly Leu Gln Ile Gly Thr Asn

180 185 190

Gly Ala Ala Thr Glu Gly Asn Lys Glu Ile Tyr Ala Asp Lys Thr Tyr

195 200 205

Gln Pro Glu Pro Gln Ile Gly Glu Ser Gln Trp Asn Glu Ala Glu Ser

210 215 220

Ser Val Ala Gly Gly Arg Val Leu Lys Lys Thr Thr Pro Met Lys Pro

225 230 235 240

Cys Tyr Gly Ser Tyr Ala Arg Pro Thr Asn Ser Asn Gly Gly Gln Gly

245 250 255

Val Met Val Glu Gln Asn Gly Lys Leu Glu Ser Gln Val Glu Met Gln

260 265 270

Phe Phe Ser Thr Ser Val Asn Ala Met Asn Glu Ala Asn Ala Ile Gln

275 280 285

Pro Lys Leu Val Leu Tyr Ser Glu Asp Val Asn Met Glu Thr Pro Asp

290 295 300

Thr His Leu Ser Tyr Lys Pro Gly Lys Ser Asp Asp Asn Ser Lys Ala

305 310 315 320

Met Leu Gly Gln Gln Ser Met Pro Asn Arg Pro Asn Tyr Ile Ala Phe

325 330 335

Arg Asp Asn Phe Ile Gly Leu Met Tyr Tyr Asn Ser Thr Gly Asn Met

340 345 350

Gly Val Leu Ala Gly Gln Ala Ser Gln Leu Asn Ala Val Val Asp Leu

355 360 365

Gln Asp Arg Asn Thr Glu Leu Ser Tyr Gln Leu Leu Leu Asp Ser Ile

370 375 380

Gly Asp Arg Thr Arg Tyr Phe Ser Met Trp Asn Gln Ala Val Asp Ser

385 390 395 400

Tyr Asp Pro Asp Val Arg Ile Ile Glu Asn His Gly Thr Glu Asp Glu

405 410 415

Leu Pro Asn Tyr Cys Phe Pro Leu Gly Gly Ile Gly Ile Thr Asp Thr

420 425 430

Phe Gln Ala Val Lys Thr Thr Ala Ala Asn Gly Asp Gln Gly Asn Thr

435 440 445

Thr Trp Gln Lys Asp Ser Thr Phe Ala Glu Arg Asn Glu Ile Gly Val

450 455 460

Gly Asn Asn Phe Ala Met Glu Ile Asn Leu Asn Ala Asn Leu Trp Arg

465 470 475 480

Asn Phe Leu Tyr Ser Asn Ile Ala Leu Tyr Leu Pro Asp Lys Leu Lys

485 490 495

Tyr Asn Pro Thr Asn Val Glu Ile Ser Asp Asn Pro Asn Thr Tyr Asp

500 505 510

Tyr Met Asn Lys Arg Val Val Ala Pro Gly Leu Val Asp Cys Tyr Ile

515 520 525

Asn Leu Gly Ala Arg Trp Ser Leu Asp Tyr Met Asp Asn Val Asn Pro

530 535 540

Phe Asn His Pro Arg His Ala Gly Leu Arg Tyr Arg Ser Met Leu Leu

545 550 555 560

Gly Asn Gly Arg Tyr Val Pro Phe His Ile Gln Val Pro Gln Lys Phe

565 570 575

Phe Ala Ile Lys Asn Leu Leu Leu Leu Pro Gly Ser Tyr Thr Tyr Glu

580 585 590

Trp Asn Phe Arg Lys Asp Val Asn Met Val Leu Gln Ser Ser Leu Gly

595 600 605

Asn Asp Leu Arg Val Asp Gly Ala Ser Ile Lys Phe Asp Ser Ile Cys

610 615 620

Leu Tyr Ala Thr Phe Phe Pro Met Ala His Asn Thr Ala Ser Thr Leu

625 630 635 640

Glu Ala Met Leu Arg Asn Asp Thr Asn Asp Gln Ser Phe Asn Asp Tyr

645 650 655

Leu Ser Ala Ala Asn Met Leu Tyr Pro Ile Pro Ala Asn Ala Thr Asn

660 665 670

Val Pro Ile Ser Ile Pro Ser Arg Asn Trp Ala Ala Phe Arg Gly Trp

675 680 685

Ala Phe Thr Arg Leu Lys Thr Lys Glu Thr Pro Ser Leu Gly Ser Gly

690 695 700

Tyr Asp Pro Tyr Tyr Thr Tyr Ser Gly Ser Ile Pro Tyr Leu Asp Gly

705 710 715 720

Thr Phe Tyr Leu Asn His Thr Phe Lys Lys Val Ala Ile Thr Phe Asp

725 730 735

Ser Ser Val Ser Trp Pro Gly Asn Asp Arg Leu Leu Thr Pro Asn Glu

740 745 750

Phe Glu Ile Lys Arg Ser Val Asp Gly Glu Gly Tyr Asn Val Ala Gln

755 760 765

Cys Asn Met Thr Lys Asp Trp Phe Leu Val Gln Met Leu Ala Asn Tyr

770 775 780

Asn Ile Gly Tyr Gln Gly Phe Tyr Ile Pro Glu Ser Tyr Lys Asp Arg

785 790 795 800

Met Tyr Ser Phe Phe Arg Asn Phe Gln Pro Met Ser Arg Gln Val Val

805 810 815

Asp Asp Thr Lys Tyr Lys Asp Tyr Gln Gln Val Gly Ile Ile His Gln

820 825 830

His Asn Asn Ser Gly Phe Val Gly Tyr Leu Ala Pro Thr Met Arg Glu

835 840 845

Gly Gln Ala Tyr Pro Ala Asn Val Pro Tyr Pro Leu Ile Gly Lys Thr

850 855 860

Ala Val Asp Ser Ile Thr Gln Lys Lys Phe Leu Cys Asp Arg Thr Leu

865 870 875 880

Trp Arg Ile Pro Phe Ser Ser Asn Phe Met Ser Met Gly Ala Leu Thr

885 890 895

Asp Leu Gly Gln Asn Leu Leu Tyr Ala Asn Ser Ala His Ala Leu Asp

900 905 910

Met Thr Phe Glu Val Asp Pro Met Asp Glu Pro Thr Leu Leu Tyr Val

915 920 925

Leu Phe Glu Val Phe Asp Val Val Arg Val His Gln Pro His Arg Gly

930 935 940

Val Ile Glu Thr Val Tyr Leu Arg Thr Pro Phe Ser Ala Gly Asn Ala

945 950 955 960

Thr Thr

<210> 5

<211> 288

<212> PROTEIN

<213> Homo sapiens

<400> 5

Met Gln Ile Pro Gln Ala Pro Trp Pro Val Val Trp Ala Val Leu Gln

1 5 10 15

Leu Gly Trp Arg Pro Gly Trp Phe Leu Asp Ser Pro Asp Arg Pro Trp

20 25 30

Asn Pro Pro Thr Phe Ser Pro Ala Leu Leu Val Val Thr Glu Gly Asp

35 40 45

Asn Ala Thr Phe Thr Cys Ser Phe Ser Asn Thr Ser Glu Ser Phe Val

50 55 60

Leu Asn Trp Tyr Arg Met Ser Pro Ser Asn Gln Thr Asp Lys Leu Ala

65 70 75 80

Ala Phe Pro Glu Asp Arg Ser Gln Pro Gly Gln Asp Cys Arg Phe Arg

85 90 95

Val Thr Gln Leu Pro Asn Gly Arg Asp Phe His Met Ser Val Val Arg

100 105 110

Ala Arg Arg Asn Asp Ser Gly Thr Tyr Leu Cys Gly Ala Ile Ser Leu

115 120 125

Ala Pro Lys Ala Gln Ile Lys Glu Ser Leu Arg Ala Glu Leu Arg Val

130 135 140

Thr Glu Arg Arg Ala Glu Val Pro Thr Ala His Pro Ser Pro Ser Pro

145 150 155 160

Arg Pro Ala Gly Gln Phe Gln Thr Leu Val Val Gly Val Val Gly Gly

165 170 175

Leu Leu Gly Ser Leu Val Leu Leu Val Trp Val Leu Ala Val Ile Cys

180 185 190

Ser Arg Ala Ala Arg Gly Thr Ile Gly Ala Arg Arg Thr Gly Gln Pro

195 200 205

Leu Lys Glu Asp Pro Ser Ala Val Pro Val Phe Ser Val Asp Tyr Gly

210 215 220

Glu Leu Asp Phe Gln Trp Arg Glu Lys Thr Pro Glu Pro Pro Val Pro

225 230 235 240

Cys Val Pro Glu Gln Thr Glu Tyr Ala Thr Ile Val Phe Pro Ser Gly

245 250 255

Met Gly Thr Ser Ser Pro Ala Arg Arg Gly Ser Ala Asp Gly Pro Arg

260 265 270

Ser Ala Gln Pro Leu Arg Pro Glu Asp Gly His Cys Ser Trp Pro Leu

275 280 285

<210> 6

<211> 288

<212> PROTEIN

<213> Mus musculus

<400> 6

Met Trp Val Arg Gln Val Pro Trp Ser Phe Thr Trp Ala Val Leu Gln

1 5 10 15

Leu Ser Trp Gln Ser Gly Trp Leu Leu Glu Val Pro Asn Gly Pro Trp

20 25 30

Arg Ser Leu Thr Phe Tyr Pro Ala Trp Leu Thr Val Ser Glu Gly Ala

35 40 45

Asn Ala Thr Phe Thr Cys Ser Leu Ser Asn Trp Ser Glu Asp Leu Met

50 55 60

Leu Asn Trp Asn Arg Leu Ser Pro Ser Asn Gln Thr Glu Lys Gln Ala

65 70 75 80

Ala Phe Cys Asn Gly Leu Ser Gln Pro Val Gln Asp Ala Arg Phe Gln

85 90 95

Ile Ile Gln Leu Pro Asn Arg His Asp Phe His Met Asn Ile Leu Asp

100 105 110

Thr Arg Arg Asn Asp Ser Gly Ile Tyr Leu Cys Gly Ala Ile Ser Leu

115 120 125

His Pro Lys Ala Lys Ile Glu Glu Ser Pro Gly Ala Glu Leu Val Val

130 135 140

Thr Glu Arg Ile Leu Glu Thr Ser Thr Arg Tyr Pro Ser Pro Ser Pro

145 150 155 160

Lys Pro Glu Gly Arg Phe Gln Gly Met Val Ile Gly Ile Met Ser Ala

165 170 175

Leu Val Gly Ile Pro Val Leu Leu Leu Leu Ala Trp Ala Leu Ala Val

180 185 190

Phe Cys Ser Thr Ser Met Ser Glu Ala Arg Gly Ala Gly Ser Lys Asp

195 200 205

Asp Thr Leu Lys Glu Glu Pro Ser Ala Ala Pro Val Pro Ser Val Ala

210 215 220

Tyr Glu Glu Leu Asp Phe Gln Gly Arg Glu Lys Thr Pro Glu Leu Pro

225 230 235 240

Thr Ala Cys Val His Thr Glu Tyr Ala Thr Ile Val Phe Thr Glu Gly

245 250 255

Leu Gly Ala Ser Ala Met Gly Arg Arg Gly Ser Ala Asp Gly Leu Gln

260 265 270

Gly Pro Arg Pro Pro Arg His Glu Asp Gly His Cys Ser Trp Pro Leu

275 280 285

<210> 7

<211> 390

<212> PROTEIN

<213> Artificial sequence

<220>

<223>PD-1 fusion protein

<400> 7

Met Trp Val Arg Gln Val Pro Trp Ser Phe Thr Trp Ala Val Leu Gln

1 5 10 15

Leu Ser Trp Gln Ser Gly Trp Leu Leu Glu Val Pro Asn Gly Pro Trp

20 25 30

Arg Ser Leu Thr Phe Tyr Pro Ala Trp Leu Thr Val Ser Glu Gly Ala

35 40 45

Asn Ala Thr Phe Thr Cys Ser Leu Ser Asn Trp Ser Glu Asp Leu Met

50 55 60

Leu Asn Trp Asn Arg Leu Ser Pro Ser Asn Gln Thr Glu Lys Gln Ala

65 70 75 80

Ala Phe Cys Asn Gly Leu Ser Gln Pro Val Gln Asp Ala Arg Phe Gln

85 90 95

Ile Ile Gln Leu Pro Asn Arg His Asp Phe His Met Asn Ile Leu Asp

100 105 110

Thr Arg Arg Asn Asp Ser Gly Ile Tyr Leu Cys Gly Ala Ile Ser Leu

115 120 125

His Pro Lys Ala Lys Ile Glu Glu Ser Pro Gly Ala Glu Leu Val Val

130 135 140

Thr Glu Arg Ile Leu Glu Thr Ser Thr Arg Tyr Pro Ser Pro Ser Pro

145 150 155 160

Lys Pro Glu Gly Arg Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro

165 170 175

Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp

180 185 190

Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp

195 200 205

Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly

210 215 220

Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn

225 230 235 240

Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Val His Gln Asp Trp

245 250 255

Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro

260 265 270

Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu

275 280 285

Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn

290 295 300

Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile

305 310 315 320

Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr

325 330 335

Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys

340 345 350

Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys

355 360 365

Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu

370 375 380

Ser Leu Ser Pro Gly Lys

385 390

<210> 8

<211> 273

<212> PROTEIN

<213> Artificial sequence

<220>

<223>PD-1 fusion protein

<400> 8

Met Trp Val Arg Gln Val Pro Trp Ser Phe Thr Trp Ala Val Leu Gln

1 5 10 15

Leu Ser Trp Gln Ser Gly Trp Leu Leu Glu Val Pro Asn Gly Pro Trp

20 25 30

Arg Ser Leu Thr Phe Tyr Pro Ala Trp Leu Thr Val Ser Glu Gly Ala

35 40 45

Asn Ala Thr Phe Thr Cys Ser Leu Ser Asn Trp Ser Glu Asp Leu Met

50 55 60

Leu Asn Trp Asn Arg Leu Ser Pro Ser Asn Gln Thr Glu Lys Gln Ala

65 70 75 80

Ala Phe Cys Asn Gly Leu Ser Gln Pro Val Gln Asp Ala Arg Phe Gln

85 90 95

Ile Ile Gln Leu Pro Asn Arg His Asp Phe His Met Asn Ile Leu Asp

100 105 110

Thr Arg Arg Asn Asp Ser Gly Ile Tyr Leu Cys Gly Ala Ile Ser Leu

115 120 125

His Pro Lys Ala Lys Ile Glu Glu Ser Pro Gly Ala Glu Leu Val Val

130 135 140

Thr Glu Arg Ile Leu Glu Thr Ser Thr Arg Tyr Pro Ser Pro Ser Pro

145 150 155 160

Lys Pro Glu Gly Arg Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro

165 170 175

Pro Val Ala Gly Gly Ser Ala Asn Ser Phe Leu Glu Glu Met Lys Lys

180 185 190

Gly His Leu Glu Arg Glu Cys Met Glu Glu Thr Cys Ser Tyr Glu Glu

195 200 205

Ala Arg Glu Val Phe Glu Asp Ser Asp Lys Thr Asn Glu Phe Trp Asn

210 215 220

Lys Tyr Lys Asp Gly Asp Gln Cys Glu Thr Ser Pro Cys Gln Asn Gln

225 230 235 240

Gly Lys Cys Lys Asp Gly Leu Gly Glu Tyr Thr Cys Thr Cys Leu Glu

245 250 255

Gly Phe Glu Gly Lys Asn Cys Glu Leu Pro Val Gly Tyr Arg Gly Gly

260 265 270

Ser

<210> 9

<211> 252

<212> PROTEIN

<213> Artificial sequence

<220>

<223>PD-1 fusion protein

<400> 9

Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Gly

1 5 10 15

Ser Ala Asn Ser Phe Leu Glu Glu Met Lys Lys Gly His Leu Glu Arg

20 25 30

Glu Cys Met Glu Glu Thr Cys Ser Tyr Glu Glu Ala Arg Glu Val Phe

35 40 45

Glu Asp Ser Asp Lys Thr Asn Glu Phe Trp Asn Lys Tyr Lys Asp Gly

50 55 60

Asp Gln Cys Glu Thr Ser Pro Cys Gln Asn Gln Gly Lys Cys Lys Asp

65 70 75 80

Gly Leu Gly Glu Tyr Thr Cys Thr Cys Leu Glu Gly Phe Glu Gly Lys

85 90 95

Asn Cys Glu Leu Pro Val Gly Tyr Arg Gly Gly Ser Gly Trp Leu Leu

100 105 110

Glu Val Pro Asn Gly Pro Trp Arg Ser Leu Thr Phe Tyr Pro Ala Trp

115 120 125

Leu Thr Val Ser Glu Gly Ala Asn Ala Thr Phe Thr Cys Ser Leu Ser

130 135 140

Asn Trp Ser Glu Asp Leu Met Leu Asn Trp Asn Arg Leu Ser Pro Ser

145 150 155 160

Asn Gln Thr Glu Lys Gln Ala Ala Phe Cys Asn Gly Leu Ser Gln Pro

165 170 175

Val Gln Asp Ala Arg Phe Gln Ile Ile Gln Leu Pro Asn Arg His Asp

180 185 190

Phe His Met Asn Ile Leu Asp Thr Arg Arg Asn Asp Ser Gly Ile Tyr

195 200 205

Leu Cys Gly Ala Ile Ser Leu His Pro Lys Ala Lys Ile Glu Glu Ser

210 215 220

Pro Gly Ala Glu Leu Val Val Thr Glu Arg Ile Leu Glu Thr Ser Thr

225 230 235 240

Arg Tyr Pro Ser Pro Ser Pro Lys Pro Glu Gly Arg

245 250

<210> 10

<211> 310

<212> PROTEIN

<213> Artificial sequence

<220>

<223>PD-1 fusion protein

<400> 10

Met Trp Val Arg Gln Val Pro Trp Ser Phe Thr Trp Ala Val Leu Gln

1 5 10 15

Leu Ser Trp Gln Ser Gly Trp Leu Leu Glu Val Pro Asn Gly Pro Trp

20 25 30

Arg Ser Leu Thr Phe Tyr Pro Ala Trp Leu Thr Val Ser Glu Gly Ala

35 40 45

Asn Ala Thr Phe Thr Cys Ser Leu Ser Asn Trp Ser Glu Asp Leu Met

50 55 60

Leu Asn Trp Asn Arg Leu Ser Pro Ser Asn Gln Thr Glu Lys Gln Ala

65 70 75 80

Ala Phe Cys Asn Gly Leu Ser Gln Pro Val Gln Asp Ala Arg Phe Gln

85 90 95

Ile Ile Gln Leu Pro Asn Arg His Asp Phe His Met Asn Ile Leu Asp

100 105 110

Thr Arg Arg Asn Asp Ser Gly Ile Tyr Leu Cys Gly Ala Ile Ser Leu

115 120 125

His Pro Lys Ala Lys Ile Glu Glu Ser Pro Gly Ala Glu Leu Val Val

130 135 140

Thr Glu Arg Ile Leu Glu Thr Ser Thr Arg Tyr Pro Ser Pro Ser Pro

145 150 155 160

Lys Pro Glu Gly Arg Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro

165 170 175

Pro Val Ala Gly Ser Lys Gly Leu Glu Ser Arg Val Ser Ala Leu Glu

180 185 190

Lys Thr Ser Gln Ile His Ser Asp Thr Ile Leu Arg Ile Thr Gln Gly

195 200 205

Leu Asp Asp Ala Asn Lys Arg Ile Ile Ala Leu Glu Gln Ser Arg Asp

210 215 220

Asp Leu Val Ala Ser Val Ser Asp Ala Gln Leu Ala Ile Ser Arg Leu

225 230 235 240

Glu Ser Ser Ile Gly Ala Leu Gln Thr Val Val Asn Gly Leu Asp Ser

245 250 255

Ser Val Thr Gln Leu Gly Ala Arg Val Gly Gln Leu Glu Thr Gly Leu

260 265 270

Ala Glu Leu Arg Val Asp His Asp Asn Leu Val Ala Arg Val Asp Thr

275 280 285

Ala Glu Arg Asn Ile Gly Ser Leu Thr Thr Glu Leu Ser Thr Leu Thr

290 295 300

Leu Arg Val Thr Ser Ile

305 310

<210> 11

<211> 599

<212> PROTEIN

<213> Artificial sequence

<220>

<223>PD-1 fusion polypeptide

<400> 11

Met Trp Val Arg Gln Val Pro Trp Ser Phe Thr Trp Ala Val Leu Gln

1 5 10 15

Leu Ser Trp Gln Ser Gly Trp Leu Leu Glu Val Pro Asn Gly Pro Trp

20 25 30

Arg Ser Leu Thr Phe Tyr Pro Ala Trp Leu Thr Val Ser Glu Gly Ala

35 40 45

Asn Ala Thr Phe Thr Cys Ser Leu Ser Asn Trp Ser Glu Asp Leu Met

50 55 60

Leu Asn Trp Asn Arg Leu Ser Pro Ser Asn Gln Thr Glu Lys Gln Ala

65 70 75 80

Ala Phe Cys Asn Gly Leu Ser Gln Pro Val Gln Asp Ala Arg Phe Gln

85 90 95

Ile Ile Gln Leu Pro Asn Arg His Asp Phe His Met Asn Ile Leu Asp

100 105 110

Thr Arg Arg Asn Asp Ser Gly Ile Tyr Leu Cys Gly Ala Ile Ser Leu

115 120 125

His Pro Lys Ala Lys Ile Glu Glu Ser Pro Gly Ala Glu Leu Val Val

130 135 140

Thr Glu Arg Ile Leu Glu Thr Ser Thr Arg Tyr Pro Ser Pro Ser Pro

145 150 155 160

Lys Pro Glu Gly Arg Ser Gly Trp Leu Leu Glu Val Pro Asn Gly Pro

165 170 175

Trp Arg Ser Leu Thr Phe Tyr Pro Ala Trp Leu Thr Val Ser Glu Gly

180 185 190

Ala Asn Ala Thr Phe Thr Cys Ser Leu Ser Asn Trp Ser Glu Asp Leu

195 200 205

Met Leu Asn Trp Asn Arg Leu Ser Pro Ser Asn Gln Thr Glu Lys Gln

210 215 220

Ala Ala Phe Cys Asn Gly Leu Ser Gln Pro Val Gln Asp Ala Arg Phe

225 230 235 240

Gln Ile Ile Gln Leu Pro Asn Arg His Asp Phe His Met Asn Ile Leu

245 250 255

Asp Thr Arg Arg Asn Asp Ser Gly Ile Tyr Leu Cys Gly Ala Ile Ser

260 265 270

Leu His Pro Lys Ala Lys Ile Glu Glu Ser Pro Gly Ala Glu Leu Val

275 280 285

Val Thr Glu Arg Ile Leu Glu Thr Ser Thr Arg Tyr Pro Ser Pro Ser

290 295 300

Pro Lys Pro Glu Gly Arg Cys Cys Val Glu Cys Pro Pro Cys Pro Ala

305 310 315 320

Pro Pro Val Ala Gly Thr Met Gly Gly Ala Ala Gly Ser Gly Ala Ala

325 330 335

Glu Ala Gly Ile Thr Gly Thr Trp Tyr Asn Gln Leu Gly Ser Thr Phe

340 345 350

Ile Val Thr Ala Gly Ala Asp Gly Ala Leu Thr Gly Thr Tyr Glu Ser

355 360 365

Ala Val Gly Asn Ala Glu Ser Arg Tyr Val Leu Thr Gly Arg Tyr Glu

370 375 380

Ser Ala Pro Ala Thr Asp Gly Ser Gly Thr Ala Leu Gly Trp Thr Val

385 390 395 400

Ala Trp Lys Asn Asn Tyr Arg Asn Ala His Ser Ala Thr Thr Trp Ser

405 410 415

Gly Gln Tyr Val Gly Gly Ala Glu Ala Arg Ile Asn Thr Gln Trp Leu

420 425 430

Leu Thr Ser Gly Thr Thr Glu Ala Asn Ala Trp Lys Ser Thr Leu Val

435 440 445

Gly His Asp Thr Phe Thr Lys Val Lys Pro Ser Ala Ala Ser Gly Ser

450 455 460

Ala Ala Gly Ser Gly Ala Ala Glu Ala Gly Ile Thr Gly Thr Trp Tyr

465 470 475 480

Asn Gln Leu Gly Ser Thr Phe Ile Val Thr Ala Gly Ala Asp Gly Ala

485 490 495

Leu Thr Gly Thr Tyr Glu Ser Ala Val Gly Asn Ala Glu Ser Arg Tyr

500 505 510

Val Leu Thr Gly Arg Tyr Glu Ser Ala Pro Ala Thr Asp Gly Ser Gly

515 520 525

Thr Ala Leu Gly Trp Thr Val Ala Trp Lys Asn Asn Tyr Arg Asn Ala

530 535 540

His Ser Ala Thr Thr Trp Ser Gly Gln Tyr Val Gly Gly Ala Glu Ala

545 550 555 560

Arg Ile Asn Thr Gln Trp Leu Leu Thr Ser Gly Thr Thr Glu Ala Asn

565 570 575

Ala Trp Lys Ser Thr Leu Val Gly His Asp Thr Phe Thr Lys Val Lys

580 585 590

Pro Ser Ala Ala Ser Gly Ser

595

<210> 12

<211> 765

<212> PROTEIN

<213> Artificial sequence

<220>

<223>PD-1 fusion polypeptide

<400> 12

Met Ser Pro Gln Arg Asp Arg Ile Asn Ala Phe Tyr Lys Asp Asn Pro

1 5 10 15

His Pro Lys Gly Ser Arg Ile Val Ile Asn Arg Glu His Leu Met Ile

20 25 30

Asp Arg Pro Tyr Val Leu Leu Ala Val Leu Phe Val Met Phe Leu Ser

35 40 45

Leu Ile Gly Leu Leu Ala Ile Ala Gly Ile Arg Leu His Arg Ala Ala

50 55 60

Ile Tyr Thr Ala Glu Ile His Lys Ser Leu Ser Thr Asn Leu Asp Val

65 70 75 80

Thr Asn Ser Ile Glu His Gln Val Lys Asp Val Leu Thr Pro Leu Phe

85 90 95

Lys Ile Ile Gly Asp Glu Val Gly Leu Arg Thr Pro Gln Arg Phe Thr

100 105 110

Asp Leu Val Lys Phe Ile Ser Asp Lys Ile Lys Phe Leu Asn Pro Asp

115 120 125

Arg Glu Tyr Asp Phe Arg Asp Leu Thr Trp Cys Ile Asn Pro Pro Glu

130 135 140

Arg Ile Lys Leu Asp Tyr Asp Gln Tyr Cys Ala Asp Val Ala Ala Glu

145 150 155 160

Glu Leu Met Asn Ala Leu Val Asn Ser Thr Leu Leu Glu Thr Arg Thr

165 170 175

Thr Asn Gln Phe Leu Ala Val Ser Lys Gly Asn Cys Ser Gly Pro Thr

180 185 190

Thr Ile Arg Gly Gln Phe Ser Asn Met Ser Leu Ser Leu Leu Asp Leu

195 200 205

Tyr Leu Gly Arg Gly Tyr Asn Val Ser Ser Ile Val Thr Met Thr Ser

210 215 220

Gln Gly Met Tyr Gly Gly Thr Tyr Leu Val Glu Lys Pro Asn Leu Ser

225 230 235 240

Ser Lys Arg Ser Glu Leu Ser Gln Leu Ser Met Tyr Arg Val Phe Glu

245 250 255

Val Gly Val Ile Arg Asn Pro Gly Leu Gly Ala Pro Val Phe His Met

260 265 270

Thr Asn Tyr Leu Glu Gln Pro Val Ser Asn Asp Leu Ser Asn Cys Met

275 280 285

Val Ala Leu Gly Glu Leu Lys Leu Ala Ala Leu Cys His Gly Glu Asp

290 295 300

Ser Ile Thr Ile Pro Tyr Gln Gly Ser Gly Lys Gly Val Ser Phe Gln

305 310 315 320

Leu Val Lys Leu Gly Val Trp Lys Ser Pro Thr Asp Met Gln Ser Trp

325 330 335

Val Pro Leu Ser Thr Asp Asp Pro Val Ile Asp Arg Leu Tyr Leu Ser

340 345 350

Ser His Arg Gly Val Ile Ala Asp Asn Gln Ala Lys Trp Ala Val Pro

355 360 365

Thr Thr Arg Thr Asp Asp Lys Leu Arg Met Glu Thr Cys Phe Gln Gln

370 375 380

Ala Cys Lys Gly Lys Ile Gln Ala Leu Cys Glu Asn Pro Glu Trp Ala

385 390 395 400

Pro Leu Lys Asp Asn Arg Ile Pro Ser Tyr Gly Val Leu Ser Val Asp

405 410 415

Leu Ser Leu Thr Val Glu Leu Lys Ile Lys Ile Ala Ser Gly Phe Gly

420 425 430

Pro Leu Ile Thr His Gly Ser Gly Met Asp Leu Tyr Lys Ser Asn His

435 440 445

Asn Asn Val Tyr Trp Leu Thr Ile Pro Pro Met Lys Asn Leu Ala Leu

450 455 460

Gly Val Ile Asn Thr Leu Glu Trp Ile Pro Arg Phe Lys Val Ser Pro

465 470 475 480

Ala Leu Phe Asn Val Pro Ile Lys Glu Ala Gly Gly Asp Cys His Ala

485 490 495

Pro Thr Tyr Leu Pro Ala Glu Val Asp Gly Asp Val Lys Leu Ser Ser

500 505 510

Asn Leu Val Ile Leu Pro Gly Gln Asp Leu Gln Tyr Val Leu Ala Thr

515 520 525

Tyr Asp Thr Ser Ala Val Glu His Ala Val Val Tyr Tyr Val Tyr Ser

530 535 540

Pro Ser Arg Ser Phe Ser Tyr Phe Tyr Pro Phe Arg Leu Pro Ile Lys

545 550 555 560

Gly Val Pro Ile Glu Leu Gln Val Glu Cys Phe Thr Trp Asp Gln Lys

565 570 575

Leu Trp Cys Arg His Phe Cys Val Leu Ala Asp Ser Glu Ser Gly Gly

580 585 590

His Ile Thr His Ser Gly Met Val Gly Met Gly Val Ser Cys Thr Val

595 600 605

Thr Arg Glu Asp Gly Thr Asn Ile Glu Gly Arg Pro Gly Trp Phe Leu

610 615 620

Asp Ser Pro Asp Arg Pro Trp Asn Pro Pro Thr Phe Ser Pro Ala Leu

625 630 635 640

Leu Val Val Thr Glu Gly Asp Asn Ala Thr Phe Thr Cys Ser Phe Ser

645 650 655

Asn Thr Ser Glu Ser Phe His Val Ile Trp His Arg Glu Ser Pro Ser

660 665 670

Gly Gln Thr Asp Thr Leu Ala Ala Phe Pro Glu Asp Arg Ser Gln Pro

675 680 685

Gly Gln Asp Cys Arg Phe Arg Val Thr Gln Leu Pro Asn Gly Arg Asp

690 695 700

Phe His Met Ser Val Val Arg Ala Arg Arg Asn Asp Ser Gly Thr Tyr

705 710 715 720

Val Cys Gly Val Ile Ser Leu Ala Pro Lys Ile Gln Ile Lys Glu Ser

725 730 735

Leu Arg Ala Glu Leu Arg Val Thr Glu Arg Arg Ala Glu Val Pro Thr

740 745 750

Ala His Pro Ser Pro Ser Pro His His His His His

755 760 765

<210> 13

<211> 385

<212> PROTEIN

<213> Artificial sequence

<220>

<223>PD-1 fusion polypeptide

<400> 13

Met Gln Ile Pro Gln Ala Pro Trp Pro Val Val Trp Ala Val Leu Gln

1 5 10 15

Leu Gly Trp Arg Pro Gly Trp Phe Leu Asp Ser Pro Asp Arg Pro Trp

20 25 30

Asn Pro Pro Thr Phe Ser Pro Ala Leu Leu Val Val Thr Glu Gly Asp

35 40 45

Asn Ala Thr Phe Thr Cys Ser Phe Ser Asn Thr Ser Glu Ser Phe Val

50 55 60

Leu Asn Trp Tyr Arg Met Ser Pro Ser Asn Gln Thr Asp Lys Leu Ala

65 70 75 80

Ala Phe Pro Glu Asp Arg Ser Gln Pro Gly Gln Asp Cys Arg Phe Arg

85 90 95

Val Thr Gln Leu Pro Asn Gly Arg Asp Phe His Met Ser Val Val Arg

100 105 110

Ala Arg Arg Asn Asp Ser Gly Thr Tyr Leu Cys Gly Ala Ile Ser Leu

115 120 125

Ala Pro Lys Ala Gln Ile Lys Glu Ser Leu Arg Ala Glu Leu Arg Val

130 135 140

Thr Glu Arg Arg Ala Glu Val Pro Thr Ala His Pro Ser Pro Ser Pro

145 150 155 160

Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro

165 170 175

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

180 185 190

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp

195 200 205

Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

210 215 220

Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Val

225 230 235 240

Val Ser Val Leu Thr Val Val His Gln Asp Trp Leu Asn Gly Lys Glu

245 250 255

Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys

260 265 270

Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

275 280 285

Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr

290 295 300

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

305 310 315 320

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu

325 330 335

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys

340 345 350

Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu

355 360 365

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

370 375 380

Lys

385

<210> 14

<211> 268

<212> PROTEIN

<213> Artificial sequence

<220>

<223>PD-1 fusion polypeptide

<400> 14

Met Gln Ile Pro Gln Ala Pro Trp Pro Val Val Trp Ala Val Leu Gln

1 5 10 15

Leu Gly Trp Arg Pro Gly Trp Phe Leu Asp Ser Pro Asp Arg Pro Trp

20 25 30

Asn Pro Pro Thr Phe Ser Pro Ala Leu Leu Val Val Thr Glu Gly Asp

35 40 45

Asn Ala Thr Phe Thr Cys Ser Phe Ser Asn Thr Ser Glu Ser Phe Val

50 55 60

Leu Asn Trp Tyr Arg Met Ser Pro Ser Asn Gln Thr Asp Lys Leu Ala

65 70 75 80

Ala Phe Pro Glu Asp Arg Ser Gln Pro Gly Gln Asp Cys Arg Phe Arg

85 90 95

Val Thr Gln Leu Pro Asn Gly Arg Asp Phe His Met Ser Val Val Arg

100 105 110

Ala Arg Arg Asn Asp Ser Gly Thr Tyr Leu Cys Gly Ala Ile Ser Leu

115 120 125

Ala Pro Lys Ala Gln Ile Lys Glu Ser Leu Arg Ala Glu Leu Arg Val

130 135 140

Thr Glu Arg Arg Ala Glu Val Pro Thr Ala His Pro Ser Pro Ser Pro

145 150 155 160

Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Gly

165 170 175

Ser Ala Asn Ser Phe Leu Glu Glu Met Lys Lys Gly His Leu Glu Arg

180 185 190

Glu Cys Met Glu Glu Thr Cys Ser Tyr Glu Glu Ala Arg Glu Val Phe

195 200 205

Glu Asp Ser Asp Lys Thr Asn Glu Phe Trp Asn Lys Tyr Lys Asp Gly

210 215 220

Asp Gln Cys Glu Thr Ser Pro Cys Gln Asn Gln Gly Lys Cys Lys Asp

225 230 235 240

Gly Leu Gly Glu Tyr Thr Cys Thr Cys Leu Glu Gly Phe Glu Gly Lys

245 250 255

Asn Cys Glu Leu Pro Val Gly Tyr Arg Gly Gly Ser

260 265

<210> 15

<211> 248

<212> PROTEIN

<213> Artificial sequence

<220>

<223>PD-1 fusion polypeptide

<400> 15

Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Gly

1 5 10 15

Ser Ala Asn Ser Phe Leu Glu Glu Met Lys Lys Gly His Leu Glu Arg

20 25 30

Glu Cys Met Glu Glu Thr Cys Ser Tyr Glu Glu Ala Arg Glu Val Phe

35 40 45

Glu Asp Ser Asp Lys Thr Asn Glu Phe Trp Asn Lys Tyr Lys Asp Gly

50 55 60

Asp Gln Cys Glu Thr Ser Pro Cys Gln Asn Gln Gly Lys Cys Lys Asp

65 70 75 80

Gly Leu Gly Glu Tyr Thr Cys Thr Cys Leu Glu Gly Phe Glu Gly Lys

85 90 95

Asn Cys Glu Leu Pro Val Gly Tyr Arg Gly Gly Ser Pro Gly Trp Phe

100 105 110

Leu Asp Ser Pro Asp Arg Pro Trp Asn Pro Pro Thr Phe Ser Pro Ala

115 120 125

Leu Leu Val Val Thr Glu Gly Asp Asn Ala Thr Phe Thr Cys Ser Phe

130 135 140

Ser Asn Thr Ser Glu Ser Phe Val Leu Asn Trp Tyr Arg Met Ser Pro

145 150 155 160

Ser Asn Gln Thr Asp Lys Leu Ala Ala Phe Pro Glu Asp Arg Ser Gln

165 170 175

Pro Gly Gln Asp Cys Arg Phe Arg Val Thr Gln Leu Pro Asn Gly Arg

180 185 190

Asp Phe His Met Ser Val Val Arg Ala Arg Arg Asn Asp Ser Gly Thr

195 200 205

Tyr Leu Cys Gly Ala Ile Ser Leu Ala Pro Lys Ala Gln Ile Lys Glu

210 215 220

Ser Leu Arg Ala Glu Leu Arg Val Thr Glu Arg Arg Ala Glu Val Pro

225 230 235 240

Thr Ala His Pro Ser Pro Ser Pro

245

<210> 16

<211> 305

<212> PROTEIN

<213> Artificial sequence

<220>

<223>PD-1 fusion polypeptide

<400> 16

Met Gln Ile Pro Gln Ala Pro Trp Pro Val Val Trp Ala Val Leu Gln

1 5 10 15

Leu Gly Trp Arg Pro Gly Trp Phe Leu Asp Ser Pro Asp Arg Pro Trp

20 25 30

Asn Pro Pro Thr Phe Ser Pro Ala Leu Leu Val Val Thr Glu Gly Asp

35 40 45

Asn Ala Thr Phe Thr Cys Ser Phe Ser Asn Thr Ser Glu Ser Phe Val

50 55 60

Leu Asn Trp Tyr Arg Met Ser Pro Ser Asn Gln Thr Asp Lys Leu Ala

65 70 75 80

Ala Phe Pro Glu Asp Arg Ser Gln Pro Gly Gln Asp Cys Arg Phe Arg

85 90 95

Val Thr Gln Leu Pro Asn Gly Arg Asp Phe His Met Ser Val Val Arg

100 105 110

Ala Arg Arg Asn Asp Ser Gly Thr Tyr Leu Cys Gly Ala Ile Ser Leu

115 120 125

Ala Pro Lys Ala Gln Ile Lys Glu Ser Leu Arg Ala Glu Leu Arg Val

130 135 140

Thr Glu Arg Arg Ala Glu Val Pro Thr Ala His Pro Ser Pro Ser Pro

145 150 155 160

Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Ser

165 170 175

Lys Gly Leu Glu Ser Arg Val Ser Ala Leu Glu Lys Thr Ser Gln Ile

180 185 190

His Ser Asp Thr Ile Leu Arg Ile Thr Gln Gly Leu Asp Asp Ala Asn

195 200 205

Lys Arg Ile Ile Ala Leu Glu Gln Ser Arg Asp Asp Leu Val Ala Ser

210 215 220

Val Ser Asp Ala Gln Leu Ala Ile Ser Arg Leu Glu Ser Ser Ile Gly

225 230 235 240

Ala Leu Gln Thr Val Val Asn Gly Leu Asp Ser Ser Val Thr Gln Leu

245 250 255

Gly Ala Arg Val Gly Gln Leu Glu Thr Gly Leu Ala Glu Leu Arg Val

260 265 270

Asp His Asp Asn Leu Val Ala Arg Val Asp Thr Ala Glu Arg Asn Ile

275 280 285

Gly Ser Leu Thr Thr Glu Leu Ser Thr Leu Thr Leu Arg Val Thr Ser

290 295 300

Ile

305

<210> 17

<211> 594

<212> PROTEIN

<213> Artificial sequence

<220>

<223>PD-1 fusion polypeptide

<400> 17

Met Gln Ile Pro Gln Ala Pro Trp Pro Val Val Trp Ala Val Leu Gln

1 5 10 15

Leu Gly Trp Arg Pro Gly Trp Phe Leu Asp Ser Pro Asp Arg Pro Trp

20 25 30

Asn Pro Pro Thr Phe Ser Pro Ala Leu Leu Val Val Thr Glu Gly Asp

35 40 45

Asn Ala Thr Phe Thr Cys Ser Phe Ser Asn Thr Ser Glu Ser Phe Val

50 55 60

Leu Asn Trp Tyr Arg Met Ser Pro Ser Asn Gln Thr Asp Lys Leu Ala

65 70 75 80

Ala Phe Pro Glu Asp Arg Ser Gln Pro Gly Gln Asp Cys Arg Phe Arg

85 90 95

Val Thr Gln Leu Pro Asn Gly Arg Asp Phe His Met Ser Val Val Arg

100 105 110

Ala Arg Arg Asn Asp Ser Gly Thr Tyr Leu Cys Gly Ala Ile Ser Leu

115 120 125

Ala Pro Lys Ala Gln Ile Lys Glu Ser Leu Arg Ala Glu Leu Arg Val

130 135 140

Thr Glu Arg Arg Ala Glu Val Pro Thr Ala His Pro Ser Pro Ser Pro

145 150 155 160

Ser Gly Trp Leu Leu Glu Val Pro Asn Gly Pro Trp Arg Ser Leu Thr

165 170 175

Phe Tyr Pro Ala Trp Leu Thr Val Ser Glu Gly Ala Asn Ala Thr Phe

180 185 190

Thr Cys Ser Leu Ser Asn Trp Ser Glu Asp Leu Met Leu Asn Trp Asn

195 200 205

Arg Leu Ser Pro Ser Asn Gln Thr Glu Lys Gln Ala Ala Phe Cys Asn

210 215 220

Gly Leu Ser Gln Pro Val Gln Asp Ala Arg Phe Gln Ile Ile Gln Leu

225 230 235 240

Pro Asn Arg His Asp Phe His Met Asn Ile Leu Asp Thr Arg Arg Asn

245 250 255

Asp Ser Gly Ile Tyr Leu Cys Gly Ala Ile Ser Leu His Pro Lys Ala

260 265 270

Lys Ile Glu Glu Ser Pro Gly Ala Glu Leu Val Val Thr Glu Arg Ile

275 280 285

Leu Glu Thr Ser Thr Arg Tyr Pro Ser Pro Ser Pro Lys Pro Glu Gly

290 295 300

Arg Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly

305 310 315 320

Thr Met Gly Gly Ala Ala Gly Ser Gly Ala Ala Glu Ala Gly Ile Thr

325 330 335

Gly Thr Trp Tyr Asn Gln Leu Gly Ser Thr Phe Ile Val Thr Ala Gly

340 345 350

Ala Asp Gly Ala Leu Thr Gly Thr Tyr Glu Ser Ala Val Gly Asn Ala

355 360 365

Glu Ser Arg Tyr Val Leu Thr Gly Arg Tyr Glu Ser Ala Pro Ala Thr

370 375 380

Asp Gly Ser Gly Thr Ala Leu Gly Trp Thr Val Ala Trp Lys Asn Asn

385 390 395 400

Tyr Arg Asn Ala His Ser Ala Thr Thr Trp Ser Gly Gln Tyr Val Gly

405 410 415

Gly Ala Glu Ala Arg Ile Asn Thr Gln Trp Leu Leu Thr Ser Gly Thr

420 425 430

Thr Glu Ala Asn Ala Trp Lys Ser Thr Leu Val Gly His Asp Thr Phe

435 440 445

Thr Lys Val Lys Pro Ser Ala Ala Ser Gly Ser Ala Ala Gly Ser Gly

450 455 460

Ala Ala Glu Ala Gly Ile Thr Gly Thr Trp Tyr Asn Gln Leu Gly Ser

465 470 475 480

Thr Phe Ile Val Thr Ala Gly Ala Asp Gly Ala Leu Thr Gly Thr Tyr

485 490 495

Glu Ser Ala Val Gly Asn Ala Glu Ser Arg Tyr Val Leu Thr Gly Arg

500 505 510

Tyr Glu Ser Ala Pro Ala Thr Asp Gly Ser Gly Thr Ala Leu Gly Trp

515 520 525

Thr Val Ala Trp Lys Asn Asn Tyr Arg Asn Ala His Ser Ala Thr Thr

530 535 540

Trp Ser Gly Gln Tyr Val Gly Gly Ala Glu Ala Arg Ile Asn Thr Gln

545 550 555 560

Trp Leu Leu Thr Ser Gly Thr Thr Glu Ala Asn Ala Trp Lys Ser Thr

565 570 575

Leu Val Gly His Asp Thr Phe Thr Lys Val Lys Pro Ser Ala Ala Ser

580 585 590

Gly Ser

<210> 18

<211> 765

<212> PROTEIN

<213> Artificial sequence

<220>

<223>PD-1 fusion polypeptide

<400> 18

Met Ser Pro Gln Arg Asp Arg Ile Asn Ala Phe Tyr Lys Asp Asn Pro

1 5 10 15

His Pro Lys Gly Ser Arg Ile Val Ile Asn Arg Glu His Leu Met Ile

20 25 30

Asp Arg Pro Tyr Val Leu Leu Ala Val Leu Phe Val Met Phe Leu Ser

35 40 45

Leu Ile Gly Leu Leu Ala Ile Ala Gly Ile Arg Leu His Arg Ala Ala

50 55 60

Ile Tyr Thr Ala Glu Ile His Lys Ser Leu Ser Thr Asn Leu Asp Val

65 70 75 80

Thr Asn Ser Ile Glu His Gln Val Lys Asp Val Leu Thr Pro Leu Phe

85 90 95

Lys Ile Ile Gly Asp Glu Val Gly Leu Arg Thr Pro Gln Arg Phe Thr

100 105 110

Asp Leu Val Lys Phe Ile Ser Asp Lys Ile Lys Phe Leu Asn Pro Asp

115 120 125

Arg Glu Tyr Asp Phe Arg Asp Leu Thr Trp Cys Ile Asn Pro Pro Glu

130 135 140

Arg Ile Lys Leu Asp Tyr Asp Gln Tyr Cys Ala Asp Val Ala Ala Glu

145 150 155 160

Glu Leu Met Asn Ala Leu Val Asn Ser Thr Leu Leu Glu Thr Arg Thr

165 170 175

Thr Asn Gln Phe Leu Ala Val Ser Lys Gly Asn Cys Ser Gly Pro Thr

180 185 190

Thr Ile Arg Gly Gln Phe Ser Asn Met Ser Leu Ser Leu Leu Asp Leu

195 200 205

Tyr Leu Gly Arg Gly Tyr Asn Val Ser Ser Ile Val Thr Met Thr Ser

210 215 220

Gln Gly Met Tyr Gly Gly Thr Tyr Leu Val Glu Lys Pro Asn Leu Ser

225 230 235 240

Ser Lys Arg Ser Glu Leu Ser Gln Leu Ser Met Tyr Arg Val Phe Glu

245 250 255

Val Gly Val Ile Arg Asn Pro Gly Leu Gly Ala Pro Val Phe His Met

260 265 270

Thr Asn Tyr Leu Glu Gln Pro Val Ser Asn Asp Leu Ser Asn Cys Met

275 280 285

Val Ala Leu Gly Glu Leu Lys Leu Ala Ala Leu Cys His Gly Glu Asp

290 295 300

Ser Ile Thr Ile Pro Tyr Gln Gly Ser Gly Lys Gly Val Ser Phe Gln

305 310 315 320

Leu Val Lys Leu Gly Val Trp Lys Ser Pro Thr Asp Met Gln Ser Trp

325 330 335

Val Pro Leu Ser Thr Asp Asp Pro Val Ile Asp Arg Leu Tyr Leu Ser

340 345 350

Ser His Arg Gly Val Ile Ala Asp Asn Gln Ala Lys Trp Ala Val Pro

355 360 365

Thr Thr Arg Thr Asp Asp Lys Leu Arg Met Glu Thr Cys Phe Gln Gln

370 375 380

Ala Cys Lys Gly Lys Ile Gln Ala Leu Cys Glu Asn Pro Glu Trp Ala

385 390 395 400

Pro Leu Lys Asp Asn Arg Ile Pro Ser Tyr Gly Val Leu Ser Val Asp

405 410 415

Leu Ser Leu Thr Val Glu Leu Lys Ile Lys Ile Ala Ser Gly Phe Gly

420 425 430

Pro Leu Ile Thr His Gly Ser Gly Met Asp Leu Tyr Lys Ser Asn His

435 440 445

Asn Asn Val Tyr Trp Leu Thr Ile Pro Pro Met Lys Asn Leu Ala Leu

450 455 460

Gly Val Ile Asn Thr Leu Glu Trp Ile Pro Arg Phe Lys Val Ser Pro

465 470 475 480

Ala Leu Phe Asn Val Pro Ile Lys Glu Ala Gly Gly Asp Cys His Ala

485 490 495

Pro Thr Tyr Leu Pro Ala Glu Val Asp Gly Asp Val Lys Leu Ser Ser

500 505 510

Asn Leu Val Ile Leu Pro Gly Gln Asp Leu Gln Tyr Val Leu Ala Thr

515 520 525

Tyr Asp Thr Ser Ala Val Glu His Ala Val Val Tyr Tyr Val Tyr Ser

530 535 540

Pro Ser Arg Ser Phe Ser Tyr Phe Tyr Pro Phe Arg Leu Pro Ile Lys

545 550 555 560

Gly Val Pro Ile Glu Leu Gln Val Glu Cys Phe Thr Trp Asp Gln Lys

565 570 575

Leu Trp Cys Arg His Phe Cys Val Leu Ala Asp Ser Glu Ser Gly Gly

580 585 590

His Ile Thr His Ser Gly Met Val Gly Met Gly Val Ser Cys Thr Val

595 600 605

Thr Arg Glu Asp Gly Thr Asn Ile Glu Gly Arg Pro Gly Trp Phe Leu

610 615 620

Asp Ser Pro Asp Arg Pro Trp Asn Pro Pro Thr Phe Ser Pro Ala Leu

625 630 635 640

Leu Val Val Thr Glu Gly Asp Asn Ala Thr Phe Thr Cys Ser Phe Ser

645 650 655

Asn Thr Ser Glu Ser Phe Val Leu Asn Trp Tyr Arg Met Ser Pro Ser

660 665 670

Asn Gln Thr Asp Lys Leu Ala Ala Phe Pro Glu Asp Arg Ser Gln Pro

675 680 685

Gly Gln Asp Cys Arg Phe Arg Val Thr Gln Leu Pro Asn Gly Arg Asp

690 695 700

Phe His Met Ser Val Val Arg Ala Arg Arg Asn Asp Ser Gly Thr Tyr

705 710 715 720

Leu Cys Gly Ala Ile Ser Leu Ala Pro Lys Ala Gln Ile Lys Glu Ser

725 730 735

Leu Arg Ala Glu Leu Arg Val Thr Glu Arg Arg Ala Glu Val Pro Thr

740 745 750

Ala His Pro Ser Pro Ser Pro His His His His His

755 760 765

<210> 19

<211> 765

<212> PROTEIN

<213> Artificial sequence

<220>

<223>PD-1 fusion polypeptide

<400> 19

Met Ser Pro Gln Arg Asp Arg Ile Asn Ala Phe Tyr Lys Asp Asn Pro

1 5 10 15

His Pro Lys Gly Ser Arg Ile Val Ile Asn Arg Glu His Leu Met Ile

20 25 30

Asp Arg Pro Tyr Val Leu Leu Ala Val Leu Phe Val Met Phe Leu Ser

35 40 45

Leu Ile Gly Leu Leu Ala Ile Ala Gly Ile Arg Leu His Arg Ala Ala

50 55 60

Ile Tyr Thr Ala Glu Ile His Lys Ser Leu Ser Thr Asn Leu Asp Val

65 70 75 80

Thr Asn Ser Ile Glu His Gln Val Lys Asp Val Leu Thr Pro Leu Phe

85 90 95

Lys Ile Ile Gly Asp Glu Val Gly Leu Arg Thr Pro Gln Arg Phe Thr

100 105 110

Asp Leu Val Lys Phe Ile Ser Asp Lys Ile Lys Phe Leu Asn Pro Asp

115 120 125

Arg Glu Tyr Asp Phe Arg Asp Leu Thr Trp Cys Ile Asn Pro Pro Glu

130 135 140

Arg Ile Lys Leu Asp Tyr Asp Gln Tyr Cys Ala Asp Val Ala Ala Glu

145 150 155 160

Glu Leu Met Asn Ala Leu Val Asn Ser Thr Leu Leu Glu Thr Arg Thr

165 170 175

Thr Asn Gln Phe Leu Ala Val Ser Lys Gly Asn Cys Ser Gly Pro Thr

180 185 190

Thr Ile Arg Gly Gln Phe Ser Asn Met Ser Leu Ser Leu Leu Asp Leu

195 200 205

Tyr Leu Gly Arg Gly Tyr Asn Val Ser Ser Ile Val Thr Met Thr Ser

210 215 220

Gln Gly Met Tyr Gly Gly Thr Tyr Leu Val Glu Lys Pro Asn Leu Ser

225 230 235 240

Ser Lys Arg Ser Glu Leu Ser Gln Leu Ser Met Tyr Arg Val Phe Glu

245 250 255

Val Gly Val Ile Arg Asn Pro Gly Leu Gly Ala Pro Val Phe His Met

260 265 270

Thr Asn Tyr Leu Glu Gln Pro Val Ser Asn Asp Leu Ser Asn Cys Met

275 280 285

Val Ala Leu Gly Glu Leu Lys Leu Ala Ala Leu Cys His Gly Glu Asp

290 295 300

Ser Ile Thr Ile Pro Tyr Gln Gly Ser Gly Lys Gly Val Ser Phe Gln

305 310 315 320

Leu Val Lys Leu Gly Val Trp Lys Ser Pro Thr Asp Met Gln Ser Trp

325 330 335

Val Pro Leu Ser Thr Asp Asp Pro Val Ile Asp Arg Leu Tyr Leu Ser

340 345 350

Ser His Arg Gly Val Ile Ala Asp Asn Gln Ala Lys Trp Ala Val Pro

355 360 365

Thr Thr Arg Thr Asp Asp Lys Leu Arg Met Glu Thr Cys Phe Gln Gln

370 375 380

Ala Cys Lys Gly Lys Ile Gln Ala Leu Cys Glu Asn Pro Glu Trp Ala

385 390 395 400

Pro Leu Lys Asp Asn Arg Ile Pro Ser Tyr Gly Val Leu Ser Val Asp

405 410 415

Leu Ser Leu Thr Val Glu Leu Lys Ile Lys Ile Ala Ser Gly Phe Gly

420 425 430

Pro Leu Ile Thr His Gly Ser Gly Met Asp Leu Tyr Lys Ser Asn His

435 440 445

Asn Asn Val Tyr Trp Leu Thr Ile Pro Pro Met Lys Asn Leu Ala Leu

450 455 460

Gly Val Ile Asn Thr Leu Glu Trp Ile Pro Arg Phe Lys Val Ser Pro

465 470 475 480

Ala Leu Phe Asn Val Pro Ile Lys Glu Ala Gly Gly Asp Cys His Ala

485 490 495

Pro Thr Tyr Leu Pro Ala Glu Val Asp Gly Asp Val Lys Leu Ser Ser

500 505 510

Asn Leu Val Ile Leu Pro Gly Gln Asp Leu Gln Tyr Val Leu Ala Thr

515 520 525

Tyr Asp Thr Ser Ala Val Glu His Ala Val Val Tyr Tyr Val Tyr Ser

530 535 540

Pro Ser Arg Leu Ser Ser Tyr Phe Tyr Pro Phe Arg Leu Pro Ile Lys

545 550 555 560

Gly Val Pro Ile Glu Leu Gln Val Glu Cys Phe Thr Trp Asp Gln Lys

565 570 575

Leu Trp Cys Arg His Phe Cys Val Leu Ala Asp Ser Glu Ser Gly Gly

580 585 590

His Ile Thr His Ser Gly Met Val Gly Met Gly Val Ser Cys Thr Val

595 600 605

Thr Arg Glu Asp Gly Thr Asn Ile Glu Gly Arg Pro Gly Trp Phe Leu

610 615 620

Asp Ser Pro Asp Arg Pro Trp Asn Pro Pro Thr Phe Ser Pro Ala Leu

625 630 635 640

Leu Val Val Thr Glu Gly Asp Asn Ala Thr Phe Thr Cys Ser Phe Ser

645 650 655

Asn Thr Ser Glu Ser Phe Val Leu Asn Trp Tyr Arg Met Ser Pro Ser

660 665 670

Asn Gln Thr Asp Lys Leu Ala Ala Phe Pro Glu Asp Arg Ser Gln Pro

675 680 685

Gly Gln Asp Cys Arg Phe Arg Val Thr Gln Leu Pro Asn Gly Arg Asp

690 695 700

Phe His Met Ser Val Val Arg Ala Arg Arg Asn Asp Ser Gly Thr Tyr

705 710 715 720

Leu Cys Gly Ala Ile Ser Leu Ala Pro Lys Ala Gln Ile Lys Glu Ser

725 730 735

Leu Arg Ala Glu Leu Arg Val Thr Glu Arg Arg Ala Glu Val Pro Thr

740 745 750

Ala His Pro Ser Pro Ser Pro His His His His His

755 760 765

<210> 20

<211> 754

<212> PROTEIN

<213> Artificial sequence

<220>

<223>PD-1 fusion polypeptide

<400> 20

Met Leu Pro Tyr Gln Asp Lys Val Gly Ala Phe Tyr Lys Asp Asn Ala

1 5 10 15

Arg Ala Asn Ser Thr Lys Leu Ser Leu Val Thr Glu Gly His Gly Gly

20 25 30

Arg Arg Pro Pro Tyr Leu Leu Phe Val Leu Leu Ile Leu Leu Val Gly

35 40 45

Ile Leu Ala Leu Leu Ala Ile Thr Gly Val Arg Phe His Gln Val Ser

50 55 60

Thr Ser Asn Met Glu Phe Ser Arg Leu Leu Lys Glu Asp Met Glu Lys

65 70 75 80

Ser Glu Ala Val His His Gln Val Ile Asp Val Leu Thr Pro Leu Phe

85 90 95

Lys Ile Ile Gly Asp Glu Ile Gly Leu Arg Leu Pro Gln Lys Leu Asn

100 105 110

Glu Ile Lys Gln Phe Ile Leu Gln Lys Thr Asn Phe Phe Asn Pro Asn

115 120 125

Arg Glu Phe Asp Phe Arg Asp Leu His Trp Cys Ile Asn Pro Pro Ser

130 135 140

Thr Val Lys Val Asn Phe Thr Asn Tyr Cys Glu Ser Ile Gly Ile Arg

145 150 155 160

Lys Ala Ile Ala Ser Ala Ala Asn Pro Ile Leu Leu Ser Ala Leu Ser

165 170 175

Gly Gly Arg Gly Asp Ile Phe Pro Pro His Arg Cys Ser Gly Ala Thr

180 185 190

Thr Ser Val Gly Lys Val Phe Pro Leu Ser Val Ser Leu Ser Met Ser

195 200 205

Leu Ile Ser Arg Thr Ser Glu Val Ile Asn Met Leu Thr Ala Ile Ser

210 215 220

Asp Gly Val Tyr Gly Lys Thr Tyr Leu Leu Val Pro Asp Asp Ile Glu

225 230 235 240

Arg Glu Phe Asp Thr Arg Glu Ile Arg Val Phe Glu Ile Gly Phe Ile

245 250 255

Lys Arg Trp Leu Asn Asp Met Pro Leu Leu Gln Thr Thr Asn Tyr Met

260 265 270

Val Leu Pro Lys Asn Ser Lys Ala Lys Val Cys Thr Ile Ala Val Gly

275 280 285

Glu Leu Thr Leu Ala Ser Leu Cys Val Glu Glu Ser Thr Val Leu Leu

290 295 300

Tyr His Asp Ser Ser Gly Ser Gln Asp Gly Ile Leu Val Val Thr Leu

305 310 315 320

Gly Ile Phe Trp Ala Thr Pro Met Asp His Ile Glu Glu Val Ile Pro

325 330 335

Val Ala His Pro Ser Met Lys Lys Ile His Ile Thr Asn His Arg Gly

340 345 350

Phe Ile Lys Asp Ser Ile Ala Thr Trp Met Val Pro Ala Leu Ala Ser

355 360 365

Glu Lys Gln Glu Glu Gln Lys Gly Cys Leu Glu Ser Ala Cys Gln Arg

370 375 380

Lys Thr Tyr Pro Met Cys Asn Gln Ala Ser Trp Glu Pro Phe Gly Gly

385 390 395 400

Arg Gln Leu Pro Ser Tyr Gly Arg Leu Thr Leu Pro Leu Asp Ala Ser

405 410 415

Val Asp Leu Gln Leu Asn Ile Ser Phe Thr Tyr Gly Pro Val Ile Leu

420 425 430

Asn Gly Asp Gly Met Asp Tyr Tyr Glu Ser Pro Leu Leu Asn Ser Gly

435 440 445

Trp Leu Thr Ile Pro Pro Lys Asp Gly Thr Ile Ser Gly Leu Ile Asn

450 455 460

Lys Ala Gly Arg Gly Asp Gln Phe Thr Val Leu Pro His Val Leu Thr

465 470 475 480

Phe Ala Pro Arg Glu Ser Ser Gly Asn Cys Tyr Leu Pro Ile Gln Thr

485 490 495

Ser Gln Ile Arg Asp Arg Asp Val Leu Ile Glu Ser Asn Ile Val Val

500 505 510

Leu Pro Thr Gln Ser Ile Arg Tyr Val Ile Ala Thr Tyr Asp Ile Ser

515 520 525

Arg Ser Asp His Ala Ile Val Tyr Tyr Val Tyr Asp Pro Ile Arg Thr

530 535 540

Ile Ser Tyr Thr His Pro Phe Arg Leu Thr Thr Lys Gly Arg Pro Asp

545 550 555 560

Phe Leu Arg Ile Glu Cys Phe Val Trp Asp Asp Asn Leu Trp Cys His

565 570 575

Gln Phe Tyr Arg Phe Glu Ala Asp Ile Ala Asn Ser Thr Thr Ser Val

580 585 590

Glu Asn Leu Val Arg Ile Arg Phe Ser Cys Asn Arg Ile Glu Gly Arg

595 600 605

Pro Gly Trp Phe Leu Asp Ser Pro Asp Arg Pro Trp Asn Pro Pro Thr

610 615 620

Phe Ser Pro Ala Leu Leu Val Val Thr Glu Gly Asp Asn Ala Thr Phe

625 630 635 640

Thr Cys Ser Phe Ser Asn Thr Ser Glu Ser Phe Val Leu Asn Trp Tyr

645 650 655

Arg Met Ser Pro Ser Asn Gln Thr Asp Lys Leu Ala Ala Phe Pro Glu

660 665 670

Asp Arg Ser Gln Pro Gly Gln Asp Cys Arg Phe Arg Val Thr Gln Leu

675 680 685

Pro Asn Gly Arg Asp Phe His Met Ser Val Val Arg Ala Arg Arg Asn

690 695 700

Asp Ser Gly Thr Tyr Leu Cys Gly Ala Ile Ser Leu Ala Pro Lys Ala

705 710 715 720

Gln Ile Lys Glu Ser Leu Arg Ala Glu Leu Arg Val Thr Glu Arg Arg

725 730 735

Ala Glu Val Pro Thr Ala His Pro Ser Pro Ser Pro His His His His

740 745 750

His His

<210> 21

<211> 385

<212> PROTEIN

<213> Artificial sequence

<220>

<223>PD-1 fusion polypeptide

<400> 21

Met Gln Ile Pro Gln Ala Pro Trp Pro Val Val Trp Ala Val Leu Gln

1 5 10 15

Leu Gly Trp Arg Pro Gly Trp Phe Leu Asp Ser Pro Asp Arg Pro Trp

20 25 30

Asn Pro Pro Thr Phe Ser Pro Ala Leu Leu Val Val Thr Glu Gly Asp

35 40 45

Asn Ala Thr Phe Thr Cys Ser Phe Ser Asn Thr Ser Glu Ser Phe His

50 55 60

Val Ile Trp His Arg Glu Ser Pro Ser Gly Gln Thr Asp Thr Leu Ala

65 70 75 80

Ala Phe Pro Glu Asp Arg Ser Gln Pro Gly Gln Asp Cys Arg Phe Arg

85 90 95

Val Thr Gln Leu Pro Asn Gly Arg Asp Phe His Met Ser Val Val Arg

100 105 110

Ala Arg Arg Asn Asp Ser Gly Thr Tyr Val Cys Gly Val Ile Ser Leu

115 120 125

Ala Pro Lys Ile Gln Ile Lys Glu Ser Leu Arg Ala Glu Leu Arg Val

130 135 140

Thr Glu Arg Arg Ala Glu Val Pro Thr Ala His Pro Ser Pro Ser Pro

145 150 155 160

Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro

165 170 175

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

180 185 190

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp

195 200 205

Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

210 215 220

Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Val

225 230 235 240

Val Ser Val Leu Thr Val Val His Gln Asp Trp Leu Asn Gly Lys Glu

245 250 255

Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys

260 265 270

Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

275 280 285

Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr

290 295 300

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

305 310 315 320

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu

325 330 335

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys

340 345 350

Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu

355 360 365

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

370 375 380

Lys

385

<210> 22

<211> 268

<212> PROTEIN

<213> Artificial sequence

<220>

<223>PD-1 fusion polypeptide

<400> 22

Met Gln Ile Pro Gln Ala Pro Trp Pro Val Val Trp Ala Val Leu Gln

1 5 10 15

Leu Gly Trp Arg Pro Gly Trp Phe Leu Asp Ser Pro Asp Arg Pro Trp

20 25 30

Asn Pro Pro Thr Phe Ser Pro Ala Leu Leu Val Val Thr Glu Gly Asp

35 40 45

Asn Ala Thr Phe Thr Cys Ser Phe Ser Asn Thr Ser Glu Ser Phe His

50 55 60

Val Ile Trp His Arg Glu Ser Pro Ser Gly Gln Thr Asp Thr Leu Ala

65 70 75 80

Ala Phe Pro Glu Asp Arg Ser Gln Pro Gly Gln Asp Cys Arg Phe Arg

85 90 95

Val Thr Gln Leu Pro Asn Gly Arg Asp Phe His Met Ser Val Val Arg

100 105 110

Ala Arg Arg Asn Asp Ser Gly Thr Tyr Val Cys Gly Val Ile Ser Leu

115 120 125

Ala Pro Lys Ile Gln Ile Lys Glu Ser Leu Arg Ala Glu Leu Arg Val

130 135 140

Thr Glu Arg Arg Ala Glu Val Pro Thr Ala His Pro Ser Pro Ser Pro

145 150 155 160

Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Gly

165 170 175

Ser Ala Asn Ser Phe Leu Glu Glu Met Lys Lys Gly His Leu Glu Arg

180 185 190

Glu Cys Met Glu Glu Thr Cys Ser Tyr Glu Glu Ala Arg Glu Val Phe

195 200 205

Glu Asp Ser Asp Lys Thr Asn Glu Phe Trp Asn Lys Tyr Lys Asp Gly

210 215 220

Asp Gln Cys Glu Thr Ser Pro Cys Gln Asn Gln Gly Lys Cys Lys Asp

225 230 235 240

Gly Leu Gly Glu Tyr Thr Cys Thr Cys Leu Glu Gly Phe Glu Gly Lys

245 250 255

Asn Cys Glu Leu Pro Val Gly Tyr Arg Gly Gly Ser

260 265

<210> 23

<211> 248

<212> PROTEIN

<213> Artificial sequence

<220>

<223>PD-1 fusion polypeptide

<400> 23

Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Gly

1 5 10 15

Ser Ala Asn Ser Phe Leu Glu Glu Met Lys Lys Gly His Leu Glu Arg

20 25 30

Glu Cys Met Glu Glu Thr Cys Ser Tyr Glu Glu Ala Arg Glu Val Phe

35 40 45

Glu Asp Ser Asp Lys Thr Asn Glu Phe Trp Asn Lys Tyr Lys Asp Gly

50 55 60

Asp Gln Cys Glu Thr Ser Pro Cys Gln Asn Gln Gly Lys Cys Lys Asp

65 70 75 80

Gly Leu Gly Glu Tyr Thr Cys Thr Cys Leu Glu Gly Phe Glu Gly Lys

85 90 95

Asn Cys Glu Leu Pro Val Gly Tyr Arg Gly Gly Ser Pro Gly Trp Phe

100 105 110

Leu Asp Ser Pro Asp Arg Pro Trp Asn Pro Pro Thr Phe Ser Pro Ala

115 120 125

Leu Leu Val Val Thr Glu Gly Asp Asn Ala Thr Phe Thr Cys Ser Phe

130 135 140

Ser Asn Thr Ser Glu Ser Phe His Val Ile Trp His Arg Glu Ser Pro

145 150 155 160

Ser Gly Gln Thr Asp Thr Leu Ala Ala Phe Pro Glu Asp Arg Ser Gln

165 170 175

Pro Gly Gln Asp Cys Arg Phe Arg Val Thr Gln Leu Pro Asn Gly Arg

180 185 190

Asp Phe His Met Ser Val Val Arg Ala Arg Arg Asn Asp Ser Gly Thr

195 200 205

Tyr Val Cys Gly Val Ile Ser Leu Ala Pro Lys Ile Gln Ile Lys Glu

210 215 220

Ser Leu Arg Ala Glu Leu Arg Val Thr Glu Arg Arg Ala Glu Val Pro

225 230 235 240

Thr Ala His Pro Ser Pro Ser Pro

245

<210> 24

<211> 305

<212> PROTEIN

<213> Artificial sequence

<220>

<223>PD-1 fusion polypeptide

<400> 24

Met Gln Ile Pro Gln Ala Pro Trp Pro Val Val Trp Ala Val Leu Gln

1 5 10 15

Leu Gly Trp Arg Pro Gly Trp Phe Leu Asp Ser Pro Asp Arg Pro Trp

20 25 30

Asn Pro Pro Thr Phe Ser Pro Ala Leu Leu Val Val Thr Glu Gly Asp

35 40 45

Asn Ala Thr Phe Thr Cys Ser Phe Ser Asn Thr Ser Glu Ser Phe His

50 55 60

Val Ile Trp His Arg Glu Ser Pro Ser Gly Gln Thr Asp Thr Leu Ala

65 70 75 80

Ala Phe Pro Glu Asp Arg Ser Gln Pro Gly Gln Asp Cys Arg Phe Arg

85 90 95

Val Thr Gln Leu Pro Asn Gly Arg Asp Phe His Met Ser Val Val Arg

100 105 110

Ala Arg Arg Asn Asp Ser Gly Thr Tyr Val Cys Gly Val Ile Ser Leu

115 120 125

Ala Pro Lys Ile Gln Ile Lys Glu Ser Leu Arg Ala Glu Leu Arg Val

130 135 140

Thr Glu Arg Arg Ala Glu Val Pro Thr Ala His Pro Ser Pro Ser Pro

145 150 155 160

Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Ser

165 170 175

Lys Gly Leu Glu Ser Arg Val Ser Ala Leu Glu Lys Thr Ser Gln Ile

180 185 190

His Ser Asp Thr Ile Leu Arg Ile Thr Gln Gly Leu Asp Asp Ala Asn

195 200 205

Lys Arg Ile Ile Ala Leu Glu Gln Ser Arg Asp Asp Leu Val Ala Ser

210 215 220

Val Ser Asp Ala Gln Leu Ala Ile Ser Arg Leu Glu Ser Ser Ile Gly

225 230 235 240

Ala Leu Gln Thr Val Val Asn Gly Leu Asp Ser Ser Val Thr Gln Leu

245 250 255

Gly Ala Arg Val Gly Gln Leu Glu Thr Gly Leu Ala Glu Leu Arg Val

260 265 270

Asp His Asp Asn Leu Val Ala Arg Val Asp Thr Ala Glu Arg Asn Ile

275 280 285

Gly Ser Leu Thr Thr Glu Leu Ser Thr Leu Thr Leu Arg Val Thr Ser

290 295 300

Ile

305

<210> 25

<211> 765

<212> PROTEIN

<213> Artificial sequence

<220>

<223>PD-1 fusion polypeptide

<400> 25

Met Ser Pro Gln Arg Asp Arg Ile Asn Ala Phe Tyr Lys Asp Asn Pro

1 5 10 15

His Pro Lys Gly Ser Arg Ile Val Ile Asn Arg Glu His Leu Met Ile

20 25 30

Asp Arg Pro Tyr Val Leu Leu Ala Val Leu Phe Val Met Phe Leu Ser

35 40 45

Leu Ile Gly Leu Leu Ala Ile Ala Gly Ile Arg Leu His Arg Ala Ala

50 55 60

Ile Tyr Thr Ala Glu Ile His Lys Ser Leu Ser Thr Asn Leu Asp Val

65 70 75 80

Thr Asn Ser Ile Glu His Gln Val Lys Asp Val Leu Thr Pro Leu Phe

85 90 95

Lys Ile Ile Gly Asp Glu Val Gly Leu Arg Thr Pro Gln Arg Phe Thr

100 105 110

Asp Leu Val Lys Phe Ile Ser Asp Lys Ile Lys Phe Leu Asn Pro Asp

115 120 125

Arg Glu Tyr Asp Phe Arg Asp Leu Thr Trp Cys Ile Asn Pro Pro Glu

130 135 140

Arg Ile Lys Leu Asp Tyr Asp Gln Tyr Cys Ala Asp Val Ala Ala Glu

145 150 155 160

Glu Leu Met Asn Ala Leu Val Asn Ser Thr Leu Leu Glu Thr Arg Thr

165 170 175

Thr Asn Gln Phe Leu Ala Val Ser Lys Gly Asn Cys Ser Gly Pro Thr

180 185 190

Thr Ile Arg Gly Gln Phe Ser Asn Met Ser Leu Ser Leu Leu Asp Leu

195 200 205

Tyr Leu Gly Arg Gly Tyr Asn Val Ser Ser Ile Val Thr Met Thr Ser

210 215 220

Gln Gly Met Tyr Gly Gly Thr Tyr Leu Val Glu Lys Pro Asn Leu Ser

225 230 235 240

Ser Lys Arg Ser Glu Leu Ser Gln Leu Ser Met Tyr Arg Val Phe Glu

245 250 255

Val Gly Val Ile Arg Asn Pro Gly Leu Gly Ala Pro Val Phe His Met

260 265 270

Thr Asn Tyr Leu Glu Gln Pro Val Ser Asn Asp Leu Ser Asn Cys Met

275 280 285

Val Ala Leu Gly Glu Leu Lys Leu Ala Ala Leu Cys His Gly Glu Asp

290 295 300

Ser Ile Thr Ile Pro Tyr Gln Gly Ser Gly Lys Gly Val Ser Phe Gln

305 310 315 320

Leu Val Lys Leu Gly Val Trp Lys Ser Pro Thr Asp Met Gln Ser Trp

325 330 335

Val Pro Leu Ser Thr Asp Asp Pro Val Ile Asp Arg Leu Tyr Leu Ser

340 345 350

Ser His Arg Gly Val Ile Ala Asp Asn Gln Ala Lys Trp Ala Val Pro

355 360 365

Thr Thr Arg Thr Asp Asp Lys Leu Arg Met Glu Thr Cys Phe Gln Gln

370 375 380

Ala Cys Lys Gly Lys Ile Gln Ala Leu Cys Glu Asn Pro Glu Trp Ala

385 390 395 400

Pro Leu Lys Asp Asn Arg Ile Pro Ser Tyr Gly Val Leu Ser Val Asp

405 410 415

Leu Ser Leu Thr Val Glu Leu Lys Ile Lys Ile Ala Ser Gly Phe Gly

420 425 430

Pro Leu Ile Thr His Gly Ser Gly Met Asp Leu Tyr Lys Ser Asn His

435 440 445

Asn Asn Val Tyr Trp Leu Thr Ile Pro Pro Met Lys Asn Leu Ala Leu

450 455 460

Gly Val Ile Asn Thr Leu Glu Trp Ile Pro Arg Phe Lys Val Ser Pro

465 470 475 480

Ala Leu Phe Asn Val Pro Ile Lys Glu Ala Gly Gly Asp Cys His Ala

485 490 495

Pro Thr Tyr Leu Pro Ala Glu Val Asp Gly Asp Val Lys Leu Ser Ser

500 505 510

Asn Leu Val Ile Leu Pro Gly Gln Asp Leu Gln Tyr Val Leu Ala Thr

515 520 525

Tyr Asp Thr Ser Ala Val Glu His Ala Val Val Tyr Tyr Val Tyr Ser

530 535 540

Pro Ser Arg Ser Phe Ser Tyr Phe Tyr Pro Phe Arg Leu Pro Ile Lys

545 550 555 560

Gly Val Pro Ile Glu Leu Gln Val Glu Cys Phe Thr Trp Asp Gln Lys

565 570 575

Leu Trp Cys Arg His Phe Cys Val Leu Ala Asp Ser Glu Ser Gly Gly

580 585 590

His Ile Thr His Ser Gly Met Val Gly Met Gly Val Ser Cys Thr Val

595 600 605

Thr Arg Glu Asp Gly Thr Asn Ile Glu Gly Arg Pro Gly Trp Phe Leu

610 615 620

Asp Ser Pro Asp Arg Pro Trp Asn Pro Pro Thr Phe Ser Pro Ala Leu

625 630 635 640

Leu Val Val Thr Glu Gly Asp Asn Ala Thr Phe Thr Cys Ser Phe Ser

645 650 655

Asn Thr Ser Glu Ser Phe His Val Ile Trp His Arg Glu Ser Pro Ser

660 665 670

Gly Gln Thr Asp Thr Leu Ala Ala Phe Pro Glu Asp Arg Ser Gln Pro

675 680 685

Gly Gln Asp Cys Arg Phe Arg Val Thr Gln Leu Pro Asn Gly Arg Asp

690 695 700

Phe His Met Ser Val Val Arg Ala Arg Arg Asn Asp Ser Gly Thr Tyr

705 710 715 720

Val Cys Gly Val Ile Ser Leu Ala Pro Lys Ile Gln Ile Lys Glu Ser

725 730 735

Leu Arg Ala Glu Leu Arg Val Thr Glu Arg Arg Ala Glu Val Pro Thr

740 745 750

Ala His Pro Ser Pro Ser Pro His His His His His

755 760 765

<210> 26

<211> 765

<212> PROTEIN

<213> Artificial sequence

<220>

<223>PD-1 fusion polypeptide

<400> 26

Met Ser Pro Gln Arg Asp Arg Ile Asn Ala Phe Tyr Lys Asp Asn Pro

1 5 10 15

His Pro Lys Gly Ser Arg Ile Val Ile Asn Arg Glu His Leu Met Ile

20 25 30

Asp Arg Pro Tyr Val Leu Leu Ala Val Leu Phe Val Met Phe Leu Ser

35 40 45

Leu Ile Gly Leu Leu Ala Ile Ala Gly Ile Arg Leu His Arg Ala Ala

50 55 60

Ile Tyr Thr Ala Glu Ile His Lys Ser Leu Ser Thr Asn Leu Asp Val

65 70 75 80

Thr Asn Ser Ile Glu His Gln Val Lys Asp Val Leu Thr Pro Leu Phe

85 90 95

Lys Ile Ile Gly Asp Glu Val Gly Leu Arg Thr Pro Gln Arg Phe Thr

100 105 110

Asp Leu Val Lys Phe Ile Ser Asp Lys Ile Lys Phe Leu Asn Pro Asp

115 120 125

Arg Glu Tyr Asp Phe Arg Asp Leu Thr Trp Cys Ile Asn Pro Pro Glu

130 135 140

Arg Ile Lys Leu Asp Tyr Asp Gln Tyr Cys Ala Asp Val Ala Ala Glu

145 150 155 160

Glu Leu Met Asn Ala Leu Val Asn Ser Thr Leu Leu Glu Thr Arg Thr

165 170 175

Thr Asn Gln Phe Leu Ala Val Ser Lys Gly Asn Cys Ser Gly Pro Thr

180 185 190

Thr Ile Arg Gly Gln Phe Ser Asn Met Ser Leu Ser Leu Leu Asp Leu

195 200 205

Tyr Leu Gly Arg Gly Tyr Asn Val Ser Ser Ile Val Thr Met Thr Ser

210 215 220

Gln Gly Met Tyr Gly Gly Thr Tyr Leu Val Glu Lys Pro Asn Leu Ser

225 230 235 240

Ser Lys Arg Ser Glu Leu Ser Gln Leu Ser Met Tyr Arg Val Phe Glu

245 250 255

Val Gly Val Ile Arg Asn Pro Gly Leu Gly Ala Pro Val Phe His Met

260 265 270

Thr Asn Tyr Leu Glu Gln Pro Val Ser Asn Asp Leu Ser Asn Cys Met

275 280 285

Val Ala Leu Gly Glu Leu Lys Leu Ala Ala Leu Cys His Gly Glu Asp

290 295 300

Ser Ile Thr Ile Pro Tyr Gln Gly Ser Gly Lys Gly Val Ser Phe Gln

305 310 315 320

Leu Val Lys Leu Gly Val Trp Lys Ser Pro Thr Asp Met Gln Ser Trp

325 330 335

Val Pro Leu Ser Thr Asp Asp Pro Val Ile Asp Arg Leu Tyr Leu Ser

340 345 350

Ser His Arg Gly Val Ile Ala Asp Asn Gln Ala Lys Trp Ala Val Pro

355 360 365

Thr Thr Arg Thr Asp Asp Lys Leu Arg Met Glu Thr Cys Phe Gln Gln

370 375 380

Ala Cys Lys Gly Lys Ile Gln Ala Leu Cys Glu Asn Pro Glu Trp Ala

385 390 395 400

Pro Leu Lys Asp Asn Arg Ile Pro Ser Tyr Gly Val Leu Ser Val Asp

405 410 415

Leu Ser Leu Thr Val Glu Leu Lys Ile Lys Ile Ala Ser Gly Phe Gly

420 425 430

Pro Leu Ile Thr His Gly Ser Gly Met Asp Leu Tyr Lys Ser Asn His

435 440 445

Asn Asn Val Tyr Trp Leu Thr Ile Pro Pro Met Lys Asn Leu Ala Leu

450 455 460

Gly Val Ile Asn Thr Leu Glu Trp Ile Pro Arg Phe Lys Val Ser Pro

465 470 475 480

Ala Leu Phe Asn Val Pro Ile Lys Glu Ala Gly Gly Asp Cys His Ala

485 490 495

Pro Thr Tyr Leu Pro Ala Glu Val Asp Gly Asp Val Lys Leu Ser Ser

500 505 510

Asn Leu Val Ile Leu Pro Gly Gln Asp Leu Gln Tyr Val Leu Ala Thr

515 520 525

Tyr Asp Thr Ser Ala Val Glu His Ala Val Val Tyr Tyr Val Tyr Ser

530 535 540

Pro Ser Arg Leu Ser Ser Tyr Phe Tyr Pro Phe Arg Leu Pro Ile Lys

545 550 555 560

Gly Val Pro Ile Glu Leu Gln Val Glu Cys Phe Thr Trp Asp Gln Lys

565 570 575

Leu Trp Cys Arg His Phe Cys Val Leu Ala Asp Ser Glu Ser Gly Gly

580 585 590

His Ile Thr His Ser Gly Met Val Gly Met Gly Val Ser Cys Thr Val

595 600 605

Thr Arg Glu Asp Gly Thr Asn Ile Glu Gly Arg Pro Gly Trp Phe Leu

610 615 620

Asp Ser Pro Asp Arg Pro Trp Asn Pro Pro Thr Phe Ser Pro Ala Leu

625 630 635 640

Leu Val Val Thr Glu Gly Asp Asn Ala Thr Phe Thr Cys Ser Phe Ser

645 650 655

Asn Thr Ser Glu Ser Phe Val Leu Asn Trp Tyr Arg Met Ser Pro Ser

660 665 670

Asn Gln Thr Asp Lys Leu Ala Ala Phe Pro Glu Asp Arg Ser Gln Pro

675 680 685

Gly Gln Asp Cys Arg Phe Arg Val Thr Gln Leu Pro Asn Gly Arg Asp

690 695 700

Phe His Met Ser Val Val Arg Ala Arg Arg Asn Asp Ser Gly Thr Tyr

705 710 715 720

Leu Cys Gly Ala Ile Ser Leu Ala Pro Lys Ala Gln Ile Lys Glu Ser

725 730 735

Leu Arg Ala Glu Leu Arg Val Thr Glu Arg Arg Ala Glu Val Pro Thr

740 745 750

Ala His Pro Ser Pro Ser Pro His His His His His

755 760 765

<210> 27

<211> 754

<212> PROTEIN

<213> Artificial sequence

<220>

<223>PD-1 fusion polypeptide

<400> 27

Met Leu Pro Tyr Gln Asp Lys Val Gly Ala Phe Tyr Lys Asp Asn Ala

1 5 10 15

Arg Ala Asn Ser Thr Lys Leu Ser Leu Val Thr Glu Gly His Gly Gly

20 25 30

Arg Arg Pro Pro Tyr Leu Leu Phe Val Leu Leu Ile Leu Leu Val Gly

35 40 45

Ile Leu Ala Leu Leu Ala Ile Thr Gly Val Arg Phe His Gln Val Ser

50 55 60

Thr Ser Asn Met Glu Phe Ser Arg Leu Leu Lys Glu Asp Met Glu Lys

65 70 75 80

Ser Glu Ala Val His His Gln Val Ile Asp Val Leu Thr Pro Leu Phe

85 90 95

Lys Ile Ile Gly Asp Glu Ile Gly Leu Arg Leu Pro Gln Lys Leu Asn

100 105 110

Glu Ile Lys Gln Phe Ile Leu Gln Lys Thr Asn Phe Phe Asn Pro Asn

115 120 125

Arg Glu Phe Asp Phe Arg Asp Leu His Trp Cys Ile Asn Pro Pro Ser

130 135 140

Thr Val Lys Val Asn Phe Thr Asn Tyr Cys Glu Ser Ile Gly Ile Arg

145 150 155 160

Lys Ala Ile Ala Ser Ala Ala Asn Pro Ile Leu Leu Ser Ala Leu Ser

165 170 175

Gly Gly Arg Gly Asp Ile Phe Pro Pro His Arg Cys Ser Gly Ala Thr

180 185 190

Thr Ser Val Gly Lys Val Phe Pro Leu Ser Val Ser Leu Ser Met Ser

195 200 205

Leu Ile Ser Arg Thr Ser Glu Val Ile Asn Met Leu Thr Ala Ile Ser

210 215 220

Asp Gly Val Tyr Gly Lys Thr Tyr Leu Leu Val Pro Asp Asp Ile Glu

225 230 235 240

Arg Glu Phe Asp Thr Arg Glu Ile Arg Val Phe Glu Ile Gly Phe Ile

245 250 255

Lys Arg Trp Leu Asn Asp Met Pro Leu Leu Gln Thr Thr Asn Tyr Met

260 265 270

Val Leu Pro Lys Asn Ser Lys Ala Lys Val Cys Thr Ile Ala Val Gly

275 280 285

Glu Leu Thr Leu Ala Ser Leu Cys Val Glu Glu Ser Thr Val Leu Leu

290 295 300

Tyr His Asp Ser Ser Gly Ser Gln Asp Gly Ile Leu Val Val Thr Leu

305 310 315 320

Gly Ile Phe Trp Ala Thr Pro Met Asp His Ile Glu Glu Val Ile Pro

325 330 335

Val Ala His Pro Ser Met Lys Lys Ile His Ile Thr Asn His Arg Gly

340 345 350

Phe Ile Lys Asp Ser Ile Ala Thr Trp Met Val Pro Ala Leu Ala Ser

355 360 365

Glu Lys Gln Glu Glu Gln Lys Gly Cys Leu Glu Ser Ala Cys Gln Arg

370 375 380

Lys Thr Tyr Pro Met Cys Asn Gln Ala Ser Trp Glu Pro Phe Gly Gly

385 390 395 400

Arg Gln Leu Pro Ser Tyr Gly Arg Leu Thr Leu Pro Leu Asp Ala Ser

405 410 415

Val Asp Leu Gln Leu Asn Ile Ser Phe Thr Tyr Gly Pro Val Ile Leu

420 425 430

Asn Gly Asp Gly Met Asp Tyr Tyr Glu Ser Pro Leu Leu Asn Ser Gly

435 440 445

Trp Leu Thr Ile Pro Pro Lys Asp Gly Thr Ile Ser Gly Leu Ile Asn

450 455 460

Lys Ala Gly Arg Gly Asp Gln Phe Thr Val Leu Pro His Val Leu Thr

465 470 475 480

Phe Ala Pro Arg Glu Ser Ser Gly Asn Cys Tyr Leu Pro Ile Gln Thr

485 490 495

Ser Gln Ile Arg Asp Arg Asp Val Leu Ile Glu Ser Asn Ile Val Val

500 505 510

Leu Pro Thr Gln Ser Ile Arg Tyr Val Ile Ala Thr Tyr Asp Ile Ser

515 520 525

Arg Ser Asp His Ala Ile Val Tyr Tyr Val Tyr Asp Pro Ile Arg Thr

530 535 540

Ile Ser Tyr Thr His Pro Phe Arg Leu Thr Thr Lys Gly Arg Pro Asp

545 550 555 560

Phe Leu Arg Ile Glu Cys Phe Val Trp Asp Asp Asn Leu Trp Cys His

565 570 575

Gln Phe Tyr Arg Phe Glu Ala Asp Ile Ala Asn Ser Thr Thr Ser Val

580 585 590

Glu Asn Leu Val Arg Ile Arg Phe Ser Cys Asn Arg Ile Glu Gly Arg

595 600 605

Pro Gly Trp Phe Leu Asp Ser Pro Asp Arg Pro Trp Asn Pro Pro Thr

610 615 620

Phe Ser Pro Ala Leu Leu Val Val Thr Glu Gly Asp Asn Ala Thr Phe

625 630 635 640

Thr Cys Ser Phe Ser Asn Thr Ser Glu Ser Phe His Val Ile Trp His

645 650 655

Arg Glu Ser Pro Ser Gly Gln Thr Asp Thr Leu Ala Ala Phe Pro Glu

660 665 670

Asp Arg Ser Gln Pro Gly Gln Asp Cys Arg Phe Arg Val Thr Gln Leu

675 680 685

Pro Asn Gly Arg Asp Phe His Met Ser Val Val Arg Ala Arg Arg Asn

690 695 700

Asp Ser Gly Thr Tyr Val Cys Gly Val Ile Ser Leu Ala Pro Lys Ile

705 710 715 720

Gln Ile Lys Glu Ser Leu Arg Ala Glu Leu Arg Val Thr Glu Arg Arg

725 730 735

Ala Glu Val Pro Thr Ala His Pro Ser Pro Ser Pro His His His His

740 745 750

His His

<210> 28

<211> 959

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Hexon polypeptide

<400> 28

Met Ala Thr Pro Ser Met Met Pro Gln Trp Ser Tyr Met His Ile Ser

1 5 10 15

Gly Gln Asp Ala Ser Glu Tyr Leu Ser Pro Gly Leu Val Gln Phe Ala

20 25 30

Arg Ala Thr Glu Thr Tyr Phe Ser Leu Asn Asn Lys Phe Arg Asn Pro

35 40 45

Thr Val Ala Pro Thr His Asp Val Thr Thr Asp Arg Ser Gln Arg Leu

50 55 60

Thr Leu Arg Phe Ile Pro Val Asp Arg Glu Asp Thr Ala Tyr Ser Tyr

65 70 75 80

Lys Ala Arg Phe Thr Leu Ala Val Gly Asp Asn Arg Val Leu Asp Met

85 90 95

Ala Ser Thr Tyr Phe Asp Ile Arg Gly Val Leu Asp Arg Gly Pro Thr

100 105 110

Phe Lys Pro Tyr Ser Gly Thr Ala Tyr Asn Ala Leu Ala Pro Lys Gly

115 120 125

Ala Pro Asn Ser Cys Glu Trp Asp Glu Asp Asp Thr Gln Val Gln Val

130 135 140

Ala Ala Glu Asp Asp Gln Asp Asp Asp Glu Glu Glu Glu Gln Leu Pro

145 150 155 160

Gln Gln Arg Asn Gly Lys Lys Thr His Val Tyr Ala Gln Ala Pro Phe

165 170 175

Ala Gly Glu Ala Ile Asn Lys Asn Gly Leu Gln Ile Gly Thr Asn Gly

180 185 190

Ala Ala Thr Glu Gly Asn Lys Glu Ile Tyr Ala Asp Lys Thr Tyr Gln

195 200 205

Pro Glu Pro Gln Ile Gly Glu Ser Gln Trp Asn Glu Ala Glu Ser Ser

210 215 220

Val Ala Gly Gly Arg Val Leu Lys Lys Thr Thr Pro Met Lys Pro Cys

225 230 235 240

Tyr Gly Ser Tyr Ala Arg Pro Thr Asn Ser Asn Gly Gly Gln Gly Val

245 250 255

Met Val Glu Gln Asn Gly Lys Leu Glu Ser Gln Val Glu Met Gln Phe

260 265 270

Phe Ser Thr Ser Val Asn Ala Met Asn Glu Ala Asn Ala Ile Gln Pro

275 280 285

Lys Leu Val Leu Tyr Ser Glu Asp Val Asn Met Glu Thr Pro Asp Thr

290 295 300

His Leu Ser Tyr Lys Pro Gly Lys Ser Asp Asp Asn Ser Lys Ala Met

305 310 315 320

Leu Gly Gln Gln Ser Met Pro Asn Arg Pro Asn Tyr Ile Ala Phe Arg

325 330 335

Asp Asn Phe Ile Gly Leu Met Tyr Tyr Asn Ser Thr Gly Asn Met Gly

340 345 350

Val Leu Ala Gly Gln Ala Ser Gln Leu Asn Ala Val Val Asp Leu Gln

355 360 365

Asp Arg Asn Thr Glu Leu Ser Tyr Gln Leu Leu Leu Asp Ser Ile Gly

370 375 380

Asp Arg Thr Arg Tyr Phe Ser Met Trp Asn Gln Ala Val Asp Ser Tyr

385 390 395 400

Asp Pro Asp Val Arg Ile Ile Glu Asn His Gly Thr Glu Asp Glu Leu

405 410 415

Pro Asn Tyr Cys Phe Pro Leu Gly Gly Ile Gly Val Thr Asp Thr Tyr

420 425 430

Gln Ala Ile Lys Ala Thr Asn Gly Asn Gly Gly Ala Thr Thr Trp Ala

435 440 445

Gln Asp Asn Thr Phe Ala Glu Arg Asn Glu Ile Gly Val Gly Asn Asn

450 455 460

Phe Ala Met Glu Ile Asn Leu Asn Ala Asn Leu Trp Arg Asn Phe Leu

465 470 475 480

Tyr Ser Asn Ile Ala Leu Tyr Leu Pro Asp Lys Leu Lys Tyr Asn Pro

485 490 495

Thr Asn Val Glu Ile Ser Asp Asn Pro Asn Thr Tyr Asp Tyr Met Asn

500 505 510

Lys Arg Val Val Ala Pro Gly Leu Val Asp Cys Tyr Ile Asn Leu Gly

515 520 525

Ala Arg Trp Ser Leu Asp Tyr Met Asp Asn Val Asn Pro Phe Asn His

530 535 540

His Arg Asn Ala Gly Leu Arg Tyr Arg Ser Met Leu Leu Gly Asn Gly

545 550 555 560

Arg Tyr Val Pro Phe His Ile Gln Val Pro Gln Lys Phe Phe Ala Ile

565 570 575

Lys Asn Leu Leu Leu Leu Pro Gly Ser Tyr Thr Tyr Glu Trp Asn Phe

580 585 590

Arg Lys Asp Val Asn Met Val Leu Gln Ser Ser Leu Gly Asn Asp Leu

595 600 605

Arg Val Asp Gly Ala Ser Ile Lys Phe Asp Ser Ile Cys Leu Tyr Ala

610 615 620

Thr Phe Phe Pro Met Ala His Asn Thr Ala Ser Thr Leu Glu Ala Met

625 630 635 640

Leu Arg Asn Asp Thr Asn Asp Gln Ser Phe Asn Asp Tyr Leu Ser Ala

645 650 655

Ala Asn Met Leu Tyr Pro Ile Pro Ala Asn Ala Thr Asn Val Pro Ile

660 665 670

Ser Ile Pro Ser Arg Asn Trp Ala Ala Phe Arg Gly Trp Ala Phe Thr

675 680 685

Arg Leu Lys Thr Lys Glu Thr Pro Ser Leu Gly Ser Gly Tyr Asp Pro

690 695 700

Tyr Tyr Thr Tyr Ser Gly Ser Ile Pro Tyr Leu Asp Gly Thr Phe Tyr

705 710 715 720

Leu Asn His Thr Phe Lys Lys Val Ala Ile Thr Phe Asp Ser Ser Val

725 730 735

Ser Trp Pro Gly Asn Asp Arg Leu Leu Thr Pro Asn Glu Phe Glu Ile

740 745 750

Lys Arg Ser Val Asp Gly Glu Gly Tyr Asn Val Ala Gln Cys Asn Met

755 760 765

Thr Lys Asp Trp Phe Leu Val Gln Met Leu Ala Asn Tyr Asn Ile Gly

770 775 780

Tyr Gln Gly Phe Tyr Ile Pro Glu Ser Tyr Lys Asp Arg Met Tyr Ser

785 790 795 800

Phe Phe Arg Asn Phe Gln Pro Met Ser Arg Gln Val Val Asp Asp Thr

805 810 815

Lys Tyr Lys Asp Tyr Gln Gln Val Gly Ile Ile His Gln His Asn Asn

820 825 830

Ser Gly Phe Val Gly Tyr Leu Ala Pro Thr Met Arg Glu Gly Gln Ala

835 840 845

Tyr Pro Ala Asn Val Pro Tyr Pro Leu Ile Gly Lys Thr Ala Val Asp

850 855 860

Ser Ile Thr Gln Lys Lys Phe Leu Cys Asp Arg Thr Leu Trp Arg Ile

865 870 875 880

Pro Phe Ser Ser Asn Phe Met Ser Met Gly Ala Leu Thr Asp Leu Gly

885 890 895

Gln Asn Leu Leu Tyr Ala Asn Ser Ala His Ala Leu Asp Met Thr Phe

900 905 910

Glu Val Asp Pro Met Asp Glu Pro Thr Leu Leu Tyr Val Leu Phe Glu

915 920 925

Val Phe Asp Val Val Arg Val His Gln Pro His Arg Gly Val Ile Glu

930 935 940

Thr Val Tyr Leu Arg Thr Pro Phe Ser Ala Gly Asn Ala Thr Thr

945 950 955

<210> 29

<211> 962

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Hexon polypeptide

<400> 29

Met Ala Thr Pro Ser Met Met Pro Gln Trp Ser Tyr Met His Ile Ser

1 5 10 15

Gly Gln Asp Ala Ser Glu Tyr Leu Ser Pro Gly Leu Val Gln Phe Ala

20 25 30

Arg Ala Thr Glu Thr Tyr Phe Ser Leu Asn Asn Lys Phe Arg Asn Pro

35 40 45

Thr Val Ala Pro Thr His Asp Val Thr Thr Asp Arg Ser Gln Arg Leu

50 55 60

Thr Leu Arg Phe Ile Pro Val Asp Arg Glu Asp Thr Ala Tyr Ser Tyr

65 70 75 80

Lys Ala Arg Phe Thr Leu Ala Val Gly Asp Asn Arg Val Leu Asp Met

85 90 95

Ala Ser Thr Tyr Phe Asp Ile Arg Gly Val Leu Asp Arg Gly Pro Thr

100 105 110

Phe Lys Pro Tyr Ser Gly Thr Ala Tyr Asn Ala Leu Ala Pro Lys Gly

115 120 125

Ala Pro Asn Ser Cys Glu Trp Glu Gln Asn Glu Thr Ala Gln Val Asp

130 135 140

Ala Gln Glu Leu Asp Glu Glu Glu Asn Glu Ala Asn Glu Ala Gln Ala

145 150 155 160

Arg Glu Gln Glu Gln Ala Lys Lys Thr His Val Tyr Ala Gln Ala Pro

165 170 175

Leu Ser Gly Glu Ala Ile Asn Lys Asn Gly Leu Gln Ile Gly Thr Asn

180 185 190

Gly Ala Ala Thr Glu Gly Asn Lys Glu Ile Tyr Ala Asp Lys Thr Tyr

195 200 205

Gln Pro Glu Pro Gln Ile Gly Glu Ser Gln Trp Asn Glu Ala Glu Ser

210 215 220

Ser Val Ala Gly Gly Arg Val Leu Lys Lys Thr Thr Pro Met Lys Pro

225 230 235 240

Cys Tyr Gly Ser Tyr Ala Arg Pro Thr Asn Ser Asn Gly Gly Gln Gly

245 250 255

Val Met Val Glu Gln Asn Gly Lys Leu Glu Ser Gln Val Glu Met Gln

260 265 270

Phe Phe Ser Thr Ser Val Asn Ala Met Asn Glu Ala Asn Ala Ile Gln

275 280 285

Pro Lys Leu Val Leu Tyr Ser Glu Asp Val Asn Met Glu Thr Pro Asp

290 295 300

Thr His Leu Ser Tyr Lys Pro Gly Lys Ser Asp Asp Asn Ser Lys Ala

305 310 315 320

Met Leu Gly Gln Gln Ser Met Pro Asn Arg Pro Asn Tyr Ile Ala Phe

325 330 335

Arg Asp Asn Phe Ile Gly Leu Met Tyr Tyr Asn Ser Thr Gly Asn Met

340 345 350

Gly Val Leu Ala Gly Gln Ala Ser Gln Leu Asn Ala Val Val Asp Leu

355 360 365

Gln Asp Arg Asn Thr Glu Leu Ser Tyr Gln Leu Leu Leu Asp Ser Ile

370 375 380

Gly Asp Arg Thr Arg Tyr Phe Ser Met Trp Asn Gln Ala Val Asp Ser

385 390 395 400

Tyr Asp Pro Asp Val Arg Ile Ile Glu Asn His Gly Thr Glu Asp Glu

405 410 415

Leu Pro Asn Tyr Cys Phe Pro Leu Gly Gly Ile Gly Ile Thr Asp Thr

420 425 430

Phe Gln Ala Val Lys Thr Thr Ala Ala Asn Gly Asp Gln Gly Asn Thr

435 440 445

Thr Trp Gln Lys Asp Ser Thr Phe Ala Glu Arg Asn Glu Ile Gly Val

450 455 460

Gly Asn Asn Phe Ala Met Glu Ile Asn Leu Asn Ala Asn Leu Trp Arg

465 470 475 480

Asn Phe Leu Tyr Ser Asn Ile Ala Leu Tyr Leu Pro Asp Lys Leu Lys

485 490 495

Tyr Asn Pro Thr Asn Val Glu Ile Ser Asp Asn Pro Asn Thr Tyr Asp

500 505 510

Tyr Met Asn Lys Arg Val Val Ala Pro Gly Leu Val Asp Cys Tyr Ile

515 520 525

Asn Leu Gly Ala Arg Trp Ser Leu Asp Tyr Met Asp Asn Val Asn Pro

530 535 540

Phe Asn His Pro Arg His Ala Gly Leu Arg Tyr Arg Ser Met Leu Leu

545 550 555 560

Gly Asn Gly Arg Tyr Val Pro Phe His Ile Gln Val Pro Gln Lys Phe

565 570 575

Phe Ala Ile Lys Asn Leu Leu Leu Leu Pro Gly Ser Tyr Thr Tyr Glu

580 585 590

Trp Asn Phe Arg Lys Asp Val Asn Met Val Leu Gln Ser Ser Leu Gly

595 600 605

Asn Asp Leu Arg Val Asp Gly Ala Ser Ile Lys Phe Asp Ser Ile Cys

610 615 620

Leu Tyr Ala Thr Phe Phe Pro Met Ala His Asn Thr Ala Ser Thr Leu

625 630 635 640

Glu Ala Met Leu Arg Asn Asp Thr Asn Asp Gln Ser Phe Asn Asp Tyr

645 650 655

Leu Ser Ala Ala Asn Met Leu Tyr Pro Ile Pro Ala Asn Ala Thr Asn

660 665 670

Val Pro Ile Ser Ile Pro Ser Arg Asn Trp Ala Ala Phe Arg Gly Trp

675 680 685

Ala Phe Thr Arg Leu Lys Thr Lys Glu Thr Pro Ser Leu Gly Ser Gly

690 695 700

Tyr Asp Pro Tyr Tyr Thr Tyr Ser Gly Ser Ile Pro Tyr Leu Asp Gly

705 710 715 720

Thr Phe Tyr Leu Asn His Thr Phe Lys Lys Val Ala Ile Thr Phe Asp

725 730 735

Ser Ser Val Ser Trp Pro Gly Asn Asp Arg Leu Leu Thr Pro Asn Glu

740 745 750

Phe Glu Ile Lys Arg Ser Val Asp Gly Glu Gly Tyr Asn Val Ala Gln

755 760 765

Cys Asn Met Thr Lys Asp Trp Phe Leu Val Gln Met Leu Ala Asn Tyr

770 775 780

Asn Ile Gly Tyr Gln Gly Phe Tyr Ile Pro Glu Ser Tyr Lys Asp Arg

785 790 795 800

Met Tyr Ser Phe Phe Arg Asn Phe Gln Pro Met Ser Arg Gln Val Val

805 810 815

Asp Asp Thr Lys Tyr Lys Asp Tyr Gln Gln Val Gly Ile Ile His Gln

820 825 830

His Asn Asn Ser Gly Phe Val Gly Tyr Leu Ala Pro Thr Met Arg Glu

835 840 845

Gly Gln Ala Tyr Pro Ala Asn Val Pro Tyr Pro Leu Ile Gly Lys Thr

850 855 860

Ala Val Asp Ser Ile Thr Gln Lys Lys Phe Leu Cys Asp Arg Thr Leu

865 870 875 880

Trp Arg Ile Pro Phe Ser Ser Asn Phe Met Ser Met Gly Ala Leu Thr

885 890 895

Asp Leu Gly Gln Asn Leu Leu Tyr Ala Asn Ser Ala His Ala Leu Asp

900 905 910

Met Thr Phe Glu Val Asp Pro Met Asp Glu Pro Thr Leu Leu Tyr Val

915 920 925

Leu Phe Glu Val Phe Asp Val Val Arg Val His Gln Pro His Arg Gly

930 935 940

Val Ile Glu Thr Val Tyr Leu Arg Thr Pro Phe Ser Ala Gly Asn Ala

945 950 955 960

Thr Thr

<210> 30

<211> 959

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Hexon polypeptide

<400> 30

Met Ala Thr Pro Ser Met Met Pro Gln Trp Ser Tyr Met His Ile Ser

1 5 10 15

Gly Gln Asp Ala Ser Glu Tyr Leu Ser Pro Gly Leu Val Gln Phe Ala

20 25 30

Arg Ala Thr Glu Thr Tyr Phe Ser Leu Asn Asn Lys Phe Arg Asn Pro

35 40 45

Thr Val Ala Pro Thr His Asp Val Thr Thr Asp Arg Ser Gln Arg Leu

50 55 60

Thr Leu Arg Phe Ile Pro Val Asp Arg Glu Asp Thr Ala Tyr Ser Tyr

65 70 75 80

Lys Ala Arg Phe Thr Leu Ala Val Gly Asp Asn Arg Val Leu Asp Met

85 90 95

Ala Ser Thr Tyr Phe Asp Ile Arg Gly Val Leu Asp Arg Gly Pro Thr

100 105 110

Phe Lys Pro Tyr Ser Gly Thr Ala Tyr Asn Ala Leu Ala Pro Lys Gly

115 120 125

Ala Pro Asn Ser Cys Glu Trp Asp Glu Asp Asp Thr Gln Val Gln Val

130 135 140

Ala Ala Glu Asp Asp Gln Asp Asp Asp Glu Glu Glu Glu Gln Leu Pro

145 150 155 160

Gln Gln Arg Asn Gly Lys Lys Thr His Val Tyr Ala Gln Ala Pro Phe

165 170 175

Ala Gly Glu Ala Ile Asn Lys Asn Gly Leu Gln Ile Gly Thr Asn Gly

180 185 190

Ala Ala Thr Glu Gly Asn Lys Glu Ile Tyr Ala Asp Lys Thr Tyr Gln

195 200 205

Pro Glu Pro Gln Ile Gly Glu Ser Gln Trp Asn Glu Ala Glu Ser Ser

210 215 220

Val Ala Gly Gly Arg Val Leu Lys Lys Thr Thr Pro Met Lys Pro Cys

225 230 235 240

Tyr Gly Ser Tyr Ala Arg Pro Thr Asn Ser Asn Gly Gly Gln Gly Val

245 250 255

Met Val Glu Gln Asn Gly Lys Leu Glu Ser Gln Val Glu Met Gln Phe

260 265 270

Phe Ser Thr Ser Val Asn Ala Met Asn Glu Ala Asn Ala Ile Gln Pro

275 280 285

Lys Leu Val Leu Tyr Ser Glu Asp Val Asn Met Glu Thr Pro Asp Thr

290 295 300

His Leu Ser Tyr Lys Pro Gly Lys Ser Asp Asp Asn Ser Lys Ala Met

305 310 315 320

Leu Gly Gln Gln Ser Met Pro Asn Arg Pro Asn Tyr Ile Ala Phe Arg

325 330 335

Asp Asn Phe Ile Gly Leu Met Tyr Tyr Asn Ser Thr Gly Asn Met Gly

340 345 350

Val Leu Ala Gly Gln Ala Ser Gln Leu Asn Ala Val Val Asp Leu Gln

355 360 365

Asp Arg Asn Thr Glu Leu Ser Tyr Gln Leu Leu Leu Asp Ser Ile Gly

370 375 380

Asp Arg Thr Arg Tyr Phe Ser Met Trp Asn Gln Ala Val Asp Ser Tyr

385 390 395 400

Asp Pro Asp Val Arg Ile Ile Glu Asn His Gly Thr Glu Asp Glu Leu

405 410 415

Pro Asn Tyr Cys Phe Pro Leu Gly Gly Ile Gly Val Thr Asp Thr Tyr

420 425 430

Gln Ala Ile Lys Ala Thr Asn Gly Asn Gly Gly Ala Thr Thr Trp Ala

435 440 445

Gln Asp Asn Thr Phe Ala Glu Arg Asn Glu Ile Gly Val Gly Asn Asn

450 455 460

Phe Ala Met Glu Ile Asn Leu Asn Ala Asn Leu Trp Arg Asn Phe Leu

465 470 475 480

Tyr Ser Asn Ile Ala Leu Tyr Leu Pro Asp Lys Leu Lys Tyr Asn Pro

485 490 495

Thr Asn Val Glu Ile Ser Asp Asn Pro Asn Thr Tyr Asp Tyr Met Asn

500 505 510

Lys Arg Val Val Ala Pro Gly Leu Val Asp Cys Tyr Ile Asn Leu Gly

515 520 525

Ala Arg Trp Ser Leu Asp Tyr Met Asp Asn Val Asn Pro Phe Asn His

530 535 540

His Arg Asn Ala Gly Leu Arg Tyr Arg Ser Met Leu Leu Gly Asn Gly

545 550 555 560

Arg Tyr Val Pro Phe His Ile Gln Val Pro Gln Lys Phe Phe Ala Ile

565 570 575

Lys Asn Leu Leu Leu Leu Pro Gly Ser Tyr Thr Tyr Glu Trp Asn Phe

580 585 590

Arg Lys Asp Val Asn Met Val Leu Gln Ser Ser Leu Gly Asn Asp Leu

595 600 605

Arg Val Asp Gly Ala Ser Ile Lys Phe Asp Ser Ile Cys Leu Tyr Ala

610 615 620

Thr Phe Phe Pro Met Ala His Asn Thr Ala Ser Thr Leu Glu Ala Met

625 630 635 640

Leu Arg Asn Asp Thr Asn Asp Gln Ser Phe Asn Asp Tyr Leu Ser Ala

645 650 655

Ala Asn Met Leu Tyr Pro Ile Pro Ala Asn Ala Thr Asn Val Pro Ile

660 665 670

Ser Ile Pro Ser Arg Asn Trp Ala Ala Phe Arg Gly Trp Ala Phe Thr

675 680 685

Arg Leu Lys Thr Lys Glu Thr Pro Ser Leu Gly Ser Gly Tyr Asp Pro

690 695 700

Tyr Tyr Thr Tyr Ser Gly Ser Ile Pro Tyr Leu Asp Gly Thr Phe Tyr

705 710 715 720

Leu Asn His Thr Phe Lys Lys Val Ala Ile Thr Phe Asp Ser Ser Val

725 730 735

Ser Trp Pro Gly Asn Asp Arg Leu Leu Thr Pro Asn Glu Phe Glu Ile

740 745 750

Lys Arg Ser Val Asp Gly Glu Gly Tyr Asn Val Ala Gln Cys Asn Met

755 760 765

Thr Lys Asp Trp Phe Leu Val Gln Met Leu Ala Asn Tyr Asn Ile Gly

770 775 780

Tyr Gln Gly Phe Tyr Ile Pro Glu Ser Tyr Lys Asp Arg Met Tyr Ser

785 790 795 800

Phe Phe Arg Asn Phe Gln Pro Met Ser Arg Gln Val Val Asp Asp Thr

805 810 815

Lys Tyr Lys Asp Tyr Gln Gln Val Gly Ile Ile His Gln His Asn Asn

820 825 830

Ser Gly Phe Val Gly Tyr Leu Ala Pro Thr Met Arg Glu Gly Gln Ala

835 840 845

Tyr Pro Ala Asn Val Pro Tyr Pro Leu Ile Gly Lys Thr Ala Val Asp

850 855 860

Ser Ile Thr Gln Lys Lys Phe Leu Cys Asp Arg Thr Leu Trp Arg Ile

865 870 875 880

Pro Phe Ser Ser Asn Phe Met Ser Met Gly Ala Leu Thr Asp Leu Gly

885 890 895

Gln Asn Leu Leu Tyr Ala Asn Ser Ala His Ala Leu Asp Met Thr Phe

900 905 910

Glu Val Asp Pro Met Asp Glu Pro Thr Leu Leu Tyr Val Leu Phe Glu

915 920 925

Val Phe Asp Val Val Arg Val His Gln Pro His Arg Gly Val Ile Glu

930 935 940

Thr Val Tyr Leu Arg Thr Pro Phe Ser Ala Gly Asn Ala Thr Thr

945 950 955

<210> 31

<211> 132

<212> PROTEIN

<213> Artificial sequence

<220>

<223> N-terminal polypeptide E1A of adenovirus

<400> 31

Met Arg His Ile Ile Cys His Gly Gly Val Ile Thr Glu Glu Met Ala

1 5 10 15

Ala Ser Leu Leu Asp Gln Leu Ile Glu Glu Val Leu Ala Asp Asn Leu

20 25 30

Pro Pro Pro Ser His Phe Glu Pro Pro Thr Leu His Glu Leu Tyr Asp

35 40 45

Leu Asp Val Thr Ala Pro Glu Asp Pro Asn Glu Glu Ala Val Ser Gln

50 55 60

Ile Phe Pro Glu Ser Val Met Leu Ala Val Gln Glu Gly Ile Asp Leu

65 70 75 80

Phe Thr Phe Pro Pro Ala Pro Gly Ser Pro Glu Pro Pro His Leu Ser

85 90 95

Arg Gln Pro Glu Gln Pro Glu Gln Arg Ala Leu Gly Pro Val Ser Met

100 105 110

Pro Asn Leu Val Pro Glu Val Ile Asp Leu Thr Cys His Glu Ala Gly

115 120 125

Phe Pro Pro Ser

130

<210> 32

<211> 112

<212> PROTEIN

<213> Artificial sequence

<220>

<223> N-terminal E1A polypeptide

<400> 32

Met Arg His Ile Glu Glu Val Leu Ala Asp Asn Leu Pro Pro Pro Ser

1 5 10 15

His Phe Glu Pro Pro Thr Leu His Glu Leu Tyr Asp Leu Asp Val Thr

20 25 30

Ala Pro Glu Asp Pro Asn Glu Glu Ala Val Ser Gln Ile Phe Pro Glu

35 40 45

Ser Val Met Leu Ala Val Gln Glu Gly Ile Asp Leu Phe Thr Phe Pro

50 55 60

Pro Ala Pro Gly Ser Pro Glu Pro Pro His Leu Ser Arg Gln Pro Glu

65 70 75 80

Gln Pro Glu Gln Arg Ala Leu Gly Pro Val Ser Met Pro Asn Leu Val

85 90 95

Pro Glu Val Ile Asp Leu Thr Cys His Glu Ala Gly Phe Pro Pro Ser

100 105 110

<210> 33

<211> 119

<212> PROTEIN

<213> Artificial sequence

<220>

<223> N-terminal E1A polypeptide

<400> 33

Met Arg His Ile Ile Cys His Gly Gly Val Ile Thr Glu Glu Met Ala

1 5 10 15

Ala Ser Leu Leu Asp Gln Leu Ile Glu Glu Val Leu Ala Asp Asn Leu

20 25 30

Pro Pro Pro Ser His Phe Glu Pro Pro Thr Leu His Glu Leu Tyr Asp

35 40 45

Leu Asp Val Thr Ala Pro Glu Asp Pro Asn Glu Glu Ala Val Ser Gln

50 55 60

Ile Phe Pro Glu Ser Val Met Leu Ala Val Gln Glu Gly Ile Asp Leu

65 70 75 80

Phe Thr Phe Pro Pro Ala Pro Gly Ser Pro Glu Pro Pro His Leu Ser

85 90 95

Arg Gln Pro Glu Gln Pro Glu Gln Arg Ala Leu Gly Pro Val Cys His

100 105 110

Glu Ala Gly Phe Pro Pro Ser

115

<210> 34

<211> 99

<212> PROTEIN

<213> Artificial sequence

<220>

<223> N-terminal E1A polypeptide

<400> 34

Met Arg His Ile Glu Glu Val Leu Ala Asp Asn Leu Pro Pro Pro Ser

1 5 10 15

His Phe Glu Pro Pro Thr Leu His Glu Leu Tyr Asp Leu Asp Val Thr

20 25 30

Ala Pro Glu Asp Pro Asn Glu Glu Ala Val Ser Gln Ile Phe Pro Glu

35 40 45

Ser Val Met Leu Ala Val Gln Glu Gly Ile Asp Leu Phe Thr Phe Pro

50 55 60

Pro Ala Pro Gly Ser Pro Glu Pro Pro His Leu Ser Arg Gln Pro Glu

65 70 75 80

Gln Pro Glu Gln Arg Ala Leu Gly Pro Val Cys His Glu Ala Gly Phe

85 90 95

Pro Pro Ser

<210> 35

<211> 528

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Fiber polypeptide

<400> 35

Met Lys Arg Ala Arg Pro Ser Glu Asp Thr Phe Asn Pro Val Tyr Pro

1 5 10 15

Tyr Asp Thr Glu Thr Gly Pro Pro Thr Val Pro Phe Leu Thr Pro Pro

20 25 30

Phe Val Ser Pro Asn Gly Phe Gln Glu Ser Pro Pro Gly Val Leu Ser

35 40 45

Leu Arg Leu Ser Glu Pro Leu Val Thr Ser His Gly Met Leu Ala Leu

50 55 60

Lys Met Gly Ser Gly Leu Ser Leu Asp Gln Ala Gly Asn Leu Thr Ser

65 70 75 80

Asn Thr Ile Thr Val Ser Gln Pro Leu Lys Lys Thr Lys Ser Asn Ile

85 90 95

Thr Leu Glu Thr Ser Ala Pro Leu Thr Val Ser Ser Gly Ala Leu Thr

100 105 110

Met Ala Thr Thr Ser Pro Leu Val Val Ser Asp Asn Thr Leu Thr Met

115 120 125

Gln Ser Gln Ala Pro Leu Thr Val Gln Asp Ser Lys Leu Ser Ile Ala

130 135 140

Thr Lys Glu Pro Leu Thr Val Leu Asp Gly Lys Leu Ala Leu Gln Thr

145 150 155 160

Ser Ala Pro Leu Ser Ala Thr Asp Asn Asn Ala Leu Thr Ile Thr Ala

165 170 175

Ser Pro Pro Leu Thr Thr Ala Asn Gly Ser Leu Ala Val Thr Met Glu

180 185 190

Asn Pro Leu Tyr Asn Asn Asn Gly Lys Leu Gly Leu Lys Ile Gly Gly

195 200 205

Pro Leu Gln Val Ala Thr Asp Ser His Ala Leu Thr Leu Gly Thr Gly

210 215 220

Gln Gly Val Ala Val His Asn Asn Leu Leu His Thr Lys Val Thr Gly

225 230 235 240

Ala Ile Gly Phe Asp Thr Ser Gly Asn Met Glu Leu Lys Thr Gly Asp

245 250 255

Gly Leu Tyr Val Asp Ser Ala Gly Pro Asn Gln Lys Leu His Ile Asn

260 265 270

Leu Asn Thr Thr Lys Gly Leu Ala Phe Asp Asn Thr Ala Ile Thr Ile

275 280 285

Asn Ala Gly Lys Gly Leu Glu Phe Glu Thr Asp Ser Ser Asn Gly Asn

290 295 300

Pro Ile Lys Thr Lys Ile Gly Ser Gly Ile Gln Tyr Asn Thr Asn Gly

305 310 315 320

Ala Met Val Ala Lys Leu Gly Thr Gly Leu Ser Phe Asp Ser Ser Gly

325 330 335

Ala Ile Thr Met Gly Ser Ile Asn Asn Asp Arg Leu Thr Leu Trp Thr

340 345 350

Thr Pro Asp Pro Ser Pro Asn Cys Arg Ile Ala Ser Asp Lys Asp Cys

355 360 365

Lys Leu Thr Leu Ala Leu Thr Lys Cys Gly Ser Gln Ile Leu Gly Thr

370 375 380

Val Ser Ala Leu Ala Val Ser Gly Asn Met Ala Ser Ile Asn Gly Thr

385 390 395 400

Leu Ser Ser Val Asn Leu Val Leu Arg Phe Asp Asp Asn Gly Val Leu

405 410 415

Met Ser Asn Ser Ser Leu Asp Lys Gln Tyr Trp Asn Phe Arg Asn Gly

420 425 430

Asp Ser Thr Asn Gly Gln Pro Tyr Thr Tyr Ala Val Gly Phe Met Pro

435 440 445

Asn Leu Lys Ala Tyr Pro Lys Thr Gln Ser Lys Thr Ala Lys Ser Asn

450 455 460

Ile Val Ser Gln Val Tyr Leu Asn Gly Asp Lys Ser Lys Pro Leu His

465 470 475 480

Phe Thr Ile Thr Leu Asn Gly Thr Asp Glu Thr Asn Gln Val Ser Lys

485 490 495

Tyr Ser Ile Ser Phe Ser Trp Ser Trp Asn Ser Gly Gln Tyr Thr Asn

500 505 510

Asp Lys Phe Ala Thr Asn Ser Tyr Thr Phe Ser Tyr Ile Ala Gln Glu

515 520 525

<210> 36

<211> 324

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Fiber polypeptide

<400> 36

Met Lys Arg Ala Arg Pro Ser Glu Asp Thr Phe Asn Pro Val Tyr Pro

1 5 10 15

Tyr Asp Thr Glu Thr Gly Pro Pro Thr Val Pro Phe Leu Thr Pro Pro

20 25 30

Phe Val Ser Pro Asn Gly Phe Gln Glu Ser Pro Pro Gly Val Leu Thr

35 40 45

Leu Lys Cys Leu Thr Pro Leu Thr Thr Thr Gly Gly Ser Leu Gln Leu

50 55 60

Lys Val Gly Gly Gly Leu Thr Val Asp Asp Thr Asp Gly Thr Leu Gln

65 70 75 80

Glu Asn Ile Arg Ala Thr Ala Pro Ile Thr Lys Asn Asn His Ser Val

85 90 95

Glu Leu Ser Ile Gly Asn Gly Leu Glu Thr Gln Asn Asn Lys Leu Cys

100 105 110

Ala Lys Leu Gly Asn Gly Leu Lys Phe Asn Asn Gly Asp Ile Cys Ile

115 120 125

Lys Asp Ser Ile Asn Thr Leu Trp Thr Gly Ile Asn Pro Pro Pro Asn

130 135 140

Cys Gln Ile Val Glu Asn Thr Asn Thr Asn Asp Gly Lys Leu Thr Leu

145 150 155 160

Val Leu Val Lys Asn Gly Gly Leu Val Asn Gly Tyr Val Ser Leu Val

165 170 175

Gly Val Ser Asp Thr Val Asn Gln Met Phe Thr Gln Lys Thr Ala Asn

180 185 190

Ile Gln Leu Arg Leu Tyr Phe Asp Ser Ser Gly Asn Leu Leu Thr Asp

195 200 205

Glu Ser Asp Leu Lys Ile Pro Leu Lys Asn Lys Ser Ser Thr Ala Thr

210 215 220

Ser Glu Thr Val Ala Ser Ser Lys Ala Phe Met Pro Ser Thr Thr Ala

225 230 235 240

Tyr Pro Phe Asn Thr Thr Thr Arg Asp Ser Glu Asn Tyr Ile His Gly

245 250 255

Ile Cys Tyr Tyr Met Thr Ser Tyr Asp Arg Ser Leu Phe Pro Leu Asn

260 265 270

Ile Ser Ile Met Leu Asn Ser Arg Met Ile Ser Ser Asn Val Ala Tyr

275 280 285

Ala Ile Gln Phe Glu Trp Asn Leu Asn Ala Ser Glu Ser Pro Glu Ser

290 295 300

Asn Ile Ala Thr Leu Thr Thr Ser Pro Phe Phe Phe Ser Tyr Ile Thr

305 310 315 320

Glu Asp Asp Asn

<210> 37

<211> 534

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Fiber polypeptide

<400> 37

Met Lys Arg Ala Arg Pro Ser Glu Asp Thr Phe Asn Pro Val Tyr Pro

1 5 10 15

Tyr Asp Thr Glu Thr Gly Pro Pro Thr Val Pro Phe Leu Thr Pro Pro

20 25 30

Phe Val Ser Pro Asn Gly Phe Gln Glu Ser Pro Pro Gly Val Leu Ser

35 40 45

Leu Arg Leu Ser Glu Pro Leu Val Thr Ser His Gly Met Leu Ala Leu

50 55 60

Lys Met Gly Ser Gly Leu Ser Leu Asp Gln Ala Gly Asn Leu Thr Ser

65 70 75 80

Asn Thr Ile Thr Val Ser Gln Pro Leu Lys Lys Thr Lys Ser Asn Ile

85 90 95

Thr Leu Glu Thr Ser Ala Pro Leu Thr Val Ser Ser Gly Ala Leu Thr

100 105 110

Met Ala Thr Thr Ser Pro Leu Val Val Ser Asp Asn Thr Leu Thr Met

115 120 125

Gln Ser Gln Ala Pro Leu Thr Val Gln Asp Ser Lys Leu Ser Ile Ala

130 135 140

Thr Lys Glu Pro Leu Thr Val Leu Asp Gly Lys Leu Ala Leu Gln Thr

145 150 155 160

Ser Ala Pro Leu Ser Ala Thr Asp Asn Asn Ala Leu Thr Ile Thr Ala

165 170 175

Ser Pro Pro Leu Thr Thr Ala Asn Gly Ser Leu Ala Val Thr Met Glu

180 185 190

Asn Pro Leu Tyr Asn Asn Asn Gly Lys Leu Gly Leu Lys Ile Gly Gly

195 200 205

Pro Leu Gln Val Ala Thr Asp Ser His Ala Leu Thr Leu Gly Thr Gly

210 215 220

Gln Gly Val Ala Val His Asn Asn Leu Leu His Thr Lys Val Thr Gly

225 230 235 240

Ala Ile Gly Phe Asp Thr Ser Gly Asn Met Glu Leu Lys Thr Gly Asp

245 250 255

Gly Leu Tyr Val Asp Ser Ala Gly Pro Asn Gln Lys Leu His Ile Asn

260 265 270

Leu Asn Thr Thr Lys Gly Leu Ala Phe Asp Asn Thr Ala Ile Thr Ile

275 280 285

Asn Ala Gly Lys Gly Leu Glu Phe Glu Thr Asp Ser Ser Asn Gly Asn

290 295 300

Pro Ile Lys Thr Lys Ile Gly Ser Gly Ile Gln Tyr Asn Thr Asn Gly

305 310 315 320

Ala Met Val Ala Lys Leu Gly Thr Gly Leu Ser Phe Asp Ser Ser Gly

325 330 335

Ala Ile Thr Met Gly Ser Ile Asn Asn Asp Arg Leu Thr Leu Trp Thr

340 345 350

Thr Pro Asp Pro Ser Pro Asn Cys Arg Ile Ala Ser Asp Lys Asp Cys

355 360 365

Lys Leu Thr Leu Ala Leu Thr Lys Cys Gly Ser Gln Ile Leu Gly Thr

370 375 380

Val Ser Ala Leu Ala Val Ser Gly Asn Met Ala Ser Ile Asn Gly Thr

385 390 395 400

Leu Ser Ser Val Asn Leu Val Leu Arg Phe Asp Asp Asn Gly Val Leu

405 410 415

Met Ser Asn Ser Ser Leu Asp Lys Gln Tyr Trp Asn Phe Arg Asn Gly

420 425 430

Asp Ser Thr Asn Gly Gln Pro Tyr Thr Tyr Ala Val Gly Phe Met Pro

435 440 445

Asn Leu Lys Ala Tyr Pro Lys Thr Gln Ser Lys Thr Ala Lys Ser Asn

450 455 460

Ile Val Ser Gln Val Tyr Leu Asn Gly Asp Lys Ser Lys Pro Leu His

465 470 475 480

Phe Thr Ile Thr Leu Asn Gly Thr Asp Glu Thr Asn Gln Val Ser Lys

485 490 495

Tyr Ser Ile Ser Phe Ser Trp Ser Trp Asn Ser Gly Gln Tyr Thr Asn

500 505 510

Asp Lys Phe Ala Thr Asn Ser Tyr Thr Phe Ser Tyr Ile Ala Gln Glu

515 520 525

Lys Lys Lys Lys Lys Lys

530

<210> 38

<211> 425

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Fiber polypeptide

<400> 38

Met Lys Arg Ala Arg Pro Ser Glu Asp Thr Phe Asn Pro Val Tyr Pro

1 5 10 15

Tyr Asp Thr Glu Thr Gly Pro Pro Thr Val Pro Phe Leu Thr Pro Pro

20 25 30

Phe Val Ser Pro Asn Gly Phe Gln Glu Ser Pro Pro Gly Val Leu Ser

35 40 45

Leu Arg Leu Ser Glu Pro Leu Val Thr Lys Asn Gly Glu Ile Thr Leu

50 55 60

Lys Leu Gly Glu Gly Val Asp Leu Asp Ser Ser Gly Lys Leu Ile Ser

65 70 75 80

Asn Thr Ala Thr Lys Ala Ala Ala Pro Leu Ser Phe Ser Asn Asn Thr

85 90 95

Ile Ser Leu Asn Met Asp His Pro Phe Tyr Thr Lys Asp Gly Lys Leu

100 105 110

Ser Leu Gln Val Ser Pro Pro Leu Asn Ile Leu Arg Thr Ser Ile Leu

115 120 125

Asn Thr Leu Ala Leu Gly Phe Gly Ser Gly Leu Gly Leu Arg Gly Ser

130 135 140

Ala Leu Ala Val Gln Leu Val Ser Pro Leu Thr Phe Asp Thr Asp Gly

145 150 155 160

Asn Ile Lys Leu Thr Leu Asp Arg Gly Leu His Val Thr Thr Gly Asp

165 170 175

Ala Ile Glu Ser Asn Ile Ser Trp Ala Lys Gly Leu Lys Phe Glu Asp

180 185 190

Gly Ala Ile Ala Thr Asn Ile Gly Asn Gly Leu Glu Phe Gly Ser Ser

195 200 205

Ser Thr Glu Thr Gly Val Asp Asp Ala Tyr Pro Ile Gln Val Lys Leu

210 215 220

Gly Ser Gly Leu Ser Phe Asp Ser Thr Gly Ala Ile Met Ala Gly Asn

225 230 235 240

Lys Glu Asp Asp Lys Leu Thr Leu Trp Thr Thr Pro Asp Pro Ser Pro

245 250 255

Asn Cys Gln Ile Leu Ala Glu Asn Asp Ala Lys Leu Thr Leu Cys Leu

260 265 270

Thr Lys Cys Gly Ser Gln Ile Leu Ala Thr Val Ser Val Leu Val Val

275 280 285

Gly Ser Gly Asn Leu Asn Pro Ile Thr Gly Thr Val Ser Ser Ala Gln

290 295 300

Val Phe Leu Arg Phe Asp Ala Asn Gly Val Leu Leu Thr Glu His Ser

305 310 315 320

Thr Leu Lys Lys Tyr Trp Gly Tyr Arg Gln Gly Asp Ser Ile Asp Gly

325 330 335

Thr Pro Tyr Thr Asn Ala Val Gly Phe Met Pro Asn Leu Lys Ala Tyr

340 345 350

Pro Lys Ser Gln Ser Ser Thr Thr Lys Asn Asn Ile Val Gly Gln Val

355 360 365

Tyr Met Asn Gly Asp Val Ser Lys Pro Met Leu Leu Thr Ile Thr Leu

370 375 380

Asn Gly Thr Asp Asp Ser Asn Ser Thr Tyr Ser Met Ser Phe Ser Tyr

385 390 395 400

Thr Trp Thr Asn Gly Ser Tyr Val Gly Ala Thr Phe Gly Ala Asn Ser

405 410 415

Tyr Thr Phe Ser Tyr Ile Ala Gln Glu

420 425

<210> 39

<211> 432

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Fiber polypeptide

<400> 39

Met Lys Arg Ala Arg Pro Ser Glu Asp Thr Phe Asn Pro Val Tyr Pro

1 5 10 15

Tyr Asp Thr Glu Thr Gly Pro Pro Thr Val Pro Phe Leu Thr Pro Pro

20 25 30

Phe Val Ser Pro Asn Gly Phe Gln Glu Ser Pro Pro Gly Val Leu Ser

35 40 45

Leu Arg Leu Ser Glu Pro Leu Val Thr Lys Asn Gly Glu Ile Thr Leu

50 55 60

Lys Leu Gly Glu Gly Val Asp Leu Asp Ser Ser Gly Lys Leu Ile Ser

65 70 75 80

Asn Thr Ala Thr Lys Ala Ala Ala Pro Leu Ser Phe Ser Asn Asn Thr

85 90 95

Ile Ser Leu Asn Met Asp His Pro Phe Tyr Thr Lys Asp Gly Lys Leu

100 105 110

Ser Leu Gln Val Ser Pro Pro Leu Asn Ile Leu Arg Thr Ser Ile Leu

115 120 125

Asn Thr Leu Ala Leu Gly Phe Gly Ser Gly Leu Gly Leu Arg Gly Ser

130 135 140

Ala Leu Ala Val Gln Leu Val Ser Pro Leu Thr Phe Asp Thr Asp Gly

145 150 155 160

Asn Ile Lys Leu Thr Leu Asp Arg Gly Leu His Val Thr Thr Gly Asp

165 170 175

Ala Ile Glu Ser Asn Ile Ser Trp Ala Lys Gly Leu Lys Phe Glu Asp

180 185 190

Gly Ala Ile Ala Thr Asn Ile Gly Asn Gly Leu Glu Phe Gly Ser Ser

195 200 205

Ser Thr Glu Thr Gly Val Asp Asp Ala Tyr Pro Ile Gln Val Lys Leu

210 215 220

Gly Ser Gly Leu Ser Phe Asp Ser Thr Gly Ala Ile Met Ala Gly Asn

225 230 235 240

Lys Glu Asp Asp Lys Leu Thr Leu Trp Thr Thr Pro Asp Pro Ser Pro

245 250 255

Asn Cys Gln Ile Leu Ala Glu Asn Asp Ala Lys Leu Thr Leu Cys Leu

260 265 270

Thr Lys Cys Gly Ser Gln Ile Leu Ala Thr Val Ser Val Leu Val Val

275 280 285

Gly Ser Gly Asn Leu Asn Pro Ile Thr Gly Thr Val Ser Ser Ala Gln

290 295 300

Val Phe Leu Arg Phe Asp Ala Asn Gly Val Leu Leu Thr Glu His Ser

305 310 315 320

Thr Leu Lys Lys Tyr Trp Gly Tyr Arg Gln Gly Asp Ser Ile Asp Gly

325 330 335

Thr Pro Tyr Thr Asn Ala Val Gly Phe Met Pro Asn Leu Lys Ala Tyr

340 345 350

Pro Lys Ser Gln Ser Ser Thr Thr Lys Asn Asn Ile Val Gly Gln Val

355 360 365

Tyr Met Asn Gly Asp Val Ser Lys Pro Met Leu Leu Thr Ile Thr Leu

370 375 380

Asn Gly Thr Asp Asp Ser Asn Ser Thr Tyr Ser Met Ser Phe Ser Tyr

385 390 395 400

Thr Trp Thr Asn Gly Ser Tyr Val Gly Ala Thr Phe Gly Ala Asn Ser

405 410 415

Tyr Thr Phe Ser Tyr Ile Ala Gln Glu Lys Lys Lys Lys Lys Lys Lys

420 425 430

<210> 40

<211> 441

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Fiber polypeptide

<400> 40

Met Lys Arg Ala Arg Pro Ser Glu Asp Thr Phe Asn Pro Val Tyr Pro

1 5 10 15

Tyr Asp Thr Glu Thr Gly Pro Pro Thr Val Pro Phe Leu Thr Pro Pro

20 25 30

Phe Val Ser Pro Asn Gly Phe Gln Glu Ser Pro Pro Gly Val Leu Ser

35 40 45

Leu Arg Leu Ser Glu Pro Leu Val Thr Lys Asn Gly Glu Ile Thr Leu

50 55 60

Lys Leu Gly Glu Gly Val Asp Leu Asp Ser Ser Gly Lys Leu Ile Ser

65 70 75 80

Asn Thr Ala Thr Lys Ala Ala Ala Pro Leu Ser Phe Ser Asn Asn Thr

85 90 95

Ile Ser Leu Asn Met Asp His Pro Phe Tyr Thr Lys Asp Gly Lys Leu

100 105 110

Ser Leu Gln Val Ser Pro Pro Leu Asn Ile Leu Arg Thr Ser Ile Leu

115 120 125

Asn Thr Leu Ala Leu Gly Phe Gly Ser Gly Leu Gly Leu Arg Gly Ser

130 135 140

Ala Leu Ala Val Gln Leu Val Ser Pro Leu Thr Phe Asp Thr Asp Gly

145 150 155 160

Asn Ile Lys Leu Thr Leu Asp Arg Gly Leu His Val Thr Thr Gly Asp

165 170 175

Ala Ile Glu Ser Asn Ile Ser Trp Ala Lys Gly Leu Lys Phe Glu Asp

180 185 190

Gly Ala Ile Ala Thr Asn Ile Gly Asn Gly Leu Glu Phe Gly Ser Ser

195 200 205

Ser Thr Glu Thr Gly Val Asp Asp Ala Tyr Pro Ile Gln Val Lys Leu

210 215 220

Gly Ser Gly Leu Ser Phe Asp Ser Thr Gly Ala Ile Met Ala Gly Asn

225 230 235 240

Lys Glu Asp Asp Lys Leu Thr Leu Trp Thr Thr Pro Asp Pro Ser Pro

245 250 255

Asn Cys Gln Ile Leu Ala Glu Asn Asp Ala Lys Leu Thr Leu Cys Leu

260 265 270

Thr Lys Cys Gly Ser Gln Ile Leu Ala Thr Val Ser Val Leu Val Val

275 280 285

Gly Ser Gly Asn Leu Asn Pro Ile Thr Gly Thr Val Ser Ser Ala Gln

290 295 300

Val Phe Leu Arg Phe Asp Ala Asn Gly Val Leu Leu Thr Glu His Ser

305 310 315 320

Thr Leu Lys Lys Tyr Trp Gly Tyr Arg Gln Gly Asp Ser Ile Asp Gly

325 330 335

Thr Pro Tyr Thr Asn Ala Val Gly Phe Met Pro Asn Leu Lys Ala Tyr

340 345 350

Pro Lys Ser Gln Ser Ser Thr Thr Lys Asn Asn Ile Val Gly Gln Val

355 360 365

Tyr Met Asn Gly Asp Val Ser Lys Pro Met Leu Leu Thr Ile Thr Leu

370 375 380

Asn Gly Thr Asp Asp Ser Gly Gly Ser Ser Gly Lys Lys Lys Lys Lys

385 390 395 400

Lys Lys Ala Ser Gly Gly Ser Ser Thr Tyr Ser Met Ser Phe Ser Tyr

405 410 415

Thr Trp Thr Asn Gly Ser Tyr Val Gly Ala Thr Phe Gly Ala Asn Ser

420 425 430

Tyr Thr Phe Ser Tyr Ile Ala Gln Glu

435 440

<210> 41

<211> 1608

<212> DNA

<213> Artificial sequence

<220>

<223> Probazin-E1

<400> 41

tcgagcgacg gtatcgataa gcttggagct tatgatagca tcttgttctt agtctttttc

60

ttaataggga cataaagccc acaaataaaa atatgcctga agaatgggac aggcattggg

120

cattgtccat gcctagtaaa gtactccaag aacctatttg tatactagat gacacaatgt

180

tctagccaag cttggtagtc atcatgttta aacatctacc attccagtta agaaaatatg

240

atagcatctt gttcttagtc tttttcttaa tagggacata aagcccacaa ataaaaatat

300

gcctgaagaa tgggacaggc attgggcatt gtccatgcct agtaaagtac tccaagaacc

360

tatttgtata ctagatgaca caatgtcaat gtctgtgtac aactgccaac tgggatgcaa

420

gacactgccc atgccaatca tcctgaaaag cagctataaa aagcaggaag ctactctgca

480

ccttgtcagt gaggtccaga tacctccctc gagcggccgc gacgcgcagt gtatttatac

540

ccggtgagtt cctcaagagg ccactcttga gtgccagcga gtagagtttt ctcctccgag

600

ccgctccgac accgggactg aaaatgagac atattatctg ccacggaggt gttattaccg

660

aagaaatggc cgccagtctt ttggaccagc tgatcgaaga ggtactggct gataatcttc

720

cacctcctag ccattttgaa ccacctaccc ttcacgaact gtatgattta gacgtgacgg

780

cccccgaaga tcccaacgag gaggcggttt cgcagatttt tcccgagtct gtaatgttgg

840

cggtgcagga agggattgac ttattcactt ttccgccggc gcccggttct ccggagccgc

900

ctcacctttc ccggcagccc gagcagccgg agcagagagc cttgggtccg gtttctatgc

960

caaaccttgt gccggaggtg atcgatctta cctgccacga ggctggcttt ccacccagtg

1020

acgacgagga tgaagagggt gaggagtttg tgttagatta tgtggagcac cccgggcacg

1080

gttgcaggtc ttgtcattat caccggagga atacggggga cccagatatt atgtgttcgc

1140

tttgctatat gaggacctgt ggcatgtttg tctacagtaa gtgaaaatta tgggcagtcg

1200

gtgatagagt ggtgggtttg gtgtggtaat ttttttttaa tttttacagt tttgtggttt

1260

aaagaatttt gtattgtgat tttttaaaag gtcctgtgtc tgaacctgag cctgagcccg

1320

agccagaacc ggagcctgca agacctaccc ggcgtcctaa attggtgcct gctatcctga

1380

gacgcccgac atcacctgtg tctagagaat gcaatagtag tacggatagc tgtgactccg

1440

gtccttctaa cacacctcct gagatacacc cggtggtccc gctgtgcccc attaaaccag

1500

ttgccgtgag agttggtggg cgtcgccagg ctgtggaatg tatcgaggac ttgcttaacg

1560

agtctgggca acctttggac ttgagctgta aacgccccag gccataag

1608

<210> 42

<211> 967

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Hexon polypeptide

<400> 42

Met Ala Thr Pro Ser Met Met Pro Gln Trp Ser Tyr Met His Ile Ser

1 5 10 15

Gly Gln Asp Ala Ser Glu Tyr Leu Ser Pro Gly Leu Val Gln Phe Ala

20 25 30

Arg Ala Thr Glu Thr Tyr Phe Ser Leu Asn Asn Lys Phe Arg Asn Pro

35 40 45

Thr Val Ala Pro Thr His Asp Val Thr Thr Asp Arg Ser Gln Arg Leu

50 55 60

Thr Leu Arg Phe Ile Pro Val Asp Arg Glu Asp Thr Ala Tyr Ser Tyr

65 70 75 80

Lys Ala Arg Phe Thr Leu Ala Val Gly Asp Asn Arg Val Leu Asp Met

85 90 95

Ala Ser Thr Tyr Phe Asp Ile Arg Gly Val Leu Asp Arg Gly Pro Thr

100 105 110

Phe Lys Pro Tyr Ser Gly Thr Ala Tyr Asn Ala Leu Ala Pro Lys Gly

115 120 125

Ala Pro Asn Ser Cys Glu Trp Asp Glu Asp Asp Thr Gln Val Gln Val

130 135 140

Ala Ala Glu Asp Asp Gln Asp Asp Asp Ser Ser Cys Ser Ser Gly Gly

145 150 155 160

Thr Glu Glu Glu Glu Gln Leu Pro Gln Gln Arg Asn Gly Lys Lys Thr

165 170 175

His Val Tyr Ala Gln Ala Pro Phe Ala Gly Glu Ala Ile Asn Lys Asn

180 185 190

Gly Leu Gln Ile Gly Thr Asn Gly Ala Ala Thr Glu Gly Asn Lys Glu

195 200 205

Ile Tyr Ala Asp Lys Thr Tyr Gln Pro Glu Pro Gln Ile Gly Glu Ser

210 215 220

Gln Trp Asn Glu Ala Glu Ser Ser Val Ala Gly Gly Arg Val Leu Lys

225 230 235 240

Lys Thr Thr Pro Met Lys Pro Cys Tyr Gly Ser Tyr Ala Arg Pro Thr

245 250 255

Asn Ser Asn Gly Gly Gln Gly Val Met Val Glu Gln Asn Gly Lys Leu

260 265 270

Glu Ser Gln Val Glu Met Gln Phe Phe Ser Thr Ser Val Asn Ala Met

275 280 285

Asn Glu Ala Asn Ala Ile Gln Pro Lys Leu Val Leu Tyr Ser Glu Asp

290 295 300

Val Asn Met Glu Thr Pro Asp Thr His Leu Ser Tyr Lys Pro Gly Lys

305 310 315 320

Ser Asp Asp Asn Ser Lys Ala Met Leu Gly Gln Gln Ser Met Pro Asn

325 330 335

Arg Pro Asn Tyr Ile Ala Phe Arg Asp Asn Phe Ile Gly Leu Met Tyr

340 345 350

Tyr Asn Ser Thr Gly Asn Met Gly Val Leu Ala Gly Gln Ala Ser Gln

355 360 365

Leu Asn Ala Val Val Asp Leu Gln Asp Arg Asn Thr Glu Leu Ser Tyr

370 375 380

Gln Leu Leu Leu Asp Ser Ile Gly Asp Arg Thr Arg Tyr Phe Ser Met

385 390 395 400

Trp Asn Gln Ala Val Asp Ser Tyr Asp Pro Asp Val Arg Ile Ile Glu

405 410 415

Asn His Gly Thr Glu Asp Glu Leu Pro Asn Tyr Cys Phe Pro Leu Gly

420 425 430

Gly Ile Gly Val Thr Asp Thr Tyr Gln Ala Ile Lys Ala Thr Asn Gly

435 440 445

Asn Gly Gly Ala Thr Thr Trp Ala Gln Asp Asn Thr Phe Ala Glu Arg

450 455 460

Asn Glu Ile Gly Val Gly Asn Asn Phe Ala Met Glu Ile Asn Leu Asn

465 470 475 480

Ala Asn Leu Trp Arg Asn Phe Leu Tyr Ser Asn Ile Ala Leu Tyr Leu

485 490 495

Pro Asp Lys Leu Lys Tyr Asn Pro Thr Asn Val Glu Ile Ser Asp Asn

500 505 510

Pro Asn Thr Tyr Asp Tyr Met Asn Lys Arg Val Val Ala Pro Gly Leu

515 520 525

Val Asp Cys Tyr Ile Asn Leu Gly Ala Arg Trp Ser Leu Asp Tyr Met

530 535 540

Asp Asn Val Asn Pro Phe Asn His His Arg Asn Ala Gly Leu Arg Tyr

545 550 555 560

Arg Ser Met Leu Leu Gly Asn Gly Arg Tyr Val Pro Phe His Ile Gln

565 570 575

Val Pro Gln Lys Phe Phe Ala Ile Lys Asn Leu Leu Leu Leu Pro Gly

580 585 590

Ser Tyr Thr Tyr Glu Trp Asn Phe Arg Lys Asp Val Asn Met Val Leu

595 600 605

Gln Ser Ser Leu Gly Asn Asp Leu Arg Val Asp Gly Ala Ser Ile Lys

610 615 620

Phe Asp Ser Ile Cys Leu Tyr Ala Thr Phe Phe Pro Met Ala His Asn

625 630 635 640

Thr Ala Ser Thr Leu Glu Ala Met Leu Arg Asn Asp Thr Asn Asp Gln

645 650 655

Ser Phe Asn Asp Tyr Leu Ser Ala Ala Asn Met Leu Tyr Pro Ile Pro

660 665 670

Ala Asn Ala Thr Asn Val Pro Ile Ser Ile Pro Ser Arg Asn Trp Ala

675 680 685

Ala Phe Arg Gly Trp Ala Phe Thr Arg Leu Lys Thr Lys Glu Thr Pro

690 695 700

Ser Leu Gly Ser Gly Tyr Asp Pro Tyr Tyr Thr Tyr Ser Gly Ser Ile

705 710 715 720

Pro Tyr Leu Asp Gly Thr Phe Tyr Leu Asn His Thr Phe Lys Lys Val

725 730 735

Ala Ile Thr Phe Asp Ser Ser Val Ser Trp Pro Gly Asn Asp Arg Leu

740 745 750

Leu Thr Pro Asn Glu Phe Glu Ile Lys Arg Ser Val Asp Gly Glu Gly

755 760 765

Tyr Asn Val Ala Gln Cys Asn Met Thr Lys Asp Trp Phe Leu Val Gln

770 775 780

Met Leu Ala Asn Tyr Asn Ile Gly Tyr Gln Gly Phe Tyr Ile Pro Glu

785 790 795 800

Ser Tyr Lys Asp Arg Met Tyr Ser Phe Phe Arg Asn Phe Gln Pro Met

805 810 815

Ser Arg Gln Val Val Asp Asp Thr Lys Tyr Lys Asp Tyr Gln Gln Val

820 825 830

Gly Ile Ile His Gln His Asn Asn Ser Gly Phe Val Gly Tyr Leu Ala

835 840 845

Pro Thr Met Arg Glu Gly Gln Ala Tyr Pro Ala Asn Val Pro Tyr Pro

850 855 860

Leu Ile Gly Lys Thr Ala Val Asp Ser Ile Thr Gln Lys Lys Phe Leu

865 870 875 880

Cys Asp Arg Thr Leu Trp Arg Ile Pro Phe Ser Ser Asn Phe Met Ser

885 890 895

Met Gly Ala Leu Thr Asp Leu Gly Gln Asn Leu Leu Tyr Ala Asn Ser

900 905 910

Ala His Ala Leu Asp Met Thr Phe Glu Val Asp Pro Met Asp Glu Pro

915 920 925

Thr Leu Leu Tyr Val Leu Phe Glu Val Phe Asp Val Val Arg Val His

930 935 940

Gln Pro His Arg Gly Val Ile Glu Thr Val Tyr Leu Arg Thr Pro Phe

945 950 955 960

Ser Ala Gly Asn Ala Thr Thr

965

<210> 43

<211> 955

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Hexon polypeptide

<400> 43

Met Ala Thr Pro Ser Met Met Pro Gln Trp Ser Tyr Met His Ile Ser

1 5 10 15

Gly Gln Asp Ala Ser Glu Tyr Leu Ser Pro Gly Leu Val Gln Phe Ala

20 25 30

Arg Ala Thr Glu Thr Tyr Phe Ser Leu Asn Asn Lys Phe Arg Asn Pro

35 40 45

Thr Val Ala Pro Thr His Asp Val Thr Thr Asp Arg Ser Gln Arg Leu

50 55 60

Thr Leu Arg Phe Ile Pro Val Asp Arg Glu Asp Thr Ala Tyr Ser Tyr

65 70 75 80

Lys Ala Arg Phe Thr Leu Ala Val Gly Asp Asn Arg Val Leu Asp Met

85 90 95

Ala Ser Thr Tyr Phe Asp Ile Arg Gly Val Leu Asp Arg Gly Pro Thr

100 105 110

Phe Lys Pro Tyr Ser Gly Thr Ala Tyr Asn Ala Leu Ala Pro Lys Gly

115 120 125

Ala Pro Asn Ser Cys Glu Trp Asp Glu Asp Asp Thr Gln Val Gln Val

130 135 140

Ala Ala Glu Asp Asp Gln Asp Asp Asp Glu Glu Glu Glu Gln Leu Pro

145 150 155 160

Gln Gln Arg Asn Gly Lys Lys Thr His Val Tyr Ala Gln Ala Pro Phe

165 170 175

Ala Gly Glu Ala Ile Asn Lys Asn Gly Leu Gln Ile Gly Thr Asn Gly

180 185 190

Ala Ala Thr Glu Gly Asn Lys Glu Ile Tyr Ala Asp Lys Thr Tyr Gln

195 200 205

Pro Glu Pro Gln Ile Gly Glu Ser Gln Trp Asn Glu Ala Glu Ser Ser

210 215 220

Val Ala Gly Gly Arg Val Leu Lys Lys Thr Thr Pro Met Lys Pro Cys

225 230 235 240

Tyr Gly Ser Tyr Ala Arg Pro Thr Asn Ser Asn Gly Gly Gln Gly Val

245 250 255

Met Val Glu Gln Asn Gly Lys Leu Glu Ser Gln Val Glu Met Gln Phe

260 265 270

Phe Ser Thr Ser Ser Cys Ser Ser Gly Gly Thr Pro Lys Leu Val Leu

275 280 285

Tyr Ser Glu Asp Val Asn Met Glu Thr Pro Asp Thr His Leu Ser Tyr

290 295 300

Lys Pro Gly Lys Ser Asp Asp Asn Ser Lys Ala Met Leu Gly Gln Gln

305 310 315 320

Ser Met Pro Asn Arg Pro Asn Tyr Ile Ala Phe Arg Asp Asn Phe Ile

325 330 335

Gly Leu Met Tyr Tyr Asn Ser Thr Gly Asn Met Gly Val Leu Ala Gly

340 345 350

Gln Ala Ser Gln Leu Asn Ala Val Val Asp Leu Gln Asp Arg Asn Thr

355 360 365

Glu Leu Ser Tyr Gln Leu Leu Leu Asp Ser Ile Gly Asp Arg Thr Arg

370 375 380

Tyr Phe Ser Met Trp Asn Gln Ala Val Asp Ser Tyr Asp Pro Asp Val

385 390 395 400

Arg Ile Ile Glu Asn His Gly Thr Glu Asp Glu Leu Pro Asn Tyr Cys

405 410 415

Phe Pro Leu Gly Gly Ile Gly Val Thr Asp Thr Tyr Gln Ala Ile Lys

420 425 430

Ala Thr Asn Gly Asn Gly Gly Ala Thr Thr Trp Ala Gln Asp Asn Thr

435 440 445

Phe Ala Glu Arg Asn Glu Ile Gly Val Gly Asn Asn Phe Ala Met Glu

450 455 460

Ile Asn Leu Asn Ala Asn Leu Trp Arg Asn Phe Leu Tyr Ser Asn Ile

465 470 475 480

Ala Leu Tyr Leu Pro Asp Lys Leu Lys Tyr Asn Pro Thr Asn Val Glu

485 490 495

Ile Ser Asp Asn Pro Asn Thr Tyr Asp Tyr Met Asn Lys Arg Val Val

500 505 510

Ala Pro Gly Leu Val Asp Cys Tyr Ile Asn Leu Gly Ala Arg Trp Ser

515 520 525

Leu Asp Tyr Met Asp Asn Val Asn Pro Phe Asn His His Arg Asn Ala

530 535 540

Gly Leu Arg Tyr Arg Ser Met Leu Leu Gly Asn Gly Arg Tyr Val Pro

545 550 555 560

Phe His Ile Gln Val Pro Gln Lys Phe Phe Ala Ile Lys Asn Leu Leu

565 570 575

Leu Leu Pro Gly Ser Tyr Thr Tyr Glu Trp Asn Phe Arg Lys Asp Val

580 585 590

Asn Met Val Leu Gln Ser Ser Leu Gly Asn Asp Leu Arg Val Asp Gly

595 600 605

Ala Ser Ile Lys Phe Asp Ser Ile Cys Leu Tyr Ala Thr Phe Phe Pro

610 615 620

Met Ala His Asn Thr Ala Ser Thr Leu Glu Ala Met Leu Arg Asn Asp

625 630 635 640

Thr Asn Asp Gln Ser Phe Asn Asp Tyr Leu Ser Ala Ala Asn Met Leu

645 650 655

Tyr Pro Ile Pro Ala Asn Ala Thr Asn Val Pro Ile Ser Ile Pro Ser

660 665 670

Arg Asn Trp Ala Ala Phe Arg Gly Trp Ala Phe Thr Arg Leu Lys Thr

675 680 685

Lys Glu Thr Pro Ser Leu Gly Ser Gly Tyr Asp Pro Tyr Tyr Thr Tyr

690 695 700

Ser Gly Ser Ile Pro Tyr Leu Asp Gly Thr Phe Tyr Leu Asn His Thr

705 710 715 720

Phe Lys Lys Val Ala Ile Thr Phe Asp Ser Ser Val Ser Trp Pro Gly

725 730 735

Asn Asp Arg Leu Leu Thr Pro Asn Glu Phe Glu Ile Lys Arg Ser Val

740 745 750

Asp Gly Glu Gly Tyr Asn Val Ala Gln Cys Asn Met Thr Lys Asp Trp

755 760 765

Phe Leu Val Gln Met Leu Ala Asn Tyr Asn Ile Gly Tyr Gln Gly Phe

770 775 780

Tyr Ile Pro Glu Ser Tyr Lys Asp Arg Met Tyr Ser Phe Phe Arg Asn

785 790 795 800

Phe Gln Pro Met Ser Arg Gln Val Val Asp Asp Thr Lys Tyr Lys Asp

805 810 815

Tyr Gln Gln Val Gly Ile Ile His Gln His Asn Asn Ser Gly Phe Val

820 825 830

Gly Tyr Leu Ala Pro Thr Met Arg Glu Gly Gln Ala Tyr Pro Ala Asn

835 840 845

Val Pro Tyr Pro Leu Ile Gly Lys Thr Ala Val Asp Ser Ile Thr Gln

850 855 860

Lys Lys Phe Leu Cys Asp Arg Thr Leu Trp Arg Ile Pro Phe Ser Ser

865 870 875 880

Asn Phe Met Ser Met Gly Ala Leu Thr Asp Leu Gly Gln Asn Leu Leu

885 890 895

Tyr Ala Asn Ser Ala His Ala Leu Asp Met Thr Phe Glu Val Asp Pro

900 905 910

Met Asp Glu Pro Thr Leu Leu Tyr Val Leu Phe Glu Val Phe Asp Val

915 920 925

Val Arg Val His Gln Pro His Arg Gly Val Ile Glu Thr Val Tyr Leu

930 935 940

Arg Thr Pro Phe Ser Ala Gly Asn Ala Thr Thr

945 950 955

<210> 44

<211> 964

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Hexon polypeptide

<400> 44

Met Ala Thr Pro Ser Met Met Pro Gln Trp Ser Tyr Met His Ile Ser

1 5 10 15

Gly Gln Asp Ala Ser Glu Tyr Leu Ser Pro Gly Leu Val Gln Phe Ala

20 25 30

Arg Ala Thr Glu Thr Tyr Phe Ser Leu Asn Asn Lys Phe Arg Asn Pro

35 40 45

Thr Val Ala Pro Thr His Asp Val Thr Thr Asp Arg Ser Gln Arg Leu

50 55 60

Thr Leu Arg Phe Ile Pro Val Asp Arg Glu Asp Thr Ala Tyr Ser Tyr

65 70 75 80

Lys Ala Arg Phe Thr Leu Ala Val Gly Asp Asn Arg Val Leu Asp Met

85 90 95

Ala Ser Thr Tyr Phe Asp Ile Arg Gly Val Leu Asp Arg Gly Pro Thr

100 105 110

Phe Lys Pro Tyr Ser Gly Thr Ala Tyr Asn Ala Leu Ala Pro Lys Gly

115 120 125

Ala Pro Asn Ser Cys Glu Trp Asp Glu Asp Asp Thr Gln Val Gln Val

130 135 140

Ala Ala Glu Asp Asp Gln Asp Asp Asp Ser Ser Gly Gly Thr Glu Glu

145 150 155 160

Glu Glu Gln Leu Pro Gln Gln Arg Asn Gly Lys Lys Thr His Val Tyr

165 170 175

Ala Gln Ala Pro Phe Ala Gly Glu Ala Ile Asn Lys Asn Gly Leu Gln

180 185 190

Ile Gly Thr Asn Gly Ala Ala Thr Glu Gly Asn Lys Glu Ile Tyr Ala

195 200 205

Asp Lys Thr Tyr Gln Pro Glu Pro Gln Ile Gly Glu Ser Gln Trp Asn

210 215 220

Glu Ala Glu Ser Ser Val Ala Gly Gly Arg Val Leu Lys Lys Thr Thr

225 230 235 240

Pro Met Lys Pro Cys Tyr Gly Ser Tyr Ala Arg Pro Thr Asn Ser Asn

245 250 255

Gly Gly Gln Gly Val Met Val Glu Gln Asn Gly Lys Leu Glu Ser Gln

260 265 270

Val Glu Met Gln Phe Phe Ser Thr Ser Val Asn Ala Met Asn Glu Ala

275 280 285

Asn Ala Ile Gln Pro Lys Leu Val Leu Tyr Ser Glu Asp Val Asn Met

290 295 300

Glu Thr Pro Asp Thr His Leu Ser Tyr Lys Pro Gly Lys Ser Asp Asp

305 310 315 320

Asn Ser Lys Ala Met Leu Gly Gln Gln Ser Met Pro Asn Arg Pro Asn

325 330 335

Tyr Ile Ala Phe Arg Asp Asn Phe Ile Gly Leu Met Tyr Tyr Asn Ser

340 345 350

Thr Gly Asn Met Gly Val Leu Ala Gly Gln Ala Ser Gln Leu Asn Ala

355 360 365

Val Val Asp Leu Gln Asp Arg Asn Thr Glu Leu Ser Tyr Gln Leu Leu

370 375 380

Leu Asp Ser Ile Gly Asp Arg Thr Arg Tyr Phe Ser Met Trp Asn Gln

385 390 395 400

Ala Val Asp Ser Tyr Asp Pro Asp Val Arg Ile Ile Glu Asn His Gly

405 410 415

Thr Glu Asp Glu Leu Pro Asn Tyr Cys Phe Pro Leu Gly Gly Ile Gly

420 425 430

Val Thr Asp Thr Tyr Gln Ala Ile Lys Ala Thr Asn Gly Asn Gly Gly

435 440 445

Ala Thr Thr Trp Ala Gln Asp Asn Thr Phe Ala Glu Arg Asn Glu Ile

450 455 460

Gly Val Gly Asn Asn Phe Ala Met Glu Ile Asn Leu Asn Ala Asn Leu

465 470 475 480

Trp Arg Asn Phe Leu Tyr Ser Asn Ile Ala Leu Tyr Leu Pro Asp Lys

485 490 495

Leu Lys Tyr Asn Pro Thr Asn Val Glu Ile Ser Asp Asn Pro Asn Thr

500 505 510

Tyr Asp Tyr Met Asn Lys Arg Val Val Ala Pro Gly Leu Val Asp Cys

515 520 525

Tyr Ile Asn Leu Gly Ala Arg Trp Ser Leu Asp Tyr Met Asp Asn Val

530 535 540

Asn Pro Phe Asn His His Arg Asn Ala Gly Leu Arg Tyr Arg Ser Met

545 550 555 560

Leu Leu Gly Asn Gly Arg Tyr Val Pro Phe His Ile Gln Val Pro Gln

565 570 575

Lys Phe Phe Ala Ile Lys Asn Leu Leu Leu Leu Pro Gly Ser Tyr Thr

580 585 590

Tyr Glu Trp Asn Phe Arg Lys Asp Val Asn Met Val Leu Gln Ser Ser

595 600 605

Leu Gly Asn Asp Leu Arg Val Asp Gly Ala Ser Ile Lys Phe Asp Ser

610 615 620

Ile Cys Leu Tyr Ala Thr Phe Phe Pro Met Ala His Asn Thr Ala Ser

625 630 635 640

Thr Leu Glu Ala Met Leu Arg Asn Asp Thr Asn Asp Gln Ser Phe Asn

645 650 655

Asp Tyr Leu Ser Ala Ala Asn Met Leu Tyr Pro Ile Pro Ala Asn Ala

660 665 670

Thr Asn Val Pro Ile Ser Ile Pro Ser Arg Asn Trp Ala Ala Phe Arg

675 680 685

Gly Trp Ala Phe Thr Arg Leu Lys Thr Lys Glu Thr Pro Ser Leu Gly

690 695 700

Ser Gly Tyr Asp Pro Tyr Tyr Thr Tyr Ser Gly Ser Ile Pro Tyr Leu

705 710 715 720

Asp Gly Thr Phe Tyr Leu Asn His Thr Phe Lys Lys Val Ala Ile Thr

725 730 735

Phe Asp Ser Ser Val Ser Trp Pro Gly Asn Asp Arg Leu Leu Thr Pro

740 745 750

Asn Glu Phe Glu Ile Lys Arg Ser Val Asp Gly Glu Gly Tyr Asn Val

755 760 765

Ala Gln Cys Asn Met Thr Lys Asp Trp Phe Leu Val Gln Met Leu Ala

770 775 780

Asn Tyr Asn Ile Gly Tyr Gln Gly Phe Tyr Ile Pro Glu Ser Tyr Lys

785 790 795 800

Asp Arg Met Tyr Ser Phe Phe Arg Asn Phe Gln Pro Met Ser Arg Gln

805 810 815

Val Val Asp Asp Thr Lys Tyr Lys Asp Tyr Gln Gln Val Gly Ile Ile

820 825 830

His Gln His Asn Asn Ser Gly Phe Val Gly Tyr Leu Ala Pro Thr Met

835 840 845

Arg Glu Gly Gln Ala Tyr Pro Ala Asn Val Pro Tyr Pro Leu Ile Gly

850 855 860

Lys Thr Ala Val Asp Ser Ile Thr Gln Lys Lys Phe Leu Cys Asp Arg

865 870 875 880

Thr Leu Trp Arg Ile Pro Phe Ser Ser Asn Phe Met Ser Met Gly Ala

885 890 895

Leu Thr Asp Leu Gly Gln Asn Leu Leu Tyr Ala Asn Ser Ala His Ala

900 905 910

Leu Asp Met Thr Phe Glu Val Asp Pro Met Asp Glu Pro Thr Leu Leu

915 920 925

Tyr Val Leu Phe Glu Val Phe Asp Val Val Arg Val His Gln Pro His

930 935 940

Arg Gly Val Ile Glu Thr Val Tyr Leu Arg Thr Pro Phe Ser Ala Gly

945 950 955 960

Asn Ala Thr Thr

<210> 45

<211> 952

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Hexon polypeptide

<400> 45

Met Ala Thr Pro Ser Met Met Pro Gln Trp Ser Tyr Met His Ile Ser

1 5 10 15

Gly Gln Asp Ala Ser Glu Tyr Leu Ser Pro Gly Leu Val Gln Phe Ala

20 25 30

Arg Ala Thr Glu Thr Tyr Phe Ser Leu Asn Asn Lys Phe Arg Asn Pro

35 40 45

Thr Val Ala Pro Thr His Asp Val Thr Thr Asp Arg Ser Gln Arg Leu

50 55 60

Thr Leu Arg Phe Ile Pro Val Asp Arg Glu Asp Thr Ala Tyr Ser Tyr

65 70 75 80

Lys Ala Arg Phe Thr Leu Ala Val Gly Asp Asn Arg Val Leu Asp Met

85 90 95

Ala Ser Thr Tyr Phe Asp Ile Arg Gly Val Leu Asp Arg Gly Pro Thr

100 105 110

Phe Lys Pro Tyr Ser Gly Thr Ala Tyr Asn Ala Leu Ala Pro Lys Gly

115 120 125

Ala Pro Asn Ser Cys Glu Trp Asp Glu Asp Asp Thr Gln Val Gln Val

130 135 140

Ala Ala Glu Asp Asp Gln Asp Asp Asp Glu Glu Glu Glu Gln Leu Pro

145 150 155 160

Gln Gln Arg Asn Gly Lys Lys Thr His Val Tyr Ala Gln Ala Pro Phe

165 170 175

Ala Gly Glu Ala Ile Asn Lys Asn Gly Leu Gln Ile Gly Thr Asn Gly

180 185 190

Ala Ala Thr Glu Gly Asn Lys Glu Ile Tyr Ala Asp Lys Thr Tyr Gln

195 200 205

Pro Glu Pro Gln Ile Gly Glu Ser Gln Trp Asn Glu Ala Glu Ser Ser

210 215 220

Val Ala Gly Gly Arg Val Leu Lys Lys Thr Thr Pro Met Lys Pro Cys

225 230 235 240

Tyr Gly Ser Tyr Ala Arg Pro Thr Asn Ser Asn Gly Gly Gln Gly Val

245 250 255

Met Val Glu Gln Asn Gly Lys Leu Glu Ser Gln Val Glu Met Gln Phe

260 265 270

Phe Ser Thr Ser Ser Gly Gly Thr Pro Lys Leu Val Leu Tyr Ser Glu

275 280 285

Asp Val Asn Met Glu Thr Pro Asp Thr His Leu Ser Tyr Lys Pro Gly

290 295 300

Lys Ser Asp Asp Asn Ser Lys Ala Met Leu Gly Gln Gln Ser Met Pro

305 310 315 320

Asn Arg Pro Asn Tyr Ile Ala Phe Arg Asp Asn Phe Ile Gly Leu Met

325 330 335

Tyr Tyr Asn Ser Thr Gly Asn Met Gly Val Leu Ala Gly Gln Ala Ser

340 345 350

Gln Leu Asn Ala Val Val Asp Leu Gln Asp Arg Asn Thr Glu Leu Ser

355 360 365

Tyr Gln Leu Leu Leu Asp Ser Ile Gly Asp Arg Thr Arg Tyr Phe Ser

370 375 380

Met Trp Asn Gln Ala Val Asp Ser Tyr Asp Pro Asp Val Arg Ile Ile

385 390 395 400

Glu Asn His Gly Thr Glu Asp Glu Leu Pro Asn Tyr Cys Phe Pro Leu

405 410 415

Gly Gly Ile Gly Val Thr Asp Thr Tyr Gln Ala Ile Lys Ala Thr Asn

420 425 430

Gly Asn Gly Gly Ala Thr Thr Trp Ala Gln Asp Asn Thr Phe Ala Glu

435 440 445

Arg Asn Glu Ile Gly Val Gly Asn Asn Phe Ala Met Glu Ile Asn Leu

450 455 460

Asn Ala Asn Leu Trp Arg Asn Phe Leu Tyr Ser Asn Ile Ala Leu Tyr

465 470 475 480

Leu Pro Asp Lys Leu Lys Tyr Asn Pro Thr Asn Val Glu Ile Ser Asp

485 490 495

Asn Pro Asn Thr Tyr Asp Tyr Met Asn Lys Arg Val Val Ala Pro Gly

500 505 510

Leu Val Asp Cys Tyr Ile Asn Leu Gly Ala Arg Trp Ser Leu Asp Tyr

515 520 525

Met Asp Asn Val Asn Pro Phe Asn His His Arg Asn Ala Gly Leu Arg

530 535 540

Tyr Arg Ser Met Leu Leu Gly Asn Gly Arg Tyr Val Pro Phe His Ile

545 550 555 560

Gln Val Pro Gln Lys Phe Phe Ala Ile Lys Asn Leu Leu Leu Leu Pro

565 570 575

Gly Ser Tyr Thr Tyr Glu Trp Asn Phe Arg Lys Asp Val Asn Met Val

580 585 590

Leu Gln Ser Ser Leu Gly Asn Asp Leu Arg Val Asp Gly Ala Ser Ile

595 600 605

Lys Phe Asp Ser Ile Cys Leu Tyr Ala Thr Phe Phe Pro Met Ala His

610 615 620

Asn Thr Ala Ser Thr Leu Glu Ala Met Leu Arg Asn Asp Thr Asn Asp

625 630 635 640

Gln Ser Phe Asn Asp Tyr Leu Ser Ala Ala Asn Met Leu Tyr Pro Ile

645 650 655

Pro Ala Asn Ala Thr Asn Val Pro Ile Ser Ile Pro Ser Arg Asn Trp

660 665 670

Ala Ala Phe Arg Gly Trp Ala Phe Thr Arg Leu Lys Thr Lys Glu Thr

675 680 685

Pro Ser Leu Gly Ser Gly Tyr Asp Pro Tyr Tyr Thr Tyr Ser Gly Ser

690 695 700

Ile Pro Tyr Leu Asp Gly Thr Phe Tyr Leu Asn His Thr Phe Lys Lys

705 710 715 720

Val Ala Ile Thr Phe Asp Ser Ser Val Ser Trp Pro Gly Asn Asp Arg

725 730 735

Leu Leu Thr Pro Asn Glu Phe Glu Ile Lys Arg Ser Val Asp Gly Glu

740 745 750

Gly Tyr Asn Val Ala Gln Cys Asn Met Thr Lys Asp Trp Phe Leu Val

755 760 765

Gln Met Leu Ala Asn Tyr Asn Ile Gly Tyr Gln Gly Phe Tyr Ile Pro

770 775 780

Glu Ser Tyr Lys Asp Arg Met Tyr Ser Phe Phe Arg Asn Phe Gln Pro

785 790 795 800

Met Ser Arg Gln Val Val Asp Asp Thr Lys Tyr Lys Asp Tyr Gln Gln

805 810 815

Val Gly Ile Ile His Gln His Asn Asn Ser Gly Phe Val Gly Tyr Leu

820 825 830

Ala Pro Thr Met Arg Glu Gly Gln Ala Tyr Pro Ala Asn Val Pro Tyr

835 840 845

Pro Leu Ile Gly Lys Thr Ala Val Asp Ser Ile Thr Gln Lys Lys Phe

850 855 860

Leu Cys Asp Arg Thr Leu Trp Arg Ile Pro Phe Ser Ser Asn Phe Met

865 870 875 880

Ser Met Gly Ala Leu Thr Asp Leu Gly Gln Asn Leu Leu Tyr Ala Asn

885 890 895

Ser Ala His Ala Leu Asp Met Thr Phe Glu Val Asp Pro Met Asp Glu

900 905 910

Pro Thr Leu Leu Tyr Val Leu Phe Glu Val Phe Asp Val Val Arg Val

915 920 925

His Gln Pro His Arg Gly Val Ile Glu Thr Val Tyr Leu Arg Thr Pro

930 935 940

Phe Ser Ala Gly Asn Ala Thr Thr

945 950

<210> 46

<211> 1033

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Hexon polypeptide

<400> 46

Met Ala Thr Pro Ser Met Met Pro Gln Trp Ser Tyr Met His Ile Ser

1 5 10 15

Gly Gln Asp Ala Ser Glu Tyr Leu Ser Pro Gly Leu Val Gln Phe Ala

20 25 30

Arg Ala Thr Glu Thr Tyr Phe Ser Leu Asn Asn Lys Phe Arg Asn Pro

35 40 45

Thr Val Ala Pro Thr His Asp Val Thr Thr Asp Arg Ser Gln Arg Leu

50 55 60

Thr Leu Arg Phe Ile Pro Val Asp Arg Glu Asp Thr Ala Tyr Ser Tyr

65 70 75 80

Lys Ala Arg Phe Thr Leu Ala Val Gly Asp Asn Arg Val Leu Asp Met

85 90 95

Ala Ser Thr Tyr Phe Asp Ile Arg Gly Val Leu Asp Arg Gly Pro Thr

100 105 110

Phe Lys Pro Tyr Ser Gly Thr Ala Tyr Asn Ala Leu Ala Pro Lys Gly

115 120 125

Ala Pro Asn Ser Cys Glu Trp Asp Glu Asp Asp Thr Gln Val Gln Val

130 135 140

Ala Ala Glu Asp Asp Gln Asp Asp Asp Ser Thr Gly Glu Ile Pro Ala

145 150 155 160

Pro Leu Ala Gly Thr Val Ser Lys Ile Leu Val Lys Glu Gly Asp Thr

165 170 175

Val Lys Ala Gly Gln Thr Val Leu Val Leu Glu Ala Met Lys Met Glu

180 185 190

Thr Glu Ile Asn Ala Pro Thr Asp Gly Lys Val Glu Lys Val Leu Val

195 200 205

Lys Glu Arg Asp Ala Val Gln Gly Gly Gln Gly Leu Ile Lys Ile Gly

210 215 220

Gly Gly Thr Glu Glu Glu Glu Gln Leu Pro Gln Gln Arg Asn Gly Lys

225 230 235 240

Lys Thr His Val Tyr Ala Gln Ala Pro Phe Ala Gly Glu Ala Ile Asn

245 250 255

Lys Asn Gly Leu Gln Ile Gly Thr Asn Gly Ala Ala Thr Glu Gly Asn

260 265 270

Lys Glu Ile Tyr Ala Asp Lys Thr Tyr Gln Pro Glu Pro Gln Ile Gly

275 280 285

Glu Ser Gln Trp Asn Glu Ala Glu Ser Ser Val Ala Gly Gly Arg Val

290 295 300

Leu Lys Lys Thr Thr Pro Met Lys Pro Cys Tyr Gly Ser Tyr Ala Arg

305 310 315 320

Pro Thr Asn Ser Asn Gly Gly Gln Gly Val Met Val Glu Gln Asn Gly

325 330 335

Lys Leu Glu Ser Gln Val Glu Met Gln Phe Phe Ser Thr Ser Val Asn

340 345 350

Ala Met Asn Glu Ala Asn Ala Ile Gln Pro Lys Leu Val Leu Tyr Ser

355 360 365

Glu Asp Val Asn Met Glu Thr Pro Asp Thr His Leu Ser Tyr Lys Pro

370 375 380

Gly Lys Ser Asp Asp Asn Ser Lys Ala Met Leu Gly Gln Gln Ser Met

385 390 395 400

Pro Asn Arg Pro Asn Tyr Ile Ala Phe Arg Asp Asn Phe Ile Gly Leu

405 410 415

Met Tyr Tyr Asn Ser Thr Gly Asn Met Gly Val Leu Ala Gly Gln Ala

420 425 430

Ser Gln Leu Asn Ala Val Val Asp Leu Gln Asp Arg Asn Thr Glu Leu

435 440 445

Ser Tyr Gln Leu Leu Leu Asp Ser Ile Gly Asp Arg Thr Arg Tyr Phe

450 455 460

Ser Met Trp Asn Gln Ala Val Asp Ser Tyr Asp Pro Asp Val Arg Ile

465 470 475 480

Ile Glu Asn His Gly Thr Glu Asp Glu Leu Pro Asn Tyr Cys Phe Pro

485 490 495

Leu Gly Gly Ile Gly Val Thr Asp Thr Tyr Gln Ala Ile Lys Ala Thr

500 505 510

Asn Gly Asn Gly Gly Ala Thr Thr Trp Ala Gln Asp Asn Thr Phe Ala

515 520 525

Glu Arg Asn Glu Ile Gly Val Gly Asn Asn Phe Ala Met Glu Ile Asn

530 535 540

Leu Asn Ala Asn Leu Trp Arg Asn Phe Leu Tyr Ser Asn Ile Ala Leu

545 550 555 560

Tyr Leu Pro Asp Lys Leu Lys Tyr Asn Pro Thr Asn Val Glu Ile Ser

565 570 575

Asp Asn Pro Asn Thr Tyr Asp Tyr Met Asn Lys Arg Val Val Ala Pro

580 585 590

Gly Leu Val Asp Cys Tyr Ile Asn Leu Gly Ala Arg Trp Ser Leu Asp

595 600 605

Tyr Met Asp Asn Val Asn Pro Phe Asn His His Arg Asn Ala Gly Leu

610 615 620

Arg Tyr Arg Ser Met Leu Leu Gly Asn Gly Arg Tyr Val Pro Phe His

625 630 635 640

Ile Gln Val Pro Gln Lys Phe Phe Ala Ile Lys Asn Leu Leu Leu Leu

645 650 655

Pro Gly Ser Tyr Thr Tyr Glu Trp Asn Phe Arg Lys Asp Val Asn Met

660 665 670

Val Leu Gln Ser Ser Leu Gly Asn Asp Leu Arg Val Asp Gly Ala Ser

675 680 685

Ile Lys Phe Asp Ser Ile Cys Leu Tyr Ala Thr Phe Phe Pro Met Ala

690 695 700

His Asn Thr Ala Ser Thr Leu Glu Ala Met Leu Arg Asn Asp Thr Asn

705 710 715 720

Asp Gln Ser Phe Asn Asp Tyr Leu Ser Ala Ala Asn Met Leu Tyr Pro

725 730 735

Ile Pro Ala Asn Ala Thr Asn Val Pro Ile Ser Ile Pro Ser Arg Asn

740 745 750

Trp Ala Ala Phe Arg Gly Trp Ala Phe Thr Arg Leu Lys Thr Lys Glu

755 760 765

Thr Pro Ser Leu Gly Ser Gly Tyr Asp Pro Tyr Tyr Thr Tyr Ser Gly

770 775 780

Ser Ile Pro Tyr Leu Asp Gly Thr Phe Tyr Leu Asn His Thr Phe Lys

785 790 795 800

Lys Val Ala Ile Thr Phe Asp Ser Ser Val Ser Trp Pro Gly Asn Asp

805 810 815

Arg Leu Leu Thr Pro Asn Glu Phe Glu Ile Lys Arg Ser Val Asp Gly

820 825 830

Glu Gly Tyr Asn Val Ala Gln Cys Asn Met Thr Lys Asp Trp Phe Leu

835 840 845

Val Gln Met Leu Ala Asn Tyr Asn Ile Gly Tyr Gln Gly Phe Tyr Ile

850 855 860

Pro Glu Ser Tyr Lys Asp Arg Met Tyr Ser Phe Phe Arg Asn Phe Gln

865 870 875 880

Pro Met Ser Arg Gln Val Val Asp Asp Thr Lys Tyr Lys Asp Tyr Gln

885 890 895

Gln Val Gly Ile Ile His Gln His Asn Asn Ser Gly Phe Val Gly Tyr

900 905 910

Leu Ala Pro Thr Met Arg Glu Gly Gln Ala Tyr Pro Ala Asn Val Pro

915 920 925

Tyr Pro Leu Ile Gly Lys Thr Ala Val Asp Ser Ile Thr Gln Lys Lys

930 935 940

Phe Leu Cys Asp Arg Thr Leu Trp Arg Ile Pro Phe Ser Ser Asn Phe

945 950 955 960

Met Ser Met Gly Ala Leu Thr Asp Leu Gly Gln Asn Leu Leu Tyr Ala

965 970 975

Asn Ser Ala His Ala Leu Asp Met Thr Phe Glu Val Asp Pro Met Asp

980 985 990

Glu Pro Thr Leu Leu Tyr Val Leu Phe Glu Val Phe Asp Val Val Arg

995 1000 1005

Val His Gln Pro His Arg Gly Val Ile Glu Thr Val Tyr Leu Arg

1010 1015 1020

Thr Pro Phe Ser Ala Gly Asn Ala Thr Thr

1025 1030

<210> 47

<211> 1021

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Hexon polypeptide

<400> 47

Met Ala Thr Pro Ser Met Met Pro Gln Trp Ser Tyr Met His Ile Ser

1 5 10 15

Gly Gln Asp Ala Ser Glu Tyr Leu Ser Pro Gly Leu Val Gln Phe Ala

20 25 30

Arg Ala Thr Glu Thr Tyr Phe Ser Leu Asn Asn Lys Phe Arg Asn Pro

35 40 45

Thr Val Ala Pro Thr His Asp Val Thr Thr Asp Arg Ser Gln Arg Leu

50 55 60

Thr Leu Arg Phe Ile Pro Val Asp Arg Glu Asp Thr Ala Tyr Ser Tyr

65 70 75 80

Lys Ala Arg Phe Thr Leu Ala Val Gly Asp Asn Arg Val Leu Asp Met

85 90 95

Ala Ser Thr Tyr Phe Asp Ile Arg Gly Val Leu Asp Arg Gly Pro Thr

100 105 110

Phe Lys Pro Tyr Ser Gly Thr Ala Tyr Asn Ala Leu Ala Pro Lys Gly

115 120 125

Ala Pro Asn Ser Cys Glu Trp Asp Glu Asp Asp Thr Gln Val Gln Val

130 135 140

Ala Ala Glu Asp Asp Gln Asp Asp Asp Glu Glu Glu Glu Gln Leu Pro

145 150 155 160

Gln Gln Arg Asn Gly Lys Lys Thr His Val Tyr Ala Gln Ala Pro Phe

165 170 175

Ala Gly Glu Ala Ile Asn Lys Asn Gly Leu Gln Ile Gly Thr Asn Gly

180 185 190

Ala Ala Thr Glu Gly Asn Lys Glu Ile Tyr Ala Asp Lys Thr Tyr Gln

195 200 205

Pro Glu Pro Gln Ile Gly Glu Ser Gln Trp Asn Glu Ala Glu Ser Ser

210 215 220

Val Ala Gly Gly Arg Val Leu Lys Lys Thr Thr Pro Met Lys Pro Cys

225 230 235 240

Tyr Gly Ser Tyr Ala Arg Pro Thr Asn Ser Asn Gly Gly Gln Gly Val

245 250 255

Met Val Glu Gln Asn Gly Lys Leu Glu Ser Gln Val Glu Met Gln Phe

260 265 270

Phe Ser Thr Ser Thr Gly Glu Ile Pro Ala Pro Leu Ala Gly Thr Val

275 280 285

Ser Lys Ile Leu Val Lys Glu Gly Asp Thr Val Lys Ala Gly Gln Thr

290 295 300

Val Leu Val Leu Glu Ala Met Lys Met Glu Thr Glu Ile Asn Ala Pro

305 310 315 320

Thr Asp Gly Lys Val Glu Lys Val Leu Val Lys Glu Arg Asp Ala Val

325 330 335

Gln Gly Gly Gln Gly Leu Ile Lys Ile Gly Gly Gly Thr Pro Lys Leu

340 345 350

Val Leu Tyr Ser Glu Asp Val Asn Met Glu Thr Pro Asp Thr His Leu

355 360 365

Ser Tyr Lys Pro Gly Lys Ser Asp Asp Asn Ser Lys Ala Met Leu Gly

370 375 380

Gln Gln Ser Met Pro Asn Arg Pro Asn Tyr Ile Ala Phe Arg Asp Asn

385 390 395 400

Phe Ile Gly Leu Met Tyr Tyr Asn Ser Thr Gly Asn Met Gly Val Leu

405 410 415

Ala Gly Gln Ala Ser Gln Leu Asn Ala Val Val Asp Leu Gln Asp Arg

420 425 430

Asn Thr Glu Leu Ser Tyr Gln Leu Leu Leu Asp Ser Ile Gly Asp Arg

435 440 445

Thr Arg Tyr Phe Ser Met Trp Asn Gln Ala Val Asp Ser Tyr Asp Pro

450 455 460

Asp Val Arg Ile Ile Glu Asn His Gly Thr Glu Asp Glu Leu Pro Asn

465 470 475 480

Tyr Cys Phe Pro Leu Gly Gly Ile Gly Val Thr Asp Thr Tyr Gln Ala

485 490 495

Ile Lys Ala Thr Asn Gly Asn Gly Gly Ala Thr Thr Trp Ala Gln Asp

500 505 510

Asn Thr Phe Ala Glu Arg Asn Glu Ile Gly Val Gly Asn Asn Phe Ala

515 520 525

Met Glu Ile Asn Leu Asn Ala Asn Leu Trp Arg Asn Phe Leu Tyr Ser

530 535 540

Asn Ile Ala Leu Tyr Leu Pro Asp Lys Leu Lys Tyr Asn Pro Thr Asn

545 550 555 560

Val Glu Ile Ser Asp Asn Pro Asn Thr Tyr Asp Tyr Met Asn Lys Arg

565 570 575

Val Val Ala Pro Gly Leu Val Asp Cys Tyr Ile Asn Leu Gly Ala Arg

580 585 590

Trp Ser Leu Asp Tyr Met Asp Asn Val Asn Pro Phe Asn His His Arg

595 600 605

Asn Ala Gly Leu Arg Tyr Arg Ser Met Leu Leu Gly Asn Gly Arg Tyr

610 615 620

Val Pro Phe His Ile Gln Val Pro Gln Lys Phe Phe Ala Ile Lys Asn

625 630 635 640

Leu Leu Leu Leu Pro Gly Ser Tyr Thr Tyr Glu Trp Asn Phe Arg Lys

645 650 655

Asp Val Asn Met Val Leu Gln Ser Ser Leu Gly Asn Asp Leu Arg Val

660 665 670

Asp Gly Ala Ser Ile Lys Phe Asp Ser Ile Cys Leu Tyr Ala Thr Phe

675 680 685

Phe Pro Met Ala His Asn Thr Ala Ser Thr Leu Glu Ala Met Leu Arg

690 695 700

Asn Asp Thr Asn Asp Gln Ser Phe Asn Asp Tyr Leu Ser Ala Ala Asn

705 710 715 720

Met Leu Tyr Pro Ile Pro Ala Asn Ala Thr Asn Val Pro Ile Ser Ile

725 730 735

Pro Ser Arg Asn Trp Ala Ala Phe Arg Gly Trp Ala Phe Thr Arg Leu

740 745 750

Lys Thr Lys Glu Thr Pro Ser Leu Gly Ser Gly Tyr Asp Pro Tyr Tyr

755 760 765

Thr Tyr Ser Gly Ser Ile Pro Tyr Leu Asp Gly Thr Phe Tyr Leu Asn

770 775 780

His Thr Phe Lys Lys Val Ala Ile Thr Phe Asp Ser Ser Val Ser Trp

785 790 795 800

Pro Gly Asn Asp Arg Leu Leu Thr Pro Asn Glu Phe Glu Ile Lys Arg

805 810 815

Ser Val Asp Gly Glu Gly Tyr Asn Val Ala Gln Cys Asn Met Thr Lys

820 825 830

Asp Trp Phe Leu Val Gln Met Leu Ala Asn Tyr Asn Ile Gly Tyr Gln

835 840 845

Gly Phe Tyr Ile Pro Glu Ser Tyr Lys Asp Arg Met Tyr Ser Phe Phe

850 855 860

Arg Asn Phe Gln Pro Met Ser Arg Gln Val Val Asp Asp Thr Lys Tyr

865 870 875 880

Lys Asp Tyr Gln Gln Val Gly Ile Ile His Gln His Asn Asn Ser Gly

885 890 895

Phe Val Gly Tyr Leu Ala Pro Thr Met Arg Glu Gly Gln Ala Tyr Pro

900 905 910

Ala Asn Val Pro Tyr Pro Leu Ile Gly Lys Thr Ala Val Asp Ser Ile

915 920 925

Thr Gln Lys Lys Phe Leu Cys Asp Arg Thr Leu Trp Arg Ile Pro Phe

930 935 940

Ser Ser Asn Phe Met Ser Met Gly Ala Leu Thr Asp Leu Gly Gln Asn

945 950 955 960

Leu Leu Tyr Ala Asn Ser Ala His Ala Leu Asp Met Thr Phe Glu Val

965 970 975

Asp Pro Met Asp Glu Pro Thr Leu Leu Tyr Val Leu Phe Glu Val Phe

980 985 990

Asp Val Val Arg Val His Gln Pro His Arg Gly Val Ile Glu Thr Val

995 1000 1005

Tyr Leu Arg Thr Pro Phe Ser Ala Gly Asn Ala Thr Thr

1010 1015 1020

<210> 48

<211> 1015

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Hexon polypeptide

<400> 48

Met Ala Thr Pro Ser Met Met Pro Gln Trp Ser Tyr Met His Ile Ser

1 5 10 15

Gly Gln Asp Ala Ser Glu Tyr Leu Ser Pro Gly Leu Val Gln Phe Ala

20 25 30

Arg Ala Thr Glu Thr Tyr Phe Ser Leu Asn Asn Lys Phe Arg Asn Pro

35 40 45

Thr Val Ala Pro Thr His Asp Val Thr Thr Asp Arg Ser Gln Arg Leu

50 55 60

Thr Leu Arg Phe Ile Pro Val Asp Arg Glu Asp Thr Ala Tyr Ser Tyr

65 70 75 80

Lys Ala Arg Phe Thr Leu Ala Val Gly Asp Asn Arg Val Leu Asp Met

85 90 95

Ala Ser Thr Tyr Phe Asp Ile Arg Gly Val Leu Asp Arg Gly Pro Thr

100 105 110

Phe Lys Pro Tyr Ser Gly Thr Ala Tyr Asn Ala Leu Ala Pro Lys Gly

115 120 125

Ala Pro Asn Ser Cys Glu Trp Asp Glu Asp Asp Thr Gln Val Gln Val

130 135 140

Ala Ala Glu Asp Asp Gln Asp Asp Asp Glu Glu Glu Glu Gln Leu Pro

145 150 155 160

Gln Gln Arg Asn Gly Lys Lys Thr His Val Tyr Ala Gln Ala Pro Phe

165 170 175

Ala Gly Glu Ala Ile Asn Lys Asn Gly Leu Gln Ile Gly Thr Asn Gly

180 185 190

Ala Ala Thr Glu Gly Asn Lys Glu Ile Tyr Ala Asp Lys Thr Tyr Gln

195 200 205

Pro Glu Pro Gln Ile Gly Glu Ser Gln Trp Asn Glu Ala Glu Ser Ser

210 215 220

Val Ala Gly Gly Arg Val Leu Lys Lys Thr Thr Pro Met Lys Pro Cys

225 230 235 240

Tyr Gly Ser Tyr Ala Arg Pro Thr Asn Ser Asn Gly Gly Gln Gly Val

245 250 255

Met Val Glu Gln Asn Gly Lys Leu Glu Ser Gln Val Glu Met Gln Phe

260 265 270

Phe Ser Thr Ser Ser Ser Asn Phe Thr Arg Glu Gly Asn Val Thr Tyr

275 280 285

Lys Glu Glu Met Asp Lys Val Lys Asn Cys Ser Phe Asn Val Thr Thr

290 295 300

Gly Ile Arg Asp Lys Lys Gln Lys Val Asn Ala Leu Phe Tyr Arg Leu

305 310 315 320

Asp Ile Thr Pro Leu Asp Glu Asn Asn Asn Asn Ser Ser Glu Tyr Arg

325 330 335

Leu Ile Asn Ser Gly Gly Thr Pro Lys Leu Val Leu Tyr Ser Glu Asp

340 345 350

Val Asn Met Glu Thr Pro Asp Thr His Leu Ser Tyr Lys Pro Gly Lys

355 360 365

Ser Asp Asp Asn Ser Lys Ala Met Leu Gly Gln Gln Ser Met Pro Asn

370 375 380

Arg Pro Asn Tyr Ile Ala Phe Arg Asp Asn Phe Ile Gly Leu Met Tyr

385 390 395 400

Tyr Asn Ser Thr Gly Asn Met Gly Val Leu Ala Gly Gln Ala Ser Gln

405 410 415

Leu Asn Ala Val Val Asp Leu Gln Asp Arg Asn Thr Glu Leu Ser Tyr

420 425 430

Gln Leu Leu Leu Asp Ser Ile Gly Asp Arg Thr Arg Tyr Phe Ser Met

435 440 445

Trp Asn Gln Ala Val Asp Ser Tyr Asp Pro Asp Val Arg Ile Ile Glu

450 455 460

Asn His Gly Thr Glu Asp Glu Leu Pro Asn Tyr Cys Phe Pro Leu Gly

465 470 475 480

Gly Ile Gly Val Thr Asp Thr Tyr Gln Ala Ile Lys Ala Thr Asn Gly

485 490 495

Asn Gly Gly Ala Thr Thr Trp Ala Gln Asp Asn Thr Phe Ala Glu Arg

500 505 510

Asn Glu Ile Gly Val Gly Asn Asn Phe Ala Met Glu Ile Asn Leu Asn

515 520 525

Ala Asn Leu Trp Arg Asn Phe Leu Tyr Ser Asn Ile Ala Leu Tyr Leu

530 535 540

Pro Asp Lys Leu Lys Tyr Asn Pro Thr Asn Val Glu Ile Ser Asp Asn

545 550 555 560

Pro Asn Thr Tyr Asp Tyr Met Asn Lys Arg Val Val Ala Pro Gly Leu

565 570 575

Val Asp Cys Tyr Ile Asn Leu Gly Ala Arg Trp Ser Leu Asp Tyr Met

580 585 590

Asp Asn Val Asn Pro Phe Asn His His Arg Asn Ala Gly Leu Arg Tyr

595 600 605

Arg Ser Met Leu Leu Gly Asn Gly Arg Tyr Val Pro Phe His Ile Gln

610 615 620

Val Pro Gln Lys Phe Phe Ala Ile Lys Asn Leu Leu Leu Leu Pro Gly

625 630 635 640

Ser Tyr Thr Tyr Glu Trp Asn Phe Arg Lys Asp Val Asn Met Val Leu

645 650 655

Gln Ser Ser Leu Gly Asn Asp Leu Arg Val Asp Gly Ala Ser Ile Lys

660 665 670

Phe Asp Ser Ile Cys Leu Tyr Ala Thr Phe Phe Pro Met Ala His Asn

675 680 685

Thr Ala Ser Thr Leu Glu Ala Met Leu Arg Asn Asp Thr Asn Asp Gln

690 695 700

Ser Phe Asn Asp Tyr Leu Ser Ala Ala Asn Met Leu Tyr Pro Ile Pro

705 710 715 720

Ala Asn Ala Thr Asn Val Pro Ile Ser Ile Pro Ser Arg Asn Trp Ala

725 730 735

Ala Phe Arg Gly Trp Ala Phe Thr Arg Leu Lys Thr Lys Glu Thr Pro

740 745 750

Ser Leu Gly Ser Gly Tyr Asp Pro Tyr Tyr Thr Tyr Ser Gly Ser Ile

755 760 765

Pro Tyr Leu Asp Gly Thr Phe Tyr Leu Asn His Thr Phe Lys Lys Val

770 775 780

Ala Ile Thr Phe Asp Ser Ser Val Ser Trp Pro Gly Asn Asp Arg Leu

785 790 795 800

Leu Thr Pro Asn Glu Phe Glu Ile Lys Arg Ser Val Asp Gly Glu Gly

805 810 815

Tyr Asn Val Ala Gln Cys Asn Met Thr Lys Asp Trp Phe Leu Val Gln

820 825 830

Met Leu Ala Asn Tyr Asn Ile Gly Tyr Gln Gly Phe Tyr Ile Pro Glu

835 840 845

Ser Tyr Lys Asp Arg Met Tyr Ser Phe Phe Arg Asn Phe Gln Pro Met

850 855 860

Ser Arg Gln Val Val Asp Asp Thr Lys Tyr Lys Asp Tyr Gln Gln Val

865 870 875 880

Gly Ile Ile His Gln His Asn Asn Ser Gly Phe Val Gly Tyr Leu Ala

885 890 895

Pro Thr Met Arg Glu Gly Gln Ala Tyr Pro Ala Asn Val Pro Tyr Pro

900 905 910

Leu Ile Gly Lys Thr Ala Val Asp Ser Ile Thr Gln Lys Lys Phe Leu

915 920 925

Cys Asp Arg Thr Leu Trp Arg Ile Pro Phe Ser Ser Asn Phe Met Ser

930 935 940

Met Gly Ala Leu Thr Asp Leu Gly Gln Asn Leu Leu Tyr Ala Asn Ser

945 950 955 960

Ala His Ala Leu Asp Met Thr Phe Glu Val Asp Pro Met Asp Glu Pro

965 970 975

Thr Leu Leu Tyr Val Leu Phe Glu Val Phe Asp Val Val Arg Val His

980 985 990

Gln Pro His Arg Gly Val Ile Glu Thr Val Tyr Leu Arg Thr Pro Phe

995 1000 1005

Ser Ala Gly Asn Ala Thr Thr

1010 1015

<210> 49

<211> 987

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Hexon polypeptide

<400> 49

Met Ala Thr Pro Ser Met Met Pro Gln Trp Ser Tyr Met His Ile Ser

1 5 10 15

Gly Gln Asp Ala Ser Glu Tyr Leu Ser Pro Gly Leu Val Gln Phe Ala

20 25 30

Arg Ala Thr Glu Thr Tyr Phe Ser Leu Asn Asn Lys Phe Arg Asn Pro

35 40 45

Thr Val Ala Pro Thr His Asp Val Thr Thr Asp Arg Ser Gln Arg Leu

50 55 60

Thr Leu Arg Phe Ile Pro Val Asp Arg Glu Asp Thr Ala Tyr Ser Tyr

65 70 75 80

Lys Ala Arg Phe Thr Leu Ala Val Gly Asp Asn Arg Val Leu Asp Met

85 90 95

Ala Ser Thr Tyr Phe Asp Ile Arg Gly Val Leu Asp Arg Gly Pro Thr

100 105 110

Phe Lys Pro Tyr Ser Gly Thr Ala Tyr Asn Ala Leu Ala Pro Lys Gly

115 120 125

Ala Pro Asn Ser Cys Glu Trp Asp Glu Asp Asp Thr Gln Val Gln Val

130 135 140

Ala Ala Glu Asp Asp Gln Asp Asp Asp Glu Glu Glu Glu Gln Leu Pro

145 150 155 160

Gln Gln Arg Asn Gly Lys Lys Thr His Val Tyr Ala Gln Ala Pro Phe

165 170 175

Ala Gly Glu Ala Ile Asn Lys Asn Gly Leu Gln Ile Gly Thr Asn Gly

180 185 190

Ala Ala Thr Glu Gly Asn Lys Glu Ile Tyr Ala Asp Lys Thr Tyr Gln

195 200 205

Pro Glu Pro Gln Ile Gly Glu Ser Gln Trp Asn Glu Ala Glu Ser Ser

210 215 220

Val Ala Gly Gly Arg Val Leu Lys Lys Thr Thr Pro Met Lys Pro Cys

225 230 235 240

Tyr Gly Ser Tyr Ala Arg Pro Thr Asn Ser Asn Gly Gly Gln Gly Val

245 250 255

Met Val Glu Gln Asn Gly Lys Leu Glu Ser Gln Val Glu Met Gln Phe

260 265 270

Phe Ser Thr Ser Ser Gln Ala Glu Pro Asp Arg Ala His Tyr Asn Ile

275 280 285

Val Thr Phe Cys Cys Lys Cys Asp Gln Leu Leu Arg Arg Glu Val Tyr

290 295 300

Asp Phe Ala Phe Arg Asp Leu Ser Gly Gly Thr Pro Lys Leu Val Leu

305 310 315 320

Tyr Ser Glu Asp Val Asn Met Glu Thr Pro Asp Thr His Leu Ser Tyr

325 330 335

Lys Pro Gly Lys Ser Asp Asp Asn Ser Lys Ala Met Leu Gly Gln Gln

340 345 350

Ser Met Pro Asn Arg Pro Asn Tyr Ile Ala Phe Arg Asp Asn Phe Ile

355 360 365

Gly Leu Met Tyr Tyr Asn Ser Thr Gly Asn Met Gly Val Leu Ala Gly

370 375 380

Gln Ala Ser Gln Leu Asn Ala Val Val Asp Leu Gln Asp Arg Asn Thr

385 390 395 400

Glu Leu Ser Tyr Gln Leu Leu Leu Asp Ser Ile Gly Asp Arg Thr Arg

405 410 415

Tyr Phe Ser Met Trp Asn Gln Ala Val Asp Ser Tyr Asp Pro Asp Val

420 425 430

Arg Ile Ile Glu Asn His Gly Thr Glu Asp Glu Leu Pro Asn Tyr Cys

435 440 445

Phe Pro Leu Gly Gly Ile Gly Val Thr Asp Thr Tyr Gln Ala Ile Lys

450 455 460

Ala Thr Asn Gly Asn Gly Gly Ala Thr Thr Trp Ala Gln Asp Asn Thr

465 470 475 480

Phe Ala Glu Arg Asn Glu Ile Gly Val Gly Asn Asn Phe Ala Met Glu

485 490 495

Ile Asn Leu Asn Ala Asn Leu Trp Arg Asn Phe Leu Tyr Ser Asn Ile

500 505 510

Ala Leu Tyr Leu Pro Asp Lys Leu Lys Tyr Asn Pro Thr Asn Val Glu

515 520 525

Ile Ser Asp Asn Pro Asn Thr Tyr Asp Tyr Met Asn Lys Arg Val Val

530 535 540

Ala Pro Gly Leu Val Asp Cys Tyr Ile Asn Leu Gly Ala Arg Trp Ser

545 550 555 560

Leu Asp Tyr Met Asp Asn Val Asn Pro Phe Asn His His Arg Asn Ala

565 570 575

Gly Leu Arg Tyr Arg Ser Met Leu Leu Gly Asn Gly Arg Tyr Val Pro

580 585 590

Phe His Ile Gln Val Pro Gln Lys Phe Phe Ala Ile Lys Asn Leu Leu

595 600 605

Leu Leu Pro Gly Ser Tyr Thr Tyr Glu Trp Asn Phe Arg Lys Asp Val

610 615 620

Asn Met Val Leu Gln Ser Ser Leu Gly Asn Asp Leu Arg Val Asp Gly

625 630 635 640

Ala Ser Ile Lys Phe Asp Ser Ile Cys Leu Tyr Ala Thr Phe Phe Pro

645 650 655

Met Ala His Asn Thr Ala Ser Thr Leu Glu Ala Met Leu Arg Asn Asp

660 665 670

Thr Asn Asp Gln Ser Phe Asn Asp Tyr Leu Ser Ala Ala Asn Met Leu

675 680 685

Tyr Pro Ile Pro Ala Asn Ala Thr Asn Val Pro Ile Ser Ile Pro Ser

690 695 700

Arg Asn Trp Ala Ala Phe Arg Gly Trp Ala Phe Thr Arg Leu Lys Thr

705 710 715 720

Lys Glu Thr Pro Ser Leu Gly Ser Gly Tyr Asp Pro Tyr Tyr Thr Tyr

725 730 735

Ser Gly Ser Ile Pro Tyr Leu Asp Gly Thr Phe Tyr Leu Asn His Thr

740 745 750

Phe Lys Lys Val Ala Ile Thr Phe Asp Ser Ser Val Ser Trp Pro Gly

755 760 765

Asn Asp Arg Leu Leu Thr Pro Asn Glu Phe Glu Ile Lys Arg Ser Val

770 775 780

Asp Gly Glu Gly Tyr Asn Val Ala Gln Cys Asn Met Thr Lys Asp Trp

785 790 795 800

Phe Leu Val Gln Met Leu Ala Asn Tyr Asn Ile Gly Tyr Gln Gly Phe

805 810 815

Tyr Ile Pro Glu Ser Tyr Lys Asp Arg Met Tyr Ser Phe Phe Arg Asn

820 825 830

Phe Gln Pro Met Ser Arg Gln Val Val Asp Asp Thr Lys Tyr Lys Asp

835 840 845

Tyr Gln Gln Val Gly Ile Ile His Gln His Asn Asn Ser Gly Phe Val

850 855 860

Gly Tyr Leu Ala Pro Thr Met Arg Glu Gly Gln Ala Tyr Pro Ala Asn

865 870 875 880

Val Pro Tyr Pro Leu Ile Gly Lys Thr Ala Val Asp Ser Ile Thr Gln

885 890 895

Lys Lys Phe Leu Cys Asp Arg Thr Leu Trp Arg Ile Pro Phe Ser Ser

900 905 910

Asn Phe Met Ser Met Gly Ala Leu Thr Asp Leu Gly Gln Asn Leu Leu

915 920 925

Tyr Ala Asn Ser Ala His Ala Leu Asp Met Thr Phe Glu Val Asp Pro

930 935 940

Met Asp Glu Pro Thr Leu Leu Tyr Val Leu Phe Glu Val Phe Asp Val

945 950 955 960

Val Arg Val His Gln Pro His Arg Gly Val Ile Glu Thr Val Tyr Leu

965 970 975

Arg Thr Pro Phe Ser Ala Gly Asn Ala Thr Thr

980 985

<210> 50

<211> 959

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Hexon polypeptide

<400> 50

Met Ala Thr Pro Ser Met Met Pro Gln Trp Ser Tyr Met His Ile Ser

1 5 10 15

Gly Gln Asp Ala Ser Glu Tyr Leu Ser Pro Gly Leu Val Gln Phe Ala

20 25 30

Arg Ala Thr Glu Thr Tyr Phe Ser Leu Asn Asn Lys Phe Arg Asn Pro

35 40 45

Thr Val Ala Pro Thr His Asp Val Thr Thr Asp Arg Ser Gln Arg Leu

50 55 60

Thr Leu Arg Phe Ile Pro Val Asp Arg Glu Asp Thr Ala Tyr Ser Tyr

65 70 75 80

Lys Ala Arg Phe Thr Leu Ala Val Gly Asp Asn Arg Val Leu Asp Met

85 90 95

Ala Ser Thr Tyr Phe Asp Ile Arg Gly Val Leu Asp Arg Gly Pro Thr

100 105 110

Phe Lys Pro Tyr Ser Gly Thr Ala Tyr Asn Ala Leu Ala Pro Lys Gly

115 120 125

Ala Pro Asn Ser Cys Glu Trp Asp Glu Asp Asp Thr Gln Val Gln Val

130 135 140

Ala Ala Glu Asp Asp Gln Asp Asp Asp Glu Glu Glu Glu Gln Leu Pro

145 150 155 160

Gln Gln Arg Asn Gly Lys Lys Thr His Val Tyr Ala Gln Ala Pro Phe

165 170 175

Ala Gly Glu Ala Ile Asn Lys Asn Gly Leu Gln Ile Gly Thr Asn Gly

180 185 190

Ala Ala Thr Glu Gly Asn Lys Glu Ile Tyr Ala Asp Lys Thr Tyr Gln

195 200 205

Pro Glu Pro Gln Ile Gly Glu Ser Gln Trp Asn Glu Ala Glu Ser Ser

210 215 220

Val Ala Gly Gly Arg Val Leu Lys Lys Thr Thr Pro Met Lys Pro Cys

225 230 235 240

Tyr Gly Ser Tyr Ala Arg Pro Thr Asn Ser Asn Gly Gly Gln Gly Val

245 250 255

Met Val Glu Gln Asn Gly Lys Leu Glu Ser Gln Val Glu Met Gln Phe

260 265 270

Phe Ser Thr Ser Val Asn Ala Met Asn Glu Ala Asn Ala Ile Gln Pro

275 280 285

Lys Leu Val Leu Tyr Ser Glu Asp Val Asn Met Glu Thr Pro Asp Thr

290 295 300

His Leu Ser Tyr Lys Pro Gly Lys Ser Asp Asp Asn Ser Lys Ala Met

305 310 315 320

Leu Gly Gln Gln Ser Met Pro Asn Arg Pro Asn Tyr Ile Ala Phe Arg

325 330 335

Asp Asn Phe Ile Gly Leu Met Tyr Tyr Asn Ser Thr Gly Asn Met Gly

340 345 350

Val Leu Ala Gly Gln Ala Ser Gln Leu Asn Ala Val Val Asp Leu Gln

355 360 365

Asp Arg Asn Thr Glu Leu Ser Tyr Gln Leu Leu Leu Asp Ser Ile Gly

370 375 380

Asp Arg Thr Arg Tyr Phe Ser Met Trp Asn Gln Ala Val Asp Ser Tyr

385 390 395 400

Asp Pro Asp Val Arg Ile Ile Glu Asn His Gly Thr Glu Asp Glu Leu

405 410 415

Pro Asn Tyr Cys Phe Pro Leu Gly Gly Ile Gly Val Thr Asp Thr Tyr

420 425 430

Gln Ala Ile Lys Ala Thr Asn Gly Asn Gly Gly Ala Thr Thr Trp Ala

435 440 445

Gln Asp Asn Thr Phe Ala Glu Arg Asn Glu Ile Gly Val Gly Asn Asn

450 455 460

Phe Ala Met Glu Ile Asn Leu Asn Ala Asn Leu Trp Arg Asn Phe Leu

465 470 475 480

Tyr Ser Asn Ile Ala Leu Tyr Leu Pro Asp Lys Leu Lys Tyr Asn Pro

485 490 495

Thr Asn Val Glu Ile Ser Asp Asn Pro Asn Thr Tyr Asp Tyr Met Asn

500 505 510

Lys Arg Val Val Ala Pro Gly Leu Val Asp Cys Tyr Ile Asn Leu Gly

515 520 525

Ala Arg Trp Ser Leu Asp Tyr Met Asp Asn Val Asn Pro Phe Asn His

530 535 540

His Arg Asn Ala Gly Leu Arg Tyr Arg Ser Met Leu Leu Gly Asn Gly

545 550 555 560

Arg Tyr Val Pro Phe His Ile Gln Val Pro Gln Lys Phe Phe Ala Ile

565 570 575

Lys Asn Leu Leu Leu Leu Pro Gly Ser Tyr Thr Tyr Glu Trp Asn Phe

580 585 590

Arg Lys Asp Val Asn Met Val Leu Gln Ser Ser Leu Gly Asn Asp Leu

595 600 605

Arg Val Asp Gly Ala Ser Ile Lys Phe Asp Ser Ile Cys Leu Tyr Ala

610 615 620

Thr Phe Phe Pro Met Ala His Asn Thr Ala Ser Thr Leu Glu Ala Met

625 630 635 640

Leu Arg Asn Asp Thr Asn Asp Gln Ser Phe Asn Asp Tyr Leu Ser Ala

645 650 655

Ala Asn Met Leu Tyr Pro Ile Pro Ala Asn Ala Thr Asn Val Pro Ile

660 665 670

Ser Ile Pro Ser Arg Asn Trp Ala Ala Phe Arg Gly Trp Ala Phe Thr

675 680 685

Arg Leu Lys Thr Lys Glu Thr Pro Ser Leu Gly Ser Gly Tyr Asp Pro

690 695 700

Tyr Tyr Thr Tyr Ser Gly Ser Ile Pro Tyr Leu Asp Gly Thr Phe Tyr

705 710 715 720

Leu Asn His Thr Phe Lys Lys Val Ala Ile Thr Phe Asp Ser Ser Val

725 730 735

Ser Trp Pro Gly Asn Asp Arg Leu Leu Thr Pro Asn Glu Phe Glu Ile

740 745 750

Lys Arg Ser Val Asp Gly Glu Gly Tyr Asn Val Ala Gln Cys Asn Met

755 760 765

Thr Lys Asp Trp Phe Leu Val Gln Met Leu Ala Asn Tyr Asn Ile Gly

770 775 780

Tyr Gln Gly Phe Tyr Ile Pro Glu Ser Tyr Lys Asp Arg Met Tyr Ser

785 790 795 800

Phe Phe Arg Asn Phe Gln Pro Met Ser Arg Gln Val Val Asp Asp Thr

805 810 815

Lys Tyr Lys Asp Tyr Gln Gln Val Gly Ile Ile His Gln His Asn Asn

820 825 830

Ser Gly Phe Val Gly Tyr Leu Ala Pro Thr Met Arg Glu Gly Gln Ala

835 840 845

Tyr Pro Ala Asn Val Pro Tyr Pro Leu Ile Gly Lys Thr Ala Val Asp

850 855 860

Ser Ile Thr Gln Lys Lys Phe Leu Cys Asp Arg Thr Leu Trp Arg Ile

865 870 875 880

Pro Phe Ser Ser Asn Phe Met Ser Met Gly Ala Leu Thr Asp Leu Gly

885 890 895

Gln Asn Leu Leu Tyr Ala Asn Ser Ala His Ala Leu Asp Met Thr Phe

900 905 910

Glu Val Asp Pro Met Asp Glu Pro Thr Leu Leu Tyr Val Leu Phe Glu

915 920 925

Val Phe Asp Val Val Arg Val His Gln Pro His Arg Gly Val Ile Glu

930 935 940

Thr Val Tyr Leu Arg Thr Pro Phe Ser Ala Gly Asn Ala Thr Thr

945 950 955

<210> 51

<211> 962

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Hexon polypeptide

<400> 51

Met Ala Thr Pro Ser Met Met Pro Gln Trp Ser Tyr Met His Ile Ser

1 5 10 15

Gly Gln Asp Ala Ser Glu Tyr Leu Ser Pro Gly Leu Val Gln Phe Ala

20 25 30

Arg Ala Thr Glu Thr Tyr Phe Ser Leu Asn Asn Lys Phe Arg Asn Pro

35 40 45

Thr Val Ala Pro Thr His Asp Val Thr Thr Asp Arg Ser Gln Arg Leu

50 55 60

Thr Leu Arg Phe Ile Pro Val Asp Arg Glu Asp Thr Ala Tyr Ser Tyr

65 70 75 80

Lys Ala Arg Phe Thr Leu Ala Val Gly Asp Asn Arg Val Leu Asp Met

85 90 95

Ala Ser Thr Tyr Phe Asp Ile Arg Gly Val Leu Asp Arg Gly Pro Thr

100 105 110

Phe Lys Pro Tyr Ser Gly Thr Ala Tyr Asn Ala Leu Ala Pro Lys Gly

115 120 125

Ala Pro Asn Ser Cys Glu Trp Glu Gln Asn Glu Thr Ala Gln Val Asp

130 135 140

Ala Gln Glu Leu Asp Glu Glu Glu Asn Glu Ala Asn Glu Ala Gln Ala

145 150 155 160

Arg Glu Gln Glu Gln Ala Lys Lys Thr His Val Tyr Ala Gln Ala Pro

165 170 175

Leu Ser Gly Glu Ala Ile Asn Lys Asn Gly Leu Gln Ile Gly Thr Asn

180 185 190

Gly Ala Ala Thr Glu Gly Asn Lys Glu Ile Tyr Ala Asp Lys Thr Tyr

195 200 205

Gln Pro Glu Pro Gln Ile Gly Glu Ser Gln Trp Asn Glu Ala Glu Ser

210 215 220

Ser Val Ala Gly Gly Arg Val Leu Lys Lys Thr Thr Pro Met Lys Pro

225 230 235 240

Cys Tyr Gly Ser Tyr Ala Arg Pro Thr Asn Ser Asn Gly Gly Gln Gly

245 250 255

Val Met Val Glu Gln Asn Gly Lys Leu Glu Ser Gln Val Glu Met Gln

260 265 270

Phe Phe Ser Thr Ser Val Asn Ala Met Asn Glu Ala Asn Ala Ile Gln

275 280 285

Pro Lys Leu Val Leu Tyr Ser Glu Asp Val Asn Met Glu Thr Pro Asp

290 295 300

Thr His Leu Ser Tyr Lys Pro Gly Lys Ser Asp Asp Asn Ser Lys Ala

305 310 315 320

Met Leu Gly Gln Gln Ser Met Pro Asn Arg Pro Asn Tyr Ile Ala Phe

325 330 335

Arg Asp Asn Phe Ile Gly Leu Met Tyr Tyr Asn Ser Thr Gly Asn Met

340 345 350

Gly Val Leu Ala Gly Gln Ala Ser Gln Leu Asn Ala Val Val Asp Leu

355 360 365

Gln Asp Arg Asn Thr Glu Leu Ser Tyr Gln Leu Leu Leu Asp Ser Ile

370 375 380

Gly Asp Arg Thr Arg Tyr Phe Ser Met Trp Asn Gln Ala Val Asp Ser

385 390 395 400

Tyr Asp Pro Asp Val Arg Ile Ile Glu Asn His Gly Thr Glu Asp Glu

405 410 415

Leu Pro Asn Tyr Cys Phe Pro Leu Gly Gly Ile Gly Ile Thr Asp Thr

420 425 430

Phe Gln Ala Val Lys Thr Thr Ala Ala Asn Gly Asp Gln Gly Asn Thr

435 440 445

Thr Trp Gln Lys Asp Ser Thr Phe Ala Glu Arg Asn Glu Ile Gly Val

450 455 460

Gly Asn Asn Phe Ala Met Glu Ile Asn Leu Asn Ala Asn Leu Trp Arg

465 470 475 480

Asn Phe Leu Tyr Ser Asn Ile Ala Leu Tyr Leu Pro Asp Lys Leu Lys

485 490 495

Tyr Asn Pro Thr Asn Val Glu Ile Ser Asp Asn Pro Asn Thr Tyr Asp

500 505 510

Tyr Met Asn Lys Arg Val Val Ala Pro Gly Leu Val Asp Cys Tyr Ile

515 520 525

Asn Leu Gly Ala Arg Trp Ser Leu Asp Tyr Met Asp Asn Val Asn Pro

530 535 540

Phe Asn His Pro Arg His Ala Gly Leu Arg Tyr Arg Ser Met Leu Leu

545 550 555 560

Gly Asn Gly Arg Tyr Val Pro Phe His Ile Gln Val Pro Gln Lys Phe

565 570 575

Phe Ala Ile Lys Asn Leu Leu Leu Leu Pro Gly Ser Tyr Thr Tyr Glu

580 585 590

Trp Asn Phe Arg Lys Asp Val Asn Met Val Leu Gln Ser Ser Leu Gly

595 600 605

Asn Asp Leu Arg Val Asp Gly Ala Ser Ile Lys Phe Asp Ser Ile Cys

610 615 620

Leu Tyr Ala Thr Phe Phe Pro Met Ala His Asn Thr Ala Ser Thr Leu

625 630 635 640

Glu Ala Met Leu Arg Asn Asp Thr Asn Asp Gln Ser Phe Asn Asp Tyr

645 650 655

Leu Ser Ala Ala Asn Met Leu Tyr Pro Ile Pro Ala Asn Ala Thr Asn

660 665 670

Val Pro Ile Ser Ile Pro Ser Arg Asn Trp Ala Ala Phe Arg Gly Trp

675 680 685

Ala Phe Thr Arg Leu Lys Thr Lys Glu Thr Pro Ser Leu Gly Ser Gly

690 695 700

Tyr Asp Pro Tyr Tyr Thr Tyr Ser Gly Ser Ile Pro Tyr Leu Asp Gly

705 710 715 720

Thr Phe Tyr Leu Asn His Thr Phe Lys Lys Val Ala Ile Thr Phe Asp

725 730 735

Ser Ser Val Ser Trp Pro Gly Asn Asp Arg Leu Leu Thr Pro Asn Glu

740 745 750

Phe Glu Ile Lys Arg Ser Val Asp Gly Glu Gly Tyr Asn Val Ala Gln

755 760 765

Cys Asn Met Thr Lys Asp Trp Phe Leu Val Gln Met Leu Ala Asn Tyr

770 775 780

Asn Ile Gly Tyr Gln Gly Phe Tyr Ile Pro Glu Ser Tyr Lys Asp Arg

785 790 795 800

Met Tyr Ser Phe Phe Arg Asn Phe Gln Pro Met Ser Arg Gln Val Val

805 810 815

Asp Asp Thr Lys Tyr Lys Asp Tyr Gln Gln Val Gly Ile Ile His Gln

820 825 830

His Asn Asn Ser Gly Phe Val Gly Tyr Leu Ala Pro Thr Met Arg Glu

835 840 845

Gly Gln Ala Tyr Pro Ala Asn Val Pro Tyr Pro Leu Ile Gly Lys Thr

850 855 860

Ala Val Asp Ser Ile Thr Gln Lys Lys Phe Leu Cys Asp Arg Thr Leu

865 870 875 880

Trp Arg Ile Pro Phe Ser Ser Asn Phe Met Ser Met Gly Ala Leu Thr

885 890 895

Asp Leu Gly Gln Asn Leu Leu Tyr Ala Asn Ser Ala His Ala Leu Asp

900 905 910

Met Thr Phe Glu Val Asp Pro Met Asp Glu Pro Thr Leu Leu Tyr Val

915 920 925

Leu Phe Glu Val Phe Asp Val Val Arg Val His Gln Pro His Arg Gly

930 935 940

Val Ile Glu Thr Val Tyr Leu Arg Thr Pro Phe Ser Ala Gly Asn Ala

945 950 955 960

Thr Thr

<---

Claims (12)

1. A multivalent programmed death protein 1 (PD-L1) ligand binding compound comprising a plurality of amino acid chains, wherein each amino acid chain contains at least one programmed death protein 1 (PD-1) polypeptide, wherein each amino acid chain comprising at least at least one programmed death protein 1 (PD-1) polypeptide is associated with a recombinant adenovirus (Ad) and a plurality of amino acid chains are located on a recombinant Ad envelope polypeptide and wherein the recombinant Ad contains hexon Ad capsid polypeptides of strain Ad6 and at least one hypervariable region polypeptide ( HVR) hexon capsid from Ad strain Ad57. 2. The multivalent programmed death protein 1 (PD-L1) ligand binding compound according to claim 1, wherein the Ad capsid hexon polypeptides of strain Ad6 contain HVR polypeptides 1-7 from Ad strain Ad57. 3. The multivalent programmed death protein 1 (PD-L1) ligand binding compound according to claim 1, wherein the Ad capsid hexon polypeptides of strain Ad6 contain HVR 2-6 polypeptides from Ad strain Ad57. 4. The multivalent programmed death protein 1 (PD-L1) ligand binding compound according to claim 1, wherein the programmed death protein 1 (PD-1) polypeptide is human PD-1 or mouse PD-1. 5. The multivalent programmed death protein 1 (PD-L1) ligand binding compound of claim 1, wherein the recombinant Ad is further modified to contain heterologous polypeptides selected from targeting polypeptides, therapeutic polypeptides and antigens. 6. The multivalent programmed death protein 1 (PD-L1) ligand binding compound of claim 5, wherein the targeting polypeptide is selected from measles virus hemagglutinin (MVH) polypeptide, measles virus fusion polypeptide (MVF) and vesicular stomatitis virus glycoprotein (VSVG) polypeptide ). 7. The multivalent programmed death protein ligand 1 (PD-L1) binding compound according to claim 5, wherein the therapeutic polypeptide is selected from a 4-1BB ligand polypeptide (4-1BBL), an OX40 ligand polypeptide (OX40L), a CD40 ligand polypeptide (CD40L ) and granulocyte-macrophage colony-stimulating factor polypeptide (GM-CSF). 8. The multivalent programmed death protein ligand 1 (PD-L1) binding compound of claim 1, wherein the PD-1 polypeptide is fused to the vitamin K-dependent gamma-carboxyglutamine domain of a single-chain anti-factor X antibody polypeptide (GLA domain of the FX polypeptide). 9. The multivalent programmed death protein 1 (PD-L1) ligand binding compound of claim 1, wherein each amino acid chain comprises a targeting molecule selected from a measles virus hemagglutinin (MVH) polypeptide, a measles virus fusion (MVF) polypeptide, and a glycoprotein polypeptide vesicular stomatitis virus (VSVG). 10. The multivalent programmed death protein ligand 1 (PD-L1) binding compound of claim 1, wherein each amino acid chain comprises one or more therapeutic polypeptides selected from a 4-1BB ligand polypeptide (4-1BBL), an OX40 ligand polypeptide ( OX40L), CD40 ligand polypeptide (CD40L) and granulocyte-macrophage colony-stimulating factor (GM-CSF) polypeptide. 11. A pharmaceutical composition for the treatment or prevention of malignant neoplasms that are positive for PD-L1 expression, comprising a multivalent programmed death protein 1 (PD-L1) ligand binding compound according to claim 1 and a pharmaceutically acceptable carrier. 12. Use of the multivalent programmed death protein 1 (PD-L1) ligand binding compound according to any one of claims 1 to 10 in the preparation of a medicament for use in the treatment of a cancer in an individual in need thereof, wherein the cancer cells of said cancer are positive regarding PD-L1 expression.

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