Fusion proteins of antibodies, or single chain variable fragments thereof, targeting oncoproteins inside cells with cancer cell penetrating peptides, and uses thereof

Abstract

The present invention relates to: an antibody targeting a mutation of KRAS as an intracellular oncoprotein; or a fusion protein in which a cancer cell-penetrating peptide is linked to a single-chain variable fragment of the antibody by a genetic expression method or a chemical bonding method; and a tumor therapeutic use thereof. The fusion protein thus produced has the benefit of maximizing an anti-tumor or anticancer effect of the antibody or the single-chain variable fragment of the antibody targeting an intracellular oncoprotein or an oncomutant protein by effectively invading tumor cells.

Description

TECHNICAL FIELD

[0001] The present application relates to a fusion protein comprising an intracellular oncogenic protein targeting antibody or a single chain variable fragment thereof and a cancer cell penetrating peptide and uses thereof; more particularly, to a fusion protein in which a cancer cell penetrating peptide is linked to an antibody or a single chain variable fragment thereof targeting a mutation in an intracellular oncogenic protein KRAS by a genetic expression or chemical conjugation method; and uses thereof for treating a tumor. BACKGROUND

[0002] A tumor is caused by a genetic mutation in a normal cell due to activation of an oncogenic pathway or inhibition of a tumor suppressor pathway (1). Among various oncogenic factors, RAS mutant proteins are the most well-known oncogenic factors and occur in about 30% of all cancer patients. RAS mutant proteins bind to GTP and remain activated, leading to proliferation and growth of tumor cells. RAS can be classified into three types: HRAS, NRAS, and KRAS. KRAS mutations are mainly found in non-small cell lung cancer, colorectal cancer, and pancreatic cancer, HRAS mutations are found in bladder cancer, kidney cancer, and thyroid cancer, and NRAS mutations are mainly found in melanoma, hepatocellular carcinoma, and hematological malignancies (2). KRAS gene mutations account for 86% of cancers caused by RAS mutations, and are known to be found in more than 98% of pancreatic cancer, more than 53% of colorectal cancer, and more than 30% of lung adenocarcinoma (3).

[0003] KRAS mutations are known to be closely related to the resistance mechanism of anti-EGFR therapeutic agents such as gefitinib, erlotinib, and cetuximab targeting the epidermal growth factor receptor (EGFR). Even when EGFR inhibitory anticancer drugs are used, they have no effect at all in the state in which KRAS downstream of EGFR remains activated due to a mutation therein. In fact, after determining that a patient has no KRAS mutation, an anti-EGFR monoclonal therapeutic antibody can be administered in colorectal cancer.

[0004] Antibody drugs have been extensively studied as an anticancer treatment means. For example, trastuzumab is a monoclonal antibody against a Her2 protein and is used as a treatment method for breast cancer (4), and rituximab is a monoclonal antibody against CD20 and is used as a treatment method for B-cell malignant lymphoma (5). Antibodies have excellent inhibitory effects on targets, but because they cannot pass through a cell membrane due to their high molecular weight, it is difficult to obtain the inhibitory effects of antibodies when the target of the antibody is in a cell.

[0005] Prostate cancer is the third most common male cancer in the world, and in the United States, it is the most common male cancer and its cancer-specific mortality rate is second only to lung cancer (7). Local prostate cancer can be cured by surgery or radiotherapy, but chemical castration is mainly used as a standard treatment method for advanced or metastatic prostate cancer. After it was proven that prostate cancer is a testosterone-dependent cancer, artificial castration was established as the primary treatment method for advanced or metastatic prostate cancer, and castration therapy for various types of prostate cancer has been used in clinical practice (8). In metastatic prostate cancer, castration shows effects such as disease progression and alleviation of accompanying symptoms, but it is known that only 75% of patients consistently respond to treatment when castration is performed for more than 18 months. Castration shows a relatively good response at the beginning, but as time passes, prostate cancer cells do not undergo apoptosis due to androgen blockade, but are transformed into castration-resistant prostate cancer (CRPC) cells that are no longer responsive to hormone therapy (9).

[0006] CRPC is defined as a condition in which the testicles do not produce testosterone, so that the testosterone in the blood is reduced, and the prostate-specific antigen (PSA) does not decrease even after the administration of all drugs that act through the androgen receptor is stopped, and a significant increase in PSA or radiological progression occurs. If CRPC is not treated, the average survival period is less than 12 months, and through various treatments, the average survival period of metastatic prostate cancer patients is less than 3 years, and in the case of local invasive disease, the average survival period is only 4.5 years (10).

[0007] It is known that the androgen receptor (AR) in CRPC is reactivated through various mechanisms such as AR overexpression, mutation, hypersensitivity, and androgen synthesis in tumors (11).

[0008] Abiraterone (an FDA-approved CRPC treatment drug) is an androgen synthesis inhibitor, and enzalutamide (MDV3100) (a second-generation androgen receptor antagonist) completely inhibits the binding of androgens to androgen receptors, thereby preventing the movement of androgen receptors into the nucleus and binding to target genes. Since most prostate cancers that relapse after abiraterone or enzalutamide treatment are PSA-positive and androgen receptors are reactivated, CRPC patients resistant to abiraterone or enzalutamide require new androgen receptor inhibition therapy (12). The androgen receptor, a member of the steroid receptor superfamily, is a ligand-dependent transcription factor that regulates the expression of genes affected by androgens. In order for the transcription factor to enter the target gene, the transcription factor must enter the nucleus, so if the transcription factor can be maintained only in the cytoplasm, the transcription function can be blocked. Therefore, the most important step is to control the transport of androgen receptors to the nucleus. Androgen-sensitive receptors remain in the cytoplasm in the absence of androgens, but enter the nucleus and activate target genes in the presence of androgens. However, in CRPC cells, androgen receptors continue to activate target genes because they remain in the nucleus even in the absence of androgens (13). Therefore, if any substance can inhibit the transport of androgen receptors to the nucleus, it can be used as an effective therapeutic drug for CRPC tumors.

[0009] It is known that oncogenic proteins such as KRAS and androgen receptors play an important role in the development of human cancer, but antibody therapeutics targeting these oncogenic proteins have not been clinically used. The reason is that antibodies against RAS and androgen receptors distributed in cells cannot pass through the cell membrane due to their high molecular weight. It is known that a single-chain variable fragment (scFv) smaller than an antibody has a smaller molecular weight than an antibody and does not have an Fc region that causes immune rejection, so it can be a substitute for a general antibody. However, since the scFv itself has a molecular weight of 25 kDa or more, unlike low-molecular-weight drugs, the scFv is difficult to freely pass through the cell membrane, and thus it is difficult to obtain a large inhibitory effect as expected.

[0010] Therefore, the present inventors made a great effort to solve the above-described problems existing in the related art, and as a result, have tried to fuse an antibody or a single-chain variable fragment thereof against an intracellular oncogenic protein with a cancer cell-penetrating functional peptide so as to be able to penetrate cancer tissues. As a fusion method, a genetic expression method or a chemical conjugation method was used, and the present inventors have found through in vitro experiments and animal experiments that a cell-penetrating fusion protein produced by this method can effectively inhibit the proliferation and growth of cancer cells highly expressing an oncogenic protein, thereby completing the present invention.

[0011] Related Art

[0012] Non-Patent Literature

[0013] (Zhang J, Chen YH, Lu Q. Pro-oncogenic and anti-oncogenic pathways: opportunities and challenges of cancer therapy. Future Oncol. 2010 Apr;6(4):587-603.)

[0014] (Cox AD, Fesik SW, Kimmelman AC, Luo J, Der CJ. Drugging the undruggable RAS: mission possible? Nat Rev Drug Discov 2014; 13: 828-51.)

[0015] (RAS oncogenes: weaving a tumorigenic web. Pylayeva-Gupta Y, Grabocka E, Bar-Sagi D. Nat Rev Cancer. 2011 Oct 13; 11(11):761-74.)

[0016] (Trastuzumab (herceptin) for the medical treatment of breast cancer. Bayoudh L, Afrit M, Daldoul O, Zarrad M, Boussen H. Tunis Med. 2012 Jan; 90(1):6-12.)

[0017] (Evolution of anti-CD20 monoclonal antibody therapeutics in oncology. Oflazoglu E, Audoly LP. MAbs. 2010 Jan-Feb; 2(1): 14-9.)

[0018] (Non-patent document 6) Demarest SJ, Glaser SM. Antibody therapeutics, antibody engineering, and the merits of protein stability. Curr Opin Drug Discov Devel. 2008; 11(5): 675-687.

[0019] (Non-patent document 7) Jemal A, Clegg LX, Ward E, Ries LA, Wu X, Jamison PM et al. Annual report to the nation on the status of cancer, 1975-2001, with a special feature regarding survival. Cancer 2004; 101: 3-27.

[0020] (Non-patent document 8) Yagoda A, Petrylak D. Cytotoxic chemotherapy for advanced hormone-resistant prostate cancer. Cancer 1993; 71: 1098.

[0021] (Non-patent document 9) Rini BI, Small EJ. Hormone-refractory prostate cancer. Curr Treat Opt Oncol 2002; 3: 437-46.

[0022] (Non-patent document 10) Sella A, Yarom N, Zisman A, Kovel S. Paclitaxel, estramustine and carboplatin combination chemotherapy after initial docetaxel-based chemotherapy in castration resistant prostate cancer. Oncology 2009; 76: 442.

[0023] (Non-patent document 11) Chen CD, Welsbie DS, Tran C, Baek SH, Chen R, Vessella R, et al. Molecular determinants of resistance to antiandrogen therapy. Nat Med 2004;10:33-9.

[0024] (Non-patent document 12) Boudadi K, Antonarakis ES. Resistance to novel antiandrogen therapies in metastatic castration-resistant prostate cancer. Clin Med Insights Oncol 2016;10:1-9.

[0025] (Non-patent document 13) Zhang L, Johnson M, Le KH, Sato M, Ilagan R, Iyer M, et al. Interrogating androgen receptor function in recurrent prostate cancer. Cancer Res 2003;63:4552-60.

[0026] (Non-patent document 14) Lars Kober, Christoph Zehe, Juergen Bode, Biotechnology and Bioengineering, Volume 110, Issue 4, April 2013, Optimized signal peptides for the development of high expressing CHO cell lines. SUMMARY

[0027] An object of the present application is to provide a fusion protein in which the therapeutic effect on tumors is significantly improved.

[0028] Another object of the present application is to provide a pharmaceutical composition for treating tumors, which comprises the fusion protein as an active ingredient.

[0029] Still another object of the present application is to provide a method for preventing or treating tumors, which comprises the step of administering the fusion protein.

[0030] Still another object of the present application is to provide use of the fusion protein for preventing or treating a tumor.

[0031] Still another object of the present application is to provide use of the fusion protein in the manufacture of a medicament for treating a tumor.

[0032] To achieve the above object, the present application provides a fusion protein in which (i) an antibody or a single chain variable fragment thereof targeting an intracellular oncogenic protein or oncogenic mutant protein is linked to (ii) a cancer cell penetrating peptide.

[0033] The present application also provides a nucleic acid encoding the fusion protein.

[0034] The present application also provides a recombinant vector into which the nucleic acid has been introduced.

[0035] The present application also provides a recombinant cell into which the recombinant vector has been introduced.

[0036] The present application also provides a method for producing the fusion protein, the method comprising the steps of:

[0037] (a) expressing the fusion protein by culturing the recombinant cell; and

[0038] (b) recovering the expressed fusion protein.

[0039] The present application also provides a pharmaceutical composition for treating a tumor, the pharmaceutical composition comprising the fusion protein as an active ingredient.

[0040] The present application also provides a method for preventing or treating a tumor, the method comprising administering the fusion protein.

[0041] The present application also provides use of the fusion protein for preventing or treating a tumor.

[0042] The present application also provides use of the fusion protein in the manufacture of a medicament for treating a tumor. BRIEF DESCRIPTION OF DRAWINGS

[0043] Figure 1 is a schematic diagram showing an outline of the present application.

[0044] Figure 2 Expression vectors are shown, each of which comprises a cancer cell penetrating KRAS mutant antibody or scFv.

[0045] FIG. 3 shows the results of SDS-PAGE and immunoblot analysis of cancer cell penetrating KRAS mutant antibodies and scFv after expression and purification.

[0046] Figure 4Results of SDS-PAGE and immunoblot analysis of cancer cell-penetrating KRAS mutant antibody and scFv produced by chemical conjugation method are shown.

[0047] Fig. 5 shows results of evaluation of cancer cell-specific killing effect of cancer cell-penetrating KRAS mutant antibody and scFv.

[0048] Figure 6 Results of immunoblot analysis for evaluation of active KRAS (mutant KRAS) expression inhibitory activity of SEQ ID NOs: 20 to 23, each connected by GGGGS linker, are shown.

[0049] Figure 7 Results of immunoblot analysis for evaluation of active KRAS (mutant KRAS) expression inhibitory activity of SEQ ID NOs: 90 to 93, each connected by GGGGSGGGGSGGGGS linker, are shown.

[0050] Figure 8 Results of analysis of the ability of SEQ ID NOs: 20 to 23 to penetrate cancer cells are shown.

[0051] Figure 9 Results of evaluation of the distribution of SEQ ID NOs: 2 and 23 in cancer tissues in a tumor xenograft animal model are shown.

[0052] Figure 10 Fluorescence intensity of cancer tissues in a tumor xenograft animal model is shown.

[0053] Figure 11 Tumor inhibitory effect of SEQ ID NOs: 2 and 23 is shown.

[0054] Figure 12 Changes in tumor volume formed by H358 after euthanasia and images of cancer tissues are shown.

[0055] Figure 13 Results of immunoblot analysis for evaluation of active KRAS (mutant KRAS) expression inhibitory activity of SEQ ID NOs: 80 to 83 produced by chemical conjugation are shown.

[0056] Figure 14 Results of evaluation of cancer cell-specific killing effect of SEQ ID NOs: 80 to 83 produced by chemical conjugation are shown.

[0057] Figure 15 Results of evaluation of cancer cell-penetrating ability of SEQ ID NOs: 80 to 83 produced by chemical conjugation are shown.

[0058] Figure 16 Results showing evaluation of distribution to cancer tissue in tumor xenograft animal models are shown.

[0059] Figure 17 Fluorescence intensity of cancer tissue in tumor xenograft animal models is shown.

[0060] Figure 18 Tumor suppression of SEQ ID NO: 3 and 83 is shown.

[0061] Figure 19 Changes in tumor volume and images of cancer tissue formed by H358 after euthanasia are shown. DETAILED DESCRIPTION

[0062] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Generally, the term used in the specification and the experimental methods described below are well known and commonly used in the art.

[0063] The present inventors noted the fact that even if antibodies or single chain variable fragments thereof targeting intracellular oncogenic proteins or oncogenic mutant proteins reach cancer cells, they cannot sufficiently act on cancer cells. Accordingly, the present inventors produced a fusion protein in which an antibody or single chain variable fragment thereof is linked to a cancer cell penetrating peptide. To produce the fusion protein, a linker is introduced into the N- or C-terminus of an antibody or single chain variable fragment thereof targeting intracellular oncogenic proteins or oncogenic mutant proteins, and a cancer cell penetrating peptide is linked thereto. This fusion protein is produced by either of two different methods: a genetic expression method and a chemical conjugation method. In a cancer cell killing experiment using the fusion protein produced as described above, it was confirmed that the fusion protein comprising a cancer cell penetrating peptide exhibited a significantly better cancer cell-specific action than the antibody or single chain variable fragment thereof targeting intracellular oncogenic proteins or oncogenic mutant proteins alone.

[0064] Accordingly, in one aspect, the present application relates to a fusion protein in which (i) an antibody or single chain variable fragment thereof targeting intracellular oncogenic proteins or oncogenic mutant proteins is linked to (ii) a cancer cell penetrating peptide.

[0065] In the present application, the intracellular oncogenic protein or oncogenic mutant protein can be KRAS or androgen receptor, but is not limited thereto.

[0066] In the present application, the antibody or single chain variable fragment thereof can be selected from SEQ ID NOs: 1 to 3, but is not limited thereto.

[0067] In one embodiment, the single chain variable fragment can be a KRAS mutant scFv represented by the following SEQ ID NO: 1.

[0068] SEQ ID NO: 1

[0069] MAWVWTLLFLMAAAQSIQA EWIKQRPGQGLEWIGVIHPNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSDIEL TQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK LEIKRGSENLYFQGGSGKPIPNPLLGLDSTGGSGGSHHHHHH

[0070] Here, MAWVWTLLFLMAAAQSIQA is the signal peptide, and the amino acid sequence following the signal peptide is a variable heavy chain (V... H The region GGGGSGKGGSGGGGSGGGGS is the linker, and the amino acid sequence following the linker is a variable light chain (V). L The signal peptide can also be replaced by a known sequence (such as MTRLTVLALLAGLLASSRA or MKWVTFISLLFLFSSAYS).

[0071] In another embodiment, the single-stranded variable fragment may be a KRAS mutant scFv represented by the following SEQ ID NO: 2.

[0072] SEQ ID NO: 2

[0073] MAQVKLQESGPELVRPGTSVKVSCKASGYAFTNYLI EWIKQRPGQGLEWIGVIHPNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGGGGSGGGGSDIELTQ SPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK LEIKRLEHHHHHH

[0074] MAQVKLQESGPELVRPGTSVKVSCKASGYAFTNYLI is a signal peptide, and the amino acid sequence following the signal peptide is a variable heavy chain (V...). Hregion, GGGGS is a linker, and the amino acid sequence after the linker is a variable heavy chain (V L ) region.

[0075] In another embodiment, the single chain variable fragment can be a KRAS mutant scFv represented by the following SEQ ID NO: 3.

[0076] SEQ ID NO: 3

[0077] MAWVWTLLFLMAAAQSIQA EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKC LEIKRGSENLYFQGGSGKPIPNPLLGLDSTGGSGGSHHHHHH

[0078] SEQ ID NO: 3 is identical to SEQ ID NO: 1, except that a cysteine is introduced after V L .

[0079] In the present application, the cancer cell penetrating peptide can be selected from the group consisting of CCPP1 (H4K), CCPP2 (H4P), CCPP3 (LMWP), and CCPP4 (hBD3-3), but is not limited thereto. The cancer cell penetrating peptide can be selected from the group consisting of SEQ ID NO: 86 to SEQ ID NO: 89, but is not limited thereto.

[0080] In the present application, the cancer cell penetrating peptide can be connected to the N-terminus or C-terminus of the antibody or the single chain variable fragment thereof via a linker.

[0081] In the present application, the linker serves to provide a space so that the cancer cell penetrating peptide and the antibody or the single chain variable fragment thereof can individually form a functional structure when connected together. The linker is preferably a peptide linker comprising an appropriate combination of amino acids G and S. Here, more preferably, the number of amino acid residues G is from 3 to 20, and the number of amino acid residues S is from 1 to 5, but the present application is not limited thereto. Most preferably, the linker is GGGGS or GGGGSGGGGSGGGGS.

[0082] In the present application, when the linker and the cancer cell penetrating peptide are linked together, they can be represented by the amino acid sequence of any one of SEQ ID NO: 4 to SEQ ID NO: 11, but are not limited thereto. In this case, the fusion peptide represented by any one of the amino acid sequences of SEQ ID NO: 4 to 7 is preferably linked to the C-terminus of the single chain variable fragment, and the fusion peptide represented by the amino acid sequence of any one of SEQ ID NO: 8 to 11 is preferably linked to the N-terminus of the single chain variable fragment. In another embodiment, the fusion peptide represented by the amino acid sequence of any one of SEQ ID NO: 4 to 11 can be further bound to the linker connecting the heavy chain and the light chain of the single chain fragment of the antibody. On the other hand, when the fusion peptide represented by any one of the amino acid sequences of SEQ ID NO: 4 to 11 is linked to the antibody, it is preferably linked to the sugar structure present in the Fc region.

[0083] In the present application, the antibody or the single chain variable fragment thereof and the cancer cell penetrating peptide can be linked together by a chemical conjugation method. In this case, the fusion protein can be represented by the amino acid sequence of any one of SEQ ID NO: 68 to 83, but is not limited thereto.

[0084] In the present application, the fusion protein can be a fusion protein in which the cancer cell penetrating peptide selected from any one of SEQ ID NO: 86 to 89 is linked to the lysine or cysteine residue of the single chain variable fragment selected from any one of SEQ ID NO: 1 to 3 via a linker.

[0085] In the present application, the linker can be CGGGGG or CGGGGGSSGGGGG.

[0086] In the present application, the antibody or the single chain variable fragment thereof and the cancer cell penetrating peptide can be linked together by a gene expression method and expressed. In this case, the fusion protein can be expressed in Escherichia coli (E. coli) Escherichia coli ) and can be represented by the amino acid sequence of any one of SEQ ID NO: 12 to 27, but is not limited thereto. In addition, the fusion protein can be expressed in mammalian cells and can be represented by the amino acid sequence of any one of SEQ ID NO: 28 to SEQ ID NO: 59 and SEQ ID NO: 90 to SEQ ID NO: 93, but is not limited thereto.

[0087] For example, the cancer cell-penetrating peptide and the linker can be connected to the N-terminus of the single-chain variable fragment (any one of SEQ ID NO: 1 to SEQ ID NO: 3) targeting the KRAS mutant protein, or the cancer cell-penetrating peptide and the linker can be connected to the N-terminus of the single-chain variable fragment. Various embodiments related thereto are specifically described in Tables 2 to 5 in the following Examples.

[0088] In the present application, the fusion protein can be represented by any one of the amino acid sequences of SEQ ID NO: 12 to SEQ ID NO: 59, SEQ ID NO: 68 to SEQ ID NO: 83, and SEQ ID NO: 90 to SEQ ID NO: 93.

[0089] In the present application, the fusion protein can be produced by a gene expression method or a chemical conjugation method, but the production method is not limited thereto.

[0090] When the fusion protein is produced by the gene expression method, a vector that expresses both (i) an antibody or a single-chain variable fragment thereof targeting an intracellular oncogenic protein or an oncogenic mutant protein and (ii) a cancer cell-penetrating peptide can be used. In this case, the vector for expression can further contain a linker sequence between the antibody or the single-chain variable fragment thereof and the cancer cell-penetrating peptide. pET vectors, pcDNA 3.4, pcDNA3.1, pcDNA3.1-TOPO, pcDNA3.4-TOPO, pSecTag vectors, etc. can be used, but the vector is not limited thereto. In the present application, expression is performed in Escherichia coli using a pET vector, and expression is performed in mammalian cells using a pcDNA3.1-TOPO or pcDNA3.4-TOPO vector. E. coli ) and expression is performed in mammalian cells using a pcDNA3.1-TOPO or pcDNA3.4-TOPO vector.

[0091] Therefore, in another aspect, the present application relates to a nucleic acid encoding the fusion protein.

[0092] The present application also relates to a recombinant vector into which the nucleic acid has been introduced.

[0093] The present application also relates to a recombinant cell into which the recombinant vector has been introduced.

[0094] In the present application, the recombinant cell can be Escherichia coli or a mammalian cell, but is not limited thereto.

[0095] Examples of preferred mammalian cell lines that can be used in the present application include, but are not limited to, Chinese hamster ovary (CHO) cells, human embryonic kidney cells (HEK293), etc.

[0096] The present application also relates to a method for producing a fusion protein, the method comprising the steps of: (a) expressing a fusion protein by culturing a recombinant cell; and (b) recovering the expressed fusion protein.

[0097] On the other hand, in the present application, an antibody or a single chain variable fragment thereof targeting an intracellular oncogenic protein or an oncogenic mutant protein can be expressed and purified, and then a cancer cell penetrating peptide can be introduced thereto by a chemical conjugation method.

[0098] The cancer cell penetrating peptide can be linked to the antibody or the single chain variable fragment thereof via a linker and a cross-linking agent. As the linker, any linker can be used as long as it can provide a space capable of forming a functional structure. For example, the linker can be a peptide linker of natural and / or synthetic origin. The peptide linker of natural and / or synthetic origin can include an amino acid chain consisting of 1 to 50 amino acids, and can contain a repeated amino acid sequence or a sequence of a naturally occurring polypeptide such as a polypeptide having a hinge function. In another embodiment, the peptide linker amino acid sequence can be a "synthetic linker amino acid sequence" designated as being rich in glycine, glutamine, and / or serine residues. These residues can be arranged in a small repeating unit of up to five amino acids, and the small repeating unit can be repeated to form a multimeric unit. At the amino terminal end and / or the carboxyl terminal end of the multimeric unit, up to six additional arbitrary naturally occurring amino acids can be added. Other synthetic peptide linkers can consist of a single amino acid repeated between 10 to 20 times, and can contain up to six additional arbitrary naturally occurring amino acids at the amino terminal end and / or the carboxyl terminal end. On the other hand, the linker can be in a form in which the amino acid is chemically modified. For example, the linker can be in a form of Fmoc-6-aminohexanoic acid bound as an Fmoc-(9-fluorenylmethoxycarbonyl) blocking group, but the linker is not limited thereto.

[0099] In some embodiments of the present application, the fusion peptide in which the cancer cell penetrating peptide and the linker are linked together can be represented by the amino acid sequence of any one of SEQ ID NOs: 60 to 67. In this case, the thiol group of the cysteine located at the N-terminal end of the cancer cell penetrating peptide can be linked to the amine group of the lysine in the linker portion of SEQ ID NO: 1 or the amine group of the lysine in the exposed portion of the three-dimensional structure.

[0100] In this case, examples of the cross-linking agent that can be used include, but are not limited to, 1,4-bis-maleimidobutane (BMB), 1,11-bis-maleimidotetraethyleneglycol (BM[PEO]4), 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC), succinimidyl-4-[N-maleimidomethyl cyclohexane-1-carboxyl-[6-aminohexanoate]] (SMCC) and its sulfonate (sulfo-SMCC), 6-[3-(2-pyridyldithio)-propionamido]hexanoic acid succinimidyl ester (SPDP) and its sulfonate (sulfo-SPDP), m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS) and its sulfonate (sulfo-MBS), [4-(p-maleimidophenyl)butyric acid succinimidyl ester] (SMPB) and its sulfonate (sulfo-SMPB), and the like.

[0101] Further, the cross-linking agent that can be linked to the thiol of the cysteine at the N-terminus of the fusion peptide represented by any one of the amino acid sequences represented by SEQ ID NOs: 60 to 67 after reduction of the thiol of the cysteine at the C-terminus of SEQ ID NO: 3 can be tris(2-carboxyethyl)phosphine (TCEP), 5,50-dithiobis-(2-nitrobenzoic acid) (DTNB), or the like, but is not limited thereto.

[0102] In the present application, the method of producing a fusion protein using a chemical conjugation method includes the following steps:

[0103] (a) activating the amine group of a lysine present in a linker portion or an exposed portion of an antibody or a single chain variable fragment thereof that targets an intracellular oncogenic protein or an oncogenic mutant protein with a cross-linking agent;

[0104] (b) linking the cancer cell penetrating peptide to the antibody or the single chain variable fragment thereof activated with the cross-linking agent; and

[0105] (c) purifying the fusion protein in which the cancer cell penetrating peptide is linked to the antibody or the single chain variable fragment thereof.

[0106] Alternatively, step (a) of the above method can be a step of reducing the thiol of a cysteine residue of an antibody or a single chain variable fragment thereof that targets an intracellular oncogenic protein or an oncogenic mutant protein.

[0107] The fusion protein produced by the chemical conjugation method can be represented by any one of the amino acid sequences of SEQ ID NOs: 68 to 83, but is not limited thereto.

[0108] In one embodiment of the present application, the fusion protein can be one in which the cancer cell penetrating peptide is linked to a lysine or cysteine residue of an antibody or single chain variable fragment thereof targeting an intracellular oncogenic protein or oncogenic mutant protein via a cross-linking agent.

[0109] Preferably, the fusion protein can be one in which the fusion peptide selected from the group consisting of SEQ ID NOs: 60 to 67 is chemically bound to a lysine or cysteine residue of any one of the single chain variable fragments selected from the group consisting of SEQ ID NOs: 1 to 3 via a cross-linking agent.

[0110] In the case where the residue is lysine, the cross-linking agent can be selected from the group consisting of 1,4-bis-maleimidobutane (BMB), 1,11-bis-maleimidotetraethyleneglycol (BM[PEO]4), 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC), succinimidyl-4-[N-maleimidomethyl cyclohexane-1-carboxyl-[6-aminohexanoate]] (SMCC) and its sulfonate (sulfo-SMCC), 6-[3-(2-pyridyldithio)-propionamido]hexanoic acid succinimidyl ester (SPDP) and its sulfonate (sulfo-SPDP), m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS) and its sulfonate (sulfo-MBS), and [4-(p-maleimidophenyl)butyric acid succinimidyl ester] (SMPB) and its sulfonate (sulfo-SMPB).

[0111] On the other hand, when the residue is cysteine, the cross-linking agent can be tris(2-carboxyethyl)phosphine (TCEP) or 5,50-dithiobis-(2-nitrobenzoic acid) (DTNB).

[0112] KRAS mutant protein according to the present application is known as an important factor in tumorigenesis, which accelerates the growth of cancer cells, but is present inside the cell membrane, and thus an antibody or single chain variable fragment thereof targeting KRAS mutant protein does not have a significant inhibitory effect on KRAS mutant protein. However, as a result of evaluating the tumor growth inhibitory effect of KRAS mutant targeting antibody or single chain variable fragment thereof after introducing a cancer cell penetrating peptide therein, it was confirmed according to the results of in vitro experiments and animal experiments that the KRAS mutant targeting antibody or single chain variable fragment having cancer cell penetrating ability had a significant effect of inhibiting the growth of tumor cells.

[0113] Accordingly, in another aspect, the present application relates to a pharmaceutical composition for treating a tumor, the pharmaceutical composition comprising the fusion protein as an active ingredient.

[0114] In the present application, the tumor can be at least one selected from the group consisting of non-small cell lung cancer, colorectal cancer, pancreatic cancer, bladder cancer, kidney cancer, thyroid cancer, breast cancer, colon cancer, liver cancer, brain tumor, skin cancer, melanoma, colorectal cancer, prostate cancer, and blood cancer, but is not limited thereto.

[0115] In the present application, the pharmaceutical composition can be formulated in any one selected from the group consisting of a liquid formulation (e.g., for injection) such as an injection, a formulation for oral administration, an aqueous solution, a suspension, and an emulsion, a capsule, a granule, a tablet, and a formulation for mucosal administration, but is not limited thereto. These formulations can be prepared by a conventional method for formulation in the art or by a method disclosed in Remington's Pharmaceutical Science (latest edition), Mack Publishing Company, Easton Pa, and can vary depending on each disease or component.

[0116] On the other hand, the pharmaceutical composition of the present application can further comprise at least one pharmaceutically acceptable carrier in addition to the antibody or single chain variable fragment thereof having the ability to penetrate cancer cells and target KRAS mutant proteins. The pharmaceutically acceptable carrier can be at least one selected from the group consisting of saline, sterile water, Ringer's solution, buffered saline, a glucose solution, a maltodextrin solution, glycerol, ethanol, and a combination thereof.

[0117] If necessary, the pharmaceutical composition of the present application can further comprise a pharmaceutically acceptable adjuvant. The adjuvant can be one or more selected from the group consisting of an excipient, a diluent, a dispersant, a buffer, an antimicrobial preservative, a bacteriostatic agent, a surfactant, a binder, a lubricant, an antioxidant, a thickening agent, and a viscosity adjusting agent, but is not limited thereto.

[0118] The pharmaceutical composition according to the present application can be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally, or topically) according to the desired method, and its dose can vary depending on the body weight, age, sex, health status, and diet of the patient, the time of administration, the mode of administration, the excretion rate, and the severity of the disease and can be determined by an expert.

[0119] In one embodiment of the present application, the single dose of the peptide can be 1 μg / kg to 100 mg / kg, preferably 5 μg / kg to 50 mg / kg, and can be administered once a day or 1 to 3 times a week, but the dose and the frequency of administration are not limited thereto.

[0120] In another aspect, the present application relates to a method for preventing or treating a tumor, the method comprising the step of administering the fusion protein.

[0121] In another aspect, the present application relates to the use of the fusion protein for the prevention or treatment of a tumor.

[0122] In another aspect, the present application relates to the use of the fusion protein for the manufacture of a medicament for the treatment of a tumor.

[0123] EMBODIMENT

[0124] Hereinafter, the present application will be described in more detail with reference to examples. These examples are for the purpose of illustration only and it will be apparent to those of ordinary skill in the art that the scope of the present application should not be construed as being limited to these examples.

[0125] Example 1: Preparation of scFv penetrating cancer cells and targeting KRAS mutant by protein expression method

[0126] Example 1-1 : Expression of scFv penetrating cancer cells and targeting KRAS mutants in E. coli

[0127] KRAS mutant scFv expression vectors each containing a KRAS mutant scFv and a cancer cell penetrating peptide were prepared by a PCR method using specific primers. The KRAS mutant scFv used in the present application is any one of SEQ ID NOs: 1 to 3.

[0128] KRAS mutant scFv (SEQ ID NO: 1):

[0129] MAWVWTLLFLMAAAQSIQA EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK LEIKRGSENLYFQGGSGKPIPNPLLGLDSTGGSGGSHHHHHH

[0130] KRAS mutant scFv (SEQ ID NO: 2):

[0131] MAQVKLQESGPELVRPGTSVKVSCKASGYAFTNYLIEWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK LEIKRLEHHHHHH

[0132] KRAS mutant scFv (SEQ ID NO: 3):

[0133] MAWVWTLLFLMAAAQSIQA EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK C LEIKRGSENLYFQGGSGKPIPNPLLGLDSTGGSGGSHHHHHH

[0134] pET 21a(+) DNA was digested with Nde I and Xho I, and pMX DNA was also digested with Nde I and Xho I. The 767 bp Kras scFv gene fragment was recovered by the agarose electroelution method. To insert the gene fragment into the pET 21a(+) vector, the gene was amplified by PCR. PCR was performed using a forward primer (5'-AAGGAGATATACATATGATGGCATGGGTTTGGAC-3') and a reverse primer (5'-AGCCCGAAGGGAATTCATTTGCAGATACAAAGTGTTTTTAGAGTTG-3'). Each of the digested vector and insert were mixed together at a ratio of 0.5 μg : 1.0 μg, and ligated together using T4 DNA ligase in a ligation buffer containing 500 mM Tris-HCl, 100 mM MgCl2, 200 mM DTT, and 10 mM ATP at 4ºC for 16 to 18 hours.

[0135] The recombinant DNA solution was transformed into E. coli DH5a and plated on LB + Amp solid medium, and transformants were selected.

[0136] E. coli Rosetta TM 2(DE3) Singles TM (Novagen) strains were inoculated into 20 ml of LB containing 10 pg / ml of ampicillin antibiotic and incubated at 37°C overnight. One ml of culture was diluted in 200 ml of fresh LB containing the same concentration of antibiotic. Expression of KRAS scFv was induced by adding IPTG to the cells to a final concentration of 1 mM at an OD600 value of 0.8, followed by overnight incubation. The cells were harvested by centrifugation and lysed using 10 ml of lysis buffer (50 mM NaH2P04, 300 mM NaCl, pH 8.0). After sonication of the cells, the proteins were separated into total, soluble and insoluble fractions by centrifugation at 12,000 g and 4°C for 10 minutes, and analyzed by SDS-PAGE (S.I. Choi et al., Protein solubility and folding enhancement by interaction with RNA, PLoS ONE 3 (2008) e2677).

[0137] The isolated KRAS scFv was separated and purified by nickel chromatography (HisTrap TM Excel kit, GE Healthcare). Next, the nickel column was equilibrated with a buffer [50 mM NaH2P04 (pH 8.0), 300 mM NaCl], and then a protein aqueous solution in the same buffer was applied to the column. After washing with the above buffer containing 20 mM imidazole, the protein was separated using a buffer containing imidazole gradually increasing from 50 mM to 500 mM. The separated protein was collected and dialyzed using a desalting column (PD-10 column, GE Healthcare).

[0138] The linker and cancer cell penetrating peptide sequences used in the present application are shown in Table 1 below.

[0139] [Table 1]

[0140] Type and position of the linker Cancer cell penetrating peptide (CCPP) Amino acid sequences of the linker and the cancer cell penetrating peptide SEQ ID NO Linking part of the scFv N-terminal of the CCPP - GGGGS CCPP-1 (H4K) GGGGS-HRRCNKNNKKR 4 C-terminal CCPP-2 (H4P) GGGGS-HRRCNPNNKKR 5 C-terminal CCPP-3 (LMWP) GGGGS-VSRRRRRRGGRRRR 6 C-terminal CCPP-4 (hBD3-3) GGGGS-GKCSTRGRKCCRRKK 7 C-terminal C-terminal of the CCPP - GGGGS CCPP-1 (H4K) HRRCNKNNKKR-GGGGS 8 N-terminal CCPP-2 (H4P) HRRCNPNNKKR-GGGGS 9 N-terminal CCPP-3 (LMWP) VSRRRRRRGGRRRR-GGGGS 10 N-terminal CCPP-4 (hBD3-3) GKCSTRGRKCCRRKK-GGGGS 11 N-terminal

[0141] Table 2 below shows the amino acid sequences of fusion proteins for E. coli expression in which the cancer cell penetrating peptide and KRAS mutant scFv are linked together.

[0142] [Table 2]

[0143] Fusion protein Amino acid sequence SEQ ID NO SEQ ID NO 1 - GGGGS - H4K MAWVWTLLFLMAAAQSIQA EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKGGGGS - HRRCNKNNKKR - LEIKRGSENLYFQGGSGKPIPN PLLGLDSTGGSGGSHHHHHH ]]> ​ 12 SEQ ID NO 1 - GGGGS - H4P MAWVWTLLFLMAAAQSIQA EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKGGGGS HRRCNPNNKKR LEIKRGSENLYFQGGSGKPIPN PLLGLDSTGGSGGSHHHHHH ]]> ​ 13 SEQ ID NO 1 - GGGGS - LMWP MAWVWTLLFLMAAAQSIQA LEIKRGSENLYFQGGSGKPIPN PLLGLDSTGGSGGSHHHHHH MAWVWTLLFLMAAAQSIQA LEIKRG SENLYFQGGSGKPIPNPLLGLD STGGSGGSHHHHHH EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKGGGGSVSRRRRRRGGRRRRR SEQ ID NO 1 - GGGGS-hBD3-3 MAWVWTLLFLMAAAQSIQA LEIK RG SENLYFQGGSGKPIPNPLLGLD STGGSGGSHHHHHH MAWVWTLLFLMAAAQSIQA LEIK RG SENLYFQGGSGKPIPNPLLGLD STGGSGGSHHHHHH H4K-GGGGS-SEQ ID NO 1 MAWVWTLLFLMAAAQSIQA LEIK RG SENLYFQGGSGKPIPNPLLGLD STGGSGGSHHHHHH ]]> ​ 14 MAWVWTLLFLMAAAQSIQA LEIK RG SENLYFQGGSGKPIPNPLLGLD STGGSGGSHHHHHH H4P-GGGGS-SEQ ID NO 1 MAWVWTLLFLMAAAQSIQA LEIK RG SENLYFQGGSGKPIPNPLLGLD STGGSGGSHHHHHH EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKGGGGS-GKCSTRGRKCCRRKK- MAWVWTLLFLMAAAQSIQA LEIK RG SENLYFQGGSGKPIPNPLLGLD STGGSGGSHHHHHH LMWP-GGGGS-SEQ ID NO 1 MAWVWTLLFLMAAAQSIQA LEIK RG SENLYFQGGSGKPIPNPLLGLD STGGSGGSHHHHHH MAWVWTLLFLMAAAQSIQA LEIK RG SENLYFQGGSGKPIPNPLLGLD STGGSGGSHHHHHH ]]> ​ 15 hBD3-3-GGGGS-SEQ ID NO 1 MAWVWTLLFLMAAAQSIQA LEIK RG SENLYFQGGSGKPIPNPLLGLD STGGSGGSHHHHHH MAWVWTLLFLMAAAQSIQA LEIK RG SENLYFQGGSGKPIPNPLLGLD STGGSGGSHHHHHH - HRRCNKNNKKR - GGGGS - EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK SEQ ID NO 2 - GGGGS-H4K MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI LEIKRLEHHHHHH MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI LEIKRLEHHHHHH SEQ ID NO 2 - GGGGS-H4P MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI LEIKRLEHHHHHH ]]> ​ 16 MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI LEIKRLEHHHHHH SEQ ID NO 2 - GGGGS-LMWP MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI LEIKRLEHHHHHH - HRRCNPNNKKR - GGGGS - EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI LEIKRLEHHHHHH SEQ ID NO 2 - GGGGS-hBD3-3 MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI LEIKRLEHHHHHH MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI LEIKRLEHHHHHH ]]> ​ 17 H4K-GGGGS-SEQ ID NO 2 MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI LEIKRLEHHHHHH MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI LEIKRLEHHHHHH - GGGGS-EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK H4P-GGGGS-SEQ ID NO 2 MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI LEIKRLEHHHHHH MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI LEIKRLEHHHHHH LMWP-GGGGS-SEQ ID NO 2 MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI LEIKRLEHHHHHH ]]> ​ 18 MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI LEIKRLEHHHHHH hBD3-3-GGGGS-SEQ ID NO 2 MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI LEIKRLEHHHHHH -GKCSTRGRKCCRRKK -GGGGS -EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK ​ ​ ​ ]]> ​ 19 ​ ​ ​ EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKGGGGS-HRRCNKNNKKR - ​ ]]> ​ 20 ​ ​ ​ EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK - GGGGS -HRRCNPNNKKR - ​ ]]> ​ 21 ​ ​ ​ EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKGGGGS VSRRRRRRGGRRRRR ​ ]]> ​ 22 ​ ​ ​ EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKGGGGS -GKCSTRGRKCCRRKK - ​ ]]> ​ 23 ​ ​ ​ HRRCNKNNKKR - GGGGS -EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK ​ ​ ]]> ​ 24 ​ ​ ​ HRRCNPNNKKR - GGGGS -EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK ​ ​ ]]> ​ 25 ​ ​ ​ VSRRRRRRGGRRRR - GGGGS-EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK ​ ​ ]]> ​ 26 ​ MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI - LE IKRLEHHHHHHH Example 1-2: Expression of scFv penetrating cancer cells and targeting KRAS mutants in mammalian cells GKCSTRGRKCCRRKK -GGGGS -EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK Fusion protein Amino acid sequence ]]> ​ 27

[0144] Figure 3(A) shows the results of SDS-PAGE and immunoblot analysis of purified KRAS mutant-targeting antibodies or single-chain variable fragments of SEQ ID NOs: 1, 2, and 3.

[0145] SEQ ID NO

[0146] KRAS mutant scFv expression vectors (each containing a KRAS mutant scFv and a cancer cell-penetrating peptide) for use in mammalian cells were constructed by a PCR method using specific primers.

[0147] Table 3 below shows the amino acid sequences of fusion proteins in which a cancer cell-penetrating peptide is linked to a KRAS mutant scFv, and each is cloned into a pcDNA3.4-TOPO vector for mammalian cell expression.

[0148] [Table 3]

[0149] SEQ ID NO 1-GGGGS-H4K MAWVWTLLFLMAAAQSIQA LEIKRGSENLYFQGGSGKPIPN PLLGLDSTGGSGGSHHHHHH SEQ ID NO 1-GGGGS-H4P MAWVWTLLFLMAAAQSIQA LEIKRGSENLYFQGGSGKPIPN PLLGLDSTGGSGGSHHHHHH SEQ ID NO 1-GGGGS-LMWP EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKGGGGS - HRRCNKNNKKR - MAWVWTLLFLMAAAQSIQA LEIKRG SENLYFQGGSGKPIPNPLLGLD STGGSGGSHHHHHH SEQ ID NO 1-GGGGS-hBD3-3 ]]> ​ 28 MAWVWTLLFLMAAAQSIQA LEIK RGSENLYFQGGSGKPIPNPLLG LDSTGGSGGSHHHHHH H4K-GGGGS-SEQ ID NO 1 EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKGGGGS HRRCNPNNKKR MAWVWTLLFLMAAAQSIQA LEIKRGSENL YFQGGSGKPIPNPLLGLDSTGG SGGSHHHHHH H4P-GGGGS-SEQ ID NO 1 ]]> ​ 29 MAWVWTLLFLMAAAQSIQA LEIKRGSENL YFQGGSGKPIPNPLLGLDSTGG SGGSHHHHHH LMWP-GGGGS-SEQ ID NO 1 EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKGGGGSVSRRRRRRGGRRRRR MAWVWTLLFLMAAAQSIQA LEIKRGSE NLYFQGGSGKPIPNPLLGLDST GGSGGSHHHHHH hBD3-3-GGGGS-SEQ ID NO1 MAWVWTLLFLMAAAQSIQA LE IKRGSENLYFQGGSGKPIPNPL LGLDSTGGSGGSHHHHHH ]]> ​ 30 SEQ ID NO 2-GGGGS-H4K MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI LEIKRLEHHHHHH EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKGGGGS-GKCSTRGRKCCRRKK- SEQ ID NO 2-GGGGS-H4P MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI LEIKRLEHHHHHH SEQ ID NO 2-GGGGS-LMWP ]]> ​ 31 MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI LEIKRLEHHHHHH ​ - HRRCNKNNKKR - GGGGS - EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK ​ ​ ​ ]]> ​ 32 ​ ​ - HRRCNPNNKKR - GGGGS - EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK ​ ​ ​ ]]> ​ 33 ​ ​ - GGGGS-EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK ​ ​ ​ ]]> ​ 34 ​ ​ -GKCSTRGRKCCRRKK -GGGGS -EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK ​ ​ ​ ]]> ​ 35 ​ ​ ​ EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKGGGGS-HRRCNKNNKKR - ​ ]]> ​ 36 ​ ​ ​ EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKGGGGS-HRRCNPNNKKR - ​ ]]> ​ 37 ​ ​ ​ EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKGGGGS VSRRRRRRGGRRRRR ​ ]]> ​ 38 SEQ ID NO 2 - GGGGS-hBD3-3 MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKGGGGS -GKCSTRGRKCCRRKK - LEIKRLEHHHHHH ]]> ​ 39 H4K-GGGGS-SEQ ID NO 2 MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI - HRRCNKNNKKR - GGGGS - EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK LEIKRLEHHH HHH ]]> ​ 40 H4P-GGGGS-SEQ ID NO 2 MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI - HRRCNPNNKKR - GGGGS -EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK LEIKRLEHHH HHH ]]> ​ 41 LMWP-GGGGS-SEQ ID NO 2 MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI- VSRRRRRRGGRRRR - GGGGS-EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK LEIKRLEH HHHHH ]]> ​ 42 hBD3-3-GGGGS-SEQ ID NO2 MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI - GKCSTRGRKCCRRKK -GGGGS -EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK LE IKRLEHHHHHHH ]]> ​ 43

[0150] Table 4 below shows the amino acid sequences of fusion proteins in which a cancer cell-penetrating peptide is linked to a KRAS mutant scFv, and each is cloned into a pcDNA3.1-TOPO vector for mammalian cell expression.

[0151] [Table 4]

[0152] Fusion protein Amino acid sequence SEQ ID NO SEQ ID NO 1 - GGGGS-H4K MAWVWTLLFLMAAAQSIQA EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKGGGGS HRRCNKNNKKR LEIKRGSENLYFQGGSGKPIPN PLLGLDSTGGSGGSHHHHHH ]]> ​ 44 SEQ ID NO 1 - GGGGS-H4P MAWVWTLLFLMAAAQSIQA EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKGGGGS HRRCNPNNKKR LEIKRGSENLYFQGGSGKPIPN PLLGLDSTGGSGGSHHHHHH ]]> ​ 45 SEQ ID NO 1 - GGGGS-LMWP MAWVWTLLFLMAAAQSIQA EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKGGGGSVSRRRRRRGGRRRRR LEIKRG SENLYFQGGSGKPIPNPLLGLD STGGSGGSHHHHHH ]]> ​ 46 SEQ ID NO 1 - GGGGS-hBD3-3 MAWVWTLLFLMAAAQSIQA EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKGGGGS-GKCSTRGRKCCRRKK- LEIK RGSENLYFQGGSGKPIPNPLLG LDSTGGSGGSHHHHHH ]]> ​ 47 H4K-GGGGS-SEQ ID NO 1 MAWVWTLLFLMAAAQSIQA - HRRCNKNNKKR - GGGGS - EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK LEIKRGSENL YFQGGSGKPIPNPLLGLDSTGG SGGSHHHHHH ]]> ​ 48 H4P-GGGGS-SEQ ID NO 1 MAWVWTLLFLMAAAQSIQA - HRRCNPNNKKR - GGGGS - EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK LEIKRGSENL YFQGGSGKPIPNPLLGLDSTGG SGGSHHHHHH ]]> ​ 49 LMWP-GGGGS-SEQ ID NO 1 MAWVWTLLFLMAAAQSIQA - GGGGS-EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK LEIKRGSE NLYFQGGSGKPIPNPLLGLDST GGSGGSHHHHHH ]]> ​ 50 hBD3-3-GGGGS-SEQ ID NO1 MAWVWTLLFLMAAAQSIQA -GKCSTRGRKCCRRKK -GGGGS -EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK LE IKRGSENLYFQGGSGKPIPNPL LGLDSTGGSGGSHHHHHH ]]> ​ 51 SEQ ID NO 2-GGGGS-H4K MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKGGGGS-HRRCNKNNKKR - LEIKRLEHHHHHH ]]> ​ 52 SEQ ID NO 2-GGGGS-H4P <![CDATA[ MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK - GGGGS -HRRCNPNNKKR - LEIKRLEHHHHHH ]]> 53 SEQ ID NO 2-GGGGS-LMWP MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKGGGGS VSRRRRRRGGRRRRRR LEIKRLEHHHHHH ]]> ​ 54 SEQ ID NO 2-GGGGS-hBD3-3 MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKGGGGS -GKCSTRGRKCCRRKK - LEIKRLEHHHHHH ]]> ​ 55 H4K-GGGGS-SEQ ID NO 2 MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI - HRRCNKNNKKR - GGGGS -EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK LEIKRLEHHH HHH ]]> ​ 56 H4P-GGGGS-SEQ ID NO 2 MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI - HRRCNPNNKKR - GGGGS -EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK LEIKRLEHHH HHH ]]> ​ 57 LMWP-GGGGS-SEQ ID NO 2 MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI- VSRRRRRRGGRRRR - GGGGS-EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK LEIKRLEH HHHHH ]]> ​ 58 hBD3-3-GGGGS-SEQ ID NO2 MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI - GKCSTRGRKCCRRKK -GGGGS -EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGKGGSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK LE IKRLEHHHHHHH ]]> ​ 59

[0153] Table 5 below shows the amino acid sequences of fusion proteins in which a cancer cell-penetrating peptide is linked to a KRAS mutant scFv, and each is cloned into a pcDNA3.4-TOPO for mammalian cell expression.

[0154] [Table 5]

[0155] Fusion protein Amino acid sequence SEQ ID NO SEQ ID NO 2-GGGGSGGGGSGGGGS -H4K MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTKGGGGSGGGGSGGGGS -HRRCNKNNKKR - LEIKRLEHHHHHH ]]> ​ 90 SEQ ID NO 2-GGGGSGGGGSGGGGS -H4P MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK–GGGGSGGGGSGGGGS -HRRCNPNNKKR - LEIKRLEHHHHHH ]]> ​ 91 SEQ ID NO 2-GGGGSGGGGSGGGGS -LMWP MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK –GGGGSGGGGSGGGGS -VSRRRRRRGGRRRR- LEIKRLEHHHHHH ]]> ​ 92 SEQ ID NO 2-GGGGSGGGGSGGGGS -hBD3-3 MAQVKLQESGPELVRPGTSVK VSCKASGYAFTNYLI EWIKQRPGQGLEWIGVIHPGNGGTNYNENFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCASGNDGSYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSDIELTQSPSSLTVTAGEKVTMSCKSSQSLLNSGDQKIYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSVSGTDFTLTISSVQAEDLAVYYCQNDYNYPYTFGGGTK –GGGGSGGGGSGGGGS -GKCSTRGRKCCRRKK - LEIKRLEHHHHHH ]]> ​ 93

[0156] pcDNA3.1-TOPO or pcDNA3.4-TOPO were digested with EcoR I and BamH I. The 900-960 bp KRAS mutant scFv gene fragments were recovered by the agarose electroelution method. To insert the gene fragments into pcDNA3.1-TOPO and pcDNA3.4-TOPO vectors, the gene was amplified by PCR while synthesizing BamH I and EcoR I restriction enzyme sites. Each digested vector and insert were mixed together in a ratio of 4 μl : 4 μl and ligated together using T4 DNA ligase in a ligation buffer containing 500 mM Tris-HCl, 100 mM MgCl2, 200 mM DTT, and 10 mM ATP at 4ºC for 16 to 18 hours.

[0157] The prepared vectors were transformed into the Chinese hamster ovary cell line Expi-CHO-S and the human embryonic kidney cell line Expi-293F by the inventors. The Chinese hamster ovary cell line ExpiCHO-S was obtained from Thermo Fisher Scientific (USA) and cultured in ExpiCHO Expression Medium (GIBCO, USA).

[0158] Next, the ExpiCHO-S cell line (3 X 10 8 cells) was added to 50 mL of medium in a 250 mL disposable conical flask. 160 μL of ExpiFectamine CHO reagent (Gibco, Cat. No. A20130) diluted in 1.84 mL of Opti-MEM was added to 2 mL of Opti-MEM containing 50 μg of plasmid, then incubated at room temperature for 5 minutes. Next, the dilution was added evenly to the prepared Chinese hamster ovary cells, then incubated at 37ºC in an 8% CO2incubator for 18 hours, then 300 μL of ExpiCHO Enhancer and 12 mL of ExpiCHO Feed were added thereto and incubated for 5 days.

[0159] The human embryonic kidney cell line Expi-293F was obtained from Thermo Fisher Scientific (USA) and cultured in Expi293 Expression Medium (GIBCO, USA). Next, the Expi-293F cell line (1.50 X 10 7One cell) was added to 50 mL of media in a 250 mL disposable conical. 160 μΐ^of ExpiFectamine 293 reagent (Gibco, Cat# A14525) diluted in 2.8 mL of Opti-MEM was added to 3 mL of Opti-MEM containing 50 μg of plasmid and then incubated statically for 20 minutes at room temperature. Next, the dilution was added evenly to the prepared human embryonic kidney cells and then incubated at 37ºC in an 8% C02incubator for 18 hours before adding 300 μΐ^of ExpiFectamine 293 transfection enhancer 1 and 3 mL of ExpiFectamine 293 transfection enhancer 2 to it and incubating for 3 days.

[0160] Six days post transduction, the media was collected, sterilized, and then purified using FPLC. After separation and purification using a nickel chromatography column (HisTrap TM excel, GE Healthcare) the nickel column was equilibrated with a buffer [20 mM sodium phosphate, 0.5 M NaCl, pH 7.4] before applying the media to the column. After washing with the above buffer containing 25 mM imidazole, the protein was isolated by gradually increasing the imidazole to 125 mM. After removing the imidazole by desalting, the isolated protein was stored.

[0161] Figure 3 shows the results of SDS-PAGE and immunoblot analysis of antibodies and scFvs that penetrate cancer cells and target KRAS mutants expressed and purified in pcDNA3.4-TOPO (Figure 3(B)) and pcDNA3.1-TOPO (Figure 3(C)).

[0162] Example 2: Production of cancer cell penetrating peptides and anti-KRAS mutant single chain variable fragments (scFvs) by chemical conjugation

[0163] Example 2-1 : Synthesis of cell penetrating peptides

[0164] The following Table 6 shows the amino acid sequences of the cancer cell penetrating peptides and linkers used in the chemical conjugation method.

[0165] [Table 6]

[0166] Linker type Cancer cell penetrating peptide (CCPP) Sequence and SEQ ID NO of linkers and cancer cell penetrating peptides CGGGGG CCPP1 (H4K) 60: CGGGGG-HRRCNKNNKKR CCPP2 (H4P) 61: CGGGGG-HRRCNPNNKKR CCPP3 (LMWP) 62: CGGGGG-VSRRRRRRGGRRRR CCPP4 (hBD3-3) 63: CGGGGG-GKCSTRGRKCCRRKK CGGGGGSSGGGGG CCPP1 (H4K) 64: CGGGGGSSGGGGG-RKKNNKNCRRH CCPP2 (H4P) 65: CGGGGGSSGGGGG-RKKNNPNCRRH CCPP3 (LMWP) 66: CGGGGGSSGGGGG-RRRRGGRRRRRRSV CCPP4 (hBD3-3) 67: CGGGGGSSGGGGG-GKCSTRGRKCCRRKK

[0167] The amino acids and reagents used in the synthesis were purchased from GL Biochem and Sigma-Aldrich. Peptides were synthesized from the C-terminus using an F-moc solid-state chemical synthesis method in a reaction vessel. Specifically, peptides were synthesized using Rink amide MBHA resin (0.678 mmol / g, 100 to 200 mesh) bound with Fmoc-(9-fluorenylmethoxycarbonyl) as the blocking group. After placing 1 g of Rink amide MBHA resin in the reaction vessel, the resin was swollen with DMF, and then the Fmoc group was removed using a 20% piperidine / DMF solution. Depending on the C-terminal sequence, 0.5 M amino acid solution (solvent: dimethylformamide, DMF), 1.0 M DIPEA (solvent: dimethylformamide and n-methylpyrrolidone, DMF and NMP), and 0.5 M HBTU (solvent: dimethylformamide, DMF) were added in 5, 10, and 5 equivalents, respectively, and the reaction was carried out under a nitrogen atmosphere for 1 to 2 hours. After each deprotection and coupling step, the mixture was washed three times with DMF and methanol. Deprotection was performed even after coupling the last amino acid to remove the Fmoc group.

[0168] The synthesis was confirmed using the ninhydrin assay. The resin, confirmed by the assay to be completely synthesized, was vacuum dried, and trifluoroacetic acid (TFA) was added to it at a rate of 10 ml / g resin to cut the mixture, followed by shaking for 3 hours. The mixture containing the dissolved peptides was then separated from the resin by filtration. The peptides were crystallized into a solid state by placing the filtered solution in cold ether or by adding excess cold ether directly to the TFA mixture containing the dissolved peptides, and the peptides were collected by centrifugation. At this point, the TFA mixture was completely removed by washing several times with ether and centrifugation. The peptides thus obtained were then vacuum dried.

[0169] The synthesized peptides were isolated and purified by high-performance liquid chromatography (Shimadzu, Japan). C0.05 18 The column (diameter: 4.6 mm) was analyzed by running 0.1% TFA / H₂O and 0.1% TFA / acetonitrile at a flow rate of 1 ml / min in gradients from 5% to 45%, with the UV detector wavelength set to 220 nm. Purification was performed using a column (diameter: 50 mm) at a flow rate of 50 ml / min under the same solvent and detection wavelength conditions. The molecular weight of the purified peptides was analyzed by mass spectrometry.

[0170] Example 2-2: Chemical conjugation of cancer cell penetrating peptides to linker portion of KRAS mutant targeting scFv and purification Figure 4

[0171] KRAS scFv of SEQ ID NO: 1 was dissolved in 1 mL PBS buffer (pH 8.3). 2.12 mg of SPDP (3-(2-pyridyldithio)propionic acid succinimidyl ester, ThermoFisher, 21857) was dissolved in 120 µL DMSO (dimethylsulfoxide). 40 µL of the SPDP solution was added to the KRAS scFv solution and allowed to react for 1 hour under light-protected conditions, and this addition and reaction process was repeated three times. Alternatively, 2 mg of Sulfo-SMCC (4-[N-maleimidomethyl] cyclohexane-1-carboxylate succinimidyl ester, ThermoFisher, 22322) was dissolved in PBS. 40 µL of the Sulfo-SMCC solution was added to the KRAS scFv solution and allowed to react for 1 hour under light-protected conditions, and this addition and reaction process was repeated three times.

[0172] After the reaction was complete, desalting was performed using a PD-10 desalting column (GE Healthcare), and 3.5 mL of pyridyl dithiol-activated KRAS scFv was obtained. To the pyridyl dithiol-activated KRAS scFv, a solution of each of 2.5 mg of Cys-G5-H4K (SEQ ID NO: 60), Cys-G5-H4P (SEQ ID NO: 61), Cys-G5-LMWP (SEQ ID NO: 62), Cys-G5-hBD3-3 (SEQ ID NO: 63), Cys-G5S2G5-H4K (SEQ ID NO: 64), Cys-G5S2G5-H4P (SEQ ID NO: 65), Cys-G5S2G5-LMWP (SEQ ID NO: 66), and Cys-G5S2G5-hBD3-3 (SEQ ID NO: 67) in 300 µL of triple distilled water was added, and each mixture was adjusted to pH 8.3 using 1 M tris buffer (pH = 9). Each mixture was allowed to react overnight at 4ºC under light-protected conditions. KRAS mutant scFv on which each cancer cell penetrating peptide was bound was purified by FPLC (Akta Pure, GE Healthcare) using a heparin column (HiTrap, GE Healthcare).

[0173] SEQ ID NO: 1 was dissolved in 1 mL of PBS buffer (pH 7.4). 2.98 mg of sulfo-SMCC (4-(N-maleimidomethyl) cyclohexane-1-carboxylate sulfosuccinimidyl ester, ThermoFisher, Cat. No. 22322) was dissolved in 300 μL of triple distilled water. 100 μL of the sulfo-SMCC solution was added to the scFv solution and allowed to react for 30 minutes under light shielding conditions, and this addition and reaction process was repeated three times. After completion of the reaction, desalting was performed using a PD-10 desalting column (GE Healthcare, 17-0851-01), and 3.5 mL of maleimide-activated scFv was obtained. To the maleimide-activated scFv, 2.5 mg of each of a solution of Cys-G5-H4K (SEQ ID NO: 60), Cys-G5-H4P (SEQ ID NO: 61), Cys-G5-LMWP (SEQ ID NO: 62), Cys-G5-hBD3-3 (SEQ ID NO: 63), Cys-G5S2G5-H4K (SEQ ID NO: 64), Cys-G5S2G5-H4P (SEQ ID NO: 65), Cys-G5S2G5-LMWP (SEQ ID NO: 66), and Cys-G5S2G5-hBD3-3 (SEQ ID NO: 67) in 300 μL of PBS buffer (pH 8.3) was added, and each mixture was adjusted to pH 7.0 to 7.5. Each mixture was allowed to react overnight at 4ºC under light shielding conditions. The scFv to which each cancer cell penetrating peptide was bound was purified by FPLC (Akta Pure, GE healthcare) using a heparin column (HiTrap, GE Healthcare).

[0174] Example 2-3: Chemical conjugation of cancer cell penetrating peptides to C-terminus of KRAS mutant scFv and purification Results of SDS-PAGE and immunoblot analysis of the cancer cell-penetrating and KRAS mutant-targeting antibody or single-chain variable fragment produced by the chemical conjugation method are shown.

[0175] The molecular weight of the KRAS mutant scFv of SEQ ID NO: 1 was about 30 kDa, and the molecular weight of the KRAS mutant scFv to which the cancer cell penetrating peptide was bound was measured to be about 32 kDa. In addition, for the scFv of SEQ ID NO: 1, the molecular weight of the band bound to the KRAS antibody in the immunoblot was measured to be 30 kDa, and for the scFv to which the cancer cell penetrating peptide was bound, the molecular weight was measured to be about 32 kDa, indicating that the peptide was bound to the scFv.

[0176] No.

[0177] The KRAS mutant scFv of SEQ ID NO: 3 was allowed to react with 20 molar equivalents of tris(2-carboxyethyl)phosphine (TCEP) at room temperature for 3 hours to form free thiol groups. 100 molar equivalents of 5,50-dithiobis-(2-nitrobenzoic acid) (DTNB) were added to the KRAS mutant scFv solution and reacted at room temperature for 1 hour to bind to the free thiol groups. The excess DTNB that was not bound to the KRAS mutant scFv was removed by FPLC (Akta Pure, GE healthcare) using a heparin column (HiTrap, GE Healthcare). Thereafter, the KRAS mutant scFv-DTNB conjugate was allowed to react with 10 molar equivalents of each cancer cell penetrating peptide (SEQ ID NOs: 60 to 67) at room temperature for 1 hour. The scFv on which each cancer cell penetrating peptide was bound was purified by FPLC (Akta Pure, GE healthcare) using a heparin column (HiTrap, GE Healthcare).

[0178] Table 7 below shows the amino acid sequences of fusion proteins in which a cancer cell penetrating peptide was linked to a KRAS mutant scFv and produced by a chemical conjugation method.

[0179] [Table 7]

[0180]

[0181] Example 3: Evaluation of inhibitory activity against cancer cells having different KRAS mutant expression levels

[0182] To evaluate the cancer cell-specific growth inhibitory activity of the antibodies and scFvs that penetrate cancer cells and target KRAS mutants according to the present application, H358 cells (cancer cells expressing KRAS mutants) or human dermal fibroblasts (normal cells) were seeded at 5 x 10 3The density of 1 cell / well was dispensed into a 96-well plate, and then cultured in RPMI medium 1640 (Gibco) for 24 hours. The cells were treated with each scFv (SEQ ID NOs: 1 and 28 to 35) at a concentration of 5 μM. Cytotoxicity was measured using a CCK-8 (Cell Counting Kit-8, CK04, Dojindo Lab.). 48 hours after treatment with each scFv, the medium was removed, and 100 μl of CCK-8 solution was dispensed into fresh RPMI medium 1640. After 1 hour and 30 minutes, the absorbance at 450 nm was measured.

[0183] As a result, as shown in FIG. 5, it was confirmed that SEQ ID NOs: 28 to 31, 68, and 69 showed IC 50 values (concentration at which 50% of cells were killed), and for SEQ ID NO: 1, an IC 50 value was not measured even at a concentration of 5 μM. This is considered because SEQ ID NO: 1 did not show cancer cell killing due to the inability to penetrate cancer cells, but SEQ ID NOs: 28 to 31, 68, and 69 exhibited cytotoxicity due to the ability to penetrate cancer cells. For human dermal fibroblasts, which are normal cells, SEQ ID NOs: 28 to 31, 68, and 69 showed cell viability of 80% or more at all concentrations. This means that the scFv conjugate that penetrates cancer cells and targets KRAS mutants has cancer cell-specific cytotoxicity.

[0184] Example 4: Evaluation of the activity of antibodies and scFvs that penetrate cancer cells and target KRAS mutants

[0185] 1.0 x 10 6H358 cells (ATCC, CRL-5807) were seeded at 50,000 cells / well in a 6-well plate and incubated for 24 hours in RPMI medium 1640 (1X) (Gibco, 22400-089) and then starved for 16 hours. Cells were treated with 50 ng / mL EGF (epidermal growth factor, R&D Systems, 236-EG) for 10 minutes and with 1 mM of each variant. After 24 hours, the medium was removed, each well was washed with PBS and the cells were harvested and centrifuged at 1,500 rpm for 3 minutes. 150 mL of 1x lysis buffer (1X Lysis / Binding / Washing Buffer, 11524S) was added to the cells and mixed and kept on ice for 5 minutes. The supernatant was separated by centrifugation at 4ºC at 16,000 rpm for 15 minutes. The protein concentration was measured by BCA assay (Thermo Scientific, 23227). Washed with 400 mL of 1x lysis buffer and centrifuged at 6000 x g for 15 seconds twice.

[0186] To isolate mutant KRAS, 80 pg of GST-Raf1-RBD was added using the Active GTPase Kit (Cell Signaling, 11860S) and 500 pg of protein was distributed into spin cups. After incubation at 4ºC for 1 hour, the columns were centrifuged at 6000 x g for 15 seconds, then transferred to a fresh tube and 400 mL of 1x lysis buffer was added to it. After centrifugation at 6000 x g for 15 seconds, the column was transferred to a fresh tube and 50 mL of 2x SDS buffer (containing 200 pi of 5X SDS sample loading dye and 300 pi of water) was added to it and reacted for 2 minutes. After centrifugation at 6000 x g for 2 minutes, the resulting solution was kept at 100ºC for 7 minutes. The proteins were separated by 11% SDS-PAGE electrophoresis (20 pi of protein loading amount per lane) and transferred to a nitrocellulose membrane. To measure ERK1 / 2 as a downstream signal of KRAS, 30 pg of protein was electrophoresed by 11% SDS-PAGE and transferred to a nitrocellulose membrane. The membrane was blocked with 5% skim milk in T-TBS for 1 hour and incubated with primary antibody (diluted at 1 : 1000 in T-TBS) at 4ºC overnight. The membrane was washed with T-TBS 3 times for 10 minutes each and incubated with secondary antibody (diluted at 1 : 3000 in T-TBS) at room temperature for 1 hour. The membrane was washed with T-TBS 3 times for 10 minutes each, incubated with ECL chemiluminescence substrate (Thermo, 34580) and visualized with Amersham imager 680 (GE). Information on the antibodies used is shown in Table 8 below.

[0187] [Table 8]

[0188] Antibody name Manufacturer Catalog No. GST 1 Abcam Ab19256 Active KRAS 2 Santa Cruz SC-30 RAS (E4K9L) 3 Cell Signaling p-B-Raf (S445) 91054S 4 Cell Signaling p-MEK1 / 2 (S221) 2696S 5 Cell Signaling p-Erk1 / 2 2338S 6 Santa Cruz SC-7383 GAPDH 7 Cell Signaling Goat anti-rabbit IgG 2118S 8 BETHYL A120-101P Goat anti-mouse IgG 9 BETHYL A90-116P Figure 6

[0189] In SEQ ID NO: 20 to 23, the linker between the KRAS mutant targeting antibody or scFv and the cell-penetrating peptide is GGGGS, and in SEQ ID NO: 90 to 93, the linker is GGGGSGGGGSGGGGS.

[0190] As a result, Figure 7 As shown, SEQ ID NO: 20 to 23 all confirmed that compared with EGF treatment alone, they reduced KRAS activity. Furthermore, it can be seen that pERK1 / 2, a downstream signal of KRAS, was also reduced by SEQ ID NO: 20 to 23.

[0191] like Figure 8 As shown, SEQ ID NO: 90 to 93 all confirmed that treatment with EGF alone reduced KRAS activity. Furthermore, it can be seen that pERK1 / 2, a downstream signal of KRAS, was also reduced by SEQ ID NO: 91 to 92.

[0192] Example 5: Evaluation of the cell penetration ability of antibodies and scFv that penetrate cancer cells and target KRAS mutants.

[0193] To test the cancer cell penetration ability of the antibody or scFv, 1 x 10 5 H358 cells were collected, and after 48 hours, each of SEQ ID NO: 20 to 23 was added to them at a concentration of 500 nM, and the cells were cultured for 30 minutes. The cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton-X100 for 10 minutes, and blocked with 2% BSA. KRAS in the cells was reacted with anti-RAS antibody (Origene, cst53270) at a dilution of 1:1,000, and with secondary antibody anti-rabbit Alexa Fluor 488 (Invitrogen, A27034) at a dilution of 1:2,000. Each of SEQ ID NO: 20 to 23 was reacted with V5-Tag antibody (Origene, CST 13202) at a dilution of 1:1,000, and with anti-mouse Alexa Fluor 555 (Invitrogene, A28180) at a dilution of 1:1,000. Cell nuclei were stained with 0.1 μg / mL DAPI (Thermofisher, R37606). The penetration of SEQ ID NO: 20 to 23 into cells was measured using confocal laser scanning microscopy.

[0194] As shown in FIG. 6, it can be seen that, except for SEQ ID NO: 2, all of SEQ ID NOs: 20 to 23 penetrate the cells. In addition, it can be seen that SEQ ID NOs: 20 to 23 are co-localized with KRAS present in the cancer cells. Thus, it can be seen that SEQ ID NOs: 20 to 23 can penetrate the cells and bind to KRAS present in the cells. Figure 9

[0195] Example 6: Evaluation of distribution of antibodies and scFvs penetrating cancer cells and targeting KRAS mutants in cancer tissues in a tumor xenograft animal model

[0196] A mixture of 1 x 10 6 cells and 100 μl of Matrigel (BD Bioscience, San Diego, CA, USA) was transplanted into the thigh of each 5-6-week-old female Balb / c nude mouse (5-6 weeks old; Japan SLC Inc., Hamamatsu, Japan) to form a tumor having a volume of 100 mm 3 Each of Cy5.5-labeled SEQ ID NO: 2, Cy5.5-labeled SEQ ID NO: 23, and Cy5.5 was intraperitoneally injected at a dose of 20 μg / mouse, and after 24 hours, the distribution of fluorescence in each major organ and cancer tissue was observed.

[0197] As shown in FIG. 8, it can be seen that SEQ ID NO: 23 is mainly distributed in the cancer tissue 24 hours after injection, and a part thereof is excreted by the kidney. Figure 10 In addition, as shown in FIG. 9, it can be seen that SEQ ID NO: 23 has the highest fluorescence intensity in the cancer tissue when the fluorescence intensity is measured.

[0198] Figure 11

[0199] Example 7: Evaluation of tumor inhibitory activity of antibodies and scFvs penetrating cancer cells and targeting KRAS mutants in a tumor xenograft animal model

[0200] A mixture of 1 x 10 6 cells and 100 μl of Matrigel (BD Bioscience, San Diego, CA, USA) was transplanted into the thigh of each 5-6-week-old female Balb / c nude mouse (5-6 weeks old; Japan SLC Inc., Hamamatsu, Japan) to form a tumor having a volume of 100 mm 3 ​​​tumors. Each of SEQ ID NO: 2 and SEQ ID NO: 23 was injected intraperitoneally at a dose of 1 mg / kg twice a week for 30 days. The volume of the tumor was measured with a vernier caliper at 3 to 4-day intervals, and on day 30, the mice were euthanized, and the tumors were excised and photographed.

[0201] As shown in FIG. 8, SEQ ID NO: 2 did not show tumor inhibition, but SEQ ID NO: 23 showed excellent tumor inhibition. This proves that SEQ ID NO: 23, which penetrates cancer tissue, effectively inhibits the growth of tumor cells. Example 8-1: Evaluation of inhibitory activity of antibodies and scFv penetrating cancer cells and targeting KRAS mutants on active KRAS (mutant KRAS) expression and 12 As shown in FIG. 8, SEQ ID NO: 2 did not show tumor inhibition, but SEQ ID NO: 23 showed excellent tumor inhibition. This proves that SEQ ID NO: 23, which penetrates cancer tissue, effectively inhibits the growth of tumor cells.

[0202] Example 8: Effect of fusion proteins of SEQ ID NOs: 80 to 83 produced by chemical conjugation

[0203] No. Antibody name

[0204] One.0 x 10 6 H358 cells (ATCC, CRL-5807) were seeded and cultured in RPMI medium 1640 (1X) (Gibco, 22400-089) for 24 hours, and then starved for 16 hours. The cells were treated with 50 ng / mL EGF (epidermal growth factor, R&D Systems, 236-EG) for 10 minutes, and treated with 1 μΜ of each variant. The cells were cultured at 37ºC for 24 hours. After 24 hours, the medium was removed, each well was washed with PBS, and the cells were harvested and centrifuged at 1,500 rpm for 3 minutes. 150 μL of 1x lysis buffer (1X lysis / Binding / Washing Buffer, 11524S) was added to the cells and mixed therewith, and left on ice for 5 minutes. The supernatant was separated by centrifugation at 16,000 rpm for 15 minutes at 4ºC. The protein concentration was measured by a BCA assay (Thermo Scientific, 23227). Washing was performed with 400 μL of 1x lysis buffer, and centrifugation was performed at 6000 x g for 15 seconds twice.

[0205] To isolate mutant KRAS, an active GTPase kit (Cell Signaling, 11860S) was used adding 80 pg of GST-Raf1-RBD and 500 pg of protein was distributed into spin cups. After incubation at 4°C for 1 hour, the columns were centrifuged at 6000 x g for 15 seconds, then transferred to a fresh tube and 400 pL of lx lysis buffer was added to it. After centrifugation at 6000 x g for 15 seconds, the column was transferred to a fresh tube and 50 pL of 2x SDS buffer (containing 200 mΐ of 5X SDS sample loading dye and 300 mΐ of water) was added to it and reacted for 2 minutes. After centrifugation at 6000 x g for 2 minutes, the resulting solution was heated at 100°C for 7 minutes. The proteins were separated by 11% SDS-PAGE electrophoresis (20 mΐ of protein loading amount per lane) and transferred to a nitrocellulose membrane. To measure ERK1 / 2 as a downstream signal of KRAS, 30 pg of protein was electrophoresed by 11% SDS-PAGE and transferred to a nitrocellulose membrane. The membrane was blocked with 5% skim milk in T-TBS for 1 hour and incubated with a primary antibody (diluted at 1 : 1000 in T-TBS) at 4°C overnight. The membrane was washed with T-TBS 3 times for 10 minutes each, and incubated with a secondary antibody (diluted at 1 : 3000 in T-TBS) at room temperature for 1 hour. The membrane was washed with T-TBS 3 times for 10 minutes each, incubated with ECL chemiluminescent substrate (Thermo, 34580), and visualized with an Amersham imager 680 (GE). Information on the antibodies used is shown in Table 9 below.

[0206] [Table 9] Antibodies used

[0207] Manufacturer Catalog No. GST Abcam 1 Ab19256 Active KRAS Santa Cruz 2 SC-30 RAS (E4K9L) Cell Signaling 3 p-B-Raf (S445) Cell Signaling 91054S 4 p-MEK1 / 2 (S221) Cell Signaling 2696S 5 p-Erk1 / 2 Santa Cruz 2338S 6 SC-7383 GAPDH Cell Signaling 7 Goat anti-rabbit IgG BETHYL 2118S 8 A120-101P Goat anti-mouse IgG BETHYL 9 A90-116P Figure 13 Example 8-2: Evaluation of inhibitory activity of antibodies and scFv penetrating cancer cells and targeting KRAS mutants against cancer cells having different KRAS mutant expression levels

[0208] SEQ ID NOs: 80 to 83 were produced by linking a cell penetrating peptide to a KRAS mutant targeting antibody or scFv via chemical conjugation. As shown in Figure 14 Figure 6, it was confirmed that all of SEQ ID NOs: 80 to 83 reduced active KRAS compared to treatment with EGF alone. In addition, it can be seen that pERK1 / 2 as a downstream signal of KRAS was also reduced by SEQ ID NOs: 80 to 83.

[0209] ​ ​

[0210] To evaluate the cancer cell-specific growth inhibitory activity of the antibodies and scFvs that penetrate cancer cells and target KRAS mutants according to the present application, H358 cells (cancer cells expressing KRAS mutants) or human dermal fibroblasts (normal cells) were assigned to a 96-well plate at a density of 5 x 10 3 cells / well, and then cultured in RPMI medium 1640 (Gibco) for 24 hours. The cells were treated with each scFv (SEQ ID NO: 3 and 80 to 83) at a concentration of 0 to 5 μM. Cell toxicity was measured using CCK-8 (Cell Counting Kit-8, CK04, Dojindo Lab.). Cell toxicity was measured using CCK-8 (Cell Counting Kit-8, CK04, Dojindo Lab.). 48 hours after treatment with each scFv, the culture medium was removed, and 100 μl of CCK-8 solution was assigned to fresh RPMI medium 1640. After 1 hour and 30 minutes, the absorbance at 450 nm was measured.

[0211] As a result, as shown in ​ 50 50 This is considered because SEQ ID NO: 3 did not show cancer cell killing due to the inability to penetrate cancer cells, but SEQ ID NO: 80 to 83 exhibited cytotoxicity due to the ability to penetrate cancer cells. Human dermal fibroblasts, which are normal cells, showed 80% or more cell viability at all concentrations. This means that the scFv conjugate that penetrates cancer cells and targets KRAS mutants has cancer cell-specific cytotoxicity.

[0212] Example 8-3: Evaluation of cell penetration ability of antibodies and scFv that penetrate cancer cells and target KRAS mutants

[0213] To test the cancer cell penetration ability of the antibodies or scFvs, 1 x 10 5 ​​H358 cells, and after 48 hours, each of SEQ ID NOs: 80 to 83 was added thereto at a concentration of 500 nM, and the cells were cultured for 30 minutes. The cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton-X100 for 10 minutes, and blocked with 2% BSA. KRAS in the cells was reacted with an anti-RAS antibody (Origene, cst53270) at a dilution ratio of 1 : 1,000, and with a secondary antibody, anti-rabbit Alexa Fluor 488 (Invitrogen, A27034) at a dilution ratio of 1 : 2,000. Each of SEQ ID NOs: 80 to 83 was reacted with a V5-Tag antibody (Origene, cst 13202) at a dilution ratio of 1 : 1,000, and with an anti-mouse Alexa Fluor 555 (Invitrogene, A28180) at a dilution ratio of 1 : 1,000. The nuclei were stained with 0.1 μg / mL DAPI (Thermofisher, R37606). The penetration of the cells by SEQ ID NOs: 80 to 83 was measured with a confocal laser scanning microscope.

[0214] As a result, as shown in Figure 15 FIG. 9, it can be seen that, except for SEQ ID NO: 3, all of SEQ ID NOs: 80 to 83 penetrated the cells. In addition, it can be seen that SEQ ID NOs: 80 to 83 were colocalized with KRAS present in the cancer cells. Thus, it can be seen that SEQ ID NOs: 80 to 83 can penetrate the cells and bind to KRAS present in the cells.

[0215] Example 8-4: Evaluation of distribution of antibodies and scFv that penetrate cancer cells and target KRAS mutants in cancer tissues in tumor xenograft animal models Figure 16

[0216] A mixture of 1 x 10 6 H358 cells and 100 μl of Matrigel (BD Bioscience, San Diego, CA, USA) was transplanted into the thigh of each 5-6-week-old female Balb / c nude mouse (5-6 weeks old; Japan SLC Inc., Hamamatsu, Japan) to form a tumor having a volume of 100 mm 3 Each of Cy5.5-labeled SEQ ID NO: 3, Cy5.5-labeled SEQ ID NO: 83, and Cy5.5 was intraperitoneally injected at a dose of 20 μg / mouse, and after 24 hours, the distribution of the distributed fluorescence in each of the major organs and cancer tissues was observed.

[0217] As a result, as shown in Figure 17As shown, it can be seen that SEQ ID NO: 83 is mainly distributed in the cancer tissue 24 hours after injection, and a part thereof is excreted by the kidney.

[0218] In addition, as Example 8-5: Evaluation of tumor inhibition activity of antibodies and scFv that penetrate cancer cells and target KRAS mutants in tumor xenograft animal models shown, it can be seen that SEQ ID NO: 83 has the highest fluorescence intensity in the cancer tissue when the fluorescence intensity is measured.

[0219] Figure 18 ​

[0220] A mixture of 1 x 10 6 H358 cells and 100 µl of Matrigel (BD Bioscience, San Diego, CA, USA) was transplanted into the thigh of each 5-6-week-old female Balb / c nude mouse (5-6 weeks old; Japan SLC Inc., Hamamatsu, Japan) to form a tumor having a volume of 100 mm 3 Each of SEQ ID NO: 3 and SEQ ID NO: 83 was intraperitoneally injected at a dose of 1 mg / kg twice a week for 30 days. The volume of the tumor was measured at intervals of 3 to 4 days with a vernier caliper, and on the 30th day, the mice were euthanized, and the tumor was excised and photographed.

[0221] As shown in ​ and 19 , it was confirmed that SEQ ID NO: 3 had no tumor inhibitory effect, but SEQ ID NO: 83 had an excellent tumor inhibitory effect. This proves that SEQ ID NO: 83, which penetrates the cancer tissue, effectively inhibits the growth of tumor cells.

[0222] Although the present application has been described in detail with reference to specific features, it is apparent that the description is merely a description of preferred embodiments of the present application and does not limit the scope of the present application. Therefore, the substantial scope of the present application will be defined by the appended claims and their equivalents.

[0223] Industrial applicability

[0224] An intracellular oncoprotein or a mutant thereof acts as an important factor in tumor pathogenesis. In the present application, an intracellular oncoprotein or an oncomutant-targeting antibody or a single chain variable fragment thereof, which inhibits the function of an intracellular oncoprotein or a mutant thereof by binding thereto, is used. In order to enhance the accessibility of the antibody or the single chain variable fragment thereof to cancer cells, a cancer cell-penetrating peptide is linked to the antibody or the single chain variable fragment thereof. The fusion protein produced as described above can effectively penetrate tumor cells, thereby maximizing the anti-tumor or anticancer effect of the antibody or the single chain variable fragment thereof targeting the intracellular oncoprotein or the oncomutant.

[0225] SEQUENCE LISTING Free Text

[0226] Electronic filing attached. <110> Nanosmart Biomedical Engineering Co., Ltd. <120> Antibodies targeting oncoproteins in cells, or fusion proteins of single-chain variable fragments thereof and cancer cell-penetrating peptides, and uses thereof <130> PP-B2659 <160> 93 <170> KoPatentIn 3.0 <210> 1 <211> 271 <212> PRT <213> Artificial Sequence <220> <223> KRAS mutant scfv <400> 1 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gin Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gin Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 100 105 110 Gly Gly Ser Gly Gly Gly Gly Ser Asp He Glu Leu Thr Gin Ser Pro 115 120 125 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 130 135 140 Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys He Tyr Leu Thr 145 150 155 160 Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu He Tyr Trp 165 170 175 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 180 185 190 Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Val Gin Ala Glu Asp 195 200 205 Leu Ala Val Tyr Tyr Cys Gin Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 210 215 220 Gly Gly Gly Thr Lys Leu Glu lie Lys Arg Gly Ser Glu Asn Leu Tyr 225 230 235 240 Phe Gin Gly Gly Ser Gly Lys Pro lie Pro Asn Pro Leu Leu Gly Leu 245 250 255 Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His His His 260 265 270 <210> 2 <211> 254 <212> PRT <213> Artificial Sequence <220> <223> KRAS mutant scfv <400> 2 Met Ala Gin Val Lys Leu Gin Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu 35 40 45 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 50 55 60 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 65 70 75 80 Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 85 90 95 Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro Ser Ser Leu Thr 130 135 140 Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser 145 150 155 160 Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu Thr Trp Tyr Gln Gln 165 170 175 Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg 180 185 190 Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val Ser Gly Thr Asp 195 200 205 Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr 210 215 220 Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe Gly Gly Gly Thr 225 230 235 240 Lys Leu Glu Ile Lys Arg Leu Glu His His His His His His His 245 250 <210> 3 <211> 272 <212> PRT <213> Artificial Sequence <220> <223> KRAS mutant scfv <400> 3 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 100 105 110 Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro 115 120 125 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 130 135 140 Ser Ser Gln Ser Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu Thr 145 150 155 160 Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp 165 170 175 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 180 185 190 Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp 195 200 205 Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 210 215 220 Gly Gly Gly Thr Lys Cys Leu Glu Ile Lys Arg Gly Ser Glu Asn Leu 225 230 235 240 Tyr Phe Gln Gly Gly Ser Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly 245 250 255 Leu Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His His His 260 265 270 <210> 4 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 4 Gly Gly Gly Gly Ser His Arg Arg Cys Asn Lys Asn Asn Lys Lys Arg 1 5 10 15 <210> 5 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 5 Gly Gly Gly Gly Ser His Arg Arg Cys Asn Pro Asn Asn Lys Lys Arg 1 5 10 15 <210> 6 <211> 19 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 6 Gly Gly Gly Gly Ser Val Ser Arg Arg Arg Arg Arg Arg Gly Gly Arg 1 5 10 15 Arg Arg Arg <210> 7 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 7 Gly Gly Gly Gly Ser Gly Lys Cys Ser Thr Arg Gly Arg Lys Cys Cys 1 5 10 15 Arg Arg Lys Lys 20 <210> 8 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 8 His Arg Arg Cys Asn Lys Asn Asn Lys Lys Arg Gly Gly Gly Gly Ser 1 5 10 15 <210> 9 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 9 His Arg Arg Cys Asn Pro Asn Asn Lys Lys Arg Gly Gly Gly Gly Ser 1 5 10 15 <210> 10 <211> 19 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 10 Val Ser Arg Arg Arg Arg Arg Arg Gly Gly Arg Arg Arg Arg Gly Gly 1 5 10 15 Gly Gly Ser <210> 11 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 11 Gly Lys Cys Ser Thr Arg Gly Arg Lys Cys Cys Arg Arg Lys Lys Gly 1 5 10 15 Gly Gly Gly Ser 20 <210> 12 <211> 287 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 12 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gin Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 100 105 110 Gly Gly Ser Gly Gly Gly Gly Ser Asp He Glu Leu Thr Gin Ser Pro 115 120 125 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 130 135 140 Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys He Tyr Leu Thr 145 150 155 160 Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu He Tyr Trp 165 170 175 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 180 185 190 Ser Gly Thr Asp Phe Thr Leu Thr He Ser Ser Val Gin Ala Glu Asp 195 200 205 Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 210 215 220 Gly Gly Gly Thr Lys Gly Gly Gly Gly Ser His Arg Arg Cys Asn Lys 225 230 235 240 Asn Asn Lys Lys Arg Leu Glu Ile Lys Arg Gly Ser Glu Asn Leu Tyr 245 250 255 Phe Gln Gly Gly Ser Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu 260 265 270 Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His His His 275 280 285 <210> 13 <211> 287 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 13 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 100 105 110 Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro 115 120 125 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 130 135 140 Ser Ser Gln Ser Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu Thr 145 150 155 160 Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp 165 170 175 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 180 185 190 Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp 195 200 205 Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 210 215 220 Gly Gly Gly Thr Lys Gly Gly Gly Gly Ser His Arg Arg Cys Asn Pro 225 230 235 240 Asn Asn Lys Lys Arg Leu Glu Ile Lys Arg Gly Ser Glu Asn Leu Tyr 245 250 255 Phe Gln Gly Gly Ser Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu 260 265 270 Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His His His 275 280 285 <210> 14 <211> 290 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 14 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 100 105 110 Gly Gly Ser Gly Gly Gly Gly Ser Asp lie Glu Leu Thr Gin Ser Pro 115 120 125 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 130 135 140 Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys lie Tyr Leu Thr 145 150 155 160 Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu lie Tyr Trp 165 170 175 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 180 185 190 Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Val Gin Ala Glu Asp 195 200 205 Leu Ala Val Tyr Tyr Cys Gin Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 210 215 220 Gly Gly Gly Thr Lys Gly Gly Gly Gly Ser Val Ser Arg Arg Arg Arg 225 230 235 240 Arg Arg Gly Gly Arg Arg Arg Arg Leu Glu Ile Lys Arg Gly Ser Glu 245 250 255 Asn Leu Tyr Phe Gln Gly Gly Ser Gly Lys Pro Ile Pro Asn Pro Leu 260 265 270 Leu Gly Leu Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His 275 280 285 His His 290 <210> 15 <211> 291 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 15 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val lie His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gin Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gin Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 100 105 110 Gly Gly Ser Gly Gly Gly Gly Ser Asp lie Gin Leu Thr Gin Ser Pro 115 120 125 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 130 135 140 Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys lie Tyr Leu Thr 145 150 155 160 Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu lie Tyr Trp 165 170 175 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 180 185 190 Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Val Gin Ala Glu Asp 195 200 205 Leu Ala Val Tyr Tyr Cys Gin Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 210 215 220 Gly Gly Gly Thr Lys Gly Gly Gly Gly Ser Gly Lys Cys Ser Thr Arg 225 230 235 240 Gly Arg Lys Cys Cys Arg Arg Lys Lys Leu Glu lie Lys Arg Gly Ser 245 250 255 Glu Asn Leu Tyr Phe Gin Gly Gly Ser Gly Lys Pro lie Pro Asn Pro 260 265 270 Leu Leu Gly Leu Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His 275 280 285 His His His 290 <210> 16 <211> 287 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 16 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala His Arg Arg Cys Asn Lys Asn Asn Lys Lys Arg Gly Gly 20 25 30 Gly Gly Ser Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 35 40 45 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 50 55 60 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 65 70 75 80 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 85 90 95 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gin Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp He Glu Leu Thr Gin Ser Pro Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys He Tyr Leu Thr Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu He Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val Ser Gly Thr Asp Phe Thr Leu Thr He Ser Ser Val Gin Ala Glu Asp Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys​​​​​​​Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 225 230 235 240 Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Gly Ser Glu Asn Leu Tyr 245 250 255 Phe Gln Gly Gly Ser Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu 260 265 270 Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His His His 275 280 285 <210> 17 <211> 287 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 17 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala His Arg Arg Cys Asn Pro Asn Asn Lys Lys Arg Gly Gly 20 25 30 Gly Gly Ser Glu Trp lie Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp 35 40 45 Gly Gly Ser Gly Gly Gly Gly Ser Gly lie Glu Leu Thr Gin Ser Pro 50 55 60 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gin Gin Gin 65 70 75 80 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gin Gin Gin Gin Gin Gin Gin Gin 85 90 95 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gin Gin Gin Gin Gin Gin Gin Gin 100 105 110 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gin Gin Gin Gin Gin Gin Gin Gin 115 120 125 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gin Gin Gin Gin Gin Gin Gin Gin 130 135 140 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gin Gin Gin Gin Gin Gin Gin Gin 145 150 155 160 ​​​​​​​​Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys He Tyr Leu Thr 165 170 175 Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu He Tyr Trp 180 185 190 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 195 200 205 Ser Gly Thr Asp Phe Thr Leu Thr He Ser Ser Val Gin Ala Glu Asp 210 215 220 Leu Ala Val Tyr Tyr Cys Gin Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 225 230 235 240 Gly Gly Gly Thr Lys Leu Glu He Lys Arg Gly Ser Glu Asn Leu Tyr 245 250 255 Phe Gin Gly Gly Ser Gly Lys Pro He Pro Asn Pro Leu Leu Gly Leu 260 265 270 Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His His His His 275 280 285 <210> 18 <211> 290 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 18 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Val Ser Arg Arg Arg Arg Arg Arg Gly Gly Arg Arg Arg 20 25 30 Arg Gly Gly Gly Gly Ser Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly 35 40 45 Leu Glu Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr 50 55 60 Asn Glu Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser 65 70 75 80 Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala 85 90 95 Val Tyr Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly 115 120 125 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Glu Leu Thr 130 135 140 Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met 145 150 155 160 Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser Gly Asp Gln Lys Ile 165 170 175 Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu 180 185 190 Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr 195 200 205 Gly Ser Val Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln 210 215 220 Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro 225 230 235 240 Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Gly Ser Glu 245 250 255 Asn Leu Tyr Phe Gln Gly Gly Ser Gly Lys Pro Ile Pro Asn Pro Leu 260 265 270 Leu Gly Leu Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His 275 280 285 His His 290 <210> 19 <211> 291 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 19 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Gly Lys Cys Ser Thr Arg Gly Arg Lys Cys Cys Arg Arg 20 25 30 Lys Lys Gly Gly Gly Gly Ser Glu Trp Ile Lys Gln Arg Pro Gly Gln 35 40 45 Gly Leu Glu Trp lie Gly Val lie His Pro Gly Asn Gly Gly Thr Asn 50 55 60 Tyr Asn Glu Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser 65 70 75 80 Ser Ser Thr Ala Tyr Met Gin Leu Ser Ser Leu Thr Ser Asp Asp Ser 85 90 95 Ala Val Tyr Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gin 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly 115 120 125 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp lie Glu Leu 130 135 140 Thr Gin Ser Pro Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr 145 150 155 160 Met Ser Cys Lys Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys 165 170 175 Ile Tyr Leu Thr Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu 180 185 190 Leu Ile Tyr Trp Ala Ser Thr Arg Gin Ser Gly Val Pro Asp Arg Phe 195 200 205 Thr Gly Ser Val Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val 210 215 220 Gln Ala Gin Asp Leu Ala Val Tyr Tyr Cys Gin Asn Asp Tyr Asn Tyr 225 230 235 240 Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Gly Ser 245 250 255 Glu Asn Leu Tyr Phe Gin Gin Gly Ser Gly Lys Pro Ile Pro Asn Pro 260 265 270 Leu Leu Gly Leu Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His 275 280 285 His His His 290 <210> 20 <211> 270 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 20 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu 35 40 45 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 50 55 60 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 65 70 75 80 Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 85 90 95 Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro Ser Ser Leu Thr 130 135 140 Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser 145 150 155 160 Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu Thr Trp Tyr Gln Gln 165 170 175 Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg 180 185 190 Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val Ser Gly Thr Asp 195 200 205 Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr 210 215 220 Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe Gly Gly Gly Thr 225 230 235 240 Lys Gly Gly Gly Gly Ser His Arg Arg Cys Asn Lys Asn Asn Lys Lys 245 250 255 Arg Leu Glu Ile Lys Arg Leu Glu His His His His His His 260 265 270 <210> 21 <211> 270 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 21 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu 35 40 45 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 50 55 60 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 65 70 75 80 Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 85 90 95 Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro Ser Ser Leu Thr 130 135 140 Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser 145 150 155 160 Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu Thr Trp Tyr Gln Gln 165 170 175 Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg 180 185 190 Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val Ser Gly Thr Asp 195 200 205 Phe Thr Leu Thr lie Ser Ser Val Gin Ala Glu Asp Leu Ala Val Tyr 210 215 220 Tyr Cys Gin Asp Tyr Asn Tyr Pro Tyr Thr Phe Gly Gly Gly Thr 225 230 235 240 Lys Gly Gly Gly Gly Ser His Arg Arg Cys Asn Pro Asn Asn Lys Lys 245 250 255 Arg Leu Glu lie Lys Arg Leu Glu His His His His His His 260 265 270 <210> 22 <211> 273 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 22 Met Ala Gin Val Lys Leu Gin Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile Glu Trp Ile Lys Gin Arg Pro Gly Gin Gly Leu Glu 35 40 45 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 50 55 60 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 65 70 75 80 Ala Tyr Met Gin Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 85 90 95 Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gin Gly Thr Thr 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Asp Ile Glu Leu Thr Gin Ser Pro Ser Ser Leu Thr 130 135 140 Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gin Ser 145 150 155 160 Leu Leu Asn Ser Gly Asp Gin Lys He Tyr Leu Thr Trp Tyr Gin Gin 165 170 175 Lys Pro Gly Gin Pro Pro Lys Leu Leu He Tyr Trp Ala Ser Thr Arg 180 185 190 Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val Ser Gly Thr Asp 195 200 205 Phe Thr Leu Thr He Ser Ser Val Gin Ala Glu Asp Leu Ala Val Tyr 210 215 220 Tyr Cys Gin Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe Gly Gly Gly Thr 225 230 235 240 Lys Gly Gly Gly Gly Ser Val Ser Arg Arg Arg Arg Arg Arg Gly Gly 245 250 255 Arg Arg Arg Arg Leu Glu He Lys Arg Leu Glu His His His His His 260 265 270 His <210> 23 <211> 274 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 23 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu 35 40 45 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 50 55 60 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 65 70 75 80 Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 85 90 95 Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro Ser Ser Leu Thr 130 135 140 Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser 145 150 155 160 Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu Thr Trp Tyr Gln Gln 165 170 175 Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg 180 185 190 Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val Ser Gly Thr Asp 195 200 205 Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr 210 215 220 Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe Gly Gly Gly Thr 225 230 235 240 Lys Gly Gly Gly Gly Ser Gly Lys Cys Ser Thr Arg Gly Arg Lys Cys 245 250 255 Cys Arg Arg Lys Lys Leu Glu lie Lys Arg Leu Glu His His His His 260 265 270 His His <210> 24 <211> 275 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 24 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile His Arg Arg Cys Asn Lys Asn Asn Lys Lys Arg Gly 35 40 45 Gly Gly Gly Ser Glu Trp lie Lys Gln Arg Pro Gly Gln Gly Leu Glu 50 55 60 Trp lie Gly Val lie His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 65 70 75 80 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met Gin Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gin Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp He Gin Leu Thr Gin Ser Pro Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys He Tyr Leu Thr Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu He Tyr ​​​​​​​​Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser 210 215 220 Val Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu 225 230 235 240 Asp Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr 245 250 255 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Leu Glu His His His 260 265 270 His His His 275 <210> 25 <211> 275 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 25 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile His Arg Arg Cys Asn Pro Asn Asn Lys Lys Arg Gly 35 40 45 Gly Gly Gly Ser Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu 50 55 60 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 65 70 75 80 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 85 90 95 Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 100 105 110 Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr 115 120 125 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly 130 135 140 Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser 145 150 155 160 Pro Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys 165 170 175 Lys Ser Ser Gln Ser Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu 180 185 190 Thr Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr 195 200 205 Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser 210 215 220 Val Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu 225 230 235 240 Asp Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr 245 250 255 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Leu Glu His His His 260 265 270 His His His 275 <210> 26 <211> 278 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 26 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile Val Ser Arg Arg Arg Arg Arg Arg Gly Gly Arg Arg 35 40 45 Arg Arg Gly Gly Gly Gly Ser Glu Trp Ile Lys Gln Arg Pro Gly Gln 50 55 60 Gly Leu Glu Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn 65 70 75 80 Tyr Asn Glu Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser 85 90 95 Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser 100 105 110 Ala Val Tyr Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gin 115 120 125 Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly 130 135 140 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Glu Leu 145 150 155 160 Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr 165 170 175 Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser Gly Asp Gin Lys 180 185 190 Ile Tyr Leu Thr Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu 195 200 205 Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe 210 215 220 Thr Gly Ser Val Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val 225 230 235 240 Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr 245 250 255 Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Leu Glu 260 265 270 His His His His His His 275 <210> 27 <211> 280 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 27 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile Gly Lys Cys Ser Thr Arg Gly Arg Lys Cys Cys Arg 35 40 45 Arg Lys Lys Gly Gly Gly Gly Ser Glu Trp Ile Lys Gln Arg Pro Gly 50 55 60 Gln Gly Leu Glu Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr 65 70 75 80 Asn Tyr Asn Glu Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys 85 90 95 Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp 100 105 110 Ser Ala Val Tyr Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly 115 120 125 Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys 130 135 140 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Glu 145 150 155 160 Leu Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val 165 170 175 Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser Gly Asp Gln 180 185 190 Lys Ile Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys 195 200 205 Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg 210 215 220 Phe Thr Gly Ser Val Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser 225 230 235 240 Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn 245 250 255 Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Leu 260 265 270 Glu His His His His His His His 275 280 <210> 28 <211> 287 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 28 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 151015 Ile Gin Ala Glu Trp lie Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp 202530 Ile Gly Val lie His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 354045 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 505060 Tyr Met Gin Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65707580 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gin Gly Thr Thr Val 859595 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 100105110 Gly Gly Ser Gly Gly Gly Gly Ser Asp lie Glu Leu Thr Gin Ser Pro 115120125 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 130135140 Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys He Tyr Leu Thr 145 150 155 160 Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu He Tyr Trp 165 170 175 Ala Ser Thr Arg Gin Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 180 185 190 Ser Gly Thr Asp Phe Thr Leu Thr He Ser Ser Val Gin Ala Gin Asp 195 200 205 Leu Ala Val Tyr Tyr Cys Gin Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 210 215 220 Gly Gly Gly Thr Lys Gly Gly Gly Gly Ser His Arg Arg Cys Asn Lys 225 230 235 240 Asn Gin Lys Lys Arg Leu Gin He Lys Arg Gly Ser Gin Asn Leu Tyr 245 250 255 Phe Gin Gly Gly Ser Gly Lys Pro He Pro Asn Pro Leu Leu Gly Leu 260 265 270 Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His His His 275 280 285 <210> 29 <211> 287 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 29 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gin Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 100 105 110 Gly Gly Ser Gly Gly Gly Gly Ser Asp He Glu Leu Thr Gin Ser Pro 115 120 125 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 130 135 140 Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys He Tyr Leu Thr 145 150 155 160 Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu He Tyr Trp 165 170 175 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 180 185 190 Ser Gly Thr Asp Phe Thr Leu Thr He Ser Ser Val Gin Ala Glu Asp 195 200 205 Leu Ala Val Tyr Tyr Cys Gin Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 210 215 220 Gly Gly Gly Thr Lys Gly Gly Gly Gly Ser His Arg Arg Cys Asn Pro 225 230 235 240 Asn Asn Lys Lys Arg Leu Glu Ile Lys Arg Gly Ser Glu Asn Leu Tyr 245 250 255 Phe Gin Gly Gly Ser Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu 260 265 270 Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His His His 275 280 285 <210> 30 <211> 290 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 30 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gin Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 100 105 110 Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro 115 120 125 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 130 135 140 Ser Ser Gln Ser Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu Thr 145 150 155 160 Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp 165 170 175 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 180 185 190 Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp 195 200 205 Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 210 215 220 Gly Gly Gly Thr Lys Gly Gly Gly Gly Ser Val Ser Arg Arg Arg Arg Arg 225 230 235 240 Arg Arg Gly Gly Arg Arg Arg Arg Leu Glu Ile Lys Arg Gly Ser Glu 245 250 255 Asn Leu Tyr Phe Gln Gly Gly Ser Gly Lys Pro Ile Pro Asn Pro Leu 260 265 270 Leu Gly Leu Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His 275 280 285 His His 290 <210> 31 <211> 291 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 31 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 100 105 110 Gly Gly Ser Gly Gly Gly Gly Ser Asp lie Glu Leu Thr Gin Ser Pro 115 120 125 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 130 135 140 Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys lie Tyr Leu Thr 145 150 155 160 Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu lie Tyr Trp 165 170 175 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 180 185 190 Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Val Gin Ala Glu Asp 195 200 205 Leu Ala Val Tyr Tyr Cys Gin Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 210 215 220 Gly Gly Gly Thr Lys Gly Gly Gly Gly Ser Gly Lys Cys Ser Thr Arg 225 230 235 240 Gly Arg Lys Cys Cys Arg Arg Lys Lys Leu Glu Ile Lys Arg Gly Ser 245 250 255 Glu Asn Leu Tyr Phe Gln Gly Gly Ser Gly Lys Pro Ile Pro Asn Pro 260 265 270 Leu Leu Gly Leu Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His 275 280 285 His His His 290 <210> 32 <211> 287 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 32 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala His Arg Arg Cys Asn Lys Asn Asn Lys Lys Arg Gly Gly 20 25 30 Gly Gly Ser Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 35 40 45 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 50 55 60 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 65 70 75 80 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 85 90 95 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro 130 135 140 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 145 150 155 160 Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys He Tyr Leu Thr 165 170 175 Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu He Tyr Trp 180 185 190 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 195 200 205 Ser Gly Thr Asp Phe Thr Leu Thr He Ser Ser Val Gin Ala Glu Asp 210 215 220 Leu Ala Val Tyr Tyr Cys Gin Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 225 230 235 240 Gly Gly Gly Thr Lys Leu Glu He Lys Arg Gly Ser Glu Asn Leu Tyr 245 250 255 Phe Gin Gly Gly Ser Gly Lys Pro He Pro Asn Pro Leu Leu Gly Leu 260 265 270 Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His His His His 275 280 285 <210> 33 <211> 287 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 33 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala His Arg Arg Cys Asn Pro Asn Asn Lys Lys Arg Gly Gly 20 25 30 Gly Gly Ser Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 35 40 45 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 50 55 60 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 65 70 75 80 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 85 90 95 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro 130 135 140 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 145 150 155 160 Ser Ser Gln Ser Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu Thr 165 170 175 Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp 180 185 190 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 195 200 205 Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp 210 215 220 Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 225 230 235 240 Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Gly Ser Glu Asn Leu Tyr 245 250 255 Phe Gln Gly Gly Ser Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu 260 265 270 Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His His His 275 280 285 <210> 34 <211> 290 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 34 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Val Ser Arg Arg Arg Arg Arg Arg Gly Gly Arg Arg Arg 20 25 30 Arg Gly Gly Gly Gly Ser Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly 35 40 45 Leu Glu Trp lie Gly Val lie His Pro Gly Asn Gly Gly Thr Asn Tyr 50 55 60 Asn Glu Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser 65 70 75 80 Ser Thr Ala Tyr Met Gin Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala 85 90 95 Val Tyr Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gin Gly 100 105 110 Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly 115 120 125 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp lie Glu Leu Thr 130 135 140 Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met 145 150 155 160 Ser Cys Lys Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys lie 165 170 175 Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu 180 185 190 Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr 195 200 205 Gly Ser Val Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln 210 215 220 Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro 225 230 235 240 Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Gly Ser Glu 245 250 255 Asn Leu Tyr Phe Gln Gly Gly Ser Gly Lys Pro Ile Pro Asn Pro Leu 260 265 270 Leu Gly Leu Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His 275 280 285 His His 290 <210> 35 <211> 291 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 35 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Gly Lys Cys Ser Thr Arg Gly Arg Lys Cys Cys Arg Arg 20 25 30 Lys Lys Gly Gly Gly Gly Ser Glu Trp Ile Lys Gln Arg Pro Gly Gln 35 40 45 Gly Leu Glu Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn 50 55 60 Tyr Asn Glu Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser 65 70 75 80 Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser 85 90 95 Ala Val Tyr Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly 115 120 125 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp lie Glu Leu 130 135 140 Thr Gin Ser Pro Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr 145 150 155 160 Met Ser Cys Lys Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys 165 170 175 lie Tyr Leu Thr Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu 180 185 190 Leu lie Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe 195 200 205 Thr Gly Ser Val Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Val 210 215 220 Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gin Asn Asp Tyr Asn Tyr 225 230 235 240 Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Gly Ser 245 250 255 Glu Asn Leu Tyr Phe Gln Gly Gly Ser Gly Lys Pro Ile Pro Asn Pro 260 265 270 Leu Leu Gly Leu Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His 275 280 285 His His His 290 <210> 36 <211> 270 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 36 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu 35 40 45 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 50 55 60 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 65 70 75 80 Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 85 90 95 Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro Ser Ser Leu Thr 130 135 140 Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser 145 150 155 160 Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu Thr Trp Tyr Gln Gln 165 170 175 Lys Pro Gly Gln Pro Pro Lys Leu Leu lie Tyr Trp Ala Ser Thr Arg 180 185 190 Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val Ser Gly Thr Asp 195 200 205 Phe Thr Leu Thr lie Ser Ser Val Gin Ala Glu Asp Leu Ala Val Tyr 210 215 220 Tyr Cys Gin Asp Tyr Asn Tyr Pro Tyr Thr Phe Gly Gly Gly Thr 225 230 235 240 Lys Gly Gly Gly Gly Ser His Arg Arg Cys Asn Lys Asn Asn Lys Lys 245 250 255 Arg Leu Glu lie Lys Arg Leu Glu His His His His His His 260 265 270 <210> 37 <211> 270 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 37 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu 35 40 45 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 50 55 60 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 65 70 75 80 Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 85 90 95 Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro Ser Ser Leu Thr 130 135 140 Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser 145 150 155 160 Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu Thr Trp Tyr Gln Gln 165 170 175 Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg 180 185 190 Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val Ser Gly Thr Asp 195 200 205 Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr 210 215 220 Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe Gly Gly Gly Thr 225 230 235 240 Lys Gly Gly Gly Gly Ser His Arg Arg Cys Asn Pro Asn Asn Lys Lys 245 250 255 Arg Leu Glu Ile Lys Arg Leu Glu His His His His His His 260 265 270 <210> 38 <211> 273 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 38 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu 35 40 45 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 50 55 60 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 65 70 75 80 Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 85 90 95 Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro Ser Ser Leu Thr 130 135 140 Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser 145 150 155 160 Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu Thr Trp Tyr Gln Gln 165 170 175 Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg 180 185 190 Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val Ser Gly Thr Asp 195 200 205 Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr 210 215 220 Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe Gly Gly Gly Thr 225 230 235 240 Lys Gly Gly Gly Gly Ser Val Ser Arg Arg Arg Arg Arg Arg Arg Gly Gly 245 250 255 Arg Arg Arg Arg Leu Glu Ile Lys Arg Leu Glu His His His His His 260 265 270 His <210> 39 <211> 274 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 39 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu 35 40 45 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 50 55 60 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 65 70 75 80 Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 85 90 95 Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro Ser Ser Leu Thr 130 135 140 Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser 145 150 155 160 Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu Thr Trp Tyr Gln Gln 165 170 175 Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg 180 185 190 Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val Ser Gly Thr Asp 195 200 205 Phe Thr Leu Thr Ile Ser Ser Val Gin Ala Glu Asp Leu Ala Val Tyr 210 215 220 Tyr Cys Gin Asp Tyr Asn Tyr Pro Tyr Thr Phe Gly Gly Gly Thr 225 230 235 240 Lys Gly Gly Gly Gly Ser Gly Lys Cys Ser Thr Arg Gly Arg Lys Cys 245 250 255 Cys Arg Arg Lys Lys Leu Glu Ile Lys Arg Leu Glu His His His His 260 265 270 His His <210> 40 <211> 275 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 40 Met Ala Gin Val Lys Leu Gin Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile His Arg Arg Cys Asn Lys Asn Asn Lys Lys Arg Gly 35 40 45 Gly Gly Gly Ser Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu 50 55 60 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 65 70 75 80 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 85 90 95 Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 100 105 110 Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr 115 120 125 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly 130 135 140 Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser 145 150 155 160 Pro Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys 165 170 175 Lys Ser Ser Gln Ser Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu 180 185 190 Thr Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr 195 200 205 Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser 210 215 220 Val Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu 225 230 235 240 Asp Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr 245 250 255 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Leu Glu His His His 260 265 270 His His His 275 <210> 41 <211> 275 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 41 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile His Arg Arg Cys Asn Pro Asn Asn Lys Lys Arg Gly 35 40 45 Gly Gly Gly Ser Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu 50 55 60 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 65 70 75 80 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 85 90 95 Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 100 105 110 Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gin Gly Thr Thr 115 120 125 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly 130 135 140 Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp He Glu Leu Thr Gin Ser 145 150 155 160 Pro Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys 165 170 175 Lys Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys He Tyr Leu 180 185 190 Thr Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu He Tyr 195 200 205 Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser 210 215 220 Val Ser Gly Thr Asp Phe Thr Leu Thr He Ser Ser Val Gin Ala Glu 225 230 235 240 Asp Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr 245 250 255 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Leu Glu His His His 260 265 270 His His His 275 <210> 42 <211> 278 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 42 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile Val Ser Arg Arg Arg Arg Arg Arg Gly Gly Arg Arg 35 40 45 Arg Arg Gly Gly Gly Gly Ser Glu Trp Ile Lys Gln Arg Pro Gly Gln 50 55 60 Gly Leu Glu Trp lie Gly Val lie His Pro Gly Asn Gly Gly Thr Asn 65 70 75 80 Tyr Asn Glu Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser 85 90 95 Ser Ser Thr Ala Tyr Met Gin Leu Ser Ser Leu Thr Ser Asp Asp Ser 100 105 110 Ala Val Tyr Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gin 115 120 125 Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly 130 135 140 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp lie Glu Leu 145 150 155 160 Thr Gin Ser Pro Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr 165 170 175 Met Ser Cys Lys Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys 180 185 190 Ile Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu 195 200 205 Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe 210 215 220 Thr Gly Ser Val Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val 225 230 235 240 Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr 245 250 255 Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Leu Glu 260 265 270 His His His His His His 275 <210> 43 <211> 280 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 43 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile Gly Lys Cys Ser Thr Arg Gly Arg Lys Cys Cys Arg 35 40 45 Arg Lys Lys Gly Gly Gly Gly Ser Glu Trp Ile Lys Gln Arg Pro Gly 50 55 60 Gln Gly Leu Glu Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr 65 70 75 80 Asn Tyr Asn Glu Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys 85 90 95 Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp 100 105 110 Ser Ala Val Tyr Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly 115 120 125 Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys 130 135 140 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Glu 145 150 155 160 Leu Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val 165 170 175 Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser Gly Asp Gln 180 185 190 Lys Ile Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys 195 200 205 Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg 210 215 220 Phe Thr Gly Ser Val Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser 225 230 235 240 Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn 245 250 255 Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Leu 260 265 270 Glu His His His His His His His 275 280 <210> 44 <211> 287 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 44 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 100 105 110 Gly Gly Ser Gly Gly Gly Gly Ser Asp lie Glu Leu Thr Gin Ser Pro 115 120 125 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 130 135 140 Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys lie Tyr Leu Thr 145 150 155 160 Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu lie Tyr Trp 165 170 175 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 180 185 190 Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Val Gin Ala Glu Asp 195 200 205 Leu Ala Val Tyr Tyr Cys Gin Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 210 215 220 Gly Gly Gly Thr Lys Gly Gly Gly Gly Ser His Arg Arg Cys Asn Lys 225 230 235 240 Asn Asn Lys Lys Arg Leu Glu Ile Lys Arg Gly Ser Glu Asn Leu Tyr 245 250 255 Phe Gln Gly Gly Ser Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu 260 265 270 Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His His His 275 280 285 <210> 45 <211> 287 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 45 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val lie His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gin Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gin Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 100 105 110 Gly Gly Ser Gly Gly Gly Gly Ser Asp lie Gin Leu Thr Gin Ser Pro 115 120 125 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 130 135 140 Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys lie Tyr Leu Thr 145 150 155 160 Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu lie Tyr Trp 165 170 175 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 180 185 190 Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Val Gin Ala Glu Asp 195 200 205 Leu Ala Val Tyr Tyr Cys Gin Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 210 215 220 Gly Gly Gly Thr Lys Gly Gly Gly Gly Ser His Arg Arg Cys Asn Pro 225 230 235 240 Asn Asn Lys Lys Arg Leu Glu lie Lys Arg Gly Ser Glu Asn Leu Tyr 245 250 255 Phe Gin Gly Gly Ser Gly Lys Pro lie Pro Asn Pro Leu Leu Gly Leu 260 265 270 Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His His His His 275 280 285 <210> 46 <211> 290 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 46 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 100 105 110 Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro 115 120 125 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 130 135 140 Ser Ser Gln Ser Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu Thr 145 150 155 160 Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp 165 170 175 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 180 185 190 Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp 195 200 205 Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 210 215 220 Gly Gly Gly Thr Lys Gly Gly Gly Gly Ser Val Ser Arg Arg Arg Arg 225 230 235 240 Arg Arg Gly Gly Arg Arg Arg Arg Leu Glu lie Lys Arg Gly Ser Glu 245 250 255 Asn Leu Tyr Phe Gin Gly Gly Ser Gly Lys Pro lie Pro Asn Pro Leu 260 265 270 Leu Gly Leu Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His 275 280 285 His His 290 <210> 47 <211> 291 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 47 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gin Ser 1 5 10 15 Ile Gin Ala Glu Trp lie Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp 20 25 30 Ile Gly Val lie His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gin Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gin Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 100 105 110 Gly Gly Ser Gly Gly Gly Gly Ser Asp He Gin Leu Thr Gin Ser Pro 115 120 125 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 130 135 140 Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys He Tyr Leu Thr 145 150 155 160 Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu He Tyr Trp 165 170 175 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 180 185 190 Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp 195 200 205 Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 210 215 220 Gly Gly Gly Thr Lys Gly Gly Gly Gly Ser Gly Lys Cys Ser Thr Arg 225 230 235 240 Gly Arg Lys Cys Cys Arg Arg Lys Lys Leu Glu Ile Lys Arg Gly Ser 245 250 255 Glu Asn Leu Tyr Phe Gln Gly Gly Ser Gly Lys Pro Ile Pro Asn Pro 260 265 270 Leu Leu Gly Leu Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His 275 280 285 His His His 290 <210> 48 <211> 287 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 48 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala His Arg Arg Cys Asn Lys Asn Asn Lys Lys Arg Gly Gly 20 25 30 Gly Gly Ser Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 35 40 45 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 50 55 60 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 65 70 75 80 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 85 90 95 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Gly Gly Gly Gly Ser Asp lie Glu Leu Thr Gin Ser Pro 130 135 140 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 145 150 155 160 Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys lie Tyr Leu Thr 165 170 175 Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu lie Tyr Trp 180 185 190 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 195 200 205 Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Val Gin Ala Glu Asp 210 215 220 Leu Ala Val Tyr Tyr Cys Gin Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 225 230 235 240 Gly Gly Gly Thr Lys Leu Glu lie Lys Arg Gly Ser Glu Asn Leu Tyr 245 250 255 Phe Gin Gly Gly Ser Gly Lys Pro lie Pro Asn Pro Leu Leu Gly Leu 260 265 270 Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His His His 275 280 285 <210> 49 <211> 287 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 49 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gin Ser 1 5 10 15 Ile Gin Ala His Arg Arg Cys Asn Pro Asn Asn Lys Lys Arg Gly Gly 20 25 30 Gly Gly Ser Glu Trp lie Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp 35 40 45 Ile Gly Val lie His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 50 55 60 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 65 70 75 80 Tyr Met Gin Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 85 90 95 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gin Gly Thr Thr Val 100 105 110 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Gly Gly Gly Gly Ser Asp lie Gin Leu Thr Gin Ser Pro 130 135 140 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 145 150 155 160 Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys lie Tyr Leu Thr 165 170 175 Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu lie Tyr Trp 180 185 190 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 195 200 205 Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Val Gin Ala Glu Asp 210 215 220 Leu Ala Val Tyr Tyr Cys Gin Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 225 230 235 240 Gly Gly Gly Thr Lys Leu Glu lie Lys Arg Gly Ser Glu Asn Leu Tyr 245 250 255 Phe Gin Gly Gly Ser Gly Lys Pro lie Pro Asn Pro Leu Leu Gly Leu 260 265 270 Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His His His His 275 280 285 <210> 50 <211> 290 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 50 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Val Ser Arg Arg Arg Arg Arg Arg Arg Gly Gly Arg Arg Arg 20 25 30 Arg Gly Gly Gly Gly Ser Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly 35 40 45 Leu Glu Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr 50 55 60 Asn Glu Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser 65 70 75 80 Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala 85 90 95 Val Tyr Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly 115 120 125 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Glu Leu Thr 130 135 140 Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met 145 150 155 160 Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser Gly Asp Gln Lys Ile 165 170 175 Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu 180 185 190 Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr 195 200 205 Gly Ser Val Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln 210 215 220 Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro 225 230 235 240 Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Gly Ser Glu 245 250 255 Asn Leu Tyr Phe Gin Gly Gly Ser Gly Lys Pro lie Pro Asn Pro Leu 260 265 270 Leu Gly Leu Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His 275 280 285 His His 290 <210> 51 <211> 291 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 51 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gin Ser 1 5 10 15 Ile Gin Ala Gly Lys Cys Ser Thr Arg Gly Arg Lys Cys Cys Arg Arg 20 25 30 Lys Lys Gly Gly Gly Gly Ser Glu Trp lie Lys Gin Arg Pro Gly Gin 35 40 45 Gly Leu Glu Trp lie Gly Val lie His Pro Gly Asn Gly Gly Thr Asn 50 55 60 Tyr Asn Glu Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser 65 70 75 80 Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser 85 90 95 Ala Val Tyr Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly 115 120 125 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Glu Leu 130 135 140 Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr 145 150 155 160 Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser Gly Asp Gln Lys 165 170 175 Ile Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu 180 185 190 Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe 195 200 205 Thr Gly Ser Val Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val 210 215 220 Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr 225 230 235 240 Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Gly Ser 245 250 255 Glu Asn Leu Tyr Phe Gln Gly Gly Ser Gly Lys Pro Ile Pro Asn Pro 260 265 270 Leu Leu Gly Leu Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His 275 280 285 His His His 290 <210> 52 <211> 270 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 52 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu 35 40 45 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 50 55 60 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 65 70 75 80 Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 85 90 95 Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Asp lie Glu Leu Thr Gin Ser Pro Ser Ser Leu Thr 130 135 140 Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gin Ser 145 150 155 160 Leu Leu Asn Ser Gly Asp Gin Lys lie Tyr Leu Thr Trp Tyr Gin Gin 165 170 175 Lys Pro Gly Gin Pro Pro Lys Leu Leu lie Tyr Trp Ala Ser Thr Arg 180 185 190 Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val Ser Gly Thr Asp 195 200 205 Phe Thr Leu Thr lie Ser Ser Val Gin Ala Glu Asp Leu Ala Val Tyr 210 215 220 Tyr Cys Gin Asp Asp Tyr Gin Tyr Pro Tyr Thr Phe Gly Gly Gly Thr 225 230 235 240 Lys Gly Gly Gly Gly Ser His Arg Arg Cys Gin Lys Asn Gin Lys Lys 245 250 255 Arg Leu Glu Ile Lys Arg Leu Glu His His His His His His His 260 265 270 <210> 53 <211> 270 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 53 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu 35 40 45 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 50 55 60 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 65 70 75 80 Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 85 90 95 Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro Ser Ser Leu Thr 130 135 140 Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser 145 150 155 160 Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu Thr Trp Tyr Gln Gln 165 170 175 Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg 180 185 190 Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val Ser Gly Thr Asp 195 200 205 Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr 210 215 220 Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe Gly Gly Gly Thr 225 230 235 240 Lys Gly Gly Gly Gly Ser His Arg Arg Cys Asn Pro Asn Asn Lys Lys 245 250 255 Arg Leu Glu Ile Lys Arg Leu Glu His His His His His His His 260 265 270 <210> 54 <211> 273 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 54 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu 35 40 45 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 50 55 60 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 65 70 75 80 Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 85 90 95 Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro Ser Ser Leu Thr 130 135 140 Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser 145 150 155 160 Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu Thr Trp Tyr Gln Gln 165 170 175 Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg 180 185 190 Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val Ser Gly Thr Asp 195 200 205 Phe Thr Leu Thr Ile Ser Ser Val Gin Ala Glu Asp Leu Ala Val Tyr 210 215 220 Tyr Cys Gin Asp Tyr Gin Tyr Pro Tyr Thr Phe Gly Gly Gly Thr 225 230 235 240 Lys Gly Gly Gly Gly Ser Val Ser Arg Arg Arg Arg Arg Arg Gly Gly 245 250 255 Arg Arg Arg Arg Leu Glu Ile Lys Arg Leu Glu His His His His His 260 265 270 His <210> 55 <211> 274 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 55 Met Ala Gin Val Lys Leu Gin Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu 35 40 45 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 50 55 60 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 65 70 75 80 Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 85 90 95 Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro Ser Ser Leu Thr 130 135 140 Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser 145 150 155 160 Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu Thr Trp Tyr Gln Gln 165 170 175 Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg 180 185 190 Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val Ser Gly Thr Asp 195 200 205 Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr 210 215 220 Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe Gly Gly Gly Thr 225 230 235 240 Lys Gly Gly Gly Gly Ser Gly Lys Cys Ser Thr Arg Gly Arg Lys Cys 245 250 255 Cys Arg Arg Lys Lys Leu Glu Ile Lys Arg Leu Glu His His His His 260 265 270 His His <210> 56 <211> 275 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 56 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile His Arg Arg Cys Asn Lys Asn Asn Lys Lys Arg Gly 35 40 45 Gly Gly Gly Ser Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu 50 55 60 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 65 70 75 80 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 85 90 95 Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 100 105 110 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 115 120 125 Gly Gly Gly Ser Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu 130 135 140 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 145 150 155 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 160 165 170 Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 175 180 185 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 190 195 200 Gly Gly Gly Ser Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu 205 210 215 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 220 225 230 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 235 240 245 Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 250 255 260 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 265 270 275 Gly Gly Gly Ser Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu 280 285 290 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 295 300 305 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 310 315 320 Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val TyrPhe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gin Gly Thr Thr 115 120 125 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly 130 135 140 Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp He Glu Leu Thr Gin Ser 145 150 155 160 Pro Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys 165 170 175 Lys Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys He Tyr Leu 180 185 190 Thr Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu He Tyr 195 200 205 Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser 210 215 220 Val Ser Gly Thr Asp Phe Thr Leu Thr He Ser Ser Val Gin Ala Glu 225 230 235 240 Asp Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr 245 250 255 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Leu Glu His His His 260 265 270 His His His 275 <210> 57 <211> 275 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 57 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile His Arg Arg Cys Asn Pro Asn Asn Lys Lys Arg Gly 35 40 45 Gly Gly Gly Ser Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu 50 55 60 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 65 70 75 80 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 85 90 95 Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 100 105 110 Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr 115 120 125 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly 130 135 140 Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser 145 150 155 160 Pro Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys 165 170 175 Lys Ser Ser Gln Ser Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu 180 185 190 Thr Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu lie Tyr 195 200 205 Trp Ala Ser Thr Arg Gin Ser Gly Val Pro Asp Arg Phe Thr Gly Ser 210 215 220 Val Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Val Gin Ala Gin 225 230 235 240 Asp Leu Ala Val Tyr Tyr Cys Gin Asn Asp Tyr Asn Tyr Pro Tyr Thr 245 250 255 Phe Gly Gly Gly Thr Lys Leu Glu lie Lys Arg Leu Glu His His His 260 265 270 His His His 275 <210> 58 <211> 278 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 58 Met Ala Gin Val Lys Leu Gin Gin Ser Gly Pro Gin Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile Val Ser Arg Arg Arg Arg Arg Arg Gly Gly Arg Arg 35 40 45 Arg Arg Gly Gly Gly Gly Ser Glu Trp Ile Lys Gln Arg Pro Gly Gln 50 55 60 Gly Leu Glu Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn 65 70 75 80 Tyr Asn Glu Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser 85 90 95 Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser 100 105 110 Ala Val Tyr Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln 115 120 125 Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly 130 135 140 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp lie Glu Leu 145 150 155 160 Thr Gin Ser Pro Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr 165 170 175 Met Ser Cys Lys Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys 180 185 190 Ile Tyr Leu Thr Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu 195 200 205 Leu lie Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe 210 215 220 Thr Gly Ser Val Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Val 225 230 235 240 Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gin Asn Asp Tyr Asn Tyr 245 250 255 Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu lie Lys Arg Leu Glu 260 265 270 His His His His His His 275 <210> 59 <211> 280 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 59 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile Gly Lys Cys Ser Thr Arg Gly Arg Lys Cys Cys Arg 35 40 45 Arg Lys Lys Gly Gly Gly Gly Ser Glu Trp Ile Lys Gln Arg Pro Gly 50 55 60 Gln Gly Leu Glu Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr 65 70 75 80 Asn Tyr Asn Glu Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys 85 90 95 Ser Ser Ser Thr Ala Tyr Met Gin Leu Ser Ser Leu Thr Ser Asp Asp 100 105 110 Ser Ala Val Tyr Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly 115 120 125 Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys 130 135 140 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp He Glu 145 150 155 160 Leu Thr Gin Ser Pro Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val 165 170 175 Thr Met Ser Cys Lys Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin 180 185 190 Lys He Tyr Leu Thr Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys 195 200 205 Leu Leu He Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg 210 215 220 Phe Thr Gly Ser Val Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser 225 230 235 240 Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn 245 250 255 Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Leu 260 265 270 Glu His His His His His His His 275 280 <210> 60 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 60 Cys Gly Gly Gly Gly Gly His Arg Arg Cys Asn Lys Asn Asn Lys Lys 1 5 10 15 Arg <210> 61 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 61 Cys Gly Gly Gly Gly Gly His Arg Arg Cys Asn Pro Asn Asn Lys Lys 1 5 10 15 Arg <210> 62 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 62 Cys Gly Gly Gly Gly Gly Val Ser Arg Arg Arg Arg Arg Arg Gly Gly 1 5 10 15 Arg Arg Arg Arg 20 <210> 63 <211> 21 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 63 Cys Gly Gly Gly Gly Gly Gly Lys Cys Ser Thr Arg Gly Arg Lys Cys 1 5 10 15 Cys Arg Arg Lys Lys 20 <210> 64 <211> 24 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 64 Cys Gly Gly Gly Gly Gly Ser Ser Gly Gly Gly Gly Gly Arg Lys Lys 1 5 10 15 Asn Asn Lys Asn Cys Arg Arg His 20 <210> 65 <211> 24 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 65 Cys Gly Gly Gly Gly Gly Ser Ser Gly Gly Gly Gly Gly Arg Lys Lys 1 5 10 15 Asn Asn Pro Asn Cys Arg Arg His 20 <210> 66 <211> 27 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 66 Cys Gly Gly Gly Gly Gly Ser Ser Gly Gly Gly Gly Gly Arg Arg Arg 1 5 10 15 Arg Gly Gly Arg Arg Arg Arg Arg Arg Ser Val 20 25 <210> 67 <211> 28 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 67 Cys Gly Gly Gly Gly Gly Ser Ser Gly Gly Gly Gly Gly Gly Lys Cys 1 5 10 15 Ser Thr Arg Gly Arg Lys Cys Cys Arg Arg Lys Lys 20 25 <210> 68 <211> 288 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 68 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Cys Gly Gly Gly Gly 100 105 110 Gly His Arg Arg Cys Asn Lys Asn Asn Lys Lys Arg Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp lie Glu Leu Thr Gin Ser 130 135 140 Pro Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys 145 150 155 160 Lys Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys lie Tyr Leu 165 170 175 Thr Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu lie Tyr 180 185 190 Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser 195 200 205 Val Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Val Gin Ala Glu 210 215 220 Asp Leu Ala Val Tyr Tyr Cys Gin Asn Asp Tyr Asn Tyr Pro Tyr Thr 225 230 235 240 Phe Gly Gly Gly Thr Lys Leu Glu lie Lys Arg Gly Ser Glu Asn Leu 245 250 255 Tyr Phe Gin Gly Gly Ser Gly Lys Pro lie Pro Asn Pro Leu Leu Gly 260 265 270 Leu Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His His His 275 280 285 <210> 69 <211> 288 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 69 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gin Ser 1 5 10 15 Ile Gin Ala Glu Trp lie Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp 20 25 30 Ile Gly Val lie His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gin Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gin Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Cys Gly Gly Gly Gly 100 105 110 Gly His Arg Arg Cys Asn Pro Asn Asn Lys Lys Arg Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp He Glu Leu Thr Gin Ser 130 135 140 Pro Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys 145 150 155 160 Lys Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys He Tyr Leu 165 170 175 Thr Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu He Tyr 180 185 190 Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser 195 200 205 Val Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu 210 215 220 Asp Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr 225 230 235 240 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Gly Ser Glu Asn Leu 245 250 255 Tyr Phe Gln Gly Gly Ser Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly 260 265 270 Leu Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His His His 275 280 285 <210> 70 <211> 291 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 70 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Cys Gly Gly Gly Gly 100 105 110 Gly Val Ser Arg Arg Arg Arg Arg Arg Gly Gly Arg Arg Arg Arg Gly 115 120 125 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Glu Leu 130 135 140 Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr 145 150 155 160 Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser Gly Asp Gln Lys 165 170 175 Ile Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu 180 185 190 Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe 195 200 205 Thr Gly Ser Val Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val 210 215 220 Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr 225 230 235 240 Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Gly Ser 245 250 255 Glu Asn Leu Tyr Phe Gln Gly Gly Ser Gly Lys Pro Ile Pro Asn Pro 260 265 270 Leu Leu Gly Leu Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His 275 280 285 His His His 290 <210> 71 <211> 292 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 71 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gin Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Cys Gly Gly Gly Gly 100 105 110 Gly Gly Lys Cys Ser Thr Arg Gly Arg Lys Cys Cys Arg Arg Lys Lys 115 120 125 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp He Glu 130 135 140 Leu Thr Gin Ser Pro Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val 145 150 155 160 Thr Met Ser Cys Lys Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin 165 170 175 Lys He Tyr Leu Thr Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys 180 185 190 Leu Leu He Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg 195 200 205 Phe Thr Gly Ser Val Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser 210 215 220 Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn 225 230 235 240 Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Gly 245 250 255 Ser Glu Asn Leu Tyr Phe Gln Gly Gly Ser Gly Lys Pro Ile Pro Asn 260 265 270 Pro Leu Leu Gly Leu Asp Ser Thr Gly Gly Ser Gly Gly Ser His His 275 280 285 His His His His 290 <210> 72 <211> 294 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 72 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Cys Gly Gly Gly Gly 100 105 110 Gly Ser Ser Gly Gly Gly Gly His Arg Arg Cys Asn Lys Asn Asn Lys 115 120 125 Lys Arg Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp 130 135 140 Ile Glu Leu Thr Gin Ser Pro Ser Ser Leu Thr Val Thr Ala Gly Glu 145 150 155 160 Lys Val Thr Met Ser Cys Lys Ser Ser Gin Ser Leu Leu Asn Ser Gly 165 170 175 Asp Gin Lys Ile Tyr Leu Thr Trp Tyr Gin Gin Lys Pro Gly Gin Pro 180 185 190 Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val Pro 195 200 205 Asp Arg Phe Thr Gly Ser Val Ser Gly Thr Asp Phe Thr Leu Thr Ile 210 215 220 Ser Ser Val Gin Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gin Asn Asp 225 230 235 240 Tyr Asn Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 245 250 255 Arg Gly Ser Glu Asn Leu Tyr Phe Gin Gly Gly Ser Gly Lys Pro Ile 260 265 270 Pro Asn Pro Leu Leu Gly Leu Asp Ser Thr Gly Gly Ser Gly Gly Ser 275 280 285 His His His His His His 290 <210> 73 <211> 295 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 73 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Cys Gly Gly Gly Gly 100 105 110 Gly Ser Ser Gly Gly Gly Gly Gly Arg Lys Lys Asn Asn Pro Asn Cys 115 120 125 Arg Arg His Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 130 135 140 Asp Ile Glu Leu Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly 145 150 155 160 Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 165 170 175 Gly Asp Gln Lys Ile Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln 180 185 190 Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 195 200 205 Pro Asp Arg Phe Thr Gly Ser Val Ser Gly Thr Asp Phe Thr Leu Thr 210 215 220 Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn 225 230 235 240 Asp Tyr Asn Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile 245 250 255 Lys Arg Gly Ser Glu Asn Leu Tyr Phe Gln Gly Gly Ser Gly Lys Pro 260 265 270 Ile Pro Asn Pro Leu Leu Gly Leu Asp Ser Thr Gly Gly Ser Gly Gly 275 280 285 Ser His His His His His His 290 295 <210> 74 <211> 298 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 74 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Cys Gly Gly Gly Gly 100 105 110 Gly Ser Ser Gly Gly Gly Gly Gly Arg Arg Arg Arg Gly Gly Arg Arg 115 120 125 Arg Arg Arg Arg Ser Val Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 130 135 140 Gly Gly Ser Asp lie Glu Leu Thr Gin Ser Pro Ser Ser Leu Thr Val 145 150 155 160 Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gin Ser Leu 165 170 175 Leu Asn Ser Gly Asp Gin Lys lie Tyr Leu Thr Trp Tyr Gin Gin Lys 180 185 190 Pro Gly Gin Pro Pro Lys Leu Leu lie Tyr Trp Ala Ser Thr Arg Glu 195 200 205 Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val Ser Gly Thr Asp Phe 210 215 220 Thr Leu Thr lie Ser Ser Val Gin Ala Glu Asp Leu Ala Val Tyr Tyr 225 230 235 240 Cys Gin Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys 245 250 255 Leu Glu lie Lys Arg Gly Ser Glu Asn Leu Tyr Phe Gin Gly Gly Ser 260 265 270 Gly Lys Pro lie Pro Asn Pro Leu Leu Gly Leu Asp Ser Thr Gly Gly 275 280 285 Ser Gly Gly Ser His His His His His His 290 295 <210> 75 <211> 299 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 75 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 lie Gin Ala Glu Trp lie Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp 20 25 30 lie Gly Val lie His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gin Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Cys Gly Gly Gly Gly 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Cys Gly Gly Gly Gly 100 105 110 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Cys Gly Gly Gly Gly 115 120 125 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Cys Gly Gly Gly Gly 130 135 140 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Cys Gly Gly Gly Gly 145 150 155 160 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Cys Gly Gly Gly Gly 165 170 175 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Cys Gly Gly Gly Gly 180 185 190 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Cys Gly Gly Gly Gly 195 200 205 Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val Ser Gly Thr Asp 210 215 220 Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr 225 230 235 240 Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe Gly Gly Gly Thr 245 250 255 Lys Leu Glu Ile Lys Arg Gly Ser Glu Asn Leu Tyr Phe Gln Gly Gly 260 265 270 Ser Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp Ser Thr Gly 275 280 285 Gly Ser Gly Gly Ser His His His His His His 290 295 <210> 76 <211> 289 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 76 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 100 105 110 Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro 115 120 125 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 130 135 140 Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys He Tyr Leu Thr 145 150 155 160 Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu He Tyr Trp 165 170 175 Ala Ser Thr Arg Gin Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 180 185 190 Ser Gly Thr Asp Phe Thr Leu Thr He Ser Ser Val Gin Ala Gin Asp 195 200 205 Leu Ala Val Tyr Tyr Cys Gin Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 210 215 220 Gly Gly Gly Thr Lys Cys Cys Gly Gly Gly Gly Gly His Arg Arg Cys 225 230 235 240 Asn Lys Asn Asn Lys Lys Arg Leu Gin He Lys Arg Gly Ser Gin Asn 245 250 255 Leu Tyr Phe Gin Gly Gly Ser Gly Lys Pro He Pro Asn Pro Leu Leu 260 265 270 Gly Leu Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His His 275 280 285 His <210> 77 <211> 289 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 77 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gin Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 100 105 110 Gly Gly Ser Gly Gly Gly Gly Ser Asp He Glu Leu Thr Gin Ser Pro 115 120 125 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 130 135 140 Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys He Tyr Leu Thr 145 150 155 160 Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu He Tyr Trp 165 170 175 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 180 185 190 Ser Gly Thr Asp Phe Thr Leu Thr He Ser Ser Val Gin Ala Glu Asp 195 200 205 Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 210 215 220 Gly Gly Gly Thr Lys Cys Cys Gly Gly Gly Gly Gly His Arg Arg Cys 225 230 235 240 Asn Pro Asn Asn Lys Lys Arg Leu Glu Ile Lys Arg Gly Ser Glu Asn 245 250 255 Leu Tyr Phe Gln Gly Gly Ser Gly Lys Pro Ile Pro Asn Pro Leu Leu 260 265 270 Gly Leu Asp Ser Thr Gly Gly Ser Gly Gly Ser His His His His His 275 280 285 His <210> 78 <211> 292 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 78 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 100 105 110 Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro 115 120 125 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 130 135 140 Ser Ser Gln Ser Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu Thr 145 150 155 160 Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp 165 170 175 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 180 185 190 Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp 195 200 205 Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 210 215 220 Gly Gly Gly Thr Lys Cys Cys Gly Gly Gly Gly Gly Val Ser Arg Arg 225 230 235 240 Arg Arg Arg Arg Gly Gly Arg Arg Arg Arg Leu Glu Ile Lys Arg Gly 245 250 255 Ser Glu Asn Leu Tyr Phe Gln Gly Gly Ser Gly Lys Pro Ile Pro Asn 260 265 270 Pro Leu Leu Gly Leu Asp Ser Thr Gly Gly Ser Gly Gly Ser His His 275 280 285 His His His His 290 <210> 79 <211> 293 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 79 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 100 105 110 Gly Gly Ser Gly Gly Gly Gly Ser Asp lie Glu Leu Thr Gin Ser Pro 115 120 125 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 130 135 140 Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys lie Tyr Leu Thr 145 150 155 160 Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu lie Tyr Trp 165 170 175 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 180 185 190 Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Val Gin Ala Glu Asp 195 200 205 Leu Ala Val Tyr Tyr Cys Gin Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 210 215 220 Gly Gly Gly Thr Lys Cys Cys Gly Gly Gly Gly Gly Gly Lys Cys Ser 225 230 235 240 Thr Arg Gly Arg Lys Cys Cys Arg Arg Lys Lys Leu Glu Ile Lys Arg 245 250 255 Gly Ser Glu Asn Leu Tyr Phe Gln Gly Gly Ser Gly Lys Pro Ile Pro 260 265 270 Asn Pro Leu Leu Gly Leu Asp Ser Thr Gly Gly Ser Gly Gly Ser His 275 280 285 His His His His His 290 <210> 80 <211> 295 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 80 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 100 105 110 Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro 115 120 125 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 130 135 140 Ser Ser Gln Ser Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu Thr 145 150 155 160 Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu lie Tyr Trp 165 170 175 Ala Ser Thr Arg Gin Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 180 185 190 Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Val Gin Ala Glu Asp 195 200 205 Leu Ala Val Tyr Tyr Cys Gin Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 210 215 220 Gly Gly Gly Thr Lys Cys Cys Gly Gly Gly Gly Gly Ser Ser Gly Gly 225 230 235 240 Gly Gly His Arg Arg Cys Asn Lys Asn Asn Lys Lys Arg Leu Glu lie 245 250 255 Lys Arg Gly Ser Glu Asn Leu Tyr Phe Gin Gly Gly Ser Gly Lys Pro 260 265 270 lie Pro Asn Pro Leu Leu Gly Leu Asp Ser Thr Gly Gly Ser Gly Gly 275 280 285 Ser His His His His His His 290 295 <210> 81 <211> 296 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 81 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 100 105 110 Gly Gly Ser Gly Gly Gly Gly Ser Asp lie Glu Leu Thr Gin Ser Pro 115 120 125 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 130 135 140 Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys lie Tyr Leu Thr 145 150 155 160 Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu lie Tyr Trp 165 170 175 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 180 185 190 Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Val Gin Ala Glu Asp 195 200 205 Leu Ala Val Tyr Tyr Cys Gin Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 210 215 220 Gly Ser Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly GlyGly Gly Gly Thr Lys Cys Cys Gly Gly Gly Gly Gly Gly Ser Ser Gly Gly 225 230 235 240 Gly Gly Gly Arg Lys Lys Asn Asn Pro Asn Cys Arg Arg His Leu Glu 245 250 255 Ile Lys Arg Gly Ser Glu Asn Leu Tyr Phe Gln Gly Gly Ser Gly Lys 260 265 270 Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp Ser Thr Gly Gly Ser Gly 275 280 285 Gly Ser His His His His His His 290 295 <210> 82 <211> 301 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 82 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 100 105 110 Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro 115 120 125 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 130 135 140 Ser Ser Gln Ser Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu Thr 145 150 155 160 Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp 165 170 175 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 180 185 190 Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp 195 200 205 Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 210 215 220 Gly Gly Gly Thr Lys Cys Cys Gly Gly Gly Gly Gly Ser Ser Gly Gly 225 230 235 240 Gly Gly Gly Val Ser Arg Arg Arg Arg Gly Gly Arg Arg Arg Arg Arg 245 250 255 Arg Ser Val Leu Glu Ile Lys Arg Gly Ser Glu Asn Leu Tyr Phe Gln 260 265 270 Gly Gly Ser Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp Ser 275 280 285 Thr Gly Gly Ser Gly Gly Ser His His His His His His 290 295 300 <210> 83 <211> 300 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 83 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ile Gln Ala Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp 20 25 30 Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu Asn 35 40 45 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 50 55 60 Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe 65 70 75 80 Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr Val 85 90 95 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Lys Gly Gly Ser Gly Gly 100 105 110 Gly Gly Ser Gly Gly Gly Gly Ser Asp lie Glu Leu Thr Gin Ser Pro 115 120 125 Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys 130 135 140 Ser Ser Gin Ser Leu Leu Asn Ser Gly Asp Gin Lys lie Tyr Leu Thr 145 150 155 160 Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu lie Tyr Trp 165 170 175 Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val 180 185 190 Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Val Gin Ala Glu Asp 195 200 205 Leu Ala Val Tyr Tyr Cys Gin Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe 210 215 220 Gly Gly Gly Thr Lys Cys Cys Gly Gly Gly Gly Gly Ser Ser Gly Gly 225 230 235 240 Gly Gly Gly Gly Lys Cys Ser Thr Arg Gly Arg Lys Cys Cys Arg Arg 245 250 255 Lys Lys Leu Glu Ile Lys Arg Gly Ser Glu Asn Leu Tyr Phe Gln Gly 260 265 270 Gly Ser Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp Ser Thr 275 280 285 Gly Gly Ser Gly Gly Ser His His His His His His 290 295 300 <210> 84 <211> 34 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 84 aaggagatat acatatgatg gcatgggttt ggac 34 <210> 85 <211> 46 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 85 agcccgaagg gaattcattt gcagatacaa agtgttttta gagttg 46 <400> 85 <211> 11 <400> 85 <213> Artificial Sequence <220> <223> H4K <400> 86 His Arg Arg Cys Asn Lys Asn Lys Lys Arg 1 5 10 <400> 86 <211> 11 <400> 86 <213> Artificial Sequence <220> <223> H4P <400> 87 His Arg Arg Cys Asn Pro Asn Lys Lys Arg 1 5 10 <400> 87 <211> 14 <400> 87 <213> Artificial Sequence <220> <223> LMWP <400> 88 Val Ser Arg Arg Arg Arg Arg Arg Gly Gly Arg Arg Arg Arg 1 5 10 <210> 89 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> hBD3‑3 <400> 89 Gly Lys Cys Ser Thr Arg Gly Arg Lys Cys Cys Arg Arg Lys Lys 1 5 10 15 <210> 90 <211> 280 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 90 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile Glu Trp Ile Lys Gin Arg Pro Gly Gin Gly Leu Glu 35 40 45 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 50 55 60 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 65 70 75 80 Ala Tyr Met Gin Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 85 90 95 Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gin Gly Thr Thr 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Asp Ile Gin Leu Thr Gin Ser Pro Ser Ser Leu Thr 130 135 140 Val Thr Ala Gly Gin Lys Val Thr Met Ser Cys Lys Ser Ser Gin Ser 145 150 155 160 Leu Leu Asn Ser Gly Asp Gin Lys He Tyr Leu Thr Trp Tyr Gin Gin 165 170 175 Lys Pro Gly Gin Pro Pro Lys Leu Leu He Tyr Trp Ala Ser Thr Arg 180 185 190 Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val Ser Gly Thr Asp 195 200 205 Phe Thr Leu Thr He Ser Ser Val Gin Ala Glu Asp Leu Ala Val Tyr 210 215 220 Tyr Cys Gin Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe Gly Gly Gly Thr 225 230 235 240 Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 245 250 255 His Arg Arg Cys Asn Lys Asn Asn Lys Lys Arg Leu Glu He Lys Arg 260 265 270 Leu Glu His His His His His His 275 280 <210> 91 <211> 280 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 91 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu 35 40 45 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 50 55 60 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 65 70 75 80 Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 85 90 95 Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro Ser Ser Leu Thr 130 135 140 Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser 145 150 155 160 Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu Thr Trp Tyr Gln Gln 165 170 175 Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg 180 185 190 Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val Ser Gly Thr Asp 195 200 205 Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr 210 215 220 Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe Gly Gly Gly Thr 225 230 235 240 Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 245 250 255 His Arg Arg Cys Asn Pro Asn Asn Lys Lys Arg Leu Glu Ile Lys Arg 260 265 270 Leu Glu His His His His His His 275 280 <210> 92 <211> 283 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 92 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu 35 40 45 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 50 55 60 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 65 70 75 80 Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 85 90 95 Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro Ser Ser Leu Thr 130 135 140 Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser 145 150 155 160 Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu Thr Trp Tyr Gln Gln 165 170 175 Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg 180 185 190 Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val Ser Gly Thr Asp 195 200 205 Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr 210 215 220 Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe Gly Gly Gly Thr 225 230 235 240 Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 245 250 255 Val Ser Arg Arg Arg Arg Arg Arg Gly Gly Arg Arg Arg Arg Leu Glu 260 265 270 Ile Lys Arg Leu Glu His His His His His His 275 280 <210> 93 <211> 284 <212> PRT <213> Artificial Sequence <220> <223> fusion peptide <400> 93 Met Ala Gln Val Lys Leu Gln Glu Ser Gly Pro Glu Leu Val Arg Pro 1 5 10 15 Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr 20 25 30 Asn Tyr Leu Ile Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu 35 40 45 Trp Ile Gly Val Ile His Pro Gly Asn Gly Gly Thr Asn Tyr Asn Glu 50 55 60 Asn Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 65 70 75 80 Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr 85 90 95 Phe Cys Ala Ser Gly Asn Asp Gly Ser Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro Ser Ser Leu Thr 130 135 140 Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser 145 150 155 160 Leu Leu Asn Ser Gly Asp Gln Lys Ile Tyr Leu Thr Trp Tyr Gln Gln 165 170 175 Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg 180 185 190 Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Val Ser Gly Thr Asp 195 200 205 Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr 210 215 220 Tyr Cys Gln Asn Asp Tyr Asn Tyr Pro Tyr Thr Phe Gly Gly Gly Thr 225 230 235 240 Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 245 250 255 Gly Lys Cys Ser Thr Arg Gly Arg Lys Cys Cys Arg Arg Lys Lys Leu 260 265 270 Glu Ile Lys Arg Leu Glu His His His His His His 　　 275 280

Claims

1. A fusion protein, wherein (i) an antibody or single chain variable fragment thereof targeting an intracellular oncogenic protein or oncogenic mutant protein is linked via a linker to (ii) a cancer cell penetrating peptide, wherein the fusion protein is represented by any one of the amino acid sequences of SEQ ID NO: 20 to SEQ ID NO: 23, SEQ ID NO: 28 to SEQ ID NO: 31 and SEQ ID NO: 90 to SEQ ID NO:

93.

2. A fusion protein, wherein (i) an antibody or single chain variable fragment thereof targeting an intracellular oncogenic protein or oncogenic mutant protein is linked via a linker to (ii) a cancer cell penetrating peptide, wherein the sequence of the antibody or single chain variable fragment thereof is set forth in SEQ ID NO: 1 ; wherein the sequence of the linker and the cancer cell penetrating peptide is set forth in any one of SEQ ID NO: 60 to 61, wherein the cancer cell penetrating peptide is linked via a linker to the antibody or single chain variable peptide at its lysine at position 107.

3. A fusion protein, wherein (i) an antibody or single chain variable fragment thereof targeting an intracellular oncogenic protein or oncogenic mutant protein is linked via a linker to (ii) a cancer cell penetrating peptide, wherein the sequence of the antibody or single chain variable fragment thereof is set forth in SEQ ID NO: 3; wherein the sequence of the linker and the cancer cell penetrating peptide is set forth in any one of CGGGGGSSGGGG-HRRCNKNNKKR, CGGGGGSSGGGGG-RKKNNPNCRRH, CGGGGGSSGGGGG-VSRRRRGGRRRRRRSV and CGGGGGSSGGGGG-GKCSTRGRKCCRRKK, wherein the cancer cell penetrating peptide is linked via a linker to the antibody or single chain variable peptide at its cysteine at position 230.

4. The fusion protein according to claim 1, wherein the intracellular oncogenic protein or oncogenic mutant protein is KRAS.

5. A nucleic acid encoding the fusion protein according to claim 1.

6. A recombinant vector into which the nucleic acid according to claim 5 has been introduced.

7. A recombinant cell into which the recombinant vector according to claim 6 has been introduced.

8. The recombinant cell according to claim 7, wherein the recombinant cell is E. coli or a mammalian cell.

9. A method for producing the fusion protein according to claim 1, comprising the steps of: (a) expressing the fusion protein according to claim 1 by culturing the recombinant cell according to claim 7; and (b) recovering the expressed fusion protein.

10. A pharmaceutical composition comprising the fusion protein according to any one of claims 1 to 4 as an active ingredient. ​ 11. Use of the pharmaceutical composition according to claim 10 for the manufacture of a medicament for the treatment of a tumor, wherein the tumor is at least one selected from the group consisting of non-small cell lung cancer, colorectal cancer, pancreatic cancer, bladder cancer, kidney cancer, thyroid cancer, breast cancer, colon cancer, liver cancer, brain tumor, skin cancer, melanoma, prostate cancer, and blood cancer.

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