FC fusion protein for neutralizing type 1 interferon and its use

Abstract

The present invention relates to an FC fusion protein for neutralizing type 1 interferon and its use, and more specifically, to: a dimeric polypeptide in which a monomer of an interferon receptor fragment or an antibody FC fragment is bound; a preparation method thereof; and a pharmaceutical composition comprising the same. The FC fusion protein for neutralizing type 1 interferon of the present invention blocks the binding between type 1 interferon and interferon receptor, and has an excellent ability to inhibit the signal transduction and biological activity of interferon, thereby being able to effectively treat diseases mediated by type 1 interferon.

Description

Technical Field

[0001] This application claims priority to Korean Patent Application No. 10-2018-0168801, filed on December 24, 2018, the entire description of which is incorporated herein by reference.

[0002] This invention relates to an Fc fusion protein that neutralizes type 1 interferon and its uses, and more specifically, to a dimer polypeptide comprising a monomer of an interferon receptor fragment or an antibody Fc fragment bound therein, a method for preparing the same, and a composition comprising said dimer polypeptide for the prevention or treatment of diseases associated with abnormal expression of type 1 interferon or interferon-induced genes. Background Technology

[0003] Type I interferons (IFNs) (IFN-α, IFN-β, IFN-ω, IFN-τ) are a group of structure-associated cytokines with antiviral, antitumor, and immunomodulatory effects. The human IFNα locus has two subgroups. The first subgroup contains 14 non-allelic genes and 4 pseudogenes with at least 80% identity. The second subgroup, αII or omega(ω), has 1 functional gene and 5 pseudogenes with 70% identity to the IFNα gene. The IFNα isoforms have different specific activities but exhibit the same biological spectrum and the same cellular receptor. Interferon β (IFNβ) is encoded by a single gene and shares nearly 50% identity with the IFNα gene. Interferon γ, produced by activated lymphocytes, has no identity with α / β interferons and does not respond to its receptor.

[0004] All human type I IFNs bind to a cell surface receptor (IFNα receptor, IFNAR), which is composed of two transmembrane proteins, IFNAR-1 and IFNAR-2. IFNAR-1 is essential for the high-affinity binding and varying specificity of the IFNAR complex. Although the functional differences among each type I IFN isoform have not been elucidated, it is believed that each isoform exhibits different roles in the IFNAR receptor components, potentially leading to multiple signal transduction pathways.

[0005] Early studies on the function of type I IFNs focused on innate defense against viral infections. However, recent research has identified type I IFNs as potent immunomodulatory cytokines in adaptive immune responses. In particular, type I IFNs have been shown to promote mutations in native T cells within the Th1 pathway, increase antibody production, and support the functional activity and survival of memory T cells.

[0006] Several recent studies have shown that IFN-α can enhance the maturation or activation of dendritic cells (DCs). Furthermore, increased expression of type I interferon has been described in numerous studies related to autoimmune diseases.

[0007] The most studied examples are insulin-dependent diabetes mellitus (IDDM) (Paulis et al. (1987) Lancet 2:1423), systemic lupus erythematosus (SLE) (Hooks et al. (1982) Arthritis Rheum 25:396), Sjögren's syndrome (LeeHong-Yau et al. (2013) Autoimmun Rev 12(5):558-66), inflammatory myositis (Beckler et al. (2007) Mol Med 13(1-2):59–68.), and rheumatoid arthritis (RA), in which IFN-β plays a more important role (Herzzog et al. (1988) Clin Immunol Immunopathol 48:192, Hopkins and Meggle (1988) Clin Exp Immunol 73:88, Alvin and Miller (1984) Arthritis Rheum 27:582), all of which are associated with elevated IFNα levels.

[0008] Furthermore, as a representative type of type 1 interferon, interferon-alpha has been reported to exacerbate underlying diseases in patients with psoriasis and multiple sclerosis, and to induce SLE-like symptoms in patients without prior experience with autoimmune diseases. Interferon-alpha has also been shown to induce glomerulonephritis in normal mice and accelerate the development of concomitant autoimmune disease in NZB / W mice. In addition, IFN-alpha therapy has been shown to cause adverse side effects in certain cases, including fever and neurological disturbances.

[0009] Although type 1 IFNs are diverse, and recent studies in SLE patients have shown that not only IFN-α but also IFN-β are causes of mucocutaneous dermatitis, the problem is that treatment for type 1 interferon-mediated disease has focused solely on IFN-α, such as vaccines (IFN-kinoids) or anti-IFN-α monoclonal neutralizing antibodies (Sifalimumab, Rontalizumab, AGS-009). Furthermore, these neutralizing antibodies are problematic because they do not neutralize all portions of the two different receptors that bind to interferon and only have binding capacity for IFNAR1 or IFNAR2. To address this issue, aniluzumab (MAbs. 2015 Mar-Apr; 7(2):428-439.) is being developed, which specifically binds to and neutralizes the type 1 IFN receptor IFNAR1. However, these neutralizing antibody therapies are not expected to be able to eliminate type 1 interferon from the body due to the multiple signaling mechanisms involved, and the complex signaling mechanisms, such as the non-classical pathways mediated by IFNAR1 (Nat Immunol. 2013 Sep; 14(9):901-7.) or IFNAR2 (Sci Signal. 2014 May 27; 7(327):ra50.; PLoS One. 2017; 12(8):e0182866.), rather than the classical pathways mediated by IFNAR1 and IFNAR2. Invention Details

[0011] Technical issues

[0012] The inventors of this invention completed this invention while developing a treatment for type 1 interferon-mediated diseases, because it was demonstrated that a dimeric polypeptide containing interferon receptor 1 fragments and interferon receptor 2 fragments not only binds to type 1 interferon, but also significantly inhibits the initiation of signal transduction mechanisms and the inhibition of biological activity during treatment.

[0013] Therefore, the object of the present invention is to provide

[0014] A dimer polypeptide comprising a monomer of an interferon receptor fragment or an antibody Fc fragment bound thereto,

[0015] The monomer is a polypeptide selected from (i), (ii) and (iii):

[0016] (i) A monomer containing an interferon receptor 1 (IFNAR1) fragment and an antibody Fc fragment;

[0017] (ii) A monomer containing an interferon receptor 2 (IFNAR2) fragment and an antibody Fc fragment; and

[0018] (iii) Antibody Fc fragment.

[0019] Another object of the present invention is to provide a polynucleotide encoding the said polypeptide.

[0020] Another object of the present invention is to provide a carrier containing the said polynucleotide.

[0021] Another object of the present invention is to provide a host cell transformed using the said vector.

[0022] Another object of the present invention is to provide a method for preparing the polypeptide, comprising:

[0023] (a) Provide host cells;

[0024] (b) Cultivating the provided cells; and

[0025] (c) Prepare peptides by recovering peptides from cells or culture media.

[0026] Another object of the present invention is,

[0027] A pharmaceutical composition is provided for the prevention or treatment of type 1 interferon-mediated diseases or conditions, comprising a polypeptide as an active ingredient.

[0028] Also provided is a pharmaceutical composition for the prevention or treatment of type 1 interferon-mediated diseases or conditions, comprising a polypeptide as an active ingredient.

[0029] Also provided is a pharmaceutical composition for the prevention or treatment of type 1 interferon-mediated diseases or conditions, which is mainly composed of polypeptides as active ingredients.

[0030] Another object of the present invention is to provide the use of the polypeptide in the preparation of a medicament for the prevention or treatment of type 1 interferon-mediated diseases or conditions.

[0031] Another object of the present invention is to provide a method for treating type 1 interferon-mediated diseases or conditions, comprising administering an effective amount of a composition to a subject in need, the composition comprising the polypeptide as an active ingredient.

[0032] Technical solution

[0033] Therefore, in order to achieve the objectives of this invention, this invention provides...

[0034] A dimer polypeptide comprising a monomer of an interferon receptor fragment or an antibody Fc fragment bound thereto,

[0035] The monomer is a polypeptide selected from (i), (ii) and (iii):

[0036] (ii) Monomers containing an interferon receptor 1 (IFNAR1) fragment and an antibody Fc fragment;

[0037] (ii) A monomer containing an interferon receptor 2 (IFNAR2) fragment and an antibody Fc fragment; and

[0038] (iii) Antibody Fc fragment.

[0039] To achieve another objective of the present invention, the present invention provides a polynucleotide encoding the polypeptide.

[0040] To achieve another objective of the present invention, the present invention provides a vector containing the polynucleotide.

[0041] To achieve another objective of the present invention, the present invention provides host cells transformed with the vector.

[0042] To achieve another objective of the present invention, the present invention provides a method for preparing the polypeptide, comprising:

[0043] (a) Provide host cells;

[0044] (b) Cultivating the provided cells; and

[0045] (c) Prepare peptides by recovering peptides from cells or culture media.

[0046] To achieve another objective of the present invention, the present invention

[0047] A pharmaceutical composition is provided for the prevention or treatment of type 1 interferon-mediated diseases or conditions, comprising the polypeptide as an active ingredient.

[0048] A pharmaceutical composition for the prevention or treatment of type 1 interferon-mediated diseases or conditions is also provided, comprising a polypeptide as the active ingredient.

[0049] A pharmaceutical composition for the prevention or treatment of type 1 interferon-mediated diseases or conditions is also provided, which is mainly composed of polypeptides as active ingredients.

[0050] To achieve another objective of the present invention, the present invention provides the use of the polypeptide in the preparation of a medicament for the prevention or treatment of type 1 interferon-mediated diseases or conditions.

[0051] To achieve another objective of the present invention, the present invention provides a method for treating type 1 interferon-mediated diseases or conditions, comprising administering an effective amount of a composition to a subject in need, the composition comprising the polypeptide as an active ingredient.

[0052] The present invention will now be described in detail.

[0053] Therefore, the present invention is a dimer polypeptide in which a monomer comprising an interferon receptor fragment or an antibody Fc fragment is bound.

[0054] The monomer provides polypeptides selected from (i), (ii) and (iii):

[0055] (ii) Monomers containing an interferon receptor 1 (IFNAR1) fragment and an antibody Fc fragment;

[0056] (ii) A monomer containing an interferon receptor 2 (IFNAR2) fragment and an antibody Fc fragment; and

[0057] (iii) Antibody Fc fragment.

[0058] In this specification, including the claims, the term "interferon receptor fragment" is intended to include all polypeptides having human interferon receptor activity and having an amino acid sequence wholly or partially derived from the natural human interferon receptor.

[0059] The terms “interferon receptor 1,” “IFNAR 1,” and “IFNAR-1 antigen” are used interchangeably and include variants, isotypes, species-specific isotypes of human IFNAR1, and analogs having at least one common epitope of IFNAR 1. Therefore, the peptides of the present invention may cross-react with IFNAR 1 from other non-human species or with other proteins structurally associated with human IFNAR 1 (e.g., human IFNAR-1 homologs) in certain circumstances. In other circumstances, the peptides may be completely specific to human IFNAR-1 and do not represent species-specific or other types of cross-reactivity. The complete cDNA sequence of human IFNAR 1 is available in the gene bank accession numbers XM_005260964.2, NM_000629.2, or XM_011529552.2.

[0060] The nucleotide sequence used in this invention may include the extracellular domain (28-436a.a.; P17181-1) of the sequence of IFNAR 1 isotype 1 (GenBank accession number: NM_000629.2), and may include a portion of that nucleotide sequence. The IFNAR 1 fragment of this invention is preferably an interferon receptor 1 comprising the amino acid sequence represented by SEQ ID NO: 4, more preferably an interferon receptor 1 composed of the amino acid sequence represented by SEQ ID NO: 4.

[0061] In one embodiment of the invention, the polypeptide is prepared using interferon receptor 1, which comprises the amino acid sequence represented by SEQ ID NO:4 (the polynucleotide sequence is represented by SEQ ID NO:3 - the polynucleotide sequence may include a stop codon. All subsequent DNA sequences are identical).

[0062] The terms “interferon receptor 2,” “IFNAR 2,” and “IFNAR-2 antigen” are used interchangeably and include variants, isotypes, species-specific isotypes of human IFNAR2, and analogs having at least one common epitope of IFNAR 2. Therefore, the peptides of the present invention may cross-react with IFNAR 2 from other non-human species or with other proteins structurally associated with human IFNAR 2 (e.g., human IFNAR-2 homologs) in certain circumstances. In other circumstances, the peptide may be completely specific to human IFNAR-2 and does not represent species-specific or other types of cross-reactivity. The complete cDNA sequence of human IFNAR 2 is available in the gene bank accession numbers NM_000874.4, NM_001289125.1, or NM_001289126.1. The nucleotide sequence used in this invention may include the extracellular domain (27-243a.a.; P48551-1) of the sequence of IFNAR 2 isotype 1 (GenBank accession number: NM_001289125.1), and may include a portion of that nucleotide sequence. The IFNAR 2 fragment of this invention is preferably an interferon receptor 2 comprising the amino acid sequence represented by SEQ ID NO: 6, more preferably an interferon receptor 2 composed of the amino acid sequence represented by SEQ ID NO: 6.

[0063] In one embodiment of the invention, the polypeptide is prepared using interferon receptor 2, which comprises the amino acid sequence represented by SEQ ID NO:46 (the polynucleotide sequence is represented by SEQ ID NO:5).

[0064] Furthermore, the term "peptide having all or part of the amino acid sequence derived from an interferon receptor fragment" is intended to include peptides containing all or most of the amino acid sequence of SEQ ID NO: 4 or SEQ ID NO: 6, which is the amino acid sequence of a natural interferon receptor fragment or a peptide substantially similar to such peptides.

[0065] Here, "a polypeptide comprising the major portion of the entire amino acid sequence of SEQ ID NO: 4 or SEQ ID NO: 6" is defined as a polypeptide comprising an activity equal to or greater than that of a native interferon receptor having the amino acid sequence of SEQ ID NO: 4 or SEQ ID NO: 6, or a portion of the amino acid sequence of SEQ ID NO: 4 or SEQ ID NO: 6, comprising one or more substituted amino acids, while still retaining interferon receptor activity, albeit at a low level. Furthermore, "a polypeptide substantially similar in all or most of the amino acid sequence represented by SEQ ID NO: 4 or SEQ ID NO: 6" is defined as a polypeptide comprising an interferon receptor activity equal to or greater than that of SEQ ID NO: 4 or SEQ ID NO: 6, or the entire or major portion of the amino acid sequence of SEQ ID NO: 4 or SEQ ID NO: 6, comprising one or more substituted amino acids, while still retaining interferon receptor activity, albeit at a low level.

[0066] As a polypeptide comprising the main part of the entire amino acid sequence represented by SEQ ID NO:4 or SEQ ID NO:6, there may be a lack of the N-terminal portion and / or the C-terminal portion in the polypeptide comprising the amino acid sequence represented by SEQ ID NO:4 or SEQ ID NO:6, and even if one or more amino acids are replaced, if the amino acid before the replacement is chemically identical to the replaced amino acid, for example, when alanine, which is a hydrophobic amino acid, is replaced by another hydrophobic amino acid, in particular, as a polypeptide substantially similar to all or the main part of the amino acid sequence represented by SEQ ID NO:4 or SEQ ID NO:6, it is replaced by a more hydrophobic amino acid, such as valine, leucine, or isoleucine.

[0067] In this invention, "fusion" refers to the integration of two molecules with different or the same functions or structures, which can be achieved by any physical, chemical or biological method that can bind the antibody Fc portion or individual monomers to the interferon receptor.

[0068] Fusion is preferably carried out via a linker peptide that can bind to the C-terminus of, for example, the Fc fragment of an antibody.

[0069] The “antibody Fc fragment” can be an Fc fragment of an IgA, IgM, IgE, IgD, or IgG antibody, or a modification thereof. In one embodiment, the fragment is an Fc fragment of an IgG antibody (e.g., an Fc fragment of an IgG1, IgG2a, IgG2b, IgG3, or IgG4 antibody). Furthermore, the polypeptide comprising the Fc fragment of the present invention can be partially or fully glycosylated or non-glycosylated. In addition to the Fc fragment, the polypeptide can include one or more antibody-derived regions. Furthermore, the polypeptide can include an antibody-derived antigen-binding domain, and multiple polypeptides can form an antibody or antibody-like protein.

[0070] In this specification, the amino acid residue numbers of the antibody Fc fragments are based on the Kabat numbering system commonly used in the field (Kabat et al., EU index number: Proteins of Immunological Interest 5th Ed., USDepartment of Health and Human Services, NIH Publication No. 91-3242, 1991).

[0071] According to one embodiment of the invention, it may include the wild-type Fc fragment of the invention as well as substituted Fc variants.

[0072] The wild-type Fc fragment sequence of the present invention may contain the amino acid represented by SEQ ID NO: 8, and preferably may be composed of the amino acid represented by SEQ ID NO: 8.

[0073] According to the Kabat numbering system, the substituted Fc fragment of the present invention comprises one or more amino acid substitutions selected from Q347R, K360E, D399V, F405T, and K409W.

[0074] The substituted Fc fragment of the present invention may be a fragment containing the amino acid substitutions of K360E and K409W. Preferably, it may contain the amino acid sequence represented by SEQ ID NO:12, and more preferably, it may consist of the amino acid sequence represented by SEQ ID NO:12.

[0075] The substituted Fc fragment of the present invention may be a fragment containing the amino acid substitutions of Q347R, D399V and F405T. Preferably, it may consist of the amino acid sequence represented by SEQ ID NO:14, and more preferably, it may consist of SEQ ID NO:14.

[0076] Furthermore, the antibody Fc fragment, as a monomer forming a dimer, comprises SEQ ID NO:8, SEQ ID NO:12, or SEQ ID NO:14, and a linker or other short peptide fragment may be attached to the N-terminus or C-terminus. The linker may be a connector comprising an amino acid sequence selected from SEQ ID NO:10, SEQ ID NO:34, and SEQ ID NO:53 to SEQ ID NO:76. The antibody Fc fragment, as a monomer, may consist of an amino acid sequence selected from SEQ ID NO:8, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:36, SEQ ID NO:38, and SEQ ID NO:40.

[0077] The "monomer comprising an interferon receptor 1 (IFNAR1) fragment and an antibody Fc fragment" of the present invention preferably comprises i) an interferon receptor 1 (IFNAR1) fragment comprising the amino acid sequence represented by SEQ ID NO: 4 and ii) an antibody Fc fragment comprising an amino acid sequence selected from SEQ ID NO: 8, SEQ ID NO: 12 and SEQ ID NO: 14. More preferably, it may comprise i) an interferon receptor 1 (IFNAR1) fragment comprising the amino acid sequence represented by SEQ ID NO: 4, ii) a linker comprising an amino acid sequence selected from SEQ ID NO: 10, SEQ ID NO: 34 and SEQ ID NO: 53 to SEQ ID NO: 76, and iii) an antibody Fc fragment comprising an amino acid sequence selected from SEQ ID NO: 8, SEQ ID NO: 12 and SEQ ID NO: 14. More preferably, it may be a monomer comprising or composed of amino acids selected from SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 42, SEQ ID NO: 44 and SEQ ID NO: 46.

[0078] The "monomer comprising an interferon receptor 2 (IFNAR2) fragment and an antibody Fc fragment" of the present invention preferably comprises i) an interferon receptor 2 (IFNAR2) fragment comprising the amino acid sequence represented by SEQ ID NO: 6 and ii) an antibody Fc fragment comprising an amino acid sequence selected from SEQ ID NO: 8, SEQ ID NO: 12 and SEQ ID NO: 14. More preferably, it may comprise i) an interferon receptor 2 (IFNAR2) fragment comprising the amino acid sequence represented by SEQ ID NO: 6, ii) a linker comprising an amino acid sequence selected from SEQ ID NO: 10, SEQ ID NO: 34, SEQ ID NO: 53 to SEQ ID NO: 76, and iii) an antibody Fc fragment comprising an amino acid sequence selected from SEQ ID NO: 8, SEQ ID NO: 12 and SEQ ID NO: 14. More preferably, it may be a monomer comprising or composed of amino acids selected from SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 48, SEQ ID NO: 50 and SEQ ID NO: 52.

[0079] The polypeptide of the present invention can be a polypeptide that neutralizes type I interferon, and therefore can be used as a type I interferon neutralizing antibody.

[0080] As used herein, the term "type I interferon" is intended to refer to a group of type I interferon molecules that act as ligands for IFNAR 1 and IFNAR 2 (i.e., a group of type I interferon molecules capable of binding to either IFNAR 1 or IFNAR 2). Examples of type I interferon ligands are interferon α1, 2a, 2b, 4, 5, 6, 7, 8, 10, 14, 16, 17, 21, interferon β, interferon ω, and interferon ε.

[0081] Those skilled in the art can use the above-described polypeptides without limitation when implementing the present invention.

[0082] This invention also provides a dimer polypeptide, wherein the monomer is linked to an antibody or a fragment thereof via a peptide linker. A peptide linker is a molecule that links two or more individual substances together to form a short fragment of an amino acid or amino acid analog, wherein the amino acids or amino acid-like substances are linked together by peptide bonds. Glycine, serine, and alanine are used as the main constituent amino acids; therefore, glycine-serine linkers, glycine-serine-alanine linkers, etc., can be used. Furthermore, according to a preferred embodiment of the invention, the linker may comprise or consist of the amino acid sequences represented by SEQ ID NO:8 and SEQ ID NO:10.

[0083] The polypeptides of the present invention may preferably include flexible linker sequences inserted between monomers.

[0084] This linker refers to either a naturally derived or synthetically derived peptide linker. It consists of a linear chain of amino acids, with 20 naturally occurring amino acids serving as monomeric structural units. The linker can have a repeating amino acid sequence or a sequence of a natural polypeptide, such as a polypeptide with hinge function. Since all peptide linkers can be encoded by nucleic acid molecules, they can be expressed using recombinant methods. Because the linker itself is a peptide, each monomer can be linked to the linker via peptide bonds to form a dimer.

[0085] The linker consists of amino acids linked together by peptide bonds, preferably 1 to 20 amino acids linked together by peptide bonds, in which case the amino acids are preferably selected from 20 natural amino acids. One or more of these amino acids are glycosylated as understood by those skilled in the art. Preferably, but not limited to, the 1 to 20 amino acids are selected from glycine, alanine, proline, asparagine, glutamine, and lysine.

[0086] For example, suitable linkers include cleavable linkers and non-cleavable linkers. Cleavable linkers are generally prone to cleavage under intracellular conditions. For example, suitable cleavable linkers include peptide linkers that can be cleaved by intracellular proteases, such as lysosomal proteases or endosomal proteases.

[0087] For example, the linker is a linker whose N-terminus is connected to the C-terminus of the interferon receptor. Preferably, the C-terminus connected to the interferon receptor can be directly linked to the antibody expressed by the expression vector, because the nucleotide sequence encoding the linker sequence is linked to the expression vector expressing the protein of the present invention, so that the protein expression cassette overlaps. Furthermore, the N-terminus of the Fc portion of the antibody of the present invention is connected to the C-terminus of the linker.

[0088] The peptide linkers of the present invention may be peptide linkers known in the art, but are preferably glycine-serine linkers, helical peptide linkers, or peptide linkers comprising or composed of the amino acid sequence of SEQ ID NO:8 or SEQ ID NO:10.

[0089] The linker is preferably a gly-ser linker, for example (Gly x Ser y ) z The type (x is an integer from 1 to 5, y is an integer from 1 to 2, z is an integer from 1 to 6), such as (gly4ser1)3 or (gly3ser2)3, can be A(EAAAK)nA (n = integer from 2 to 5) in the case of helical peptide linkers (or rigid helical linkers).

[0090] Preferably, the peptide linkers of the present invention can be GGGGS, (GGGGS)3, (GGGGS)n (n = 1, 2, 4), (Gly)6, (Gly)8, AEAAAAKEAAAKA, A(EAAAK)4ALEA(EAAAK)4A, (EAAAK)3, EAAAK, (EAAAK)2, (Ala-Pro) n (n is 5 to 17, length is 10 to 34aa), PAPAP, and can be a peptide linker composed of amino acid sequences selected from SEQ ID NO:10, SEQ ID NO:34 and SEQ ID NO:53 to SEQ ID NO:76.

[0091] In this invention, if necessary, the peptide linker can be appropriately constructed by overlapping the peptide linkers listed above or by combining with other linkers.

[0092] For the heterodimerization of Fc in this invention, known Fc heterodimerization techniques in the art can be used. Fc heterodimerization techniques are shown in Table 1 below.

[0093] Table 1

[0094]

[0095] The present invention also provides polypeptides comprising the amino acid sequence of SEQ ID NO:4 or SEQ ID NO:6 or all of the polypeptides.

[0096] Furthermore, the present invention also provides

[0097] (a) A monomer comprising an interferon receptor 1 fragment and an antibody Fc fragment represented by the amino acid sequence of SEQ ID NO:4;

[0098] (b) A dimer polypeptide comprising a monomer comprising an interferon receptor 2 fragment and an antibody Fc fragment represented by the amino acid sequence of SEQ ID NO:6;

[0099] The monomer (a) comprises an interferon receptor 1 fragment and an antibody Fc fragment represented by the amino acid sequence of SEQ ID NO:4, preferably comprising or consisting of an amino acid sequence selected from SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:42, SEQ ID NO:44 and SEQ ID NO:46.

[0100] The monomer (b) comprises an interferon receptor 2 fragment represented by the amino acid sequence of SEQ ID NO:6 and an antibody Fc fragment, preferably comprising or consisting of an amino acid sequence selected from SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:48, SEQ ID NO:50 and SEQ ID NO:52.

[0101] The polypeptide of the present invention is preferably an antibody that regulates type 1 interferon.

[0102] The present invention also provides a polynucleotide encoding the polypeptide.

[0103] In this invention, "polynucleotide" or "nucleic acid" refers to deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) in single-stranded or double-stranded form. Unless otherwise limited, it also includes known analogs of natural nucleotides that hybridize with nucleic acids in a manner similar to that of natural nucleotides.

[0104] The monomer comprises an interferon receptor 1 fragment and an antibody Fc fragment represented by the amino acid sequence of SEQ ID NO:4, preferably comprising or consisting of a DNA sequence selected from SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:41, SEQ ID NO:43 and SEQ ID NO:45.

[0105] The monomer comprises an interferon receptor 2 fragment represented by the amino acid sequence of SEQ ID NO:6 and an antibody Fc fragment, preferably comprising or consisting of a DNA sequence selected from SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:47, SEQ ID NO:49 and SEQ ID NO:51.

[0106] The polynucleotide can be used without limitation, as long as it encodes the polypeptide of the present invention and comprises all DNA, cDNA, and RNA sequences. It refers to a polynucleotide encoding a peptide having an amino acid sequence represented by that sequence or an amino acid sequence having at least 70% homology with that sequence, which can be isolated from nature or prepared by genetic engineering methods known in the art.

[0107] The present invention also provides a vector comprising the polynucleotide.

[0108] The vector refers to an expression vector prepared by those skilled in the art, which expresses the polypeptide of the invention by inserting the polynucleotide of the invention into the vector according to any method known in the art and using appropriate transcription / translation regulatory sequences.

[0109] The polynucleotide sequence cloned according to the present invention can be operatively linked to a suitable expression regulatory sequence, and the operatively linked gene sequence and expression regulatory sequence can be included in an expression vector (including a selection marker and an origin of replication). "Operationally linked" means that the polynucleotide sequence is linked to the expression regulatory sequence in a manner capable of gene expression. "Expression regulatory sequence" refers to a DNA sequence that regulates the expression of the operatively linked polynucleotide sequence in a specific host cell. Such regulatory sequences can include sequences selected from promoters affecting transcription, optional operon sequences regulating transcription, sequences encoding suitable mRNA liposome binding sites, and sequences controlling transcription and translation termination, etc.

[0110] The parental vector used as the expression vector is not particularly limited, and any plasmid, virus, or other medium commonly used as a host cell in the field of this invention can be used. For example, plasmids include those derived from *E. coli* (pBR322, pBR325, pUC118 and pUC119, pET-22b(+)), those derived from *Bacillus subtilis* (pUB110 and pTP5), and those derived from yeast (YEp13, YEp24, and YCp50), etc. Viruses can be animal viruses (such as retroviruses, adenoviruses, or vaccinia viruses), insect viruses (such as baculoviruses, etc.), but are not limited to these.

[0111] The present invention also provides host cells transformed using the vector.

[0112] The host cell can be selected to regulate the expression of the inserted sequence or process the gene product in a specific manner as desired. Different host cells possess characteristic and specific mechanisms for protein translation and post-translational modification. Suitable cell lines or host systems can be selected to provide the necessary modifications and processing for the expressed heterologous protein. Expression in yeast can produce a biologically active product. Expression in eukaryotic cells can increase the likelihood of "natural" folding.

[0113] Just as host cells capable of stably and continuously cloning and expressing the vector of the present invention can be any host cell known in the art, for example, Escherichia coli JM109, Escherichia coli BL21DE, Escherichia coli DH5, Escherichia coli RR1, Escherichia coli LE392, Escherichia coli B, Escherichia coli X1776 and Escherichia coli W3110, etc., can also be used as host cells. In addition, strains of Agrobacterium (such as Agrobacterium A4), bacilli (such as Bacillus subtilis), other enteric bacteria and various strains of Pseudomonas (such as Salmonella typhimurium or Serratia marcescens) can be used.

[0114] Furthermore, when the vector of the present invention is transfected into eukaryotic cells, yeast (Saccharomyces cerevisiae), insect cells, and human cells (e.g., CHO cell line (Chinese hamster ovary), Expi293, W138, BHK, COS-7, 293, HepG2, 3T3, RIN, and MDCK cell lines) can be used as host cells.

[0115] In this invention, the host cell is preferably the Expi293 or CHO cell line.

[0116] The method of transfecting host cells with this vector can be any known method and is not particularly limited. For example, transfection can be performed by gene bombardment, polycationic and receptor-mediated transfection using calcium phosphate precipitation, DEAE-dextran method, electroporation, direct microinjection, DNA-loaded liposome method, liposome-DNA complex method, cell sonication (cell ultrasound), and high-velocity microprojectile. Some of these techniques can be modified for in vivo or in vitro use.

[0117] The present invention also provides a method for producing a dimer polypeptide, comprising (a) providing a host cell, (b) culturing the provided cell, and (c) preparing the polypeptide of the present invention by recovering the polypeptide from the cell or culture medium.

[0118] Transformed cells are cultured under appropriate conditions that allow for the expression of the fusion protein (or fusion polypeptide), conditions which can be performed according to methods well known to those skilled in the art. Transformed cells can be cultured in large quantities using conventional culture methods. As the culture medium, a medium consisting of a carbon source, nitrogen source, vitamins, and minerals can be used, for example, 2XYT medium. Cell culture can be performed under normal cell culture conditions, for example, at a temperature range of 15°C to 45°C for 10 to 40 hours. To remove cells from the culture medium and recover only the medium, centrifugation or filtration can be performed. These steps can be performed by those skilled in the art as needed. The cell-free culture medium (filtrate) can be refrigerated and stored for a short period according to conventional methods to prevent loss of activity.

[0119] Fusion proteins expressed in transformed cells (or transformants) can be purified by conventional methods. For example, the fusion proteins of the present invention can be purified by using techniques such as salting out (e.g., ammonium sulfate precipitation, sodium phosphate precipitation), column chromatography, and ultrafiltration (e.g., solvent precipitation (precipitation of protein components using acetone, ethanol, etc.), dialysis, gel filtration, ion exchange, reversed-phase column chromatography, affinity chromatography) alone or in combination.

[0120] The interferon receptor of the present invention is a highly glycosylated protein containing a large amount of glycosylation during post-translational modification, which may be unsuitable for microbial culture because the corresponding sugars can cause structural stability problems. Therefore, the polypeptide of the present invention can preferably be a host cell of animal cells.

[0121] Therefore, the present invention provides a pharmaceutical composition for the prevention or treatment of type 1 interferon-mediated diseases or conditions, comprising a polypeptide as an active ingredient.

[0122] According to the present invention, "interferon type 1 mediated diseases" refers to diseases or conditions mediated by interferon type 1, or diseases associated with abnormal expression (e.g., overexpression or inhibition) of interferon-induced genes, examples including but not limited to systemic lupus erythematosus (SLE, J Immunol. 2014 Jun 15; 192(12):5459–5468.), Sjögren's syndrome (AutoimmuneRev. 2013 Mar; 12(5):558-66.), insulin-dependent diabetes mellitus (IDDM), inflammatory bowel disease (IBD, including Crohn's disease, ulcerative colitis and celiac disease), multiple sclerosis (MS), psoriasis, autoimmune thyroiditis, rheumatoid arthritis (RA, Front Immunol. 2017; 8:2007.), inflammatory myositis (Arthritis Rheum 2009; 60:181524.; Mol Med. 2007;13:5968.) and glomerulonephritis. Furthermore, the compositions of the present invention can be used to inhibit or prevent transplant rejection or to treat graft-versus-host disease (GVHD) or to treat HIV infection / AIDS.

[0123] Simultaneously, the pharmaceutical composition according to the invention can be provided by formulating the polypeptide in its pure form or in a suitable form with a pharmaceutically acceptable carrier. "Pharmaceutically acceptable" means a physiologically acceptable, non-toxic composition that, when administered to humans, generally does not cause allergic reactions such as gastrointestinal disorders, dizziness, or similar reactions. The carrier includes various solvents, dispersion media, oil-in-water or water-in-oil emulsions, aqueous compositions, liposomes, microbeads, and microparticles.

[0124] Furthermore, depending on the route of administration and the use of a suitable carrier, pharmaceutical compositions according to the present invention can be formulated. The routes of administration of the pharmaceutical compositions according to the present invention are not limited thereto, but can be administered orally or parenterally. Parenterally administration routes include a variety of routes, such as percutaneous, nasal, intraperitoneal, intramuscular, subcutaneous, or intravenous.

[0125] When the pharmaceutical composition of the present invention is for oral administration, it can be formulated into powder, granules, tablets, pills, sugar-coated pills, capsules, liquids, gels, syrups, suspensions, sheets, etc., by methods known according to the present invention and with a carrier suitable for oral administration. Examples of suitable carriers include: sugars (including lactose, glucose, sucrose, sorbitol, mannitol, xylitol, erythritol, and maltitol, etc.), starches (including corn starch, wheat starch, rice starch, and potato starch, etc.), celluloses (including cellulose, methylcellulose, sodium carboxymethylcellulose, and hydroxypropyl methylcellulose, etc.), fillers (such as gelatin, polyvinylpyrrolidone, etc.), etc. Furthermore, if desired, croscarmellose, agar, alginic acid, or sodium alginate can be added as a disintegrant. In addition, the pharmaceutical composition may also include anti-aggregating agents, lubricants, wetting agents, flavoring agents, emulsifiers, and preservatives, etc.

[0126] Furthermore, when administered parenterally, the pharmaceutical compositions of the present invention can be formulated into injections, transdermal administration, and nasal inhalation forms with a suitable carrier according to methods known in the art. In the case of injections, sterilization must be performed to prevent contamination by microorganisms (such as bacteria and fungi). Examples of suitable carriers for injections may include, but are not limited to, water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), mixtures thereof, and / or solvents or dispersion media containing vegetable oils. More preferably, as suitable carriers, Hanks' solution, Ringer's solution, isotonic solutions (such as phosphate-buffered saline (PBS) or sterile water for injection), 10% ethanol containing triethanolamine, 40% propylene glycol, and 5% glucose, etc., can be used. To prevent microbial contamination of the injections, they may also include various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc. In addition, in most cases, the injections may also contain isotonic agents, such as sugar or sodium chloride.

[0127] In the case of transdermal administration, forms include ointments, creams, lotions, gels, topical solutions, pasta, liniments, and air. As used herein, "transdermal administration" refers to the delivery of an effective amount of the active ingredient contained in a pharmaceutical composition to the skin by applying the pharmaceutical composition topically. These formulations are described in formulation manuals commonly found in medicinal chemistry.

[0128] For inhalation administration, the compounds used according to the invention can be conveniently delivered from a pressurized package or nebulizer in the form of an aerosol spray using a suitable propellant (e.g., dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gases). In the case of pressurized aerosols, the dosage unit can be determined by providing a valve for dispensing the correct amount of product. For example, gelatin capsules and pellets used in inhalers or pneumoperitoneum devices can be formulated into a powder mixture containing the compound and a suitable powder matrix (e.g., lactose or starch). Other pharmaceutically acceptable carriers can be referenced from those known in the art.

[0129] Furthermore, the pharmaceutical compositions according to the present invention may also include one or more buffer solutions (e.g., saline or PBS), carbohydrates (e.g., glucose, mannose, sucrose or dextran), stabilizers (sodium bisulfite, sodium sulfite or ascorbic acid), antioxidants, antibacterial agents, chelating agents (e.g., EDTA or glutathione), adjuvants (e.g., aluminum hydroxide), suspending agents, thickeners and / or preservatives (benzalkonium chloride, methylparaben or propylparaben and chlorobutanol).

[0130] Furthermore, the pharmaceutical compositions of the present invention can be formulated using methods known in the art to enable rapid, sustained, or delayed release of the active ingredient upon administration to mammals.

[0131] The pharmaceutical compositions formulated as described above can be administered in an effective amount via various routes, including oral, transdermal, subcutaneous, intravenous, or intramuscular. As used herein, the term "effective amount" refers to the amount of compound or extract that allows for tracking of diagnostic or therapeutic effects when administered to a patient. The dosage of the pharmaceutical composition according to the invention can be appropriately selected based on the route of administration, the target of administration, the target disease and its severity, age, sex, weight, individual differences, and disease status. Preferably, the content of the active ingredient in the pharmaceutical composition comprising the polypeptide of the invention can be varied depending on the severity of the disease; however, generally, for adults, an effective dose of 10 μg to 10 mg can be repeated multiple times daily when administered once.

[0132] Furthermore, the present invention provides a vaccine comprising a polypeptide as an active ingredient, and the vaccine according to the present invention is a vaccine for the prevention or treatment of interferon type 1 mediated diseases or conditions.

[0133] As used in this article, the term "vaccine" refers to an immunogen or antigenic substance administered to a human or animal by injection or oral administration to induce immunity in a living organism for the purpose of preventing infection. This immunogen or antigenic substance is a biological agent containing antigens that confer immunity in a living organism. In vivo immunity is largely divided into active immunity (acquired automatically after infection with a pathogen) and passive immunity (acquired through external vaccination). Autoimmunity is characterized by a long time to produce immune-related antibodies and sustained immunity. Passive immunity from vaccines provides immediate treatment for infection, but its disadvantage is poor persistence.

[0134] The vaccine compositions of the present invention may include pharmaceutically acceptable carriers. This refers to any component suitable for delivering antigenic material to a site in the body, examples including but not limited to water, saline, phosphate-buffered saline, Ringer's solution, glucose solution, serum-containing solution, Hanks' solution, other physiologically balanced aqueous solutions, oils, esters, and ethylene glycol.

[0135] The carrier of the present invention may contain suitable auxiliary components and preservatives to enhance chemical stability and isotonicity, and may include stabilizers such as trehalose, glycine, sorbitol, lactose, or monosodium glutamate (MSG) to protect the vaccine composition from temperature changes or lyophilization. The vaccine composition of the present invention may contain a suspension, such as sterile water or saline (preferably buffered saline).

[0136] The vaccine compositions of the present invention may contain any adjuvant sufficient to enhance the immune response to the immunogen. Suitable adjuvants are known in the art, and examples include, but are not limited to, aluminum salts (aluminum phosphate or aluminum hydroxide), squalene mixtures (SAF-1), muramyl peptides, saponin derivatives, mycobacterial cell wall preparations, monophospholipid A, mycoic acid derivatives, nonionic block copolymer surfactants, Quil A, cholera toxin B subunits, polyphosphazenes and their derivatives, and immunostimulatory complexes (ISCOM).

[0137] As with all other vaccine compositions, the effective immunogenic dose of the immunogen must be determined empirically. Factors to consider include the immunogenicity of the immune system, route of administration, and frequency of administration. Furthermore, adjustments can be made based on the patient's disease progression and metastasis status, formulation type, patient's age, sex, weight, health condition, diet, timing of administration, and method of administration.

[0138] The dimer polypeptides in the vaccine compositions of the present invention may be present in various concentrations in the compositions of the present invention; however, generally, the antigen material is contained in the concentration required to induce appropriate levels of antibodies in vivo.

[0139] In this invention, the term "administration" refers to the injection of a predetermined amount of substance into a patient by any suitable method. The administration route of the vaccine of this invention can be any conventional route, as long as it can reach the target tissue.

[0140] The vaccine composition of the present invention can be used to treat type 1 interferon-mediated diseases or conditions by systemic or mucosal administration.

[0141] The administration of the urogenital tract vaccine composition may include, but is not limited to, intramuscular injection, intraperitoneal injection, intradermal or subcutaneous injection, oral / dietary injection, respiratory injection, and mucosal injection into the urethra.

[0142] In addition, to improve the efficacy of the vaccine, it can be co-administered with cytokines that help activate T cells, such as IL-12, or a vaccine transfected with the genes of these cytokines can be used.

[0143] Cells containing the aforementioned dimer-type polypeptide are the active ingredient of the vaccine produced by this invention. Cell proliferation can also be depleted to increase safety, as the vaccine is administered to humans as a therapeutic vaccine. For example, to be used more safely and selectively as a cell vaccine, proliferation can be eliminated by heat treatment, radiotherapy, or mitomycin C (MMC) treatment while preserving vaccine function. For example, when using X-ray irradiation, the total radiation dose can reach 1000 to 3300 rads. Taking the mitomycin C treatment method as an example, 25–50 μg / ml of mitomycin C is added to the polypeptide, and heat treatment can be performed at 37°C for 30–60 minutes. For example, in the cell heat treatment method, heat treatment can be performed at 50°C to 65°C for 20 minutes.

[0144] Here, "treatment" refers to a clinical process designed to alter the natural processes of the treated object or cells; it can also be used to prevent clinicopathological changes. Ideal effects of treatment include suppressing the onset or recurrence of disease, alleviating symptoms, reducing any direct or indirect pathological consequences of the disease, slowing disease progression, and alleviating, improving, reducing, or improving prognosis. Furthermore, the term "prevention" refers to any action that inhibits the onset of disease or delays its progression.

[0145] In this invention, the term "comprising" has the same meaning as "including" or "characterized by," and in the composition or method according to the invention, additional components or steps not specifically mentioned in the method are not excluded. Furthermore, the term "composed of" means excluding additional elements, steps, or ingredients not separately described. The term "consistently composed of" means that, within the scope of the composition or method, in addition to the described substances or steps, it may also include substances or steps that do not substantially affect its essential properties.

[0146] The present invention also provides the use of the said polypeptide in the preparation of a medicament for the prevention or treatment of type 1 interferon-mediated diseases or conditions.

[0147] Furthermore, the present invention also provides a method for treating type 1 interferon-mediated diseases or conditions, comprising administering an effective amount of a composition to a subject in need, the composition comprising a polypeptide as an active ingredient.

[0148] The "effective amount" as described in this invention refers to the amount that, when administered to an individual, exhibits effects of improving, treating, preventing, detecting, diagnosing, inhibiting, or reducing type 1 interferon-mediated diseases or conditions, and the "subject" may be an animal, preferably an animal, including mammals, particularly humans, and cells, tissues, organs, or similar materials extracted from animals. The subject may be a patient with the need for such treatment.

[0149] The term "treatment" as used in this invention generally refers to the improvement of a disease or condition mediated by type 1 interferon, or the symptoms of a disease or condition mediated by type 1 interferon. This may include treatment, adequate prevention or improvement of the condition, and may include relief, treatment or prevention of one or most of the symptoms caused by the disease, but is not limited thereto.

[0150] Beneficial effects

[0151] The Fc-fusion protein of this invention, which neutralizes type 1 interferon, blocks the binding of type 1 interferon to the interferon receptor, effectively inhibiting the initiation of signaling mechanisms and biological activity, thereby effectively treating type 1 interferon-mediated diseases. Attached Figure Description

[0152] Appendix Figure 1 This is a schematic diagram illustrating the preparation of a type 1 interferon Fc fusion receptor protein expression vector.

[0153] Appendix Figure 2a The results show the size comparison of each protein using size exclusion chromatography.

[0154] Appendix Figure 2b To confirm the binding of the IFNAR1 / 2 heterologous Fc fusion protein to human IFNβ under both natural and reducing conditions.

[0155] Appendix Figure 2c This is a schematic diagram of the type 1 interferon Fc fusion receptor protein.

[0156] Appendix Figure 3 The results of SPR analysis of the type 1 interferon Fc fusion receptor protein.

[0157] Attached Figure Figures 4a to 4c The results were used to confirm the type 1 interferon neutralizing ability of the type 1 interferon Fc fusion receptor protein of the present invention.

[0158] Appendix Figure 4a To demonstrate the cell viability of Daudi cells treated with human interferon β-1a or type 1 interferon Fc fusion receptor protein alone.

[0159] Appendix Figure 4b To show the IC50 values ​​after treatment with human interferon β-1a or type 1 interferon Fc fusion receptor protein alone.

[0160] Appendix Figure 4c The results of treating Daudi cells with human interferon β-1a at the protein concentrations according to the present invention are shown.

[0161] Appendix Figure 5 The neutralizing ability of the IFNAR1 / 2-Fc heterodimer (4GS*3) protein of the present invention to the biological activity of type 1 interferons (IFN-α1, IFN-α2a, IFN-α2b, IFN-α5, IFN-α8, IFN-α10, IFN-β1a, IFN-ω) was confirmed, and the cell viability of Daudi cells in the group with type 1 interferon added and the group with each interferon treated with IFNAR1 / 2-Fc heterodimer (4GS*3) protein was demonstrated.

[0162] Appendix Figure 6a and 6b This is a result confirming the neutralizing ability of the IFNAR1 / 2-Fc heterodimer (4GS*3) protein of the present invention on the signaling mechanism of type 1 interferon. The results show the phosphorylation changes of STAT1 protein in Daudi cells after adding type 1 interferon (IFN-α1, IFN-α2a, IFN-α2b, IFN-α5, IFN-α8, IFN-α10, IFN-β1a, IFN-ω, IFN-ε) to each group. The interferon in each group was treated with IFNAR1 / 2-Fc heterodimer (4GS*3) protein. Detailed Implementation

[0163] The present invention will now be described in detail.

[0164] However, the following embodiments are merely illustrative of the present invention, and the content of the present invention is not limited to the following embodiments.

[0165] Example 1: Preparation of a type 1 interferon Fc fusion receptor protein expression vector

[0166] Using the amino acid sequences of human IFNAR1 (P17181, 28-436 aa.), IFNAR2 (P48552, 27-243 aa.), and IGHG (P01857, 100-330 aa.), an Fc fusion protein capable of binding to or neutralizing type 1 interferon protein was constructed. (See attached image.) Figure 1 As shown on the left, Fc is labeled with a polypeptide linker (L; IEGRMD) at the C-terminus of all Fc fusion protein expression vectors. It is linked by IFNAR1 or IFNAR2 between the C-terminus and N-terminus, including the Kozac sequence (▲) and the extracellular secretion-inducing signal sequence (S), or directly.

[0167] As attached Figure 1 As shown on the right, the sequence containing each gene is linked between the restriction enzymes NheI and XhoI. The expression vectors are named pIFNAR1-FcWT, pIFNAR2-FcWT, pIFNAR1-FcA, pIFNAR2-FcB, pIFNAR1-FcB, and pLeader-FcA, respectively.

[0168] Codon optimization was performed on genes containing a polynucleotide sequence (SEQ ID NO:1), the extracellular domain of IFNAR1 (SEQ ID NO:3), the extracellular domain of IFNAR2 (SEQ ID NO:5), a polypeptide linker (SEQ ID NO:7), and the Fc portion of human IGHG (SEQ ID NO:11), as well as variants containing a Kozac sequence and a signal sequence (SEQ ID NO:13, SEQ ID NO:15), to synthesize the gene for preparation of, for example... Figure 1 Nucleotide sequence fragments. Expression vectors were prepared using a T4 DNA ligase (RBC) between NheI and XhoI at the restriction enzyme sites of the expression vector (pOptiVec-TOPO) for each nucleotide sequence fragment.

[0169] Furthermore, the expression vector was prepared in the same manner, except that the adapter of SEQ ID NO:33 was used instead of the peptide adapter of SEQ ID NO:7. In other words, a gene containing a polynucleotide sequence (SEQ ID NO:1), the extracellular domain of IFNAR1 (SEQ ID NO:3), the extracellular domain of IFNAR2 (SEQ ID NO:5), the peptide adapter (SEQ ID NO:33), and the Fc portion of human IGHG (SEQ ID NO:11), as well as variants containing the Kozac sequence and the signal sequence (SEQ ID NO:13, SEQ ID NO:13, SEQ ID NO 15), was codon-optimized to synthesize the gene, thereby preparing... Figure 1 Nucleotide sequence fragments e to g. Expression vectors were prepared using a T4 DNA ligase (RBC) between NheI and XhoI at the restriction enzyme sites of each nucleotide sequence fragment expression vector.

[0170] Example 2: Protein expression, purification, structural confirmation, and binding confirmation

[0171] The following descriptions and purification processes are intended to produce expression vectors with attached nucleotide sequence fragments inserted for each combination. Figure 2c The protein with the structure shown.

[0172] To maintain the native post-translational modification (PTM) of each type 1 interferon receptor, all recombinant proteins were expressed using 300–600 mL of Expi293™ expression medium according to the transient expression system protocol, using the human-derived Expi293 cell line (Thermo Fisher Scientific). Depending on the final protein, one or both expression vectors were transfected into the cells, followed by the addition of enhancers 1 and 2 according to the manufacturer's protocol. The medium, cultured for 6 days, was then centrifuged at 6580 rpm for 20 minutes at 8°C and filtered through a 0.22 μm polyethylene filter (Corning). All proteins were purified in an AKTA pre-purification system (GE Healthcare Life Sciences) by filling a MabSelect SuRe (GE Healthcare Life Sciences) column with 20 mM sodium phosphate, pH 7.2, and 150 mM sodium chloride buffer at a rate of 5 mL / min to a maximum average column volume of 2 CV, and then allowing the culture medium to flow to bind to the resin.

[0173] The recombinant protein was then washed with 5 cV of 35 mM sodium phosphate, pH 7.2, 500 mM sodium chloride buffer and 1 cV of 20 mM sodium phosphate, pH 7.2 buffer, followed by elution with 100 mM citrate, pH 3.5 buffer. After 12 hours of dialysis, the protein was dialyzed three times with 4 L of 1X PBS buffer (Biosesang). The eluted protein was concentrated to 3 mL using an Amicon UltraCentrifugal filter (50K-cut off, Merck Millipore). The concentrated recombinant protein was then separated from the target protein separately on a Hiload Superdex 200 pg (GE Healthcare Life Sciences) column using the same buffer used for dialysis.

[0174] The results are as follows. Figure 2a As shown, the size of each Fc fusion protein was compared using size exclusion chromatography, and its expected size is... Figure 2c The protein structure shown.

[0175] In addition, such as Figure 2b As shown, under natural conditions (7.5% Mini- The binding of the IFNAR1 / 2 hybrid Fc fusion protein to human IFNβ-1a was confirmed under TGX™ Precase protein gel and reducing conditions (Any kD™ Mini-PROTEIN TGX™ Precase protein gel, Bio-rad).

[0176] The FC portion of the amino acid sequence of this invention includes the HINGE portion of IGHG1 (100-330 a.a.) and a dimer formed by disulfide bonds formed by amino acids 109-109 and 112-112. More specifically, due to the specific amino acid sequence variation of the CH3 portion of FC, the formation of heterodimers is mainly from homodimers to heterodimers.

[0177] Example 3: SPR analysis using Fc fusion protein

[0178] Through append Figure 1 The expression vectors of hIFNβ-1a were combined for expression / purification, and the binding affinity and dynamics of Fc fusion proteins (IFNAR2-Fc heterodimer, IFNAR1 / 2-Fc heterodimer, and IFNAR1-Fc heterodimer) in type I interferon were analyzed.

[0179] This experiment used (IFNAR1-Fc EW+-Fc RVT), (IFNAR2-Fc EW+-Fc RVT), and (IFNAR1-FcEW+IFNAR2Fc RVT) to analyze binding strength and kinetics. When expressing (IFNAR1-Fc RVT+IFNAR2-Fc EW), the homodimeric form was found to occur more frequently than (IFNAR1-Fc EW+IFNAR2-Fc RVT), so (IFNAR1-Fc RVT+IFNAR2-Fc EW) was not used.

[0180] Based on the Biacore T200 (GE Healthcare Life Sciences), the gold sensor chip was coupled with an amine-conjugated capture antibody (AffiniPure Goat Anti-Human IgG, Fcγ-specific fragment, Jackson ImmunoResearch) at 25°C, 30 μl / min, and 0.005% DPBST (DPBS / modified, Hyclone, and Tween 20, Signal) run buffer. Each analyte was tested at 180 sec in phase and 600 sec in phase.

[0181] like Figure 3As shown, curve fitting was performed based on the sensor map shapes and docking model predictions for hIFNb-1a-IFNAR2 (1:1 fitting model), hIFNb-1a-IFNAR1 / 2 (heteroligand model), and hIFNb-1a-IFNAR1 (dual-state binding model). In particular, the IFNAR1 / 2 heterologous Fc fusion protein exhibited a low KD value. (See Tables 2 and 3)

[0182] Table 2

[0183]

[0184] a Sensor maps use a one-to-one combination model Fitting

[0185] b The static equilibrium analysis was performed using various parameters over a period of 179.04 seconds.

[0186] c Sensor maps using heteroligand models Fitting

[0187] Table 3

[0188]

[0189] a Sensor image using a two-state combination model Fitting

[0190] Example 4: Confirmation of bioactivity neutralization capacity using ligands of Fc fusion protein

[0191] The following experiments were conducted to demonstrate the ligand-mediated neutralizing ability of the Fc fusion protein. Similar to Example 3, (IFNAR1-Fc EW+-Fc RVT), (IFNAR2-Fc EW+-Fc RVT), and (IFNAR1-Fc EW+IFNAR2Fc RVT) were used in this experiment.

[0192] All experiments were designed based on the antiproliferative effect of hIFN-β in Daudi cells using the Ez-cytox cell viability assay kit (Deaillab), where each receptor was highly expressed. This assay method yielded results that adequately reflect the biophysical properties of the ligands, similar to those obtained from SPR kinetic analysis.

[0193] Specifically, in a 96-well plate (SPL), 3 × 10 3The hIFN-β-1a content in cells was determined by concentrations (10 nM IFNAR1-Fc heterodimer, IFNAR2-Fc heterodimer, IFNAR1 / 2-Fc heterodimer; 6 nM IFNAR1-Fc heterodimer; IFNAR1 / 2-Fc heterodimer), followed by a 72-hour reaction in a cell culture incubator at 37°C and 5% CO2. After adding Ex-cytox reagent according to the manufacturer's protocol, an additional 3-hour reaction was performed, followed by measurements at 450 nm using a microplate reader (Genios Pro, Tecan). Nonlinear regression analysis (GraphPad Prism version 7.0 software, San Diego, California, USA) was used to compare and analyze the IC50 values. 50 Values. One-way ANOVA and Bonferroni's post-hoc multiple comparison test confirmed that the neutralizing ability of each protein in this invention was statistically significant.

[0194] As attached Figure 4a As shown, treatment with Fc fusion proteins that block the binding of IFNAR1 and IFNAR2 significantly reduced the anti-cell proliferation effect of interferon compared to treatment with fusion proteins that block IFNAR1 and IFNAR2 separately.

[0195] As attached Figure 4b As shown, the IC50 value was highest when the Fc fusion protein that blocks IFNAR1 and IFNAR2 according to the present invention was added. Therefore, it is confirmed that when IFNAR1 or IFNAR2 is blocked respectively, interferon has better anti-cell proliferation effect than cell viability.

[0196] As attached Figure 4c As shown, cell viability increased in a concentration-dependent manner when the IFNAR1 / 2-Fc heterodimer with the highest neutralizing capacity was applied.

[0197] Example 5: Confirmation of signaling mechanism activity and neutralizing ability through ligands of IFNAR1 / 2-Fc heterodimer

[0198] In the same manner as in Example 4, except for the IFNAR1 / 2-Fc heterodimer mentioned below, the ability of ligands to neutralize biological activity via the IFNAR1 / 2-Fc heterodimer was confirmed.

[0199] Specifically, this group was supplemented with 1 nM of IFN (IFN-α1, IFN-α2a, IFN-α2b, IFN-α5, IFN-α8, IFN-α10, IFN-β1a, and IFN-ω). Each interferon in this group was treated with a 10 nm concentration of type 1 interferon Fc fusion receptor protein at 3 × 10⁻⁶ m². 3 Cell populations were treated and incubated in a cell culture incubator at 37°C and 5% CO2 for 72 hours. Then, after adding Ex-cytox reagent according to the manufacturer's protocol, an additional 2 hours of reaction were performed, followed by measurement at 450 nm using a microplate reader (Genios Pro, Tecan). Nonlinear regression analysis (GraphPad Prism version 7.0 software, San Diego, California, USA) was used to compare and analyze the IC50 values. 50 value.

[0200] The results are attached. Figure 5 As shown, treatment with IFNAR1 / 2-Fc heterodimers (using 4GS*3 as linkers) that block the binding of IFNAR1 and IFNAR2 resulted in significantly superior cell viability compared to treatment with interferon alone.

[0201] These results demonstrate that it is possible to regulate excessive interferon signaling in cells by binding a dimeric polypeptide containing a monomer (monomer) of the interferon receptor fragment or antibody Fc fragment of the present invention, and thereby confirm that it will exhibit excellent pharmacological effects in patients with type 1 interferon-mediated diseases such as systemic lupus erythematosus, Sjögren's syndrome, systemic sclerosis, myositis, and rheumatoid arthritis, in which interferon-inducible genes expressed in response to excessive interferon signaling are high.

[0202] Example 6: Demonstrating the chemical signal suppression ability using ligands of IFNAR1 / 2-Fc heterodimer

[0203] All experiments were conducted to confirm changes in STAT1 and its phosphorylated pSTAT1 protein, achieved by using Western blotted type I interferon in Daudi cells, where each receptor was highly expressed.

[0204] Specifically, in 6-well plates (SPL_Class 30006), this group added IFN (IFN-α1(pbl assay science_cat#11125-1), IFN-α2a(pbl assay science_cat#11100-1), IFN-α2b(pbl assay science_cat#11105-1), IFN-α5(pbl assay science_cat#11135-1), IFN-α8(pbl assay science_cat#11115-1), IFN-α10(pbl assay science_cat#11120-1), IFN-β1a(pbl assay science_cat#11415-1), IFN-ω(pbl assay science_cat#11395-1), IFN-ε(R&D) at a concentration of 1 nM. systems_#9667-ME), and each interferon in this group was treated with IFNAR1 / 2-Fc heterodimer protein at a concentration of 10 nm at 2 × 10⁻⁶. 6Cell populations were treated and incubated at 37°C and 5% CO2 for 72 hours. Cells were then collected from plates, total protein was extracted, and determined using a BCA protein assay kit (Thermoscientific_#23227). Total protein was separated on a 10% gel by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transcribed onto a polyvinylidene fluoride membrane (BIO RAD_#1620177). The membrane was blocked for 1 hour at room temperature with 5% skim milk, 3% bovine serum albumin, 10 mmol / L Tris-HCl (pH 8.0), 150 mmol / L NaCl, and 0.05% Tween-20. The sealed membrane was treated overnight at 4°C with primary antibodies (1:3000 dilution) of STAT1 antibody (Cell Signallig_#06-501), pSTAT1 antibody (Cell Signallig_#58D6), and β-actin antibody (Santa Cruz_#SC 47778) at 4°C. Then, it was treated for 1 hour at room temperature with second-generation enzyme-labeled antibodies, goat-to-rabbit antibody (Invitrogen_#31460), and goat-to-mouse antibody (Invitrogen_#g 21040). After treatment with ECL solution (BIORAD_#1705061), the membrane was developed onto film (Agfa Healthcare_#EA8EC) using Poohung developer and fixer.

[0205] The results are attached. Figure 6a and 6b As shown, treatment with interferons (IFN-α1, IFN-α2a, IFN-α2b, IFN-α5, IFN-α8, IFN-α10, IFN-β1a) that block the binding of IFNAR1 and IFNAR2 significantly reduced pSTAT1 phosphorylation compared to the hyperphosphorylation observed with interferon alone. This indicates that the IFNAR1 / 2-Fc heterodimer binds to type 1 interferon and exhibits neutralizing ability, suggesting that interferon-related signaling in cells is inhibited.

[0206] Industrial applicability

[0207] The neutralizing Fc fusion protein type 1 interferon of the present invention has excellent industrial applicability. When the binding of type 1 interferon to the interferon receptor is blocked, it has excellent signal initiation and biological activity inhibition. Therefore, it can be used very effectively to develop therapeutic agents for the prevention or treatment of type 1 interferon-mediated diseases. sequence list <110> Seoul National University Industry-Academia Collaboration Group Lukyuan BIO Fusion Research Foundation <120> FC fusion protein for neutralizing type 1 interferon and its applications <130> OP21-0067 / PCT / CN <150> KR 10-2018-0168801 <151> 2018-12-24 <150> PCT / KR 2019 / 018233 <151> 2019-12-20 <160> 76 <170> KoPatentIn 3.0 <210> 1 <211> 69 <212> DNA <213> Artificial sequence <220> <223> Kozak-signal sequence <400> 1 gccaccatgt ggtggcgtct gtggtggctg ctgcttctgc tgttgctgct gtggcctatg 60 gtctgggcc 69 <210> 2 <211> twenty three <212> PRT <213> Artificial sequence <220> <223> Kozak-signal sequence <400> 2 Ala Thr Met Trp Trp Arg Leu Trp Trp Leu Leu Leu Leu Leu Leu Leu 1 5 10 15 Leu Trp Pro Met Val Trp Ala 20 <210> 3 <211> 1227 <212> DNA <213> Artificial sequence <220> <223> IFNAR1 extracellular domain <400> 3 aagaacctga agtcccctca gaagtggaa gtggacatca tcgacgacaa cttcatcctg 60 cggtggaaca gatccgacga gtccgtggggc aacgtgacct tcagctcga ctaccagaaa 120 accggcatgg acactggat caagctgtcc ggctgccaga acatcacctc taccaagtgc 180 aacttctcca gcctgaagct gaacgtgtac gaggaatca agctgcggat ccggccgag 240 aaagagaaca ccagcagttg gtacgaggtg ggagcttca cccctttcag aaaggcccag 300 atcggccctc ctgaagtgca cctggaagcc gaggatagg ccatcgtgat ccacatcagc 360 cccggcacca aggactctgt gatgtgggct ctcgacggcc tgtccttcac ctacagcctg 420 gtcatctgga agaactccctc cggcgtggaa gagagaatcg agaacatcta ctcccggcac 480 aagatctaca agctgagccc cgagacacc tactgcctga aagtgaaggc cgctctgctg 540 acctcctgga agatcggcgt gtactctccc gtgcactgca tcagaccac cgtggaaaac 600 gagctgcctc ctccagagaa catcgaggtg tccgtgcaga accagaacta cgtgctgaag 660 tgggactaca cctacgccaa catgacctttt caggtgcagt ggctgcacgc tttctgaag 720 cggaaccctg gcaaccacct gtacaagtgg aagcagattc ccgactgcga gaacgtgaaa 780 accacacagt gcgtgttccc tcagaacgtg ttccagaagg gcatctacct gctgagagtg 840 caggcctccg acggcaacaa caccagcttt tggagcgaag agatcaagtt cgataccgag 900 atccaggcct tcctgctgcc tccagtgttc aacatcagat ccctgtccga ctccttccac 960 atctacatcg gcgctcctaa gcagtccggc aacacccctg tgatccagga ctaccctctg 1020 atctacgaga tcatcttctg ggagaacacc tccaacgccg agcggaagat catcgagaag 1080 aaaaccgacg tgaccgtgcc taacctgaag cctctgaccg tgtactgcgt gaaggctagg 1140 gcccacacca tggacgaaaa gctgaacaag tcctccgtgt tctccgacgc cgtgtgcgaa 1200 aagaccaagc caggcaacac ctctaaa 1227 <210> 4 <211> 409 <212> PRT <213> Artificial Sequence <220> <223> IFNAR1 Extracellular Domain <400> 4 Lys Asn Leu Lys Ser Pro Gln Lys Val Glu Val Asp Ile Ile Asp Asp 1 5 10 15 Asn Phe Ile Leu Arg Trp Asn Arg Ser Asp Glu Ser Val Gly Asn Val 20 25 30 Thr Phe Ser Phe Asp Tyr Gln Lys Thr Gly Met Asp Asn Trp Ile Lys 35 40 45 Leu Ser Gly Cys Gln Asn Ile Thr Ser Thr Lys Cys Asn Phe Ser Ser 50 55 60 Leu Lys Leu Asn Val Tyr Glu Glu Ile Lys Leu Arg Ile Arg Ala Glu 65 70 75 80 Lys Glu Asn Thr Ser Ser Trp Tyr Glu Val Asp Ser Phe Thr Pro Phe 85 90 95 Arg Lys Ala Gln Ile Gly Pro Pro Glu Val His Leu Glu Ala Glu Asp 100 105 110 Lys Ala Ile Val Ile His Ile Ser Pro Gly Thr Lys Asp Ser Val Met 115 120 125 Trp Ala Leu Asp Gly Leu Ser Phe Thr Tyr Ser Leu Val Ile Trp Lys 130 135 140 Asn Ser Ser Gly Val Glu Glu Arg Ile Glu Asn Ile Tyr Ser Arg His 145 150 155 160 Lys Ile Tyr Lys Leu Ser Pro Glu Thr Thr Tyr Cys Leu Lys Val Lys 165 170 175 Ala Ala Leu Leu Thr Ser Trp Lys Ile Gly Val Tyr Ser Pro Val His 180 185 190 Cys Ile Lys Thr Thr Val Glu Asn Glu Leu Pro Pro Pro Glu Asn Ile 195 200 205 Glu Val Ser Val Gln Asn Gln Asn Tyr Val Leu Lys Trp Asp Tyr Thr 210 215 220 Tyr Ala Asn Met Thr Phe Gln Val Gln Trp Leu His Ala Phe Leu Lys 225 230 235 240 Arg Asn Pro Gly Asn His Leu Tyr Lys Trp Lys Gln Ile Pro Asp Cys 245 250 255 Glu Asn Val Lys Thr Thr Gln Cys Val Phe Pro Gln Asn Val Phe Gln 260 265 270 Lys Gly Ile Tyr Leu Leu Arg Val Gln Ala Ser Asp Gly Asn Asn Thr 275 280 285 Ser Phe Trp Ser Glu Glu Ile Lys Phe Asp Thr Glu Ile Gln Ala Phe 290 295 300 Leu Leu Pro Pro Val Phe Asn Ile Arg Ser Leu Ser Asp Ser Phe His 305 310 315 320 Ile Tyr Ile Gly Ala Pro Lys Gln Ser Gly Asn Thr Pro Val Ile Gln 325 330 335 Asp Tyr Pro Leu Ile Tyr Glu Ile Ile Phe Trp Glu Asn Thr Ser Asn 340 345 350 Ala Glu Arg Lys Ile Ile Glu Lys Lys Thr Asp Val Thr Val Pro Asn 355 360 365 Leu Lys Pro Leu Thr Val Tyr Cys Val Lys Ala Arg Ala His Thr Met 370 375 380 Asp Glu Lys Leu Asn Lys Ser Ser Val Phe Ser Asp Ala Val Cys Glu 385 390 395 400 Lys Thr Lys Pro Gly Asn Thr Ser Lys 405 <210> 5 <211> 651 <212> DNA <213> Artificial sequence <220> <223> IFNAR2 extracellular domain <400> 5 atctcctacg acagccctga ctacaccgac gagtcctgca ccttcaagat ctccctgcgg 60 aacttccggt ccatcctgtc ctgggagctg aagaaccact ccatcgtgcc cacacactac 120 accctgctgt acaccatcat gtccaagcct gaggacctga aggtggtcaa gaactgcgcc 180 aacaccacca gatctttctg cgacctgacc gatgagtggc ggtctaccca cgaggcttac 240 gtgacagtgc tggaaggctt ctccggcaat accacactgt tcagctgctc ccacaacttc 300 tggctggcca tcgacatgtc cttcgagcct ccagagttcg agatcgtggg cttcaccaac 360 cacatcaacg tgatggtcaa gttccccagc atcgtggaag aggaactcca gttcgacctg 420 agcctggtca tcgaggaaca gtccgagggc atcgtgaaga agcacaagcc cgagatcaag 480 ggcaacatgt ccggcaactt cacctacatc atcgacaagc tgatccccaa caccaactac 540 tgcgtgtccg tgtacctgga acactccgat gagcaggccg tgatcaagtc ccctctgaag 600 tgtaccctgc tgcctcctgg ccaagagtct gagtctgccg agtctgctaa a 651 <210> 6 <211> 217 <212> PRT<*** <213> Artificial Sequence <220> <223> IFNAR2 Extracellular Domain <400> 6 Ile Ser Tyr Asp Ser Pro Asp Tyr Thr Asp Glu Ser Cys Thr Phe Lys 1 5 10 15 Ile Ser Leu Arg Asn Phe Arg Ser Ile Leu Ser Trp Glu Leu Lys Asn 20 25 30 His Ser Ile Val Pro Thr His Tyr Thr Leu Leu Tyr Thr Ile Met Ser 35 40 45 It should be noted that in the provided text, there seems to be an incorrect tag format in ID=19 where "***" is present. It might need to be corrected according to the actual correct format in the original context. Lys Pro Glu Asp Leu Lys Val Val Lys Asn Cys Ala Asn Thr Thr Arg 50 55 60 Ser Phe Cys Asp Leu Thr Asp Glu Trp Arg Ser Thr His Glu Ala Tyr 65 70 75 80 Val Thr Val Leu Glu Gly Phe Ser Gly Asn Thr Thr Leu Phe Ser Cys 85 90 95 Ser His Asn Phe Trp Leu Ala Ile Asp Met Ser Phe Glu Pro Pro Glu 100 105 110 Phe Glu Ile Val Gly Phe Thr Asn His Ile Asn Val Met Val Lys Phe 115 120 125 Pro Ser Ile Val Glu Glu Glu Leu Gln Phe Asp Leu Ser Leu Val Ile 130 135 140 Glu Glu Gln Ser Glu Gly Ile Val Lys Lys His Lys Pro Glu Ile Lys 145 150 155 160 Gly Asn Met Ser Gly Asn Phe Thr Tyr Ile Ile Asp Lys Leu Ile Pro 165 170 175 Asn Thr Asn Tyr Cys Val Ser Val Tyr Leu Glu His Ser Asp Glu Gln 180 185 190 Ala Val Ile Lys Ser Pro Leu Lys Cys Thr Leu Leu Pro Pro Gly Gln 195 200 205 Glu Ser Glu Ser Ala Glu Ser Ala Lys 210 215 <210> 7 <211> 696 <212> DNA <213> Artificial sequence <220> <223> FC-WT <400> 7 cccaagagct gcgacaagac ccacacctgt cccccctgcc ctgcccctga actgctgggc 60 ggacccagcg tgttcctgtt ccccccaaag cccaaggaca ccctgatgat cagccggacc 120 cccgaagtga cctgcgtggt ggtggacgtg tcccacgagg accctgaagt gaagttcaat 180 tggtacgtgg acggcgtgga agtgcacaac gccaagacca agcccagaga ggaacagtac 240 aacagcacct accgggtggt gtccgtgctg accgtgctgc accaggactg gctgaacggc 300 aaagagtaca agtgcaaggt gtccaacaag gccctgcctg cccccatcga gaaaaccatc 360 agcaaggcca agggccagcc ccgcgagccc caggtgtaca cactgccccc cagccgggac 420 gagctgacca agaaccaggt gtccctgacc tgcctggtga aaggcttcta ccccagcgat 480 atcgccgtgg aatgggagag caacggccag cccgagaaca actacaagac caccccccct 540 gtgctggaca gcgacggctc attcttcctg tacagcaagc tgaccgtgga caagagccgg 600 tggcagcagg gcaacgtgtt cagctgcagc gtgatgcacg aggccctgca caaccactac 660 acccagaagt ccctgagcct gagccccggc aagtag 696 <210> 8 <211> 231 <212> PRT <213> Artificial sequence <220> <223> FC-WT <400> 8 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 1 5 10 15 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 65 70 75 80 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 130 135 140 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 9 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Linker <400> 9 atcgagggcc ggatggac 18 <210> 10 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Linker <400> 10 Ile Glu Gly Arg Met Asp 1 5 <210> 11 <211> 696 <212> DNA <213> Artificial Sequence <220> <223> FC-EW <400> 11 cccaagagct gcgacaagac ccacacctgt cccccctgcc ctgcccctga actgctgggc 60 ggacccagcg tgttcctgtt ccccccaaag cccaaggaca ccctgatgat cagccggacc 120 cccgaagtga cctgcgtggt ggtggacgtg tcccacgagg accctgaagt gaagttcaat 180 tggtacgtgg acggcgtgga agtgcacaac gccaagacca agcccagaga ggaacagtac 240 aacagcacct accgggtggt gtccgtgctg accgtgctgc accaggactg gctgaacggc 300 aaagagtaca agtgcaaggt gtccaacaag gccctgcctg cccccatcga gaaaaccatc 360 agcaaggcca agggccagcc ccgcgagccc caggtgtaca cactgccccc cagccgggac 420 gagctgaccg agaaccaggt gtccctgacc tgcctggtga aaggcttcta ccccagcgat 480 atcgccgtgg aatgggagag caacggccag cccgagaaca actacaagac caccccccct 540 gtgctggaca gcgacggctc attcttcctg tacagctggc tgaccgtgga caagagccgg 600 tggcagcagg gcaacgtgtt cagctgcagc gtgatgcacg aggccctgca caaccactac 660 acccagaagt ccctgagcct gagccccggc aagtag 696 <210> 12 <211> 231 <212> PRT <213> Artificial sequence <220> <223> FC-EW <400> 12 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 1 5 10 15 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 65 70 75 80 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Glu 130 135 140 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Trp Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 13 <211> 696 <212> DNA <213> Artificial sequence <220> <223> FC-RVT <400> 13 cccaagagct gcgacaagac ccacacctgt cccccctgcc ctgcccctga actgctgggc 60 ggacccagcg tgttcctgtt ccccccaaag cccaaggaca ccctgatgat cagccggacc 120 cccgaagtga cctgcgtggt ggtggacgtg tcccacgagg accctgaagt gaagttcaat 180 tggtacgtgg acggcgtgga agtgcacaac gccaagacca agcccagaga ggaacagtac 240 aacagcacct accgggtggt gtccgtgctg accgtgctgc accaggactg gctgaacggc 300 aaagagtaca agtgcaaggt gtccaacaag gccctgcctg cccccatcga gaaaaccatc 360 agcaaggcca agggccagcc ccgcgagccc cgggtgtaca cactgccccc cagccgggac 420 gagctgacca agaaccaggt gtccctgacc tgcctggtga aaggcttcta ccccagcgat 480 atcgccgtgg aatgggagag caacggccag cccgagaaca actacaagac caccccccct 540 gtgctggtca gcgacggctc attcaccctg tacagcaagc tgaccgtgga caagagccgg 600 tggcagcagg gcaacgtgtt cagctgcagc gtgatgcacg aggccctgca caaccactac 660 acccagaagt ccctgagcct gagccccggc aagtag 696 <210> 14 <211> 231 <212> PRT <213> Artificial Sequence <220> <223> FC-RVT <400> 14 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 1 5 10 15 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 65 70 75 80 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Arg Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 130 135 140 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Val Ser Asp Gly Ser Phe Thr Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 15 <211> 783 <212> DNA <213> Artificial sequence <220> <223> Linker-FC WT <400> 15 gccaccatgt ggtggcgtct gtggtggctg ctgcttctgc tgttgctgct gtggcctatg 60 gtctgggcca tcgagggccg gatggacccc aagagctgcg acaagaccca cacctgtccc 120 ccctgccctg cccctgaact gctgggcgga cccagcgtgt tcctgttccc cccaaagccc 180 aaggacaccc tgatgatcag ccggaccccc gaagtgacct gcgtggtggt ggacgtgtcc 240 cacgaggacc ctgaagtgaa gttcaattgg tacgtggacg gcgtggaagt gcacaacgcc 300 aagaccaagc ccagagagga acagtacaac agcacctacc gggtggtgtc cgtgctgacc 360 gtgctgcacc aggactggct gaacggcaaa gagtacaagt gcaaggtgtc caacaaggcc 420 ctgcctgccc ccatcgagaa aaccatcagc aaggccaagg gccagccccg cgagccccag 480 gtgtacacac tgccccccag ccgggacgag ctgaccaaga accaggtgtc cctgacctgc 540 ctggtgaaag gcttctaccc cagcgatatc gccgtggaat gggagagcaa cggccagccc 600 gagaacaact acaagaccac cccccctgtg ctggacagcg acggctcatt cttcctgtac 660 agcaagctga ccgtggacaa gagccggtgg cagcagggca acgtgttcag ctgcagcgtg 720 atgcacgagg ccctgcacaa ccactacacc cagaagtccc tgagcctgag ccccggcaag 780 tga 783 <210> 16 <211> 260 <212> PRT <213> Artificial sequence <220> <223> Linker-FC WT <400> 16 Ala Thr Met Trp Trp Arg Leu Trp Trp Leu Leu Leu Leu Leu Leu Leu 1 5 10 15 Leu Trp Pro Met Val Trp Ala Ile Glu Gly Arg Met Asp Pro Lys Ser 20 25 30 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 35 40 45 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 50 55 60 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 65 70 75 80 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 85 90 95 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 100 105 110 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 115 120 125 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 130 135 140 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 145 150 155 160 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 165 170 175 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 180 185 190 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 195 200 205 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 210 215 220 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 225 230 235 240 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 245 250 255 Ser Pro Gly Lys 260 <210> 17 <211> 783 <212> DNA <213> Artificial sequence <220> <223> Linker-FC EW <400> 17 gccaccatgt ggtggcgtct gtggtggctg ctgcttctgc tgttgctgct gtggcctatg 60 gtctgggcca tcgagggccg gatggacccc aagagctgcg acaagaccca cacctgtccc 120 ccctgccctg cccctgaact gctgggcgga cccagcgtgt tcctgttccc cccaaagccc 180 aaggacaccc tgatgatcag ccggaccccc gaagtgacct gcgtggtggt ggacgtgtcc 240 cacgaggacc ctgaagtgaa gttcaattgg tacgtggacg gcgtggaagt gcacaacgcc 300 aagaccaagc ccagagagga acagtacaac agcacctacc gggtggtgtc cgtgctgacc 360 gtgctgcacc aggactggct gaacggcaaa gagtacaagt gcaaggtgtc caacaaggcc 420 ctgcctgccc ccatcgagaa aaccatcagc aaggccaagg gccagccccg cgagccccag 480 gtgtacacac tgccccccag ccgggacgag ctgaccgaga accaggtgtc cctgacctgc 540 ctggtgaaag gcttctaccc cagcgatatc gccgtggaat gggagagcaa cggccagccc 600 gagaacaact acaagaccac cccccctgtg ctggacagcg acggctcatt cttcctgtac 660 agctggctga ccgtggacaa gagccggtgg cagcagggca acgtgttcag ctgcagcgtg 720 atgcacgagg ccctgcacaa ccactacacc cagaagtccc tgagcctgag ccccggcaag 780 tag 783 <210> 18 <211> 260 <212> PRT <213> Artificial sequence <220> <223> Linker-FC EW <400> 18 Ala Thr Met Trp Trp Arg Leu Trp Trp Leu Leu Leu Leu Leu Leu Leu 1 5 10 15 Leu Trp Pro Met Val Trp Ala Ile Glu Gly Arg Met Asp Pro Lys Ser 20 25 30 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 35 40 45 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 50 55 60 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 65 70 75 80 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 85 90 95 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 100 105 110 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 115 120 125 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 130 135 140 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 145 150 155 160 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Glu Asn Gln Val 165 170 175 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 180 185 190 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 195 200 205 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Trp Leu Thr 210 215 220 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 225 230 235 240 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 245 250 255 Ser Pro Gly Lys 260 <210> 19 <211> 783 <212> DNA <213> Artificial Sequence <220> <223> Linker-FC RVT <400> 19 gccaccatgt ggtggcgtct gtggtggctg ctgcttctgc tgttgctgct gtggcctatg 60 gtctgggcca tcgagggccg gatggacccc aagagctgcg acaagaccca cacctgtccc 120 ccctgccctg cccctgaact gctgggcgga cccagcgtgt tcctgttccc cccaaagccc 180 aaggacaccc tgatgatcag ccggaccccc gaagtgacct gcgtggtggt ggacgtgtcc 240 cacgaggacc ctgaagtgaa gttcaattgg tacgtggacg gcgtggaagt gcacaacgcc 300 aagaccaagc ccagagagga acagtacaac agcacctacc gggtggtgtc cgtgctgacc 360 gtgctgcacc aggactggct gaacggcaaa gagtacaagt gcaaggtgtc caacaaggcc 420 ctgcctgccc ccatcgagaa aaccatcagc aaggccaagg gccagccccg cgagccccgg 480 gtgtacacac tgccccccag ccgggacgag ctgaccaaga accaggtgtc cctgacctgc 540 ctggtgaaag gcttctaccc cagcgatatc gccgtggaat gggagagcaa cggccagccc 600 gagaacaact acaagaccac cccccctgtg ctggtcagcg acggctcatt caccctgtac 660 agcaagctga ccgtggacaa gagccggtgg cagcagggca acgtgttcag ctgcagcgtg 720 atgcacgagg ccctgcacaa ccactacacc cagaagtccc tgagcctgag ccccggcaag 780 tag 783 <210> 20 <211> 260 <212> PRT <213> Artificial sequence <220> <223> Linker-FC RVT <400> 20 Ala Thr Met Trp Trp Arg Leu Trp Trp Leu Leu Leu Leu Leu Leu Leu 1 5 10 15 Leu Trp Pro Met Val Trp Ala Ile Glu Gly Arg Met Asp Pro Lys Ser 20 25 30 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 35 40 45 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 50 55 60 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 65 70 75 80 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 85 90 95 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 100 105 110 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 115 120 125 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 130 135 140 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Arg 145 150 155 160 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 165 170 175 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 180 185 190 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 195 200 205 Pro Val Leu Val Ser Asp Gly Ser Phe Thr Leu Tyr Ser Lys Leu Thr 210 215 220 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 225 230 235 240 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 245 250 255 Ser Pro Gly Lys 260 <210> 21 <211> 2010 <212> DNA <213> The snowstorm <220> <223> IFNAR1-transformer-FC WT <400> 21 gccaccatgt ggtggcgtct gtggtggctg ctgcttctgc tgttgctgct gtggcctatg gtctgggcca agaacctgaa gtcccctcag aaagtggaag tggacatcat cgacgacaac ttcatcctgc ggtggaacag atccgacgag tccgtgggca acgtgacctt cagcttcgac 180 240. ccggcatgga cactggatc aagctgtccg gctgccaga catcacctct accaagtgca acttctccag cctgaagctg aacgtgtacg aggaatcaa gctgcggatc cgggccgaga aagagaacac cagcagttgg tacgaggtgg acagcttcac ccctttcaga aaggcccaga tcggccctcc tgaagtgcac ctggaagccg aggataaggc catcgtgatc 420 cacatcagcc ccggcaccaa ggactctgtg atgtgggctc tcgacggcct gtccttcacc 480 tacagcctgg tcatctgga gaactcctcc ggcgtggaag agagaatcga gaactctc tcccggcaca agatctacaa gctgagcccc gagacaacct actgcctgaa agtgaaggcc gctctgctga cctcctggaa gatcggcgtg tactctcccg tgcactgcat caagaccacc 660 gtggaaaacg agctgcctcc tccagagaac atcgaggtgt ccgtgcagaa ccagaactac 720 gtgctgaagt gggactacac ctacgccaac atgacctttc aggtgcagtg gctgcacgct 780 tttctgaagc ggaaccctgg caaccacctg tacaagtgga agcagattcc cgactgcgag 840 aacgtgaaaa ccacacagtg cgtgttccct cagaacgtgt tccagaaggg catctacctg 900 ctgagagtgc aggcctccga cggcaacaac accagctttt ggagcgaaga gatcaagttc 960 gataccgaga tccaggcctt cctgctgcct ccagtgttca acatcagatc cctgtccgac 1020 tccttccaca tctacatcgg cgctcctaag cagtccggca acacccctgt gatccaggac 1080 taccctctga tctacgagat catcttctgg gagaacacct ccaacgccga gcggaagatc 1140 atcgagaaga aaaccgacgt gaccgtgcct aacctgaagc ctctgaccgt gtactgcgtg 1200 aaggctaggg cccacaccat ggacgaaaag ctgaacaagt cctccgtgtt ctccgacgcc 1260 gtgtgcgaaa agaccaagcc aggcaacacc tctaaaatcg agggccggat ggaccccaag 1320 agctgcgaca agacccacac ctgtcccccc tgccctgccc ctgaactgct gggcggaccc 1380 agcgtgttcc tgttcccccc aaagcccaag gacaccctga tgatcagccg gacccccgaa 1440 gtgacctgcg tggtggtgga cgtgtcccac gaggaccctg aagtgaagtt caattggtac 1500 gtggacggcg tggaagtgca caacgccaag accaagccca gagaggaaca gtacaacagc 1560 acctaccggg tggtgtccgt gctgaccgtg ctgcaccagg actggctgaa cggcaaagag 1620 tacaagtgca aggtgtccaa caaggccctg cctgccccca tcgagaaaac catcagcaag 1680 gccaagggcc agccccgcga gccccaggtg tacacactgc cccccagccg ggacgagctg 1740 accaagaacc aggtgtccct gacctgcctg gtgaaaggct tctaccccag cgatatcgcc 1800 gtggaatggg agagcaacgg ccagcccgag aacaactaca agaccacccc ccctgtgctg 1860 gacagcgacg gctcattctt cctgtacagc aagctgaccg tggacaagag ccggtggcag 1920 cagggcaacg tgttcagctg cagcgtgatg cacgaggccc tgcacaacca ctacacccag 1980 aagtccctga gcctgagccc cggcaagtga 2010 <210> 22 <211> 669 <212> PRT <213> artificial sequence <220> <223> IFNAR1-connector-FC WT <400> 22 Ala Thr Met Trp Trp Arg Leu Trp Trp Leu Leu Leu Leu Leu Leu Leu 1 5 10 15 Leu Trp Pro Met Val Trp Ala Lys Asn Leu Lys Ser Pro Gln Lys Val 20 25 30 Glu Val Asp Ile Ile Asp Asp Asn Phe Ile Leu Arg Trp Asn Arg Ser 35 40 45 Asp Glu Ser Val Gly Asn Val Thr Phe Ser Phe Asp Tyr Gln Lys Thr 50 55 60 Gly Met Asp Asn Trp Ile Lys Leu Ser Gly Cys Gln Asn Ile Thr Ser 65 70 75 80 Thr Lys Cys Asn Phe Ser Ser Leu Lys Leu Asn Val Tyr Glu Glu Ile 85 90 95 Lys Leu Arg Ile Arg Ala Glu Lys Glu Asn Thr Ser Ser Trp Tyr Glu 100 105 110 Val Asp Ser Phe Thr Pro Phe Arg Lys Ala Gln Ile Gly Pro Pro Glu 115 120 125 Val His Leu Glu Ala Glu Asp Lys Ala Ile Val Ile His Ile Ser Pro 130 135 140 Gly Thr Lys Asp Ser Val Met Trp Ala Leu Asp Gly Leu Ser Phe Thr 145 150 155 160 Tyr Ser Leu Val Ile Trp Lys Asn Ser Ser Gly Val Glu Glu Arg Ile 165 170 175 Glu Asn Ile Tyr Ser Arg His Lys Ile Tyr Lys Leu Ser Pro Glu Thr 180 185 190 Thr Tyr Cys Leu Lys Val Lys Ala Ala Leu Leu Thr Ser Trp Lys Ile 195 200 205 Gly Val Tyr Ser Pro Val His Cys Ile Lys Thr Thr Val Glu Asn Glu 210 215 220 Leu Pro Pro Pro Glu Asn Ile Glu Val Ser Val Gln Asn Gln Asn Tyr 225 230 235 240 Val Leu Lys Trp Asp Tyr Thr Tyr Ala Asn Met Thr Phe Gln Val Gln 245 250 255 Trp Leu His Ala Phe Leu Lys Arg Asn Pro Gly Asn His Leu Tyr Lys 260 265 270 Trp Lys Gln Ile Pro Asp Cys Glu Asn Val Lys Thr Thr Gln Cys Val 275 280 285 Phe Pro Gln Asn Val Phe Gln Lys Gly Ile Tyr Leu Leu Arg Val Gln 290 295 300 Ala Ser Asp Gly Asn Asn Thr Ser Phe Trp Ser Glu Glu Ile Lys Phe 305 310 315 320 Asp Thr Glu Ile Gln Ala Phe Leu Leu Pro Pro Val Phe Asn Ile Arg 325 330 335 Ser Leu Ser Asp Ser Phe His Ile Tyr Ile Gly Ala Pro Lys Gln Ser 340 345 350 Gly Asn Thr Pro Val Ile Gln Asp Tyr Pro Leu Ile Tyr Glu Ile Ile 355 360 365 Phe Trp Glu Asn Thr Ser Asn Ala Glu Arg Lys Ile Ile Glu Lys Lys 370 375 380 Thr Asp Val Thr Val Pro Asn Leu Lys Pro Leu Thr Val Tyr Cys Val 385 390 395 400 Lys Ala Arg Ala His Thr Met Asp Glu Lys Leu Asn Lys Ser Ser Val 405 410 415 Phe Ser Asp Ala Val Cys Glu Lys Thr Lys Pro Gly Asn Thr Ser Lys 420 425 430 Ile Glu Gly Arg Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys 435 440 445 Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu 450 455 460 Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu 465 470 475 480 Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys 485 490 495 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 500 505 510 Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu 515 520 525 Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 530 535 540 Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 545 550 555 560 Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser 565 570 575 Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 580 585 590 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 595 600 605 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly 610 615 620 Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln 625 630 635 640 Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 645 650 655 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 660 665 <210> 23 <211> 2010 <212> DNA <213> Artificial sequence <220> <223> IFNAR1 - linker - FC EW <400> 23 gccaccatgt ggtggcgtct gtggtggctg ctgcttctgc tgttgctgct gtggcctatg 60 gtctgggcca agaacctgaa gtcccctcag aaagtggaag tggacatcat cgacgacaac 120 ttcatcctgc ggtggaacag atccgacgag tccgtgggca acgtgacctt cagcttcgac 180 taccagaaaa ccggcatgga caactggatc aagctgtccg gctgccagaa catcacctct 240 accaagtgca acttctccag cctgaagctg aacgtgtacg aggaaatcaa gctgcggatc 300 cgggccgaga agagacac cagcagttgg tacgaggtgg acagctcac ccctttcaga 360 aaggcccaga tcggccctcc tgaagtgcac ctggaagccg aggataggc catcgtgatc 420 cacatcagcc ccggcaccaa ggactgtg atgtgggctc tcgacggcct gtccttcacc 480 tacagcctgg tcatctggaa gaactcctcc ggcgtggaag agagaatcga gaacactac 540 tcccggcaca agatctacaa gctgagcccc gagacaacct actgcctgaa agtgaaggcc 600 gctctgctga cctcctggaa gatcggcgtg tactctcccg tgcactgcat caaccacc 660 gtggaaaacg agctgcctcc tccagagaac atcgaggtgt ccgtgcagaa ccagaactac 720 gtgctgaagt gggactacac ctacgccaac atgaccttc aggtgcagtg gctgcacgct 780 tttctgaagc ggaaccctgg caaccacctg tacaagtgga agcagattcc cgactgcgag 840 aacgtgaaaa cacacagtg cgtgttccct cagaacgtgt tccagaaggg catcactg 900 ctgagagtgc aggcctccga cggcaacac accagcttt ggagcgaaga gatcaagttc 960 gataccgaga tccaggcctt cctgctgcct ccagtgttca acatcagatc cctgtccgac 1020 tccttccaca tctacatcgg cgctcctaag cagtccggca acacccctgt gatccaggac 1080 taccctctga tctacgagat catcttctgg gagaacacct ccaacgccga gcggaagatc 1140 atcgagaaga aaaccgacgt gaccgtgcct aacctgaagc ctctgaccgt gtactgcgtg 1200 aaggctaggg cccacaccat ggacgaaaag ctgaacaagt cctccgtgtt ctccgacgcc 1260 gtgtgcgaaa agaccaagcc aggcaacacc tctaaaatcg agggccggat ggaccccaag 1320 agctgcgaca agacccacac ctgtcccccc tgccctgccc ctgaactgct gggcggaccc 1380 agcgtgttcc tgttcccccc aaagcccaag gacaccctga tgatcagccg gacccccgaa 1440 gtgacctgcg tggtggtgga cgtgtcccac gaggaccctg aagtgaagtt caattggtac 1500 gtggacggcg tggaagtgca caacgccaag accaagccca gagaggaaca gtacaacagc 1560 acctaccggg tggtgtccgt gctgaccgtg ctgcaccagg actggctgaa cggcaaagag 1620 tacaagtgca aggtgtccaa caaggccctg cctgccccca tcgagaaaac catcagcaag 1680 gccaagggcc agccccgcga gccccaggtg tacacactgc cccccagccg ggacgagctg 1740 accgagaacc aggtgtccct gacctgcctg gtgaaaggct tctaccccag cgatatcgcc 1800 gtggaatggg agagcaacgg ccagcccgag aacaactaca agaccacccc ccctgtgctg 1860 gacagcgacg gctcattctt cctgtacagc tggctgaccg tggacaagag ccggtggcag 1920 cagggcaacg tgttcagctg cagcgtgatg cacgaggccc tgcacaacca ctacacccag 1980 aagtccctga gcctgagccc cggcaagtag 2010 <210> 24 <211> 669 <212> PRT <213> Artificial Sequence <220> <223> IFNAR1 - Linker - FC EW <400> 24 Ala Thr Met Trp Trp Arg Leu Trp Trp Leu Leu Leu Leu Leu Leu Leu 1 5 10 15 Leu Trp Pro Met Val Trp Ala Lys Asn Leu Lys Ser Pro Gln Lys Val 20 25 30 Glu Val Asp Ile Ile Asp Asp Asn Phe Ile Leu Arg Trp Asn Arg Ser 35 40 45 Asp Glu Ser Val Gly Asn Val Thr Phe Ser Phe Asp Tyr Gln Lys Thr 50 55 60 Gly Met Asp Asn Trp Ile Lys Leu Ser Gly Cys Gln Asn Ile Thr Ser 65 70 75 80 Thr Lys Cys Asn Phe Ser Ser Leu Lys Leu Asn Val Tyr Glu Glu Ile 85 90 95 Lys Leu Arg Ile Arg Ala Glu Lys Glu Asn Thr Ser Ser Trp Tyr Glu 100 105 110 Val Asp Ser Phe Thr Pro Phe Arg Lys Ala Gln Ile Gly Pro Pro Glu 115 120 125 Val His Leu Glu Ala Glu Asp Lys Ala Ile Val Ile His Ile Ser Pro 130 135 140 Gly Thr Lys Asp Ser Val Met Trp Ala Leu Asp Gly Leu Ser Phe Thr 145 150 155 160 Tyr Ser Leu Val Ile Trp Lys Asn Ser Ser Gly Val Glu Glu Arg Ile 165 170 175 Glu Asn Ile Tyr Ser Arg His Lys Ile Tyr Lys Leu Ser Pro Glu Thr 180 185 190 Thr Tyr Cys Leu Lys Val Lys Ala Ala Leu Leu Thr Ser Trp Lys Ile 195 200 205 Gly Val Tyr Ser Pro Val His Cys Ile Lys Thr Thr Val Glu Asn Glu 210 215 220 Leu Pro Pro Pro Glu Asn Ile Glu Val Ser Val Gln Asn Gln Asn Tyr 225 230 235 240 Val Leu Lys Trp Asp Tyr Thr Tyr Ala Asn Met Thr Phe Gln Val Gln 245 250 255 Trp Leu His Ala Phe Leu Lys Arg Asn Pro Gly Asn His Leu Tyr Lys 260 265 270 Trp Lys Gln Ile Pro Asp Cys Glu Asn Val Lys Thr Thr Gln Cys Val 275 280 285 Phe Pro Gln Asn Val Phe Gln Lys Gly Ile Tyr Leu Leu Arg Val Gln 290 295 300 Ala Ser Asp Gly Asn Asn Thr Ser Phe Trp Ser Glu Glu Ile Lys Phe 305 310 315 320 Asp Thr Glu Ile Gln Ala Phe Leu Leu Pro Pro Val Phe Asn Ile Arg 325 330 335 Ser Leu Ser Asp Ser Phe His Ile Tyr Ile Gly Ala Pro Lys Gln Ser 340 345 350 Gly Asn Thr Pro Val Ile Gln Asp Tyr Pro Leu Ile Tyr Glu Ile Ile 355 360 365 Phe Trp Glu Asn Thr Ser Asn Ala Glu Arg Lys Ile Ile Glu Lys Lys 370 375 380 Thr Asp Val Thr Val Pro Asn Leu Lys Pro Leu Thr Val Tyr Cys Val 385 390 395 400 Lys Ala Arg Ala His Thr Met Asp Glu Lys Leu Asn Lys Ser Ser Val 405 410 415 Phe Ser Asp Ala Val Cys Glu Lys Thr Lys Pro Gly Asn Thr Ser Lys 420 425 430 Ile Glu Gly Arg Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys 435 440 445 Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu 450 455 460 Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu 465 470 475 480 Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys 485 490 495 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 500 505 510 Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu 515 520 525 Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 530 535 540 Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 545 550 555 560 Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser 565 570 575 Arg Asp Glu Leu Thr Glu Asn Gln Val Ser Leu Thr Cys Leu Val Lys 580 585 590 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 595 600 605 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly 610 615 620 Ser Phe Phe Leu Tyr Ser Trp Leu Thr Val Asp Lys Ser Arg Trp Gln 625 630 635 640 Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 645 650 655 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 660 665 <210> 25 <211> 2010 <212> DNA <213> Artificial Sequence <220> <223> IFNAR1 - Linker - FC RVT <400> 25 gccaccatgt gtggtggctg ctgctctgc tgttgctgct gtggcctatg 60 gtctgggcca agaacctgaa gtcccctcag aaagtggaag tggacatcat cgacgacac 120 ttcatcctgc gtggaacag atccgacgag tccgtgggca acgtgacctt cagcttcgac 180 taccagaaaa ccggcatgga caacggatc aagctgtccg gctgccagaa catcacctct 240 accaagtgca acttctccag cctgaagctg aacgtgtacg aggaatca gctgcggatc 300 cgggccgaga agagacac cagcagttgg tacgaggtgg acagctcac ccctttcaga 360 aaggcccaga tcggccctcc tgaagtgcac ctggaagccg aggataggc catcgtgatc 420 cacatcagcc ccggcaccaa ggactgtg atgtgggctc tcgacggcct gtccttcacc 480 tacagcctgg tcatctggaa gaactcctcc ggcgtggaag agagaatcga gaacactac 540 tcccggcaca agatctacaa gctgagcccc gagacaacct actgcctgaa agtgaaggcc 600 gctctgctga cctcctggaa gatcggcgtg tactctcccg tgcactgcat caaccacc 660 gtggaaaacg agctgcctcc tccagagaac atcgaggtgt ccgtgcagaa ccagaactac 720 gtgctgaagt gggactacac ctacgccaac atgacctttc aggtgcagtg gctgcacgct 780 tttctgaagc ggaaccctgg caaccacctg tacaagtgga agcagattcc cgactgcgag 840 aacgtgaaaa ccacacagtg cgtgttccct cagaacgtgt tccagaaggg catctacctg 900 ctgagagtgc aggcctccga cggcaacaac accagctttt ggagcgaaga gatcaagttc 960 gataccgaga tccaggcctt cctgctgcct ccagtgttca acatcagatc cctgtccgac 1020 tccttccaca tctacatcgg cgctcctaag cagtccggca acacccctgt gatccaggac 1080 taccctctga tctacgagat catcttctgg gagaacacct ccaacgccga gcggaagatc 1140 atcgagaaga aaaccgacgt gaccgtgcct aacctgaagc ctctgaccgt gtactgcgtg 1200 aaggctaggg cccacaccat ggacgaaaag ctgaacaagt cctccgtgtt ctccgacgcc 1260 gtgtgcgaaa agaccaagcc aggcaacacc tctaaaatcg agggccggat ggaccccaag 1320 agctgcgaca agacccacac ctgtcccccc tgccctgccc ctgaactgct gggcggaccc 1380 agcgtgttcc tgttcccccc aaagcccaag gacaccctga tgatcagccg gacccccgaa 1440 gtgacctgcg tggtggtgga cgtgtcccac gaggaccctg aagtgaagtt caattggtac 1500 gtggacggcg tggaagtgca caacgccaag accaagccca gagaggaaca gtacaacagc 1560 acctaccggg tggtgtccgt gctgaccgtg ctgcaccagg actggctgaa cggcaaagag 1620 tacaagtgca aggtgtccaa caaggccctg cctgccccca tcgagaaaac catcagcaag 1680 gccaagggcc agccccgcga gccccgggtg tacacactgc cccccagccg ggacgagctg 1740 accaagaacc aggtgtccct gacctgcctg gtgaaaggct tctaccccag cgatatcgcc 1800 gtggaatggg agagcaacgg ccagcccgag aacaactaca agaccacccc ccctgtgctg 1860 gtcagcgacg gctcattcac cctgtacagc aagctgaccg tggacaagag ccggtggcag 1920 cagggcaacg tgttcagctg cagcgtgatg cacgaggccc tgcacaacca ctacacccag 1980 aagtccctga gcctgagccc cggcaagtag 2010 <210> 26 <211> 669 <212> PRT <213> Artificial Sequence <220> <223> IFNAR1 - Linker - FC RVT <400> 26 Ala Thr Met Trp Trp Arg Leu Trp Trp Leu Leu Leu Leu Leu Leu Leu 1 5 10 15 Leu Trp Pro Met Val Trp Ala Lys Asn Leu Lys Ser Pro Gln Lys Val 20 25 30 Glu Val Asp Ile Ile Asp Asp Asn Phe Ile Leu Arg Trp Asn Arg Ser 35 40 45 Asp Glu Ser Val Gly Asn Val Thr Phe Ser Phe Asp Tyr Gln Lys Thr 50 55 60 Gly Met Asp Asn Trp Ile Lys Leu Ser Gly Cys Gln Asn Ile Thr Ser 65 70 75 80 Thr Lys Cys Asn Phe Ser Ser Leu Lys Leu Asn Val Tyr Glu Glu Ile 85 90 95 Lys Leu Arg Ile Arg Ala Glu Lys Glu Asn Thr Ser Ser Trp Tyr Glu 100 105 110 Val Asp Ser Phe Thr Pro Phe Arg Lys Ala Gln Ile Gly Pro Pro Glu 115 120 125 Val His Leu Glu Ala Glu Asp Lys Ala Ile Val Ile His Ile Ser Pro 130 135 140 Gly Thr Lys Asp Ser Val Met Trp Ala Leu Asp Gly Leu Ser Phe Thr 145 150 155 160 Tyr Ser Leu Val Ile Trp Lys Asn Ser Ser Gly Val Glu Glu Arg Ile 165 170 175 Glu Asn Ile Tyr Ser Arg His Lys Ile Tyr Lys Leu Ser Pro Glu Thr 180 185 190 Thr Tyr Cys Leu Lys Val Lys Ala Ala Leu Leu Thr Ser Trp Lys Ile 195 200 205 Gly Val Tyr Ser Pro Val His Cys Ile Lys Thr Thr Val Glu Asn Glu 210 215 220 Leu Pro Pro Pro Glu Asn Ile Glu Val Ser Val Gln Asn Gln Asn Tyr 225 230 235 240 Val Leu Lys Trp Asp Tyr Thr Tyr Ala Asn Met Thr Phe Gln Val Gln 245 250 255 Trp Leu His Ala Phe Leu Lys Arg Asn Pro Gly Asn His Leu Tyr Lys 260 265 270 Trp Lys Gln Ile Pro Asp Cys Glu Asn Val Lys Thr Thr Gln Cys Val 275 280 285 Phe Pro Gln Asn Val Phe Gln Lys Gly Ile Tyr Leu Leu Arg Val Gln 290 295 300 Ala Ser Asp Gly Asn Asn Thr Ser Phe Trp Ser Glu Glu Ile Lys Phe 305 310 315 320 Asp Thr Glu Ile Gln Ala Phe Leu Leu Pro Pro Val Phe Asn Ile Arg 325 330 335 Ser Leu Ser Asp Ser Phe His Ile Tyr Ile Gly Ala Pro Lys Gln Ser 340 345 350 Gly Asn Thr Pro Val Ile Gln Asp Tyr Pro Leu Ile Tyr Glu Ile Ile 355 360 365 Phe Trp Glu Asn Thr Ser Asn Ala Glu Arg Lys Ile Ile Glu Lys Lys 370 375 380 Thr Asp Val Thr Val Pro Asn Leu Lys Pro Leu Thr Val Tyr Cys Val 385 390 395 400 Lys Ala Arg Ala His Thr Met Asp Glu Lys Leu Asn Lys Ser Ser Val 405 410 415 Phe Ser Asp Ala Val Cys Glu Lys Thr Lys Pro Gly Asn Thr Ser Lys 420 425 430 Ile Glu Gly Arg Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys 435 440 445 Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu 450 455 460 Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu 465 470 475 480 Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys 485 490 495 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 500 505 510 Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu 515 520 525 Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 530 535 540 Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 545 550 555 560 Ala Lys Gly Gln Pro Arg Glu Pro Arg Val Tyr Thr Leu Pro Pro Ser 565 570 575 Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 580 585 590 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 595 600 605 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Val Ser Asp Gly 610 615 620 Ser Phe Thr Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln 625 630 635 640 Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 645 650 655 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 660 665 <210> 27 <211> 1434 <212> DNA <213> Artificial Sequence <220> <223> IFNAR2-Linker-FC WT <400> 27 gccaccatgt ggtggcgtct gtggtggctg ctgcttctgc tgttgctgct gtggcctatg 60 gtctgggcca tctcctacga cagccctgac tacaccgacg agtcctgcac cttcaagatc 120 tccctgcgga acttccggtc catcctgtcc tgggagctga agaaccactc catcgtgccc 180 acacactaca ccctgctgta caccatcatg tccaagcctg aggacctgaa ggtggtcaag 240 aactgcgcca acaccaccag atctttctgc gacctgaccg atgagtggcg gtctacccac 300 gaggcttacg tgacagtgct ggaaggcttc tccggcaata ccacactgtt cagctgctcc 360 cacaacttct ggctggccat cgacatgtcc ttcgagcctc cagagttcga gatcgtgggc 420 ttcaccaacc acatcaacgt gatggtcaag ttccccagca tcgtggaaga ggaactccag 480 ttcgacctga gcctggtcat cgaggaacag tccgagggca tcgtgaagaa gcacaagccc 540 gagatcaagg gcaacatgtc cggcaacttc acctacatca tcgacaagct gatcccccaac 600 accaactact gcgtgtccgt gtacctggaa cactccgatg agcaggccgt gatcaagtcc 660 cctctgaagt gtaccctgct gcctcctggc caagagtctg agtctgccga gtctgctaaa 720 atcgagggcc ggatggaccc caagagctgc gacaagaccc acacctgtcc cccctgccct 780 gcccctgaac tgctgggcgg acccagcgtg ttcctgttcc ccccaaagcc caaggacacc 840 ctgatgatca gccggacccc cgaagtgacc tgcgtggtgg tggacgtgtc ccacgaggac 900 960 cccagagagg aacagacaa cagcacctac cgggtggtgt ccgtgctgac cgtgctgcac 1020 caggactggc tgaacggcaa agagtacaag tgcaaggtgt ccaacaaggc cctgcctgcc 1080 1140 ctgcccccca gccgggacga gctgaccaag aaccaggtgt ccctgacctg cctggtgaaa 1200 ggcttctacc ccagcgatat cgccgtggaa tgggagagca acggccagcc cgagaacaac 1260 tacaagacca ccccccctgt gctggacagc gacggctcat tcttcctgta cagcaagctg 1320 accgtggaca agagccggtg gcagcagggc aacgtgttca gctgcagcgt gatgcacgag 1380 gccctgcaca accactacac ccagaagtcc ctgagcctga gccccggcaa gtga 1434 <210> 28 <211> 477 <212> PRT <213> Artificial Sequence <220> <223> IFNAR2 - Linker - FC WT <400> 28 Ala Thr Met Trp Trp Arg Leu Trp Trp Leu Leu Leu Leu Leu Leu Leu 1 5 10 15 Leu Trp Pro Met Val Trp Ala Ile Ser Tyr Asp Ser Pro Asp Tyr Thr 20 25 30 Asp Glu Ser Cys Thr Phe Lys Ile Ser Leu Arg Asn Phe Arg Ser Ile 35 40 45 Leu Ser Trp Glu Leu Lys Asn His Ser Ile Val Pro Thr His Tyr Thr 50 55 60 Leu Leu Tyr Thr Ile Met Ser Lys Pro Glu Asp Leu Lys Val Val Lys 65 70 75 80 Asn Cys Ala Asn Thr Thr Arg Ser Phe Cys Asp Leu Thr Asp Glu Trp 85 90 95 Arg Ser Thr His Glu Ala Tyr Val Thr Val Leu Glu Gly Phe Ser Gly 100 105 110 Asn Thr Thr Leu Phe Ser Cys Ser His Asn Phe Trp Leu Ala Ile Asp 115 120 125 Met Ser Phe Glu Pro Pro Glu Phe Glu Ile Val Gly Phe Thr Asn His 130 135 140 Ile Asn Val Met Val Lys Phe Pro Ser Ile Val Glu Glu Glu Leu Gln 145 150 155 160 Phe Asp Leu Ser Leu Val Ile Glu Glu Gln Ser Glu Gly Ile Val Lys 165 170 175 Lys His Lys Pro Glu Ile Lys Gly Asn Met Ser Gly Asn Phe Thr Tyr 180 185 190 Ile Ile Asp Lys Leu Ile Pro Asn Thr Asn Tyr Cys Val Ser Val Tyr 195 200 205 Leu Glu His Ser Asp Glu Gln Ala Val Ile Lys Ser Pro Leu Lys Cys 210 215 220 Thr Leu Leu Pro Pro Gly Gln Glu Ser Glu Ser Ala Glu Ser Ala Lys 225 230 235 240 Ile Glu Gly Arg Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys 245 250 255 Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu 260 265 270 Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu 275 280 285 Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys 290 295 300 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 305 310 315 320 Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu 325 330 335 Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 340 345 350 Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 355 360 365 Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser 370 375 380 Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 385 390 395 400 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 405 410 415 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly 420 425 430 Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln 435 440 445 Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 450 455 460 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 465 470 475 <210> 29 <211> 1434 <212> DNA <213> Artificial sequence <220> <223> IFNAR2 - linker - FC EW <400> 29 gccaccatgt ggtggcgtct gtggtggctg ctgcttctgc tgttgctgct gtggcctatg 60 gtctgggcca tctcctacga cagccctgac tacaccgacg agtcctgcac cttcaagatc 120 tccctgcgga acttccggtc catcctgtcc tgggagctga agaaccactc catcgtgccc 180 acacactaca ccctgctgta caccatcatg tccaagcctg aggacctgaa ggtggtcaag 240 aactgcgcca acaccaccag atctttctgc gacctgaccg atgagtggcg gtctacccac 300 gaggcttacg tgacagtgct ggaaggcttc tccggcaata ccacactgtt cagctgctcc 360 caaacttct ggctggccat cgacatgtcc ttcgagcctc cagagttcga gatcgtgggc 420 ttcaccaacc acatcaacgt gatggtcaag ttccccagca tcgtggaaga ggaactccag 480 ttcgacctga gcctggtcat cgaggaacag tccgagggca tcgtgaagaa gcacaagccc 540 gagatcaagg gcaacatgtc cggcaacttc acctacatca tcgacaagct gatcccccaac 600 accaactact gcgtgtccgt gtacctggaa cactccgatg agcaggccgt gatcaagtcc 660 cctctgaagt gtaccctgct gcctcctggc caagagtctg agtctgccga gtctgctaaa 720 atcgagggcc ggatggaccc caagagctgc gacaagaccc acacctgtcc cccctgccct 780 gcccctgaac tgctgggcgg acccagcgtg ttcctgttcc ccccaaagcc caaggacacc 840 ctgatgatca gccggacccc cgaagtgacc tgcgtggtgg tggacgtgtc ccacgaggac 900 cctgaagtga agttcaattg gtacgtggac ggcgtggaag tgcacaacgc caagaccaag 960 cccagagagg aacagtacaa cagcacctac cgggtggtgt ccgtgctgac cgtgctgcac 1020 caggactggc tgaacggcaa agagtacaag tgcaaggtgt ccaacaaggc cctgcctgcc 1080 cccatcgaga aaaccatcag caaggccaag ggccagcccc gcgagcccca ggtgtacaca 1140 ctgcccccca gccgggacga gctgaccgag aaccaggtgt ccctgacctg cctggtgaaa 1200 ggcttctacc ccagcgatat cgccgtggaa tgggagagca acggccagcc cgagaacaac 1260 tacaagacca ccccccctgt gctggacagc gacggctcat tcttcctgta cagctggctg 1320 accgtggaca agagccggtg gcagcagggc aacgtgttca gctgcagcgt gatgcacgag 1380 gccctgcaca accactacac ccagaagtcc ctgagcctga gccccggcaa gtag 1434 <210> 30 <211> 477 <212> PRT <213> Artificial Sequence <220> <223> IFNAR2 - Linker - FC EW <400> 30 Ala Thr Met Trp Trp Arg Leu Trp Trp Leu Leu Leu Leu Leu Leu Leu 1 5 10 15 Leu Trp Pro Met Val Trp Ala Ile Ser Tyr Asp Ser Pro Asp Tyr Thr 20 25 30 Asp Glu Ser Cys Thr Phe Lys Ile Ser Leu Arg Asn Phe Arg Ser Ile 35 40 45 Leu Ser Trp Glu Leu Lys Asn His Ser Ile Val Pro Thr His Tyr Thr 50 55 60 Leu Leu Tyr Thr Ile Met Ser Lys Pro Glu Asp Leu Lys Val Val Lys 65 70 75 80 Asn Cys Ala Asn Thr Thr Arg Ser Phe Cys Asp Leu Thr Asp Glu Trp 85 90 95 Arg Ser Thr His Glu Ala Tyr Val Thr Val Leu Glu Gly Phe Ser Gly 100 105 110 Asn Thr Thr Leu Phe Ser Cys Ser His Asn Phe Trp Leu Ala Ile Asp 115 120 125 Met Ser Phe Glu Pro Pro Glu Phe Glu Ile Val Gly Phe Thr Asn His 130 135 140 Ile Asn Val Met Val Lys Phe Pro Ser Ile Val Glu Glu Glu Leu Gln 145 150 155 160 Phe Asp Leu Ser Leu Val Ile Glu Glu Gln Ser Glu Gly Ile Val Lys 165 170 175 Lys His Lys Pro Glu Ile Lys Gly Asn Met Ser Gly Asn Phe Thr Tyr 180 185 190 Ile Ile Asp Lys Leu Ile Pro Asn Thr Asn Tyr Cys Val Ser Val Tyr 195 200 205 Leu Glu His Ser Asp Glu Gln Ala Val Ile Lys Ser Pro Leu Lys Cys 210 215 220 Thr Leu Leu Pro Pro Gly Gln Glu Ser Glu Ser Ala Glu Ser Ala Lys 225 230 235 240 Ile Glu Gly Arg Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys 245 250 255 Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu 260 265 270 Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu 275 280 285 Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys 290 295 300 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 305 310 315 320 Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu 325 330 335 苏氨酸 缬氨酸 亮氨酸 组氨酸 谷氨酰胺 天冬氨酸 色氨酸 亮氨酸 天冬酰胺 甘氨酸 赖氨酸 谷氨酸 酪氨酸 赖氨酸 半胱氨酸 赖氨酸 340 345 350 缬氨酸 丝氨酸 天冬酰胺 赖氨酸 丙氨酸 亮氨酸 脯氨酸 丙氨酸 脯氨酸 异亮氨酸 谷氨酸 赖氨酸 苏氨酸 异亮氨酸 丝氨酸 赖氨酸 355 360 365 丙氨酸 赖氨酸 甘氨酸 谷氨酰胺 脯氨酸 精氨酸 谷氨酸 脯氨酸 谷氨酰胺 缬氨酸 酪氨酸 苏氨酸 亮氨酸 脯氨酸 脯氨酸 丝氨酸 370 375 380 精氨酸 天冬氨酸 谷氨酸 亮氨酸 苏氨酸 谷氨酸 天冬酰胺 谷氨酰胺 缬氨酸 丝氨酸 亮氨酸 苏氨酸 半胱氨酸 亮氨酸 缬氨酸 赖氨酸 385 390 395 400 甘氨酸 苯丙氨酸 酪氨酸 脯氨酸 丝氨酸 天冬氨酸 异亮氨酸 丙氨酸 缬氨酸 谷氨酸 色氨酸 谷氨酸 丝氨酸 天冬酰胺 甘氨酸 谷氨酰胺 405 410 415 脯氨酸 谷氨酸 天冬酰胺 天冬酰胺 酪氨酸 赖氨酸 苏氨酸 苏氨酸 脯氨酸 脯氨酸 缬氨酸 亮氨酸 天冬氨酸 丝氨酸 天冬氨酸 甘氨酸 420 425 430 丝氨酸 苯丙氨酸 苯丙氨酸 亮氨酸 酪氨酸 丝氨酸 色氨酸 亮氨酸 苏氨酸 缬氨酸 天冬氨酸 赖氨酸 丝氨酸 精氨酸 色氨酸 谷氨酰胺 435 440 445 谷氨酰胺 甘氨酸 天冬酰胺 缬氨酸 苯丙氨酸 丝氨酸 半胱氨酸 丝氨酸 缬氨酸 甲硫氨酸 组氨酸 谷氨酸 丙氨酸 亮氨酸 组氨酸 天冬酰胺 450 455 460 组氨酸 酪氨酸 苏氨酸 谷氨酰胺 赖氨酸 丝氨酸 亮氨酸 丝氨酸 亮氨酸 丝氨酸 脯氨酸 甘氨酸 赖氨酸 465 470 475 <210> 31 <211> 1434 <212> DNA <213> Artificial Sequence <220> <223> IFNAR2 - Linker - FC RVT <400> 31 gccaccatgt ggtggcgtct gtggtggctg ctgcttctgc tgttgctgct gtggcctatg 60 gtctgggcca tctcctacga cagccctgac tacaccgacg agtcctgcac cttcaagatc 120 tccctgcgga acttccggtc catcctgtcc tgggagctga agaaccactc catcgtgccc 180 acacactaca ccctgctgta caccatcatg tccaagcctg aggacctgaa ggtggtcaag 240 aactgcgcca acaccaccag atctttctgc gacctgaccg atgagtggcg gtctacccac 300 gaggcttacg tgacagtgct ggaaggcttc tccggcaata ccacactgtt cagctgctcc 360 cacaacttct ggctggccat cgacatgtcc ttcgagcctc cagagttcga gatcgtgggc 420 ttcaccaacc acatcaacgt gatggtcaag ttccccagca tcgtggaaga ggaactccag 480 ttcgacctga gcctggtcat cgaggaacag tccgagggca tcgtgaagaa gcacaagccc 540 gagatcaagg gcaacatgtc cggcaacttc acctacatca tcgacaagct gatccccaac 600 accaactact gcgtgtccgt gtacctggaa cactccgatg agcaggccgt gatcaagtcc 660 cctctgaagt gtaccctgct gcctcctggc caagagtctg agtctgccga gtctgctaaa 720 atcgagggcc ggatggaccc caagagctgc gacaagaccc acacctgtcc cccctgccct 780 gcccctgaac tgctgggcgg acccagcgtg ttcctgttcc ccccaaagcc caaggacacc 840 ctgatgatca gccggacccc cgaagtgacc tgcgtggtgg tggacgtgtc ccacgaggac 900 960 cccagagagg aacagacaa cagcacctac cgggtggtgt ccgtgctgac cgtgctgcac 1020 caggactggc tgaacggcaa agagtacaag tgcaaggtgt ccaacaaggc cctgcctgcc 1080 1140 ctgcccccca gccgggacga gctgaccaag aaccaggtgt ccctgacctg cctggtgaaa 1200 ggcttctacc ccagcgatat cgccgtggaa tgggagagca acggccagcc cgagaacac 1260 tacaagacca ccccccctgt gctggtcagc gacggctcat tcaccctgta cagcaagctg 1320 accgtggaca agagccggtg gcagcagggc aacgtgttca gctgcagcgt gatgcacgag 1380 gccctgcaca accactacac ccagaagtcc ctgagcctga gccccggcaa gtag 1434 <210> 32 <211> 477 <212> PRT <213> artificial sequence <220> <223> IFNAR2-connector-FC RVT <400> 32 Ala Thr Met Trp Trp Arg Leu Trp Trp Leu Leu Leu Leu Leu Leu Leu 1 5 10 15 Leu Trp Pro Met Val Trp Ala Ile Ser Tyr Asp Ser Pro Asp Tyr Thr 20 25 30 Asp Glu Ser Cys Thr Phe Lys Ile Ser Leu Arg Asn Phe Arg Ser Ile 35 40 45 Leu Ser Trp Glu Leu Lys Asn His Ser Ile Val Pro Thr His Tyr Thr 50 55 60 Leu Leu Tyr Thr Ile Met Ser Lys Pro Glu Asp Leu Lys Val Val Lys 65 70 75 80 Asn Cys Ala Asn Thr Thr Arg Ser Phe Cys Asp Leu Thr Asp Glu Trp 85 90 95 Arg Ser Thr His Glu Ala Tyr Val Thr Val Leu Glu Gly Phe Ser Gly 100 105 110 Asn Thr Thr Leu Phe Ser Cys Ser His Asn Phe Trp Leu Ala Ile Asp 115 120 125 Met Ser Phe Glu Pro Pro Glu Phe Glu Ile Val Gly Phe Thr Asn His 130 135 140 Ile Asn Val Met Val Lys Phe Pro Ser Ile Val Glu Glu Glu Leu Gln 145 150 155 160 Phe Asp Leu Ser Leu Val Ile Glu Glu Gln Ser Glu Gly Ile Val Lys 165 170 175 Lys His Lys Pro Glu Ile Lys Gly Asn Met Ser Gly Asn Phe Thr Tyr 180 185 190 Ile Ile Asp Lys Leu Ile Pro Asn Thr Asn Tyr Cys Val Ser Val Tyr 195 200 205 Leu Glu His Ser Asp Glu Gln Ala Val Ile Lys Ser Pro Leu Lys Cys 210 215 220 Thr Leu Leu Pro Pro Gly Gln Glu Ser Glu Ser Ala Glu Ser Ala Lys 225 230 235 240 Ile Glu Gly Arg Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys 245 250 255 Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu 260 265 270 Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu 275 280 285 Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys 290 295 300 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 305 310 315 320 Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu 325 330 335 Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 340 345 350 Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 355 360 365 Ala Lys Gly Gln Pro Arg Glu Pro Arg Val Tyr Thr Leu Pro Pro Ser 370 375 380 Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 385 390 395 400 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 405 410 415 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Val Ser Asp Gly 420 425 430 Ser Phe Thr Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln 435 440 445 Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 450 455 460 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 465 470 475 <210> 33 <211> 45 <212> DNA <213> Artificial sequence <220> <223> Connector_4GS*3 <400> 33 ggcggcggag gatctggcgg aggtggaagc ggaggcggtg gatct 45 <210> 34 <211> 15 <212> PRT <213> Artificial sequence <220> <223> Connector_4GS*3 <400> 34 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 1 5 10 15 <210> 35 <211> 810 <212> DNA <213> Artificial sequence <220> <223> Connector_4GS*3-FC WT <400> 35 gccaccatgt ggtggcgtct gtggtggctg ctgcttctgc tgttgctgct gtggcctatg 60 gtctgggccg gcggcggagg atctggcgga ggtggaagcg gaggcggtgg atctcccaag 120 agctgcgaca agacccacac ctgtcccccc tgccctgccc ctgaactgct gggcggaccc 180 agcgtgttcc tgttcccccc aaagcccaag gacaccctga tgatcagccg gacccccgaa 240 gtgacctgcg tggtggtgga cgtgtcccac gaggaccctg aagtgaagtt caattggtac 300 gtggacggcg tggaagtgca caacgccaag accaagccca gagaggaaca gtacaacagc 360 acctaccggg tggtgtccgt gctgaccgtg ctgcaccagg actggctgaa cggcaaagag 420 tacaagtgca aggtgtccaa caaggccctg cctgccccca tcgagaaaac catcagcaag 480 gccaagggcc agccccgcga gccccaggtg tacacactgc cccccagccg ggacgagctg 540 accaagaacc aggtgtccct gacctgcctg gtgaaaggct tctaccccag cgatatcgcc 600 gtggaatggg agagcaacgg ccagcccgag aacaactaca agaccacccc ccctgtgctg 660 gacagcgacg gctcattctt cctgtacagc aagctgaccg tggacaagag ccggtggcag 720 cagggcaacg tgttcagctg cagcgtgatg cacgaggccc tgcacaacca ctacacccag 780 aagtccctga gcctgagccc cggcaagtga 810 <210> 36 <211> 269 <212> PRT <213> Artificial Sequence <220> <223> Linker_4GS*3-FC WT <400> 36 Ala Thr Met Trp Trp Arg Leu Trp Trp Leu Leu Leu Leu Leu Leu Leu 1 5 10 15 Leu Trp Pro Met Val Trp Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly 20 25 30 Ser Gly Gly Gly Gly Ser Pro Lys Ser Cys Asp Lys Thr His Thr Cys 35 40 45 Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu 50 55 60 Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu 65 70 75 80 Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys 85 90 95 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 100 105 110 Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu 115 120 125 Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 130 135 140 Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 145 150 155 160 Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser 165 170 175 Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 180 185 190 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 195 200 205 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly 210 215 220 Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln 225 230 235 240 Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 245 250 255 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 260 265 <210> 37 <211> 810 <212> DNA <213> Artificial Sequence <220> <223> Linker_4GS*3-FC EW <400> 37 gccaccatgt ggtggcgtct gtggtggctg ctgcttctgc tgttgctgct gtggcctatg 60 gtctgggccg gcggcggagg atctggcgga ggtggaagcg gaggcggtgg atctcccaag 120 agctgcgaca agacccacac ctgtcccccc tgccctgccc ctgaactgct gggcggaccc 180 agcgtgttcc tgttcccccc aaagcccaag gacaccctga tgatcagccg gacccccgaa 240 gtgacctgcg tggtggtgga cgtgtcccac gaggaccctg aagtgaagtt caattggtac 300 gtggacggcg tggaagtgca caacgccaag accaagccca gagaggaaca gtacaacagc 360 acctaccggg tggtgtccgt gctgaccgtg ctgcaccagg actggctgaa cggcaaagag 420 tacaagtgca aggtgtccaa caaggccctg cctgccccca tcgagaaaac catcagcaag 480 gccaagggcc agccccgcga gccccaggtg tacacactgc cccccagccg ggacgagctg 540 accgagaacc aggtgtccct gacctgcctg gtgaaaggct tctaccccag cgatatcgcc 600 gtggaatggg agagcaacgg ccagcccgag aacaactaca agaccacccc ccctgtgctg 660 gacagcgacg gctcattctt cctgtacagc tggctgaccg tggacaagag ccggtggcag 720 cagggcaacg tgttcagctg cagcgtgatg cacgaggccc tgcacaacca ctacacccag 780 aagtccctga gcctgagccc cggcaagtag 810 <210> 38 <211> 269 <212> PRT <213> Artificial Sequence <220> <223> Linker_4GS*3-FC EW <400> 38 Ala Thr Met Trp Trp Arg Leu Trp Trp Leu Leu Leu Leu Leu Leu Leu 1 5 10 15 Leu Trp Pro Met Val Trp Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly 20 25 30 Ser Gly Gly Gly Gly Ser Pro Lys Ser Cys Asp Lys Thr His Thr Cys 35 40 45 Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu 50 55 60 Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu 65 70 75 80 Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys 85 90 95 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 100 105 110 Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu 115 120 125 Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 130 135 140 Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 145 150 155 160 Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser 165 170 175 Arg Asp Glu Leu Thr Glu Asn Gln Val Ser Leu Thr Cys Leu Val Lys 180 185 190 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 195 200 205 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly 210 215 220 Ser Phe Phe Leu Tyr Ser Trp Leu Thr Val Asp Lys Ser Arg Trp Gln 225 230 235 240 Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 245 250 255 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 260 265 <210> 39 <211> 810 <212> DNA <213> Artificial sequence <220> <223> Linker_4GS*3-FC RVT <400> 39 gccaccatgt ggtggcgtct gtggtggctg ctgcttctgc tgttgctgct gtggcctatg 60 gtctgggccg gcggcggagg atctggcgga ggtggaagcg gaggcggtgg atctcccaag 120 agctgcgaca agacccacac ctgtcccccc tgccctgccc ctgaactgct gggcggaccc 180 agcgtgttcc tgttcccccc aaagcccaag gacaccctga tgatcagccg gacccccgaa 240 gtgacctgcg tggtggtgga cgtgtcccac gaggaccctg aagtgaagtt caattggtac 300 gtggacggcg tggaagtgca caacgccaag accaagccca gagaggaaca gtacaacagc 360 acctaccggg tggtgtccgt gctgaccgtg ctgcaccagg actggctgaa cggcaaagag 420 tacaagtgca aggtgtccaa caaggccctg cctgccccca tcgagaaaac catcagcaag 480 gccaagggcc agccccgcga gccccgggtg tacacactgc cccccagccg ggacgagctg 540 accaagaacc aggtgtccct gacctgcctg gtgaaaggct tctaccccag cgatatcgcc 600 gtggaatggg agagcaacgg ccagcccgag aacaactaca agaccacccc ccctgtgctg 660 gtcagcgacg gctcattcac cctgtacagc aagctgaccg tggacaagag ccggtggcag 720 cagggcaacg tgttcagctg cagcgtgatg cacgaggccc tgcacaacca ctacacccag 780 aagtccctga gcctgagccc cggcaagtag 810 <210> 40 <211> 269 <212> PRT <213> Artificial Sequence <220> <223> Linker_4GS*3-FC RVT <400> 40 Ala Thr Met Trp Trp Arg Leu Trp Trp Leu Leu Leu Leu Leu Leu Leu 1 5 10 15 Leu Trp Pro Met Val Trp Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly 20 25 30 Ser Gly Gly Gly Gly Ser Pro Lys Ser Cys Asp Lys Thr His Thr Cys 35 40 45 Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu 50 55 60 Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu 65 70 75 80 Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys 85 90 95 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 100 105 110 Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu 115 120 125 Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 130 135 140 Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 145 150 155 160 Ala Lys Gly Gln Pro Arg Glu Pro Arg Val Tyr Thr Leu Pro Pro Ser 165 170 175 Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 180 185 190 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 195 200 205 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Val Ser Asp Gly 210 215 220 Ser Phe Thr Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln 225 230 235 240 Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 245 250 255 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 260 265 <210> 41 <211> 2037 <212> DNA <213> The snowstorm <220> <223> IFNAR1-Type_4GS*3-FC WT <400> 41 gccaccatgt ggtggcgtct gtggtggctg ctgcttctgc tgttgctgct gtggcctatg gtctgggcca agaacctgaa gtcccctcag aaagtggaag tggacatcat cgacgacaac ttcatcctgc ggtggaacag atccgacgag tccgtgggca acgtgacctt cagcttcgac 180 240. ccggcatgga cactggatc aagctgtccg gctgccaga catcacctct accaagtgca acttctccag cctgaagctg aacgtgtacg aggaatcaa gctgcggatc cgggccgaga aagagaacac cagcagttgg tacgaggtgg acagcttcac ccctttcaga aaggcccaga tcggccctcc tgaagtgcac ctggaagccg aggataaggc catcgtgatc 420 cacatcagcc ccggcaccaa ggactctgtg atgtgggctc tcgacggcct gtccttcacc 480 tacagcctgg tcatctggaa gaactcctcc ggcgtggaag agaagaatcga gaacatctac 540 tcccggcaca agatctacaa gctgagcccc gagacaacct actgcctgaa agtgaaggcc 600 gctctgctga cctcctggaa gatcggcgtg tactctcccg tgcactgcat caagaccacc 660 gtggaaaacg agctgcctcc tccagagaac atcgaggtgt ccgtgcagaa ccagaactac 720 gtgctgaagt gggactacac ctacgccaac atgacctttc aggtgcagtg gctgcacgct 780 tttctgaagc ggaaccctgg caaccacctg tacaagtgga agcagattcc cgactgcgag 840 aacgtgaaaa ccacacagtg cgtgttccct cagaacgtgt tccagaaggg catctacctg 900 ctgagagtgc aggcctccga cggcaacaac accagctttt ggagcgaaga gatcaagttc 960 gataccgaga tccaggcctt cctgctgcct ccagtgttca acatcagatc cctgtccgac 1020 tccttccaca tctacatcgg cgctcctaag cagtccggca acacccctgt gatccaggac 1080 taccctctga tctacgagat catcttctgg gagaacacct ccaacgccga gcggaagatc 1140 atcgagaaga aaaccgacgt gaccgtgcct aacctgaagc ctctgaccgt gtactgcgtg 1200 aaggctaggg cccacaccat ggacgaaaag ctgaacaagt cctccgtgtt ctccgacgcc 1260 gtgtgcgaaa agaccaagcc aggcaacacc tctaaaggcg gcggaggatc tggcggaggt 1320 ggaagcggag gcggtggatc tcccaagagc tgcgacaaga cccacacctg tcccccctgc 1380 cctgcccctg aactgctggg cggacccagc gtgttcctgt tccccccaaa gcccaaggac 1440 accctgatga tcagccggac ccccgaagtg acctgcgtgg tggtggacgt gtcccacgag 1500 gaccctgaag tgaagttcaa ttggtacgtg gacggcgtgg aagtgcacaa cgccaagacc 1560 aagcccagag aggaacagta caacagcacc taccgggtgg tgtccgtgct gaccgtgctg 1620 caccaggact ggctgaacgg caaagagtac aagtgcaagg tgtccaacaa ggccctgcct 1680 gcccccatcg agaaaaccat cagcaaggcc aagggccagc cccgcgagcc ccaggtgtac 1740 acactgcccc ccagccggga cgagctgacc aagaaccagg tgtccctgac ctgcctggtg 1800 aaaggcttct accccagcga tatcgccgtg gaatgggaga gcaacggcca gcccgagaac 1860 aactacaaga ccaccccccc tgtgctggac agcgacggct cattcttcct gtacagcaag 1920 ctgaccgtgg acaagagccg gtggcagcag ggcaacgtgt tcagctgcag cgtgatgcac 1980 gaggccctgc acaaccacta cacccagaag tccctgagcc tgagccccgg caagtga 2037 <210> 42 <211> 678 <212> PRT <213> Artificial Sequence <220> <223> IFNAR1-Linker_4GS*3-FC WT <400> 42 Ala Thr Met Trp Trp Arg Leu Trp Trp Leu Leu Leu Leu Leu Leu Leu 1 5 10 15 Leu Trp Pro Met Val Trp Ala Lys Asn Leu Lys Ser Pro Gln Lys Val 20 25 30 Glu Val Asp Ile Ile Asp Asp Asn Phe Ile Leu Arg Trp Asn Arg Ser 35 40 45 Asp Glu Ser Val Gly Asn Val Thr Phe Ser Phe Asp Tyr Gln Lys Thr 50 55 60 Gly Met Asp Asn Trp Ile Lys Leu Ser Gly Cys Gln Asn Ile Thr Ser 65 70 75 80 Thr Lys Cys Asn Phe Ser Ser Leu Lys Leu Asn Val Tyr Glu Glu Ile 85 90 95 Lys Leu Arg Ile Arg Ala Glu Lys Glu Asn Thr Ser Ser Trp Tyr Glu 100 105 110 Val Asp Ser Phe Thr Pro Phe Arg Lys Ala Gln Ile Gly Pro Pro Glu 115 120 125 Val His Leu Glu Ala Glu Asp Lys Ala Ile Val Ile His Ile Ser Pro 130 135 140 Gly Thr Lys Asp Ser Val Met Trp Ala Leu Asp Gly Leu Ser Phe Thr 145 150 155 160 Tyr Ser Leu Val Ile Trp Lys Asn Ser Ser Gly Val Glu Glu Arg Ile 165 170 175 Glu Asn Ile Tyr Ser Arg His Lys Ile Tyr Lys Leu Ser Pro Glu Thr 180 185 190 Thr Tyr Cys Leu Lys Val Lys Ala Ala Leu Leu Thr Ser Trp Lys Ile 195 200 205 Gly Val Tyr Ser Pro Val His Cys Ile Lys Thr Thr Val Glu Asn Glu 210 215 220 Leu Pro Pro Pro Glu Asn Ile Glu Val Ser Val Gln Asn Gln Asn Tyr 225 230 235 240 Val Leu Lys Trp Asp Tyr Thr Tyr Ala Asn Met Thr Phe Gln Val Gln 245 250 255 Trp Leu His Ala Phe Leu Lys Arg Asn Pro Gly Asn His Leu Tyr Lys 260 265 270 Trp Lys Gln Ile Pro Asp Cys Glu Asn Val Lys Thr Thr Gln Cys Val 275 280 285 Phe Pro Gln Asn Val Phe Gln Lys Gly Ile Tyr Leu Leu Arg Val Gln 290 295 300 Ala Ser Asp Gly Asn Asn Thr Ser Phe Trp Ser Glu Glu Ile Lys Phe 305 310 315 320 Asp Thr Glu Ile Gln Ala Phe Leu Leu Pro Pro Val Phe Asn Ile Arg 325 330 335 Ser Leu Ser Asp Ser Phe His Ile Tyr Ile Gly Ala Pro Lys Gln Ser 340 345 350 Gly Asn Thr Pro Val Ile Gln Asp Tyr Pro Leu Ile Tyr Glu Ile Ile 355 360 365 Phe Trp Glu Asn Thr Ser Asn Ala Glu Arg Lys Ile Ile Glu Lys Lys 370 375 380 Thr Asp Val Thr Val Pro Asn Leu Lys Pro Leu Thr Val Tyr Cys Val 385 390 395 400 Lys Ala Arg Ala His Thr Met Asp Glu Lys Leu Asn Lys Ser Ser Val 405 410 415 Phe Ser Asp Ala Val Cys Glu Lys Thr Lys Pro Gly Asn Thr Ser Lys 420 425 430 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Pro 435 440 445 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 450 455 460 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 465 470 475 480 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 485 490 495 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 500 505 510 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 515 520 525 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 530 535 540 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 545 550 555 560 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 565 570 575 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 580 585 590 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 595 600 605 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 610 615 620 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 625 630 635 640 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 645 650 655 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 660 665 670 Ser Leu Ser Pro Gly Lys 675 <210> 43 <211> 2037 <212> DNA <213> Artificial sequence <220> <223> IFNAR1-Linker_4GS*3-FC EW <400> 43 gccaccatgt ggtggcgtct gtggtggctg ctgcttctgc tgttgctgct gtggcctatg 60 gtctgggcca agaacctgaa gtcccctcag aaagtggaag tggacatcat cgacgacac 120 ttcatcctgc gtggaacag atccgacgag tccgtgggca acgtgacctt cagcttcgac 180 taccagaaaa ccggcatgga caacggatc aagctgtccg gctgccagaa catcacctct 240 accaagtgca acttctccag cctgaagctg aacgtgtacg aggaatca gctgcggatc 300 cgggccgaga agagacac cagcagttgg tacgaggtgg acagctcac ccctttcaga 360 aaggcccaga tcggccctcc tgaagtgcac ctggaagccg aggataggc catcgtgatc 420 cacatcagcc ccggcaccaa ggactgtg atgtgggctc tcgacggcct gtccttcacc 480 tacagcctgg tcatctggaa gaactcctcc ggcgtggaag agagaatcga gaacactac 540 tcccggcaca agatctacaa gctgagcccc gagacaacct actgcctgaa agtgaaggcc 600 gctctgctga cctcctggaa gatcggcgtg tactctcccg tgcactgcat caaccacc 660 gtggaaaacg agctgcctcc tccagagaac atcgaggtgt ccgtgcagaa ccagaactac 720 gtgctgaagt gggactacac ctacgccaac atgaccttc aggtgcagtg gctgcacgct 780 tttctgaagc ggaaccctgg caaccacctg tacaagtgga agcagattcc cgactgcgag 840 aacgtgaaaa ccacacagtg cgtgttccct cagaacgtgt tccagaaggg catctacctg 900 ctgagagtgc aggcctccga cggcaacaac accagctttt ggagcgaaga gatcaagttc 960 gataccgaga tccaggcctt cctgctgcct ccagtgttca acatcagatc cctgtccgac 1020 tccttccaca tctacatcgg cgctcctaag cagtccggca acacccctgt gatccaggac 1080 taccctctga tctacgagat catcttctgg gagaacacct ccaacgccga gcggaagatc 1140 atcgagaaga aaaccgacgt gaccgtgcct aacctgaagc ctctgaccgt gtactgcgtg 1200 aaggctaggg cccacaccat ggacgaaaag ctgaacaagt cctccgtgtt ctccgacgcc 1260 gtgtgcgaaa agaccaagcc aggcaacacc tctaaaggcg gcggaggatc tggcggaggt 1320 ggaagcggag gcggtggatc tcccaagagc tgcgacaaga cccacacctg tcccccctgc 1380 cctgcccctg aactgctggg cggacccagc gtgttcctgt tccccccaaa gcccaaggac 1440 accctgatga tcagccggac ccccgaagtg acctgcgtgg tggtggacgt gtcccacgag 1500 gaccctgaag tgaagttcaa ttggtacgtg gacggcgtgg aagtgcacaa cgccaagacc 1560 aagcccagag aggaacagta caacagcacc taccgggtgg tgtccgtgct gaccgtgctg 1620 caccaggact ggctgaacgg caaagagtac aagtgcaagg tgtccaacaa ggccctgcct 1680 gcccccatcg agaaaaccat cagcaaggcc aagggccagc cccgcgagcc ccaggtgtac 1740 acactgcccc ccagccggga cgagctgacc gagaaccagg tgtccctgac ctgcctggtg 1800 aaaggcttct accccagcga tatcgccgtg gaatgggaga gcaacggcca gcccgagaac 1860 aactacaaga ccaccccccc tgtgctggac agcgacggct cattcttcct gtacagctgg 1920 ctgaccgtgg acaagagccg gtggcagcag ggcaacgtgt tcagctgcag cgtgatgcac 1980 gaggccctgc acaaccacta cacccagaag tccctgagcc tgagccccgg caagtag 2037 <210> 44 <211> 678 <212> PRT <213> Artificial Sequence <220> <223> IFNAR1-Linker_4GS*3-FC EW <400> 44 Ala Thr Met Trp Trp Arg Leu Trp Trp Leu Leu Leu Leu Leu Leu Leu 1 5 10 15 Leu Trp Pro Met Val Trp Ala Lys Asn Leu Lys Ser Pro Gln Lys Val 20 25 30 Glu Val Asp Ile Ile Asp Asp Asn Phe Ile Leu Arg Trp Asn Arg Ser 35 40 45 Asp Glu Ser Val Gly Asn Val Thr Phe Ser Phe Asp Tyr Gln Lys Thr 50 55 60 Gly Met Asp Asn Trp Ile Lys Leu Ser Gly Cys Gln Asn Ile Thr Ser 65 70 75 80 Thr Lys Cys Asn Phe Ser Ser Leu Lys Leu Asn Val Tyr Glu Glu Ile 85 90 95 Lys Leu Arg Ile Arg Ala Glu Lys Glu Asn Thr Ser Ser Trp Tyr Glu 100 105 110 Val Asp Ser Phe Thr Pro Phe Arg Lys Ala Gln Ile Gly Pro Pro Glu 115 120 125 Val His Leu Glu Ala Glu Asp Lys Ala Ile Val Ile His Ile Ser Pro 130 135 140 Gly Thr Lys Asp Ser Val Met Trp Ala Leu Asp Gly Leu Ser Phe Thr 145 150 155 160 Tyr Ser Leu Val Ile Trp Lys Asn Ser Ser Gly Val Glu Glu Arg Ile 165 170 175 Glu Asn Ile Tyr Ser Arg His Lys Ile Tyr Lys Leu Ser Pro Glu Thr 180 185 190 Thr Tyr Cys Leu Lys Val Lys Ala Ala Leu Leu Thr Ser Trp Lys Ile 195 200 205 Gly Val Tyr Ser Pro Val His Cys Ile Lys Thr Thr Val Glu Asn Glu 210 215 220 Leu Pro Pro Pro Glu Asn Ile Glu Val Ser Val Gln Asn Gln Asn Tyr 225 230 235 240 Val Leu Lys Trp Asp Tyr Thr Tyr Ala Asn Met Thr Phe Gln Val Gln 245 250 255 Trp Leu His Ala Phe Leu Lys Arg Asn Pro Gly Asn His Leu Tyr Lys 260 265 270 Trp Lys Gln Ile Pro Asp Cys Glu Asn Val Lys Thr Thr Gln Cys Val 275 280 285 Phe Pro Gln Asn Val Phe Gln Lys Gly Ile Tyr Leu Leu Arg Val Gln 290 295 300 Ala Ser Asp Gly Asn Asn Thr Ser Phe Trp Ser Glu Glu Ile Lys Phe 305 310 315 320 Asp Thr Glu Ile Gln Ala Phe Leu Leu Pro Pro Val Phe Asn Ile Arg 325 330 335 Ser Leu Ser Asp Ser Phe His Ile Tyr Ile Gly Ala Pro Lys Gln Ser 340 345 350 Gly Asn Thr Pro Val Ile Gln Asp Tyr Pro Leu Ile Tyr Glu Ile Ile 355 360 365 Phe Trp Glu Asn Thr Ser Asn Ala Glu Arg Lys Ile Ile Glu Lys Lys 370 375 380 Thr Asp Val Thr Val Pro Asn Leu Lys Pro Leu Thr Val Tyr Cys Val 385 390 395 400 Lys Ala Arg Ala His Thr Met Asp Glu Lys Leu Asn Lys Ser Ser Val 405 410 415 Phe Ser Asp Ala Val Cys Glu Lys Thr Lys Pro Gly Asn Thr Ser Lys 420 425 430 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Pro 435 440 445 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 450 455 460 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 465 470 475 480 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 485 490 495 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 500 505 510 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 515 520 525 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 530 535 540 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 545 550 555 560 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 565 570 575 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Glu Asn 580 585 590 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 595 600 605 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 610 615 620 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Trp 625 630 635 640 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 645 650 655 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 660 665 670 Ser Leu Ser Pro Gly Lys 675 <210> 45 <211> 2037 <212> DNA <213> Artificial sequence <220> <223> IFNAR1 - linker_4GS*3 - FC RVT <400> 45 gccaccatgt ggtggcgtct gtggtggctg ctgcttctgc tgttgctgct gtggcctatg 60 gtctgggcca agaacctgaa gtcccctcag aaagtggaag tggacatcat cgacgacaac 120 ttcatcctgc ggtggaacag atccgacgag tccgtgggca acgtgacctt cagcttcgac 180 taccagaaaa ccggcatgga caactggatc aagctgtccg gctgccagaa catcacctct 240 accaagtgca acttctccag cctgaagctg aacgtgtacg aggaaatcaa gctgcggatc 300 cgggccgaga aagagaacac cagcagttgg tacgaggtgg acagcttcac ccctttcaga  360 aaggcccaga tcggccctcc tgaagtgcac ctggaagccg aggataaggc catcgtgatc 420 cacatcagcc ccggcaccaa ggactctgtg atgtgggctc tcgacggcct gtccttcacc 480 tacagcctgg tcatctggaa gaactcctcc ggcgtggaag agaagaatcga gaacatctac 540 tcccggcaca agatctacaa gctgagcccc gagacaacct actgcctgaa agtgaaggcc 600 gctctgctga cctcctggaa gatcggcgtg tactctcccg tgcactgcat caagaccacc 660 gtggaaaacg agctgcctcc tccagagaac atcgaggtgt ccgtgcagaa ccagaactac 720 gtgctgaagt gggactacac ctacgccaac atgacctttc aggtgcagtg gctgcacgct 780 tttctgaagc ggaaccctgg caaccacctg tacaagtgga agcagattcc cgactgcgag 840 aacgtgaaaa ccacacagtg cgtgttccct cagaacgtgt tccagaaggg catctacctg 900 ctgagagtgc aggcctccga cggcaacaac accagctttt ggagcgaaga gatcaagttc 960 gataccgaga tccaggcctt cctgctgcct ccagtgttca acatcagatc cctgtccgac 1020 tccttccaca tctacatcgg cgctcctaag cagtccggca acacccctgt gatccaggac 1080 taccctctga tctacgagat catcttctgg gagaacacct ccaacgccga gcggaagatc 1140 your atcgaaga aaaccgacgt gaccgtgcct aacctgaagc ctctgaccgt gtactgcgtg 1200 aaggctaggg cccacaccat ggacgaaaag ctgaacaagt cctccgtgtt ctccgacgcc 1260 gtgtgcgaaa agaccaagcc aggcaacacc tctaaaggcg gcggaggatc tggcggaggt 1320 ggaagcggag gcggtggatc tcccaagagc tgcgacaaga cccacacctg tcccccctgc 1380 cctgcccctg aactgctggg cggacccagc gtgttcctgt tccccccaaa gcccaaggac 1440 accctgatga tcagccggac ccccgaagtg acctgcgtgg tggtggacgt gtcccacgag 1500 gaccctgaag tgaagttcaa ttggtacgtg gacggcgtgg aagtgcacaa cgccaagacc 1560 aagcccagag aggaacagta caacagcacc taccgggtgg tgtccgtgct gaccgtgctg 1620 caccaggact ggctgaacgg caaagagtac aagtgcaagg tgtccaacaa ggccctgcct 1680 gcccccatcg agaaaaccat cagcaaggcc aagggccagc cccgcgagcc ccgggtgtac 1740 acactgcccc ccagccggga cgagctgacc aagaaccagg tgtccctgac ctgcctggtg 1800 aaaggcttct accccagcga tatcgccgtg gaatgggaga gcaacggcca gcccgagaac 1860 aactacaaga ccaccccccc tgtgctggtc agcgacggct cattcaccct gtacagcaag 1920 ctgaccgtgg acaagagccg gtggcagcag ggcaacgtgt tcagctgcag cgtgatgcac 1980 gaggccctgc acaaccacta cacccagaag tccctgagcc tgagccccgg caagtag 2037 <210> 46 <211> 678 <212> PRT <213> Artificial Sequence <220> <223> IFNAR1 - Linker_4GS*3 - FC RVT <400> 46 Ala Thr Met Trp Trp Arg Leu Trp Trp Leu Leu Leu Leu Leu Leu Leu 1 5 10 15 Leu Trp Pro Met Val Trp Ala Lys Asn Leu Lys Ser Pro Gln Lys Val 20 25 30 Glu Val Asp Ile Ile Asp Asp Asn Phe Ile Leu Arg Trp Asn Arg Ser 35 40 45 Asp Glu Ser Val Gly Asn Val Thr Phe Ser Phe Asp Tyr Gln Lys Thr 50 55 60 Gly Met Asp Asn Trp Ile Lys Leu Ser Gly Cys Gln Asn Ile Thr Ser 65 70 75 80 Thr Lys Cys Asn Phe Ser Ser Leu Lys Leu Asn Val Tyr Glu Glu Ile 85 90 95 Lys Leu Arg Ile Arg Ala Glu Lys Glu Asn Thr Ser Ser Trp Tyr Glu 100 105 110 Val Asp Ser Phe Thr Pro Phe Arg Lys Ala Gln Ile Gly Pro Pro Glu 115 120 125 Val His Leu Glu Ala Glu Asp Lys Ala Ile Val Ile His Ile Ser Pro 130 135 140 Gly Thr Lys Asp Ser Val Met Trp Ala Leu Asp Gly Leu Ser Phe Thr 145 150 155 160 Tyr Ser Leu Val Ile Trp Lys Asn Ser Ser Gly Val Glu Glu Arg Ile 165 170 175 Glu Asn Ile Tyr Ser Arg His Lys Ile Tyr Lys Leu Ser Pro Glu Thr 180 185 190 Thr Tyr Cys Leu Lys Val Lys Ala Ala Leu Leu Thr Ser Trp Lys Ile 195 200 205 Gly Val Tyr Ser Pro Val His Cys Ile Lys Thr Thr Val Glu Asn Glu 210 215 220 Leu Pro Pro Pro Glu Asn Ile Glu Val Ser Val Gln Asn Gln Asn Tyr 225 230 235 240 Val Leu Lys Trp Asp Tyr Thr Tyr Ala Asn Met Thr Phe Gln Val Gln 245 250 255 Trp Leu His Ala Phe Leu Lys Arg Asn Pro Gly Asn His Leu Tyr Lys 260 265 270 Trp Lys Gln Ile Pro Asp Cys Glu Asn Val Lys Thr Thr Gln Cys Val 275 280 285 Phe Pro Gln Asn Val Phe Gln Lys Gly Ile Tyr Leu Leu Arg Val Gln 290 295 300 Ala Ser Asp Gly Asn Asn Thr Ser Phe Trp Ser Glu Glu Ile Lys Phe 305 310 315 320 Asp Thr Glu Ile Gln Ala Phe Leu Leu Pro Pro Val Phe Asn Ile Arg 325 330 335 Ser Leu Ser Asp Ser Phe His Ile Tyr Ile Gly Ala Pro Lys Gln Ser 340 345 350 Gly Asn Thr Pro Val Ile Gln Asp Tyr Pro Leu Ile Tyr Glu Ile Ile 355 360 365 Phe Trp Glu Asn Thr Ser Asn Ala Glu Arg Lys Ile Ile Glu Lys Lys 370 375 380 Thr Asp Val Thr Val Pro Asn Leu Lys Pro Leu Thr Val Tyr Cys Val 385 390 395 400 Lys Ala Arg Ala His Thr Met Asp Glu Lys Leu Asn Lys Ser Ser Val 405 410 415 Phe Ser Asp Ala Val Cys Glu Lys Thr Lys Pro Gly Asn Thr Ser Lys 420 425 430 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Pro 435 440 445 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 450 455 460 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 465 470 475 480 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 485 490 495 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 500 505 510 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 515 520 525 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 530 535 540 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 545 550 555 560 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 565 570 575 Pro Arg Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 580 585 590 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 595 600 605 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 610 615 620 Thr Pro Pro Val Leu Val Ser Asp Gly Ser Phe Thr Leu Tyr Ser Lys 625 630 635 640 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 645 650 655 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 660 665 670 Ser Leu Ser Pro Gly Lys 675 <210> 47 <211> 1461 <212> DNA <213> Artificial Sequence <220> <223> IFNAR2-Linker_4GS*3-FC WT <400> 47 gccaccatgt ggtggcgtct gtggtggctg ctgcttctgc tgttgctgct gtggcctatg 60 gtctgggcca tctcctacga cagccctgac tacaccgacg agtcctgcac cttcaagatc 120 tccctgcgga acttccggtc catcctgtcc tgggagctga agaaccactc catcgtgccc 180 acacactaca ccctgctgta caccatcatg tccaagcctg aggacctgaa ggtggtcaag 240 aactgcgcca acaccaccag atctttctgc gacctgaccg atgagtggcg gtctacccac 300 gaggcttacg tgacagtgct ggaaggcttc tccggcaata ccacactgtt cagctgctcc 360 caaacttct ggctggccat cgacatgtcc ttcgagcctc cagagttcga gatcgtgggc 420 ttcaccaacc acatcaacgt gatggtcaag ttccccagca tcgtggaaga ggaactccag 480 ttcgacctga gcctggtcat cgaggaacag tccgagggca tcgtgaagaa gcacaagccc 540 gagatcaagg gcaacatgtc cggcaacttc acctacatca tcgacaagct gatcccccaac 600 accaactact gcgtgtccgt gtacctggaa cactccgatg agcaggccgt gatcaagtcc 660 cctctgaagt gtaccctgct gcctcctggc caagagtctg agtctgccga gtctgctaaa 720 ggcggcggag gatctggcgg aggtggaagc ggaggcggtg gatctcccaa gagctgcgac 780 aagacccaca cctgtcccccc ctgccctgcc cctgaactgc tgggcggacc cagcgtgttc 840 ctgttccccc caaagcccaa ggacaccctg atgatcagcc ggacccccga agtgacctgc 900 gtggtggtgg acgtgtccca cgaggaccct gaagtgaagt tcaattggta cgtggacggc 960 gtggaagtgc acaacgccaa gaccaagccc agagaggaac agtacaacag cacctaccgg 1020 gtggtgtccg tgctgaccgt gctgcaccag gactggctga acggcaaaga gtacaagtgc 1080 aaggtgtcca acaaggccct gcctgccccc atcgagaaaa ccatcagcaa ggccaagggc 1140 cagccccgcg agccccaggt gtacacactg ccccccagcc gggacgagct gaccaagaac 1200 caggtgtccc tgacctgcct ggtgaaaggc ttctacccca gcgatatcgc cgtggaatgg 1260 gagagcaacg gccagcccga gaacaactac aagaccaccc cccctgtgct ggacagcgac 1320 ggctcattct tcctgtacag caagctgacc gtggacaaga gccggtggca gcagggcaac 1380 gtgttcagct gcagcgtgat gcacgaggcc ctgcacaacc actacaccca gaagtccctg 1440 agcctgagcc ccggcaagtg a 1461 <(210)> 48 <(211)> 486 <(212)> PRT [[ID=2**********]]<(213)> Artificial Sequence <220> <223> IFNAR2- link_4GS*3-FC WT <400> 48 Ala Thr Met Trp Trp Arg Leu Trp Trp Leu Leu Leu Leu Leu Leu Leu 1 5 10 15 Leu Trp Pro Met Val Trp Ala Ile Ser Tyr Asp Ser Pro Asp Tyr Thr 20 25 30 Asp Glu Ser Cys Thr Phe Lys Ile Ser Leu Arg Asn Phe Arg Ser Ile 35 40 45 Leu Ser Trp Glu Leu Lys Asn His Ser Ile Val Pro Thr His Tyr Thr 50 55 60 Leu Leu Tyr Thr Ile Met Ser Lys Pro Glu Asp Leu Lys Val Val Lys 65 70 75 80 Asn Cys Ala Asn Thr Thr Arg Ser Phe Cys Asp Leu Thr Asp Glu Trp 85 90 95 Arg Ser Thr His Glu Ala Tyr Val Thr Val Leu Glu Gly Phe Ser Gly 100 105 110 Asn Thr Thr Leu Phe Ser Cys Ser His Asn Phe Trp Leu Ala Ile Asp 115 120 125 Met Ser Phe Glu Pro Pro Glu Phe Glu Ile Val Gly Phe Thr Asn His 130 135 140 Ile Asn Val Met Val Lys Phe Pro Ser Ile Val Glu Glu Glu Leu Gln 145 150 155 160 Phe Asp Leu Ser Leu Val Ile Glu Glu Gln Ser Glu Gly Ile Val Lys 165 170 175 Lys His Lys Pro Glu Ile Lys Gly Asn Met Ser Gly Asn Phe Thr Tyr 180 185 190 Ile Ile Asp Lys Leu Ile Pro Asn Thr Asn Tyr Cys Val Ser Val Tyr 195 200 205 Leu Glu His Ser Asp Glu Gln Ala Val Ile Lys Ser Pro Leu Lys Cys 210 215 220 Thr Leu Leu Pro Pro Gly Gln Glu Ser Glu Ser Ala Glu Ser Ala Lys 225 230 235 240 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Pro 245 250 255 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 260 265 270 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 275 280 285 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 290 295 300 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 305 310 315 320 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 325 330 335 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 340 345 350 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 355 360 365 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 370 375 380 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 385 390 395 400 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 405 410 415 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 420 425 430 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 435 440 445 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 450 455 460 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 465 470 475 480 Ser Leu Ser Pro Gly Lys 485 <210> 49 <211> 1461 <212> DNA <213> Artificial sequence <220> <223> IFNAR2 - linker_4GS*3 - FC EW <400> 49 gccaccatgt ggtggcgtct gtggtggctg ctgcttctgc tgttgctgct gtggcctatg 60 gtctgggcca tctcctacga cagccctgac tacaccgacg agtcctgcac cttcaagatc 120 tccctgcgga acttccggtc catcctgtcc tgggagctga agaaccactc catcgtgccc 180 acacactaca ccctgctgta caccatcatg tccaagcctg aggacctgaa ggtggtcaag 240 aactgcgcca acaccaccag atctttctgc gacctgaccg atgagtggcg gtctacccac 300 gaggcttacg tgacagtgct ggaaggcttc tccggcaata ccacactgtt cagctgctcc 360 cacaacttct ggctggccat cgacatgtcc ttcgagcctc cagagttcga gatcgtgggc 420 ttcaccaacc acatcaacgt gatggtcaag ttccccagca tcgtggaaga ggaactccag 480 ttcgacctga gcctggtcat cgaggaacag tccgagggca tcgtgaagaa gcacaagccc 540 gagatcaagg gcaacatgtc cggcaacttc acctacatca tcgacaagct gatcccccaac 600 accaactact gcgtgtccgt gtacctggaa cactccgatg agcaggccgt gatcaagtcc 660 cctctgaagt gtaccctgct gcctcctggc caagagtctg agtctgccga gtctgctaaa 720 ggcggcggag gatctggcgg aggtggaagc ggaggcggtg gatctcccaa gagctgcgac 780 aagacccaca cctgtcccccc ctgccctgcc cctgaactgc tgggcggacc cagcgtgttc 840 ctgttcccc caaagcccaa ggacaccctg atgatcagcc ggacccccga agtgacctgc 900 gtggtggtgg acgtgtccca cgaggaccct gaagtgaagt tcaattggta cgtggacggc 960 gtggaagtgc acaacgccaa gaccaagccc agaggaac agtacaacag cacctaccgg 1020 gtggtgtccg tgctgaccgt gctgcaccag gactggctga acggcaaaga gtacaagtgc 1080 aaggtgtcca acaaggccct gcctgccccc atcgagaaaa ccatcagcaa ggccaagggc 1140 cagccccgcg agccccaggt gtacacactg ccccccagcc gggacgagct gaccgagaac 1200 caggtgtccc tgacctgcct ggtgaaaggc ttctacccca gcgatatcgc cgtggaatgg 1260 gagagcaacg gccagcccga gaacaactac aagaccaccc cccctgtgct ggacagcgac 1320 ggctcattct tcctgtacag ctggctgacc gtggacaaga gccggtggca gcagggcaac 1380 gtgttcagct gcagcgtgat gcacgaggcc ctgcacaacc actacaccca gaagtccctg 1440 agcctgagcc ccggcaagta g 1461 <210> 50 <211> 486 <212> PRT <213> Artificial Sequence <220> <223> IFNAR2-Linker_4GS*3-FC EW <400> 50 Ala Thr Met Trp Trp Arg Leu Trp Trp Leu Leu Leu Leu Leu Leu Leu 1 5 10 15 Leu Trp Pro Met Val Trp Ala Ile Ser Tyr Asp Ser Pro Asp Tyr Thr 20 25 30 Asp Glu Ser Cys Thr Phe Lys Ile Ser Leu Arg Asn Phe Arg Ser Ile 35 40 45 Leu Ser Trp Glu Leu Lys Asn His Ser Ile Val Pro Thr His Tyr Thr 50 55 60 Leu Leu Tyr Thr Ile Met Ser Lys Pro Glu Asp Leu Lys Val Val Lys 65 70 75 80 Asn Cys Ala Asn Thr Thr Arg Ser Phe Cys Asp Leu Thr Asp Glu Trp 85 90 95 Arg Ser Thr His Glu Ala Tyr Val Thr Val Leu Glu Gly Phe Ser Gly 100 105 110 Asn Thr Thr Leu Phe Ser Cys Ser His Asn Phe Trp Leu Ala Ile Asp 115 120 125 Met Ser Phe Glu Pro Pro Glu Phe Glu Ile Val Gly Phe Thr Asn His 130 135 140 Ile Asn Val Met Val Lys Phe Pro Ser Ile Val Glu Glu Glu Leu Gln 145 150 155 160 Phe Asp Leu Ser Leu Val Ile Glu Glu Gln Ser Glu Gly Ile Val Lys 165 170 175 Lys His Lys Pro Glu Ile Lys Gly Asn Met Ser Gly Asn Phe Thr Tyr 180 185 190 Ile Ile Asp Lys Leu Ile Pro Asn Thr Asn Tyr Cys Val Ser Val Tyr 195 200 205 Leu Glu His Ser Asp Glu Gln Ala Val Ile Lys Ser Pro Leu Lys Cys 210 215 220 Thr Leu Leu Pro Pro Gly Gln Glu Ser Glu Ser Ala Glu Ser Ala Lys 225 230 235 240 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Pro 245 250 255 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 260 265 270 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 275 280 285 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 290 295 300 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 305 310 315 320 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 325 330 335 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 340 345 350 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 355 360 365 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 370 375 380 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Glu Asn 385 390 395 400 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 405 410 415 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 420 425 430 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Trp 435 440 445 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 450 455 460 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 465 470 475 480 Ser Leu Ser Pro Gly Lys 485 <210> 51 <211> 1461 <212> DNA <213> Artificial Sequence <220> <223> IFNAR2-Linker_4GS*3-FC RVT <400> 51 gccaccatgt ggtggcgtct gtggtggctg ctgcttctgc tgttgctgct gtggcctatg 60 gtctgggcca tctcctacga cagccctgac tacaccgacg agtcctgcac cttcaagatc 120 tccctgcgga acttccggtc catcctgtcc tgggagctga agaaccactc catcgtgccc 180 acacactaca ccctgctgta caccatcatg tccaagcctg aggacctgaa ggtggtcaag 240 aactgcgcca acaccaccag atctttctgc gacctgaccg atgagtggcg gtctacccac 300 gaggcttacg tgacagtgct ggaaggcttc tccggcaata ccacactgtt cagctgctcc 360 caaacttct ggctggccat cgacatgtcc ttcgagcctc cagagttcga gatcgtgggc 420 ttcaccaacc acatcaacgt gatggtcaag ttccccagca tcgtggaaga ggaactccag 480 ttcgacctga gcctggtcat cgaggaacag tccgagggca tcgtgaagaa gcacaagccc 540 gagatcaagg gcaacatgtc cggcaacttc acctacatca tcgacaagct gatcccccaac 600 accaactact gcgtgtccgt gtacctggaa cactccgatg agcaggccgt gatcaagtcc 660 cctctgaagt gtaccctgct gcctcctggc caagagtctg agtctgccga gtctgctaaa 720 ggcggcggag gatctggcgg aggtggaagc ggaggcggtg gatctcccaa gagctgcgac 780 aagacccaca cctgtcccccc ctgccctgcc cctgaactgc tgggcggacc cagcgtgttc 840 ctgttccccc caaagcccaa ggacaccctg atgatcagcc ggacccccga agtgacctgc 900 gtggtggtgg acgtgtccca cgaggaccct gaagtgaagt tcaattggta cgtggacggc 960 gtggaagtgc acaacgccaa gaccaagccc agagaggaac agtacaacag cacctaccgg 1020 gtggtgtccg tgctgaccgt gctgcaccag gactggctga acggcaaaga gtacaagtgc 1080 aaggtgtcca acaaggccct gcctgccccc atcgagaaaa ccatcagcaa ggccaagggc 1140 cagccccgcg agccccgggt gtacacactg ccccccagcc gggacgagct gaccaagaac 1200 caggtgtccc tgacctgcct ggtgaaaggc ttctacccca gcgatatcgc cgtggaatgg 1260 gagagcaacg gccagcccga gaacaactac aagaccaccc cccctgtgct ggtcagcgac 1320 [[ID=​​​​​​​​​​​​​​​<223> IFNAR2- connection_4GS*3-FC RVT <400> 52 Ala Thr Met Trp Trp Arg Leu Trp Trp Leu Leu Leu Leu Leu Leu Leu 1 5 10 15 Leu Trp Pro Met Val Trp Ala Ile Ser Tyr Asp Ser Pro Asp Tyr Thr 20 25 30 Asp Glu Ser Cys Thr Phe Lys Ile Ser Leu Arg Asn Phe Arg Ser Ile 35 40 45 Leu Ser Trp Glu Leu Lys Asn His Ser Ile Val Pro Thr His Tyr Thr 50 55 60 Leu Leu Tyr Thr Ile Met Ser Lys Pro Glu Asp Leu Lys Val Val Lys 65 70 75 80 Asn Cys Ala Asn Thr Thr Arg Ser Phe Cys Asp Leu Thr Asp Glu Trp 85 90 95 Arg Ser Thr His Glu Ala Tyr Val Thr Val Leu Glu Gly Phe Ser Gly 100 105 110 Asn Thr Thr Leu Phe Ser Cys Ser His Asn Phe Trp Leu Ala Ile Asp 115 120 125 Met Ser Phe Glu Pro Pro Glu Phe Glu Ile Val Gly Phe Thr Asn His 130 135 140 Ile Asn Val Met Val Lys Phe Pro Ser Ile Val Glu Glu Glu Leu Gln 145 150 155 160 Phe Asp Leu Ser Leu Val Ile Glu Glu Gln Ser Glu Gly Ile Val Lys 165 170 175 Lys His Lys Pro Glu Ile Lys Gly Asn Met Ser Gly Asn Phe Thr Tyr 180 185 190 Ile Ile Asp Lys Leu Ile Pro Asn Thr Asn Tyr Cys Val Ser Val Tyr 195 200 205 Leu Glu His Ser Asp Glu Gln Ala Val Ile Lys Ser Pro Leu Lys Cys 210 215 220 Thr Leu Leu Pro Pro Gly Gln Glu Ser Glu Ser Ala Glu Ser Ala Lys 225 230 235 240 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Pro 245 250 255 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 260 265 270 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 275 280 285 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 290 295 300 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 305 310 315 320 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 325 330 335 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 340 345 350 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 355 360 365 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 370 375 380 Pro Arg Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 385 390 395 400 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 405 410 415 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 420 425 430 Thr Pro Pro Val Leu Val Ser Asp Gly Ser Phe Thr Leu Tyr Ser Lys 435 440 445 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 450 455 460 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 465 470 475 480 Ser Leu Ser Pro Gly Lys 485 <210> 53 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Connector_4GS <400> 53 Gly Gly Gly Gly Ser 1 5 <210> 54 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Connector_4GS*2 <400> 54 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 1 5 10 <210> 55 <211> 20 <212> PRT <213> Artificial sequence <220> <223> Connector_4GS*4 <400> 55 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 1 5 10 15 Gly Gly Gly Ser 20 <210> 56 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Connector_6G <400> 56 Gly Gly Gly Gly Gly Gly 1 5 <210> 57 <211> 8 <212> PRT <213> Artificial sequence <220> <223> Connector_8G <400> 57 Gly Gly Gly Gly Gly Gly Gly Gly 1 5 <210> 58 <211> 12 <212> PRT <213> Artificial sequence <220> <223> Connector_AEAAAKEAAAKA <400> 58 Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Ala 1 5 10 <210> 59 <211> 46 <212> PRT <213> Artificial sequence <220> <223> Linker-EAAAK complex <400> 59 Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys 1 5 10 15 Glu Ala Ala Ala Lys Ala Leu Glu Ala Glu Ala Ala Ala Lys Glu Ala 20 25 30 Ala Ala Lys Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Ala 35 40 45 <210> 60 <211> 15 <212> PRT <213> Artificial sequence <220> <223> Connector_EAAAK*3 <400> 60 Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys 1 5 10 15 <210> 61 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Connector_EAAAK <400> 61 Glu Ala Ala Ala Lys 1 5 <210> 62 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Connector_EAAAK*2 <400> 62 Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys 1 5 10 <210> 63 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Connector_AP*5 <400> 63 Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro 1 5 10 <210> 64 <211> 12 <212> PRT <213> Artificial sequence <220> <223> Connector_AP*6 <400> 64 Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro 1 5 10 <210> 65 <211> 14 <212> PRT <213> Artificial sequence <220> <223> Connector_AP*7 <400> 65 Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro 1 5 10 <210> 66 <211> 16 <212> PRT <213> Artificial sequence <220> <223> Connector_AP*8 <400> 66 Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro 1 5 10 15 <210> 67 <211> 18 <212> PRT <213> Artificial sequence <220> <223> Connector_AP*9 <400> 67 Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro 1 5 10 15 Ala Pro <210> 68 <211> 20 <212> PRT <213> Artificial sequence <220> <223> Connector_AP*10 <400> 68 Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro 1 5 10 15 Ala Pro Ala Pro 20 <210> 69 <211> twenty two <212> PRT <213> Artificial sequence <220> <223> Connector_AP*11 <400> 69 Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro 1 5 10 15 Ala Pro Ala Pro Ala Pro 20 <210> 70 <211> twenty four <212> PRT <213> Artificial sequence <220> <223> Connector_AP*12 <400> 70 Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro 1 5 10 15 Ala Pro Ala Pro Ala Pro Ala Pro 20 <210> 71 <211> 26 <212> PRT <213> Artificial sequence <220> <223> Connector_AP*13 <400> 71 Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro 1 5 10 15 Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro 20 25 <210> 72 <211> 28 <212> PRT <213> Artificial sequence <220> <223> Connector_AP*14 <400> 72 Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro 1 5 10 15 Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro 20 25 <210> 73 <211> 30 <212> PRT <213> Artificial sequence <220> <223> Connector_AP*15 <400> 73 Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro 1 5 10 15 Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro 20 25 30 <210> 74 <211> 32 <212> PRT <213> Artificial sequence <220> <223> Connector_AP*16 <400> 74 Ala For Ala For Ala For Ala For Ala For Ala For Ala For Ala For 1 5 10 15 Ala For Ala For Ala For Ala For Ala For Ala For Ala For Ala For 20 25 30 <210> 75 <211> 34 <212> PRT <213> artificial sequence <220> <223> Connection_AP*17 <400> 75 Ala For Ala For Ala For Ala For Ala For Ala For Ala For Ala For 1 5 10 15 Ala For Ala For Ala For Ala For Ala For Ala For Ala For Ala For 20 25 30 Ala Pro <210> 76 <211> 5 <212> PRT <213> artificial sequence <220> <223> connection_PAPAP <400> 76 For Ala For Ala For 1 5

Claims

1. A dimeric polypeptide, wherein a monomer comprising an interferon receptor 1 (IFNAR1) fragment and an antibody Fc fragment is combined with a monomer comprising an interferon receptor 2 (IFNAR2) fragment and an antibody Fc fragment in the dimeric polypeptide, wherein the antibody Fc fragment in the monomer comprising the IFNAR1 fragment consists of the amino acid sequence of SEQ ID NO: 12, and the antibody Fc fragment in the monomer comprising the IFNAR2 fragment consists of the amino acid sequence of SEQ ID NO:

14.

2. The dimeric polypeptide according to claim 1, wherein the dimeric polypeptide neutralizes type 1 interferon.

3. The dimeric polypeptide according to claim 1, wherein the interferon receptor 1 fragment consists of the amino acid sequence of SEQ ID NO:

4.

4. The dimeric polypeptide according to claim 1, wherein the interferon receptor 2 fragment consists of the amino acid sequence of SEQ ID NO:

6.

5. A polynucleotide encoding the dimeric polypeptide of claim 1.

6. A vector comprising the polynucleotide of claim 5.

7. A host cell transformed with the vector of claim 6.

8. A method for preparing the dimeric polypeptide of claim 1, comprising: (a) providing the host cell of claim 7; (b) culturing the provided cells; and (c) preparing the dimeric polypeptide by recovering the dimeric polypeptide from cells or culture medium.

9. A pharmaceutical composition for preventing or treating a disease or condition mediated by type 1 interferon, comprising the dimeric polypeptide of claim 1 as an active ingredient.

10. The pharmaceutical composition of claim 9, wherein the type 1 interferon-mediated disease or condition is selected from systemic lupus erythematosus, Sjögren's syndrome, systemic sclerosis, insulin-dependent diabetes mellitus (IDDM), inflammatory bowel disease (IBD), multiple sclerosis (MS), psoriasis, autoimmune thyroiditis, rheumatoid arthritis, inflammatory myositis, glomerulonephritis, HIV infection, and transplant rejection.

11. The pharmaceutical composition according to claim 9, wherein the type 1 interferon-mediated disease or condition is selected from Crohn's disease, ulcerative colitis, celiac disease, AIDS and graft-versus-host disease (GVHD).

12. Use of the dimeric polypeptide of claim 1 for the preparation of a medicament for preventing or treating a disease or condition mediated by type 1 interferon, wherein the disease or condition mediated by type 1 interferon is selected from systemic lupus erythematosus, Sjögren's syndrome, systemic sclerosis, insulin-dependent diabetes mellitus (IDDM), inflammatory bowel disease (IBD), multiple sclerosis (MS), psoriasis, autoimmune thyroiditis, rheumatoid arthritis, inflammatory myositis, glomerulonephritis, HIV infection and transplant rejection.

13. Use of the dimeric polypeptide of claim 1 for preparing a medicament for preventing or treating a disease or condition mediated by type 1 interferon, wherein the disease or condition mediated by type 1 interferon is selected from Crohn's disease, ulcerative colitis, celiac disease, AIDS and graft-versus-host disease (GVHD). 14 . The dimeric polypeptide according to claim 1 , wherein the interferon receptor 1 (IFNAR1) fragment in the monomer is connected to the antibody Fc fragment via a linker.

15. The dimeric polypeptide according to claim 1, wherein the interferon receptor 2 (IFNAR2) fragment in the monomer is connected to the antibody Fc fragment via a linker.

Images