Dual CSF1-il-10 cytokine comprising an immunoglobulin FC fragment

Abstract

The invention relates to single chain polypeptides having IL-10 and CSF1 activities and comprising an immunoglobulin Fc fragment, and to their use in therapy.

Description

[0001] DUAL CSF1-IL-10 CYTOKINE COMPRISING AN IMMUNOGLOBULIN FC FRAGMENT

[0002] Field of the Invention

[0003] The invention relates to proteins comprising a single chain polypeptide having IL-10 and CSF1 activities and a moiety increasing half-life, such as an immunoglobulin Fc fragment, and to their use in therapy.

[0004] Background of the Invention

[0005] Cytokines are proteins that are produced in the immune system and act as chemical messengers to communicate between cells of the immune system, and also with other cells of the body. Cytokines control the entire range of immune responses, including their initiation, type, potency, and duration. So much so that its unregulated production triggers a wide variety of inflammatory and autoimmune diseases, allergies, fibrosis, and can even lead to cancer. For this reason, there is great medical interest in the development and use of cytokines as drugs to treat diseases.

[0006] Interleukin-10 (IL-10) is an anti-inflammatory cytokine that plays a crucial role in regulating immune responses and maintaining immune homeostasis. IL-10 is a homodimeric cytokine that is produced as a monomer by various immune cells, including T cells, B cells, macrophages, and dendritic cells, and it exerts its effects by binding to its receptor complex, which is composed of two subunits, IL-10R1 and IL-10R2. When IL-10 binds to its receptor, IL-10R1 first recognizes and binds to IL-10 with high affinity, forming a complex that is then stabilized by the binding of IL-10R2. The binding of IL- 10 to its receptor complex leads to the activation of downstream signaling pathways.

[0007] When binds to its receptor on myeloid cells, IL-10 exerts potent anti-inflammatory activities, including inhibition of the production of pro-inflammatory mediators, expression of MHCII and costimulatory molecules. Simultaneously, activation of myeloid cells by IL-10 leads to the expression of anti-inflammatory mediators and promotes the development of tolerogenic phenotype, like M2-like anti-inflammatory macrophages and tolerogenic dendritic cells. In contrast to the strong anti-inflammatory activities on myeloid cells, IL- 10 is a growth factor for B cells and promotes plasma cell differentiation and antibody production. Similarly, IL-10 has stimulatory effects on CD8+ T cells, inducing their proliferation, IFNY production, and cytotoxicity.

[0008] In humans, there is a strong genetic association between the IL- 10 pathway and autoimmune diseases, such as inflammatory bowel disease. Loss-of-function mutations in IL-10, IL-10RA, or IL-10RB result in early-onset, therapy-resistant severe enterocolitis. Restoring IL-10RA or IL-10 expression by hematopoietic stem cell transplantation rapidlyalleviates clinical symptoms. In turn, mice deficient in IL-10 or IL-10R develop colitis spontaneously, while the administration of IL-10 in different animal models of colitis has been shown to be consistently beneficial. Finally, mice in which all cells of the organism respond to IL-10, except specific subsets of myeloid cells that includes monocytes and macrophages, also develop spontaneous colitis that is mediated by the production of the clinically validated target IL-23.

[0009] Myeloid cells exclusively express CSF1 receptor, i.e. CSF1R. Stimulation of CSF1R by its ligand, the cytokine CSF1, is essential for the differentiation, proliferation and maintenance of M2-like anti-inflammatory macrophages in the intestine, which play a key role in generating a tolerogenic environment through the production of IL-10. In autoimmune disease, the M2-like anti-inflammatory macrophages of the affected tissues, such as the mucosa of IBD patients, are replaced by inflammatory monocytes that drive disease progression.

[0010] While in preclinical studies IL-10 has shown promising results in ameliorating inflammation and tissue damage in models of colitis, clinical trials investigating the therapeutic potential of IL-10 in patients with colitis have not been successful. One possible explanation is that the delivery of IL- 10 to the site of inflammation may be a limiting factor. However, some strategies aimed to enhance the delivery of IL-10 at the site of inflammation did not show clinical benefit (AMT-101).

[0011] Another possible explanation is because the anti-inflammatory effects of IL-10 on myeloid cells are counteracted by pro-inflammatory tissue-damaging effects, including activation, differentiation, and induction of antibodies by B cells and induction of cytotoxic activities by CD8+ T cells.

[0012] To address this limitation, there is a need for IL-10-based therapeutics that can selectively activate myeloid cells while avoiding activation of other immune cell subsets. Several strategies have been proposed for developing IL-10-based therapeutics that selectively activate myeloid cells. These include fusion of IL-10 to antibodies that bind to receptors on myeloid cells, or encapsulation of IL-10 in nanocarriers (liposomes or nanoparticles) that are specifically targeted to myeloid cells

[0013] The present invention seeks to overcome or at least alleviate one or more of the deficiencies in the prior art.

[0014] Summary of the Invention

[0015] The invention is as defined by the claims.

[0016] The invention relates to a protein comprising a single chain polypeptide having IL- 10 and CSF1 activities modified to increase its half-life.The invention relates to a protein comprising a single chain polypeptide having IL-10 and CSF1 activities fused to an immunoglobulin Fc fragment, or to albumin or albumin fragment, or to an anti-albumin antibody, or anti-albumin antibody fragment, or conjugated with a poly(ethylene glycol) (PEG) molecule.

[0017] In some embodiments, the single chain polypeptide having IL-10 and CSF1 activities comprises in the N-terminus to C-terminus direction, a CSF1 domain, a peptide linker L1, and an IL-10 domain, wherein the IL-10 domain is an IL-10 monomer or a single chain dimeric IL-10, and wherein the CSF1 domain is a CSF1 monomer or a single chain dimeric CSF1.

[0018] In some embodiments, the protein comprises one single chain polypeptide having IL-10 and CSF1 activities and one immunoglobulin Fc fragment, wherein the single chain polypeptide having IL-10 and CSF1 activities is fused to the immunoglobulin Fc fragment through a peptide linker L2 connecting (i) the C-terminus of the IL-10 domain with the N-terminus of one hinge region of the Fc fragment, or (ii) the C-terminus of the one of the CH3 domains of the Fc fragment with the N-terminus of the CSF1 monomer.

[0019] In some embodiments, the protein comprises two single chain polypeptides having IL-10 and CSF1 activities and one immunoglobulin Fc fragment.

[0020] In some embodiments, the protein comprises two polypeptide chains that each comprise, in the N-terminus to C-terminus direction, a CSF1 domain which is a CSF1 monomer or a single chain dimeric CSF1, a peptide linker L1, an IL-10 domain which is an IL-10 monomer or a single chain dimeric IL-10 monomer, a peptide linker L2, and a Fc domain, wherein the two polypeptide chains dimerise through their Fc domains.

[0021] In some embodiments, the protein comprises two polypeptide chains that each comprise, in the N-terminus to C-terminus direction, a Fc domain, a peptide linker L2, a CSF1 domain which is a CSF1 monomer or a single chain dimeric CSF1, a peptide linker L1, and IL-10 domain which is an IL-10 monomer or a single chain dimeric IL-10 monomer, wherein the two polypeptide chains dimerise through their Fc domains.

[0022] In some embodiments, the CSF1 monomer is a wild-type CSF1 monomer, or a mutant thereof. In some embodiments, the CSF1 monomer is a CSF1 monomer that comprises sequence SEQ ID NO:12, SEQ ID NO: 245, or SEQ ID NO: 17, or a single chain dimeric CSF1 comprising sequence SEQ ID NO: 277, ora sequence at least 80% identical thereto, preferably that retains CSF1 activity.

[0023] In some embodiments, the CSF1 domain is a polypeptide that comprises a sequence differing from SEQ ID NO: 12, SEQ ID NO: 245, SEQ ID NO: 17, or SEQ ID NO: 277 by one or more mutations comprising:a) substitution M65Y;

[0024] b) substitutions S18I, T64H, R66V, A74D, L85V, R86Y, and V120F;

[0025] c) substitutions S18I, T64H, R66V, A74D, L85I, R86Y, and V120F; or

[0026] d) substitutions S18I, T64H, R66I, A74D, L85I, R86Y, and V120F.

[0027] In some embodiments, the IL-10 domain is an IL-10 monomer and the single chain polypeptide having IL- 10 and CSF1 activities comprises sequence SEQ ID NO: 25, or a sequence at least 80% identical thereto, preferably that retains IL-10 and CSF1 activities.

[0028] In some embodiments, the IL-10 domain is a single chain dimeric IL-10 that comprises sequence SEQ ID NO: 22, SEQ ID NO: 5, or SEQ ID NO: 1, or a sequence at least 80% identical to mentioned sequences and that retains at least the same stability, and / or at least the same level of interaction with IL-10 receptor, and the same IL-10 activity.

[0029] In some embodiments, one or more of the following conditions are met:

[0030] (i) the peptide linker L1 comprises from 5 to 50 amino acid residues;

[0031] (ii) the peptide linker L1 is a flexible peptide sequence composed of Gly and Ser residues, in different proportion;

[0032] (iii) the peptide linker L1 comprises sequence SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, or SEQ ID NO: 36;

[0033] (iv) (iv) the peptide linker L1 comprises sequence (SEQ ID NO:82)n, n being an integer from 1 to 10;

[0034] (v) the peptide linker L1 is a rigid peptide sequence composed of Glu, Ala, and Lys residues; or

[0035] (vi) the peptide linker L1 comprises sequence EAAAK (SEQ ID NO: 247), or (EAAAK; SEQ ID NO: 247)nwith n being a integer from 2 to 10.

[0036] In some embodiments, the single chain polypeptide having IL-10 and CSF1 activities comprises sequence SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 105, or any one of sequence SEQ ID NO: 264 to SEQ ID NO: 269, and SEQ ID NO: 273 to SEQ ID NO: 275, or a sequence at least 80% identical thereto, preferably that retains IL-10 and CSF1 activities.

[0037] In some embodiments, the immunoglobulin Fc fragment or Fc domain is of the isotype IgG, preferably human IgG, still preferably human lgG1 or human lgG4, optionally engineered.

[0038] In some embodiments, one or more of the following conditions are met:

[0039] (i) the peptide linker L2 comprises from 3 to 45 amino acid residues;

[0040] (ii) the peptide linker L2 is a flexible peptide sequence composed of Gly and Ser residues; (iii) the peptide linker L2 comprises sequence (GGGGS - SEQ ID NO: 82)p, wherein p is an integer from 1 to 8;(iv) the peptide linker L2 comprises sequence EAAAK (SEQ ID NO: 247), or (EAAAK; SEQ ID NO: 247)nwith n being a integer from 2 to 10;

[0041] (vi) the peptide linker L2 comprises any one of sequence SEQ ID NO: 82, SEQ ID NO: 35, SEQ ID NO: 119 to SEQ ID NO: 131, SEQ ID NO: 136, SEQ ID NO: 247, and SEQ ID NO: 248, and SEQ ID NO: 279.

[0042] In some embodiments, the protein comprises sequence SEQ ID NO: 88, SEQ ID NO: 101, SEQ ID NO: 161, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 167, SEQ ID NO: 184, SEQ ID NO: 187, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 218, SEQ ID NO: 228, SEQ ID NO: 230, SEQ ID NO: 231, SEQ ID NO: 236, SEQ ID NO:239, SEQ ID NO: 240, SEQ ID NO: 241, or a sequence at least 80% identical thereto, preferably that retains IL-10 and CSF1 activities.

[0043] In some embodiments the protein further comprises a second polypeptide comprising a second Fc domain.

[0044] In some embodiments, the protein comprises a first polypeptide comprising, or consisting of, sequence SEQ ID NO: 88, SEQ ID NO: 101, SEQ ID NO: 161, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 167, SEQ ID NO: 184, SEQ ID NO: 187, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 218, SEQ ID NO: 228, SEQ ID NO: 230, SEQ ID NO: 231, SEQ ID NO: 236, SEQ ID NO:239, SEQ ID NO: 240, SEQ ID NO: 241, or a sequence at least 80% identical thereto and that retains IL- 10 and CSF1 activities, and a second polypeptide comprising or consisting of SEQ ID NO: 109. In some embodiments, the protein comprises a first polypeptide comprising, or consisting of, sequence SEQ ID NO: 155, or a sequence at least 80% identical thereto and that retains IL-10 and CSF1 activities, and a second polypeptide comprising or consisting of SEQ ID NO: 155.

[0045] Also provided in a protein having dual IL- 10 and CSF1 activities that comprises two polypeptide chains comprising, in the N-terminus to C-terminus direction, a CSF1 monomer fused to an immunoglobulin Fc fragment, and second polypeptide chain comprises, in the N-terminus to C-terminus direction, an immunoglobulin Fc fragment fused to an IL-10 domain, wherein the IL-10 domain is an IL-10 monomer or a single chain dimeric IL-10, and wherein the two polypeptide chains dimerise through their Fc domains.

[0046] The invention further relates to a pharmaceutical composition comprising the protein according to the invention.

[0047] The invention further relates to the protein according to the invention for use as a medicament, in particular for use for treating an inflammatory disease.

[0048] Detailed description of the inventionThe inventors have designed single chain polypeptides combining IL-10 and CSF1 activities (herein called ORK1 proteins).

[0049] On one hand, IL-10 monomers pair to form a ‘swapped-domain’ dimeric IL-10 protein. In a swapped-domain dimeric protein like IL-10, the monomeric structure of the standard IL-10 helix bundle cytokine opens and embraces in an antiparallel fashion another IL-10 opened monomer generating two functional split cytokine domains together, defining two adjacent 3D domains. On the other hand, CSF1 monomers dimerise, with the two monomeric CSF1 molecules face to face and hold together by a disulphide bond.

[0050] The functional characterization of the dual single chain polypeptide having IL-10 and CSF1 activities has shown that the dual CSF1 -IL-10 cytokine inhibits activation of myeloid cells and induces differentiation of monocyte-derived macrophages that have a regulatory phenotype (M2 phenotype). Furthermore, the dual CSF1-IL-10 cytokine fused to a Fc fragment (herein called ‘ORK1-Fc’) has specificity for monocytes and does not activate CD8 T cells or B cells.

[0051] Functionally, in vivo, the dual ORK1-Fc cytokine was shown to alleviate the clinical symptoms of inflammatory disease, in a TNBS-induced colitis mouse model.

[0052] Different structural organizations were designed for the dual ORK1 / ORK1-Fc cytokines, as shown in Figures 2 and 21, and sequence variations were assayed for both the CSF1 and the IL-10 domains, as disclosed in Table 1.

[0053] Surprisingly, ORK1-Fc variants, showed an improved function both in vitro, with human PBMC, and in vivo, and improved specificity to monocytes vs T cells or B cells as shown and described in Figures (3, 5, 6, 8, 9, 12, 14) compared to ORK1_P_068 (also denoted ORK1-068) protein without IgG Fc fusion.

[0054] These single chain polypeptides combining IL-10 and CSF1 activities are advantageously provided in fusion or conjugation with a moiety increasing half-life, such as an immunoglobulin Fc fragment, albumin, or any albumin-binding domain; conjugation with poly(ethylene glycol) (PEG) or other polymer-based moieties; or any other chemical or genetic modification known to extend protein stability and circulation time in vivo.

[0055] A protein comprising a single chain polypeptide having IL-10 and CSF1 activities fused to an immunoglobulin Fc fragment, or to albumin, or albumin fragment, or to an antialbumin antibody, or anti-albumin antibody fragment, or conjugated with a poly(ethylene glycol) (PEG) molecule is thus provided according to certain aspects.

[0056] Main definitions

[0057] As used in this specification and the appended claims, the singular forms “a,” “an,” and “the” include plural references unless the context clearly dictates otherwise. Thus, forexample, “a ribonucleotide” is understood to represent one or more ribonucleotides. As such, the terms “a,” “an,” “one or more,” and “at least one” can be used interchangeably herein.

[0058] Throughout this specification and embodiments, the words “have” and “comprise,” or variations such as “has,” “having,” “comprises,” or “comprising” will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers. It is further understood that wherever embodiments are described herein with the language “comprising” or “having,” or grammatical equivalents thereof, otherwise analogous embodiments described in terms of “consisting of” and / or “consisting essentially of” are also provided.

[0059] As used herein ‘IL-10 activity” or “IL-10 activities” denotes one or more biological activities mediated by IL-10 through binding to its receptor, IL-10R. These comprise or consist of (i) the binding to IL-1 OR on myeloid cells, in particular monocytes, or (ii) antiinflammatory activities, or preferentially both. Anti-inflammatory activities include (a) the inhibition of production of pro-inflammatory mediators (e.g. pro-inflammatory cytokines such as tumour necrosis factor-α (TNF-α), IL-1, IL-12, IL-6 and granulocyte-macrophage colonystimulating factor, inflammatory enzymes such as cyclo-oxygenase 2 and inducible nitric oxide synthase, chemokines such as RANTES, membrane inflammatory protein-1α (MIP1α), IL-8, and eotaxin), and / or (b) the inhibition of expression of MHCII and costimulatory molecules by myeloid cells, and / or (c) the induction of differentiation of monocytes into tolerogenic macrophages. In some embodiments, anti-inflammatory activities comprise inhibition of production of TNF-α and / or IL-6 by activated monocytes. In some embodiments, anti-inflammatory activities comprise induction of differentiation of monocytes into tolerogenic macrophages. Tolerogenic macrophages have M2-like phenotype and can be identified by their regulatory phenotype and high levels of expression of CD206 and CD163. In some embodiments, IL-10 activities comprise or consist of binding to IL-10R on myeloid cells, in particular monocytes, inhibition of production of TNF-α and / or IL-6 by activated monocytes, and differentiation of monocytes into tolerogenic macrophages.

[0060] As used herein “CSF1 activity” or “CSF1 activities” denotes one or more biological activities mediated by activation of CSF1 receptor, CFS1-R. These comprise or consist of (i) the activation of CFS1-R at the surface of monocytes or progenitors thereof, and / or (ii) the differentiation of monocytes into macrophages, preferentially both. In some embodiments, activation of CSF1-R is triggered by binding of CSF1 to CSF1-R.

[0061] An “immunoglobulin Fc fragment” or “Fc fragment” consists of two polypeptides linked by disulfide bonds, each polypeptide, from the N-terminal to C-terminal, beingcomposed of a hinge region, a CH2 domain and a CH3 domain. The structure of the Fc fragment is nearly identical across all subtypes of human immunoglobulin. As used herein “an immunoglobulin Fc domain” or “Fc domain” denotes the monomeric polypeptide comprising immunoglobulin hinge and CH2, CH3 immunoglobulin heavy chain constant domains. Fc domain as used herein encompasses native Fc and Fc variants. The term "native Fc" as used herein refers to a molecule comprising the sequence of a non-antigen-binding fragment resulting from digestion of an antibody, or produced by other means, and that contain the hinge region. The original immunoglobulin source of the native Fc is preferably of human origin and can be any of the immunoglobulins, of isotype as IgM, I g D, IgG, IgA, or IgE, IgG has several subclasses, including, but not limited to, lgG1, lgG2, lgG3, and lgG4. The term " Fc variant" as used herein refers to a molecule or sequence that is modified from a native Fc but still comprises a binding site for the salvage receptor, FcRn (neonatal Fc receptor). Exemplary Fc variants, and their interaction with the salvage receptor, are known in the art. Thus, the term " Fc variant" can comprise a molecule or sequence that is humanized from a non-human native Fc. Furthermore, a native Fc comprises regions that can be removed because they provide structural features or biological activity that are not required for the antibody-like binding proteins of the invention. Thus, the term " Fc variant" comprises a molecule or sequence that lacks one or more native Fc sites or residues, or in which one or more Fc sites or residues has be modified, that affect or are involved in: (1) disulfide bond formation, (2) incompatibility with a selected host cell, (3) N-terminal heterogeneity upon expression in a selected host cell, (4) glycosylation, (5) interaction with complement, (6) binding to an Fc receptor other than a salvage receptor, or (7) antibody-dependent cellular cytotoxicity (ADCC).

[0062] Determining cytokine activities can be readily performed by the skilled person using conventional methods in the art, such as HEK cell reported assays, as disclosed in the examples.

[0063] Single chain polypeptide having IL-10 and CSF1 activities

[0064] A single chain polypeptide having IL-10 and CSF1 activities is disclosed. In some embodiments, the single chain polypeptide having IL-10 and CSF1 activities has reduced T cells and B cell activation capacities compared to IL-10.

[0065] In some embodiments, the single chain polypeptide having IL-10 and CSF1 activities has similar or higher affinity to CSF1 R compared to wild-type CSF1. In some embodiments, the single chain polypeptide having IL- 10 and CSF1 activities has reduced affinity to IL- 10 receptor α (IL10RA) and IL-10 receptor β (IL10RB) compared to wild-type IL-10. In some embodiments, the single chain polypeptide having IL-10 and CSF1 activities has similar orhigher affinity to CSF1R compared to wild-type CSF1 and has reduced affinity to IL-10 receptor α (IL10RA) and IL-10 receptor β (IL10RB) compared to wild-type IL-10. These properties are expected to favor selective binding of the single chain polypeptide having IL-10 and CSF1 activities to myeloid cells that exclusively express CSF1 receptor, among cells of the innate immune system, and subsequent activation thereof.

[0066] By ‘similar affinity’ it is meant herein the affinity preferably does not vary (i.e. increase or decrease) by more than a 5-fold, preferably 4-fold, 3-fold, 2-fold, or 1.5-fold factor compared to the reference level of affinity of the wild-type cytokine (CSF1).

[0067] By ‘higher’ or ‘reduced’ it is meant herein the affinity or the selectivity towards monocytes in respect to T cells or B cells activation, preferably varies (i.e. increase or decrease) by at least a 5-fold, preferably, 7-fold, 10-fold, 15-fold, 20-fold, 30-fold, 40-fold, 50-fold, 100-fold, 200-fold factor, or over compared to the reference level of affinity of the wild-type cytokine (CSF1 or IL-10 as appropriate).

[0068] The single chain polypeptide having IL-10 and CSF1 activities comprises an IL-10 domain fused to a CSF1 domain, wherein the IL-10 domain is an IL-10 monomer or a single chain dimeric IL-10, and wherein the CSF1 domain is a CSF1 monomer or single chain dimeric CSF1.

[0069] In some embodiments, the single chain polypeptide having IL-10 and CSF1 activities comprises in the N-terminus to C-terminus direction, a CSF1 monomer, a peptide linker L1, and an IL-10 domain, wherein the IL-10 domain is an IL-10 monomer or a single chain dimeric IL-10.

[0070] In some embodiments, the single chain polypeptide having IL-10 and CSF1 activities comprises in the N-terminus to C-terminus direction, a single chain dimeric CSF1, a peptide linker L1, and an IL-10 domain, wherein the IL-10 domain is an IL-10 monomer or a single chain dimeric IL-10, preferably an IL-10 monomer.

[0071] In a first aspect, the single chain polypeptide having IL- 10 and CSF1 activities is thus provided that comprises an IL-10 monomer fused to a CSF1 monomer or single chain dimeric CSF1, wherein the N-terminus of IL-10 is linked to the C-terminus of CSF1 through a linker L1.

[0072] Indeed, in this orientation, the single chain polypeptide has decreased IL-10 activity compared to wild-type IL-10, but maintain a CSF1 activity similar to wild-type CSF1.

[0073] According to this aspect, the single chain polypeptide having IL- 10 and CSF1 activities is thus comprises, in the N-terminus to C-terminus direction, a CSF1 monomer or single chain dimeric CSF1, a peptide linker L1, and an IL-10 monomer.In some embodiments the single chain dimeric IL-10 and CSF1 monomer or single chain dimeric CSF1 are linked through a linker L1 that comprises from (or consists of) 5 to 45 amino acid residues, preferably 12 to 40 amino acids, still preferably 15 to 40 amino acids, still preferably 20 to 35 amino acids, even more preferably 25 to 35 amino acids.

[0074] In some embodiments, the linker L1 bridging the C-terminus of CSF1 monomer single chain dimeric CSF1 to the N-terminus of IL-10 monomer is a (flexible) peptide sequence composed of Gly and Ser residues, in different proportions. Examples of suitable linkers comprise or consist of GGGSGGSGGSGGSGGSGGSGGSGGG (25 amino acid long, SEQ ID NO: 34), GGGGGSGGSGGSGGSGGSGGSGGSGGSGGG (30 amino acid long, SEQ ID NO: 35), or GGGGGSGGGGSGGSGGSGGSGGSGGSGGSGGSGGG (35 amino acid long, SEQ ID NO: 36). In some embodiments the L1 linker comprises or consists of a sequence (GGGGS, SEQ ID NO:82)n, n being an integer from 1 to 10.

[0075] The CSF1 monomer or single chain dimeric CSF1 is as defined in the below section. The IL-10 monomer may be wild-type mature IL-10 as shown in SEQ ID NO:11, or a mutant thereof, or a polypeptide comprising the sequence of mature IL-10 and on the N-terminal end 1, 2, 3 or more additional amino acids residues consecutively present in the signal peptide of IL-10 (i.e. A, RA or VRA).

[0076] In some embodiments, the single chain polypeptide having IL-10 and CSF1 activities comprises or consists of sequence SEQ ID NO: 25, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains IL-10 and CSF1 activities.

[0077] In a second aspect, a single chain polypeptide having IL-10 and CSF1 activities is provided that comprises a single chain dimeric IL-10 fused to a CSF1 monomer.

[0078] In some embodiments, the single chain polypeptide having IL-10 and CSF1 activities comprises a single chain (SC) dimeric IL-10-CSF1 monomer fusion protein. In some embodiments, the single chain polypeptide having IL-10 and CSF1 activities comprises a CSF1 monomer-single chain dimeric IL-10 fusion protein. In some embodiments, the single chain polypeptide having IL-10 and CSF1 activities comprises a single chain CSF1 -single chain dimeric IL-10 fusion protein.

[0079] In some embodiments, the single chain dimeric IL-10 and CSF1 monomer are linked through a linker L1 that comprises from (or consists of) 5 to 50 amino acid residues, preferably 10 to 45 amino acid residues, preferably 12 to 40 amino acids, still preferably 15 to 40 amino acids, still preferably 20 to 35 amino acids, even more preferably 25 to 35 amino acids.In some embodiments, the linker L1 bridging the C-terminus of SC dimeric IL-10 to the N-terminus of CSF1, or bridging the N-terminus of SC dimeric IL-10 to the C-terminus of CSF1, is a (flexible) peptide sequence composed of Gly and Ser residues, in different proportions. The flexible linker L1 may comprise from (or consists of) 10 to 45 amino acid residues, preferably 12 to 40 amino acids, still preferably 15 to 40 amino acids, still preferably 20 to 35 amino acids, even more preferably 25 to 35 amino acids. Examples of suitable linkers comprise or consist of GGSGGSGGSGGSGGG (15 amino acid long, SEQ ID NO: 33), GGGSGGSGGSGGSGGSGGSGGSGGG (25 amino acid long, SEQ ID NO: 34), GGGGGSGGSGGSGGSGGSGGSGGSGGSGGG (30 amino acid long, SEQ ID NO: 35), or GGGGGSGGGGSGGSGGSGGSGGSGGSGGSGGSGGG (35 amino acid long, SEQ ID NO: 36). In some embodiments the L1 linker comprises or consists of a sequence (GGGGS, SEQ ID NO:82)n, n being a integer from 1 to 10, e.g. from 3 to 6.

[0080] Preferably, when the single chain polypeptide having IL-10 and CSF1 activities comprises a CSF1 monomer-single chain dimeric IL-10 fusion protein, wherein the linker L1 bridging the C-terminus of CSF1 to the N-terminus of SC dimeric IL-10 comprises more than 15 amino acids, preferably at least 20 or 25 amino acids, and optionally no more than 40 or 35 amino acids.

[0081] In some embodiments, the linker L1 bridging the C-terminus of SC dimeric IL-10 to the N-terminus of CSF1, or bridging the N-terminus of SC dimeric IL-10 to the C-terminus of CSF1, is a (rigid) peptide sequence composed of Glu, Ala, and Lys residues. Examples of suitable linkers comprise or consist of sequence EAAAK (SEQ ID NO: 247), or (EAAAK; SEQ ID NO: 247)nwith n being a integer from 2 to 10, preferably from 3 to 6, still preferably n = 6 (sequence SEQ ID NO: 248).

[0082] CSF1 domain

[0083] As used herein, the CSF1 domain denotes a CSF1 monomer or single chain dimeric CSF1.

[0084] The CSF1 monomer incorporated into the single chain polypeptide having IL-10 and CSF1 activities may be a wild-type CSF1 monomer, a fragment, or a mutant thereof. Preferably, CSF1 is human CSF1.

[0085] A canonical sequence of human CSF1 is available from the database UniProtKB under accession number P09603-1 (entry version 234 of 27 November 2024). The sequence P09603-1 of human CSF1 includes a signal peptide of 32 amino acids (SEQ ID NO: 112) and a 522 amino acid long mature polypeptide.

[0086] In some embodiments, the CSF1 monomer is a polypeptide that comprises or consists of SEQ ID NO: 12 (amino acids E1-D150 of mature human CSF1 protein, also referred toas “short version ending in QD” or “short version ending in FAECSSQD” in Table 1), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains CSF1 activity.

[0087] In some embodiments, the CSF1 monomer is a polypeptide that comprises or consists of SEQ ID NO: 245 (amino acids E1-V151 of mature human CSF1 protein, also referred to as “short version ending in QDV” in Table 1), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains CSF1 activity.

[0088] In some embodiments, the CSF1 monomer is a polypeptide that comprises or consists of SEQ ID NO: 17 (amino acids E1-R223 of mature human CSF1 protein, also referred to as “full length”) or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains CSF1 activity.

[0089] In some embodiments, the CSF1 monomer is a polypeptide that comprises or consists of a sequence differing from SEQ ID NO: 12, SEQ ID NO: 245, or SEQ ID NO: 17 by one or more mutations (and is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO: 12, SEQ ID NO: 245, or SEQ ID NO: 17). In some embodiments, the one or more mutations in SEQ ID NO: 12, SEQ ID NO: 245, or SEQ ID NO: 17 comprise or consist of (positions indicated based on mature human CSF1 protein sequence, or by reference to any one of SEQ ID NO: 12, SEQ ID NO: 245, or SEQ ID NO: 17):

[0090] a) A substitution at one or more of positions 5, 9, 18, 51, 54, 56, 58, 64, 65, 66, 74, 78, 81, 85, 86, 93, 100, 120, 122, 132 and 140;

[0091] b) S18I;

[0092] c) F54G;

[0093] d) L85I;

[0094] e) Q58E;

[0095] f) T64H;

[0096] g) M65Y;

[0097] h) R66Y;

[0098] i) A74D;

[0099] j) V78Y;

[0100] k) Q81Y

[0101] l) N122Q;

[0102] m) L56Y;

[0103] n) V120F;

[0104] o) E5R;p) H9Q;

[0105] q) R86Y;

[0106] r) K93E;

[0107] s) K100Y;

[0108] t) W132N;

[0109] u) K51Q;

[0110] v) F54N;

[0111] w) Q58F;

[0112] x) or any combination of mutations listed in b) to w).

[0113] In some embodiments the one or more mutations in SEQ ID NO: 12, SEQ ID NO: 245, or SEQ ID NO: 17 comprise or consist of substitution M65Y, as present in variant ORK1-532 (amino acids E1-V151 of mature human CSF1 protein with M65Y mutation, as shown in SEQ ID NO: 278).

[0114] In some embodiments, the one or more mutations in SEQ ID NO: 12, SEQ ID NO: 245, or SEQ ID NO: 17 comprise or consist of (positions indicated based on mature human CSF1 protein sequence SEQ ID NO: 12, SEQ ID NO: 245, or SEQ ID NO: 17):

[0115] a) A substitution at positions i) 18, 64, 65, 66, 85, and 120; ii) 5, 18, 66, 78, 85, 86, and 120; iii) 18, 64, 66, 78, 85, 86, and 120; iv) 18, 64, 74, 85, 86, and 120; v) 18, 64, 66, 85, and 86; vi) 18, 64, 66, 74, and 85; vii) 5, 64, 66, 85, and 86; viii) 18, 64, 66, 74, 85, 86, and 120; or ix) 18, 64, 66, 74, 86, and 120;

[0116] b) S18I, T64H, M65F, R66V, L85I, R86Y, V120F;

[0117] c) E5R, S18I, T64H, R66L, L85V, R86Y, V120F;

[0118] d) S18I, T64H, R66L, V78Y, L85I, R86Y, V120F;

[0119] e) S18I, T64H, R66L, L85I, R86Y, V120F;

[0120] f) S18I, T64H, R66Y, L85I, R86Y, V120F;

[0121] g) S18I, T64H, A74D, L85I, R86Y, V120F;

[0122] h) S18I, T64H, R66I, L85I, R86Y;

[0123] i) S18I, T64H, R66V, A74D, L85I;

[0124] j) E5R, T64H, R66L, L85I, R86Y;

[0125] k) S18I, T64H, R66V, A74D, L85V, R86Y, V120F;

[0126] l) S18I, T64H, R66I, A74D, L85V, R86Y, V120F;

[0127] m) S18I, T64H, R66V, A74D, L85I, R86Y, V120F;

[0128] n) S18I, T64H, R66I, A74D, L85I, R86Y, V120F;

[0129] o) S18I, T64H, R66V, A74D, R86Y, V120F, or

[0130] p) S18I, T64H, R66V, A74D, L85Y, R86Y, V120F.In some embodiments, the one or more mutations in SEQ ID NO: 12, SEQ ID NO: 245, or SEQ ID NO: 17 comprise or consist of substitutions:

[0131] S18I, T64H, R66V, A74D, L85V, R86Y, and V120F, as present in variant ORK1-576; S18I, T64H, R66V, A74D, L85I, R86Y, and V120F, as present in variant ORK1-578; or

[0132] S18I, T64H, R66I, A74D, L85I, R86Y, and V120F, as present in variant ORK1-579. In some embodiment the CSF1 monomer comprises a sequence that differs from SEQ ID NO: 12, SEQ ID NO: 245, or SEQ ID NO: 17 by no other mutation or substitution than the one defined above.

[0133] The dimeric single chain (SC) CSF1 incorporated into the single chain polypeptide having IL-10 and CSF1 activities may be a fusion protein comprising a first CSF1 monomer, a L1.2 peptide linker, and a second CSF1 monomer. In some embodiments the second CSF1 monomer is a CSF1 circular permutant.

[0134] A circular permutant is a protein in which the original ends have been joined together and new ends are created at a different location, essentially making the sequence start from a new point while keeping the same overall parts.

[0135] In some embodiments, the linker L1.2 has the same definition as the linker L1 above. In some embodiments, the linker L1.2 consists of sequence SEQ ID NO: 26.

[0136] In some embodiments the CSF1 circular permutant consists of sequence SEQ ID NO: 276, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains CSF1 activity.

[0137] According to an embodiment, the single chain dimeric CSF1 comprises or consists of sequence SEQ ID NO: 277, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains CSF1 activity.

[0138] In some embodiments, the single chain dimeric CSF1 is a polypeptide that comprises or consists of a sequence differing from SEQ ID NO: 277 by one or more mutations (and is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO: 277). In some embodiments, the one or more mutations in SEQ ID NO: 277 comprise or consist of (positions indicated by reference to SEQ ID NO: 277) a substitution at one or more of positions 5, 9, 18, 51, 54, 56, 58, 64, 65, 66, 74, 78, 81, 85, 86, 93, 100, 120, 122, 132 and 140. The substitution or combination of substitutions may be any of the substitutions defined for the CSF1 monomer. In a particular embodiment, the single chain dimeric CSF1 is a polypeptide that comprises or consists of a sequence differing from SEQ ID NO: 277 by substitution M65Y.In some embodiment the single chain dimeric CSF1 comprises a sequence that differs from SEQ ID NO: 277 by no other mutation or substitution than the one defined above.

[0139] Single chain dimeric IL- 10

[0140] The single chain (SC) dimeric IL-10 incorporated into the single chain polypeptide having IL-10 and CSF1 activities may be a fusion protein comprising a first IL-10 monomer fragment comprising at least α-helices A F of IL-10, a peptide linker, and a second IL-10 monomer fragment comprising at least α-helices A to F of IL-10.

[0141] The folding of the SC dimeric IL-10 is such that a ‘continuous 3D IL-10 domain’ is formed and a split 3D IL-10 domain is formed, the structure is disclosed in the international patent application published under number WO2023144393 A1 (see e.g. figure 28 of WO2023144393 A1).

[0142] A polypeptide sequence of mature (without Met1) wild-type human IL-10 is shown in SEQ ID NO: 11: SPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQ

[0143] hhh hhhhhhhhhhh hhhhhh hhhhhhhhhhhhhhhhhhh HA HB AENQD PDIKAHVNSLGENLKTLRLRL RRCHRFLPCENKS KAVEQVKNAFNKL QE KGIYKAMS E FDIFINYIEAYMTMKIRN

[0144] hhh hhhhhhhhhhhhhhhhhhh hhhhhhhhhhhh hhhhhhh hhhhhhhhhhhhhhh HC HD HE HF

[0145] Positions of the helices are represented by strings of ‘h’ in the above sequence. For α-helix F, depending of the structural assignments done by different software on different crystal structures of the same molecule, the helix consistently extends up to M154, and may extends up to M156or I158(characters underlined weavy). Accordingly, it is herein considered that α-helix F is entirely incorporated as long as the IL-10 monomer fragments comprise amino acid residues extending at least up to M154.

[0146] According to an embodiment, the SC dimeric IL-10 comprises or consists of sequence X1TQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQ hhh hhhhhhhhhhh hhhhhh hhhhhhhhhhhhhhhhhhh HA HB AENQX2PD IKAH VNSLGENLKTLRLRLRRCHRFLPCENKSKAVE QVKNAFNKLQE KGI YKAMS E FD I F I NY hhh hhhhhhhhhhhhhhhhhhh hhhhhhhhhhhh hhhhhhh hhhhhh HC HD HE HF IEAYX3(L1.1) ZNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEV hhhh hhhhhhhhhhh hhhhhh hhhhhhhhhhhhhhhh

[0147] HF HA HB

[0148] V

[0149]

[0150] MPQAENQDPDIKAHVNSLGENLKTLRLRLRRX4HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIF hhhhhh hhhhhhhhhhhhhhhhhhh hhhhhhhhhhhh hhhhhhh hhh HC HD HE IN Y IE A YMTMKIRN

[0151] hhhhhhhhhhhhHF

[0152] (sequence X1-SEQ ID NO: 65-X2-SEQ ID NO: 253-X3-L1.1-Z-SEQ ID NO: 66-X4-SEQ ID NO: 254),

[0153] wherein L1.1 is a peptide linker,

[0154] wherein X1 is absent or present, and where present it consists of M or sequence SPGQG (SEQ ID NO: 81),

[0155] wherein X2 is D (the residue naturally present at position 84 of SEQ ID NO:11) or E, wherein X3 is absent or present, and where present it consists in one or more amino acids of IL-10 sequence SEQ ID NO: 11 in continuity with the preceding amino acids on its N-terminal side, optionally with a mutation to accommodate the L1.1 linker,

[0156] wherein Z is absent or present, and where present it consists one or more amino acids of the IL-10 sequence SEQ ID NO: 11 in continuity with the preceding amino acids on its C-terminal side, optionally with a mutation to accommodate the L1.1 linker, wherein X4 is C (the residue naturally present in SEQ ID NO:11), A or N, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retain at least the same stability, and / or at least the same level of interaction with IL-10 receptor.

[0157] In some embodiments, X3 represents M, MT, MTM, MTMN (SEQ ID NO: 67), or MTMKIRN (SEQ ID NO: 76) that are present in the original IL- 10 sequence in continuity with the preceding amino acids on its N-terminal side. X3 may also represent L or K, i.e. mutation compared to the original IL-10 sequence to accommodate the L1.1 linker. Accordingly, X3 is preferably selected from the group consisting of M, MT, MTM, MTMN (SEQ ID NO: 67), MTMKIRN (SEQ ID NO: 76), L and K, still preferably it consists of MTMKIRN (SEQ ID NO: 76).

[0158] In some embodiments, Z represents LP that are present in the original IL- 10 sequence in continuity with the following amino acids on its C-terminal side. Z may also represent PEFA (SEQ ID NO: 68) or ELAZ is preferably selected from the group consisting of LP, PEFA (SEQ ID NO: 68) or ELA.

[0159] In some embodiments X2 is D, and X4 is C. In some embodiments X2 is E, and X4 is N. In some embodiments X2 is D, and X4 is N.

[0160] The L1.1 linker sequence bridging the C-terminus of one IL-10 monomer fragment and the N-terminus of the other IL- 10 monomer fragment may have any suitable sequence. It preferentially comprises a sequence that defines a conformationally restrained structure, i.e. a ‘structured’ linker. In this way, the engineered linker helps to conformationally restrain the 3D domain structures in an appropriate / desired structural orientation. Such linkers may have from about 3 to about 20 amino acid residues; from about 3 to about 16 amino acid residues; from about 4 to about 12 amino acid residues, from about 4 to about 8 amino acidresidues, from about 3 to 8 amino acid residues or from about 3 to 6 amino acid residues. Beneficially, such linkers do not contain a plurality of Gly and / or Ser residues adjacent each other; for example, beneficially the linker peptide contains 5 or less adjacent Gly and / or Ser residues; suitably 3 or less adjacent Gly and / or Ser residues; 2 or less Gly and / or Ser residues; contains only isolated Gly and / or Ser residues; or in some embodiments contains no Gly and / or Ser residues. In an embodiment, the L1.1 linker comprises from 3 to 20 amino acid residues and comprises no more than 2 adjacent Gly and / or Ser residues.

[0161] According to some embodiments, the L1.1 linker comprises sequence NGGLDY (SEQ ID NO: 30), FGGLDY (SEQ ID NO: 48), YKTIT (SEQ ID NO: 31), or DKDIRDGD (SEQ ID NO: 32).

[0162] The L1.1 linker may be preceded, at its N-terminal end, by X3, i.e. amino acid residue(s) that are naturally and consecutively present at position(s) 152 up to 157 of IL- 10 as shown in SEQ ID NO: 11. Alternatively, or additionally the L1.1 linker may be followed, at its C-terminal end, by Z, i.e. amino acid residue(s) that are naturally and consecutively present at position(s) 16 up to 20 of IL- 10 as shown in SEQ ID NO: 11.

[0163] Hence, according to some embodiments, the L1.1 linker and the X3 or Z sequence at its N-terminal side or C-terminal side (X3 and Z sequences in bold have the same structure as in IL10 and they correspond to the WT IL10 sequence or could have point mutations to accommodate the linker) comprises or consists of sequence NNGGLDYL (SEQ ID NO: 71), NFGGLDYL (SEQ ID NO: 45), LYKTITPEFA (SEQ ID NO: 46), KDKDIRDGDELA (SEQ ID NO: 47), MTMKIRNNNGGLDYLP (SEQ ID NO: 77), or MTMNNGGLDYLP (SEQ ID NO: 263). Thus, in some embodiments the SC dimeric IL-10 comprises or consists of sequence X1-SEQ ID NO: 65-X2-SEQ ID NO: 253-X3- L1.1-Z-SEQ ID NO: 66-X4-SEQ ID NO: 254, wherein X3- L1.1-Z consists of SEQ ID NO: 71, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 77, or SEQ ID NO: 263, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to X1-SEQ ID NO: 65-X2-SEQ ID NO: 253-X3- L1.1-Z-SEQ ID NO: 66-X4-SEQ ID NO: 254 and that retains at least the same stability, and / or at least the same level of interaction with IL-10 receptor.

[0164] In some embodiments, the single chain dimeric IL-10 comprises or consists of:

[0165] (i) SEQ ID NO: 1,

[0166] (ii) SEQ ID NO: 2 (‘ORK10-002’),

[0167] (iii) SEQ ID NO: 3 (‘ORK10-003’),

[0168] (iv) SEQ ID NO: 4 (‘ORK10-005’),

[0169] (v) SEQ ID NO: 22 (single chain dimeric IL-10 domain (IL10-linker(G3SG4S)- IL10), in e.g. ORK1-455 and ORK1-p-482),(vi) SEQ ID NO: 5 (sclL-10 with C108N (2nd IL10 domain) of e.g. ORK1_P_494, ORK1_P_532, ORK1_P_555, ORK1_P_587, or ORK1_P_510), (vii) SEQ ID NO: 255 (sclL-10 with D84E, C108N (2nd IL10 domain), Met1, short Nter of e.g. QRK1_P_400, ORK1_P_401, ORK1_P_405),

[0170] (viii) SEQ ID NO: 256 (sclL-10 with Met1, short Nter of e.g. ORK1_P_438), (ix) SEQ ID NO: 257 (sclL-10 with C108N (2nd IL10 domain), Met1, short Nter of e.g. ORK1_P_439),

[0171] (x) SEQ ID NO: 258 (sclL-10 with D84E, Met1, short Nter of e.g. ORK1_P_441), (xi) SEQ ID NO: 259 (sclL-10 with C108N (2nd IL10 domain), short N term of e.g.

[0172] ORK1_P_473),

[0173] (xii) SEQ ID NO: 261 (sclL-10 with C108A (2nd IL10 domain), short N term of e.g.

[0174] ORK1_P_475),

[0175] (xiii) SEQ ID NO: 262 (sclL-10 with C108A (2nd IL10 domain) of e.g.

[0176] ORK1_P_476),

[0177] (xiv) or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains IL-10 activity. In some embodiments, the single chain dimeric IL-10 comprises or consists of SEQ ID NO: 22, SEQ ID NO: 5, or SEQ ID NO: 1, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains IL-10 activity.

[0178] Resulting single chain polypeptide comprising a single chain dimeric IL- 10 fused to a CSF1 monomer and having IL-10 and CSF1 activities

[0179] According to some embodiments, the IL-10 domain is an IL-10 monomer and the single chain polypeptide having IL-10 and CSF1 activities comprises or consists of sequence SEQ ID NO: 25, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains IL-10 and CSF1 activities.

[0180] According to some embodiments, the single chain polypeptide having IL-10 and CSF1 activities comprises, in the N-terminus to C-terminus direction, a CSF1 monomer comprising or consisting of sequence SEQ ID NO:12, SEQ ID NO: 245, or SEQ ID NO: 17, or a single chain dimeric CSF1 comprising or consisting of sequence SEQ ID NO: 277, a peptide linker L1 as defined above, and a single chain dimeric IL-10 that comprises or consists of sequence X1 -SEQ ID NO: 65-X2-SEQ ID NO: 253-X3-(L1.1)-Z-SEQ ID NO: 66-X4-SEQ ID NO: 254, wherein X3-(L1.1)-Z consists of SEQ ID NO: 71, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 77, or SEQ ID NO: 263, or a sequence at least80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains IL- 10 and CSF1 activities.

[0181] According to some embodiments, the single chain polypeptide having IL-10 and CSF1 activities comprises or consists of sequence SEQ ID NO: 14, or SEQ ID NO: 19, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains IL-10 and CSF1 activities.

[0182] In some embodiments, the single chain polypeptide having IL-10 and CSF1 activities comprises or consists of a sequence as present in the sequences shown in Table 1, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains IL-10 and CSF1 activities.

[0183] According to some embodiments, the single chain polypeptide having IL-10 and CSF1 activities comprises or consists of sequence as present in any one of the ORK1 constructs disclosed in Tablel, in particular in any one of ORK1-P-405, ORK1-P-494, ORK1-P-513, ORK1-P-515, ORK1-P-532, ORK1-P-555, ORK1-P-556, ORK1-P-563, ORK1-P-564, ORK1-P-566, ORK1-P-576, ORK1-P-578, ORK1-P-579, ORK1-P-587, ORK1-P-590, ORK1-P-591, ORK1-P-592, ORK1-P-510, and ORK1-P-512.

[0184] For instance, the single chain polypeptide having IL-10 and CSF1 activities comprises or consists of sequence SEQ ID NO: 79 (as e.g. in ORK1_P_485), SEQ ID NO: 80, SEQ ID NO: 105 (as in e.g. ORK1_P_455 or ORK1_P_482), SEQ ID NO: 264 (as in e.g. ORK1_P_494, and ORK1_P_513), SEQ ID NO: 265 (as in e.g. ORK1_P_555, and ORK1_P_556), SEQ ID NO: 266 (as in e.g. ORK1_P_532, and ORK1_P_589), SEQ ID NO: 267 (as in e.g. ORK1_P_587), SEQ ID NO: 268 (as in e.g. ORK1_P_515), SEQ ID NO: 269 (as in e.g. ORK1_P_512), SEQ ID NO: 270 (as in e.g. ORK1_P_563,. ORK1_P_564, and ORK1_P_566), SEQ ID NO: 273 (as in e.g. ORK1_P_576), SEQ ID NO: 274 (as in e.g. ORK1_P_578), SEQ ID NO: 275 (as in e.g. ORK1_P_579), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains IL-10 and CSF1 activities.

[0185] According to some embodiments, the single chain polypeptide having IL-10 and CSF1 activities comprises or consists of sequence SEQ ID NO: 264, SEQ ID NO: 265, SEQ ID NO: 266, SEQ ID NO: 267, SEQ ID NO: 268, SEQ ID NO: 269 or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains IL- 10 and CSF1 activities.Protein comprising the single chain polypeptide having IL-10 and CSF1 activities and a moiety increasing half-life

[0186] The invention thus relates to a protein comprising a single chain polypeptide having IL-10 and CSF1 activities fused to an immunoglobulin Fc fragment, or to albumin, or albumin fragment, or to an anti-albumin antibody, or anti-albumin antibody fragment, or conjugated with a poly(ethylene glycol) (PEG) molecule.

[0187] In some embodiments, in the protein, the single chain polypeptide having IL-10 and CSF1 activities comprises in the N-terminus to C-terminus direction, a CSF1 domain which is a CSF1 monomer or a single chain dimeric CSF1, a peptide linker L1, and an IL-10 domain which is an IL-10 monomer or a single chain dimeric IL-10. In some aspects the IL-10 domain is a single chain dimeric IL-10.

[0188] The single chain polypeptide having IL-10 and CSF1 activities, IL-10 monomer, single chain dimeric IL-10, CSF1 monomer, single chain dimeric CSF1, and peptide linker L1 are as disclosed above.

[0189] In some embodiments, the IL-10 domain is an IL-10 monomer as defined above. In some embodiments, the single chain polypeptide having IL-10 and CSF1 activities comprises or consists for instance sequence SEQ ID NO: 25, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% identical thereto and that retains IL- 10 and CSF1 activities.

[0190] In some embodiments, the IL-10 domain is a single chain dimeric IL-10 as defined above. In some embodiments, the single chain dimeric IL-10 comprises or consists of sequence X1-SEQ ID NO: 65-X2-SEQ ID NO: 253-X3-(L1.1)-Z-SEQ ID NO: 66-X4-SEQ ID NO: 254, as defined above, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retain at least the same stability, and / or at least the same level of interaction with IL- 10 receptor, as defined above.

[0191] In some embodiments, the CSF1 monomer comprises or consists of SEQ ID NO: 12, SEQ ID NO: 245, or SEQ ID NO: 17, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains CSF1 activity, as define above. In some embodiments, the single chain CSF1 comprises or consists of SEQ ID NO:246, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains CSF1 activity, as define above.

[0192] In some embodiments, the single chain polypeptide having IL-10 and CSF1 activities comprises or consists of a sequence as present in the sequences shown in Table 1. According to some embodiments, the single chain polypeptide having IL-10 and CSF1activities comprises or consists of sequence as present in any one of the ORK1 constructs disclosed in Tablel, in particular in any one of ORK1-P-405, ORK1-P-494, ORK1-P-513, ORK1-P-515, ORK1-P-532, ORK1-P-555, ORK1-P-556, ORK1-P-563, ORK1-P-564, ORK1-P-566, ORK1-P-576, ORK1-P-578, ORK1-P-579, ORK1-P-587, ORK1-P-590, ORK1-P-591, ORK1-P-592, ORK1-P-510, and ORK1-P-512.

[0193] In some embodiments, the single chain polypeptide having IL-10 and CSF1 activities comprises or consists of sequence SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 105, or any one of SEQ ID NO: 264 to SEQ ID NO: 269 and SEQ ID NO: 273 to SEQ ID NO: 275, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains IL-10 and CSF1 activities, as defined above.

[0194] In some embodiments, the protein comprises a single chain polypeptide having IL-10 and CSF1 activities and an immunoglobulin Fc fragment, preferably a human immunoglobulin Fc fragment. In some embodiments, the immunoglobulin Fc fragment is a human lgG1 or lgG4 Fc fragment. In some embodiments, the protein comprises IL-10 domain, CSF1 -domain, L1 and L2 linkers, and dimerized Fc fragment structurally organized as shown in any one of the protein formats ORK1_Fc_01 to ORK1_Fc_12 shown in Figures 2 and 21.

[0195] In some embodiments, the protein comprises a single chain polypeptide having IL-10 and CSF1 activities and albumin or a fragment thereof, preferably human albumin, or a fragment thereof. In some embodiments, the protein comprises a single chain polypeptide having IL-10 and CSF1 activities and an anti-albumin antibody, or anti-albumin antibody fragment.

[0196] In some embodiments, the protein comprises a single chain polypeptide having IL-10 and CSF1 activities conjugated to PEG. Process for protein pegylation has been reviewed for instance by Pfister and Morbidelli (Journal of Controlled Release, Volume 180, 2014, Pages 134-149).

[0197] In some embodiments, the protein comprises or consists of any one of the sequences identified in the column “Protein sequence 1 ID” in Table 1. Where appropriate the protein comprises any one of the sequences identified in the column “Protein sequence 1 ID” and “Protein sequence 2 ID” in Table 1.

[0198] According to some embodiments, the protein comprises “Protein sequence 1 ID” as present in any one of the ORK1 constructs disclosed in Tablel, in particular in any one of ORK1-P-405, ORK1-P-494, ORK1-P-513, ORK1-P-515, ORK1-P-532, ORK1-P-555, ORK1-P-556, ORK1-P-563, ORK1-P-564, ORK1-P-566, ORK1-P-576, ORK1-P-578,ORK1-P-579, ORK1-P-587, ORK1-P-590, ORK1-P-591, ORK1-P-592, ORK1-P-510, and ORK1-P-512.

[0199] Protein comprising one single chain polypeptide having IL- 10 and CSF1 activities and one immunoglobulin Fc fragment

[0200] In some embodiments, the protein comprises one SC polypeptide having IL-10 and CSF1 activities and one immunoglobulin Fc fragment.

[0201] In this aspect, the protein comprises a first polypeptide comprising the SC polypeptide having dual IL-10 and CSF1 activities fused to a first immunoglobulin Fc fragment (Fc1) through a peptide linker L2, and a second polypeptide comprising a second immunoglobulin Fc fragment (Fc2).

[0202] In some embodiments, the protein comprises one SC polypeptide having IL-10 and CSF1 activities and one immunoglobulin Fc fragment, wherein the SC polypeptide having IL-10 and CSF1 activities is fused to the immunoglobulin Fc fragment through a peptide linker L2 connecting the C-terminus of the IL-10 domain with the N-terminus of one hinge region of the Fcfragment. In some aspects the IL-10 domain is a SC dimeric IL-10. In some aspects the CSF1 domain is a SC dimeric CSF1.

[0203] According to some embodiments the protein then comprises two polypeptide chains comprising or consisting of formula (I) (or format ORK1_Fc_05 as shown in Figures 2 and 21):

[0204] i) CSF1 monomer-L1-IL-10 domain-L2-Fc1, wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and

[0205] ii) Fc2, wherein Fc2 is a Fc domain comprising hinge-CH2-CH3,

[0206] wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains. In some embodiments, the protein comprises two polypeptide chains comprising or consisting of (format ORK1_Fc_11 as shown in Figure 21):

[0207] i) dimeric single chain CSF1-L1-IL-10 domain-L2-Fc1, wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and

[0208] ii) Fc2, wherein Fc2 is a Fc domain comprising hinge-CH2-CH3,

[0209] wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains. In some embodiments, the IL-10 domain is a dimeric single chain IL-10 polypeptide.

[0210] In some embodiments, the protein comprises one SC polypeptide having IL-10 and CSF1 activities and one immunoglobulin Fc fragment, wherein the SC polypeptide having IL-10 and CSF1 activities is fused to the immunoglobulin Fc fragment through a peptidelinker L2 connecting the C-terminus of the one of the CH3 domains of the Fc fragment with the N-terminus of the CSF1 monomer. In some aspects the IL-10 domain is a SC dimeric IL- 10. According to these embodiments the protein then comprises two polypeptide chains comprising or consisting of formula (II) (or format ORK1_Fc_06 as shown in Figures 2 and 21):

[0211] i) Fc1-L2-CSF1 monomer-L1-IL-10 domain, wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and

[0212] ii) Fc2, wherein Fc2 is a Fc domain comprising hinge-CH2-CH3,

[0213] wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains.

[0214] In some embodiments, the protein comprises one SC polypeptide having IL-10 and CSF1 activities and one immunoglobulin Fc fragment, wherein the SC polypeptide having IL-10 and CSF1 activities is fused to the immunoglobulin Fc fragment through a peptide linker L2 connecting the C-terminus of the one of the CH3 domains of the Fc fragment with the N-terminus of the IL-10 domain. In some aspects the IL-10 domain is a SC dimeric IL-10. According to these embodiments the protein then comprises two polypeptide chains comprising or consisting of:

[0215] i) Fc1-L2-IL-10 domain-CSF1 monomer-L1, wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and

[0216] ii) Fc2, wherein Fc2 is a Fc domain comprising hinge-CH2-CH3,

[0217] wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains. In some embodiments, the IL-10 domain is a dimeric single chain IL-10 polypeptide (format ORK1_Fc_09 as shown in Figures 21).

[0218] Further disclosed is a protein having dual CSF1 and IL-10 activities. In some embodiments, the protein comprises two polypeptide chains comprising or consisting of (format ORK1_Fc_08):

[0219] i) CSF1 monomer-L2-Fc1, wherein Fc1 is a Fc domain comprising hinge-CH2- CH3, and

[0220] ii) IL-10 domain -L2-Fc2, wherein Fc2 is a Fc domain comprising hinge-CH2- CH3,

[0221] wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains. The linkers L2 in i) and ii) are defined independently, and may be identical or different. CSF1 monomer, IL-10 domain, L1, L2, Fc1 and F2 are as defined in the present disclosure.

[0222] In some embodiments, the IL-10 domain is a dimeric single chain IL-10.In some embodiments, the linker L2 connecting the single chain polypeptide having IL-10 and CSF1 activities and the immunoglobulin Fc fragment is preferably a peptide sequence that comprises from, or consists of, 3 to 45 amino acid residues, preferably 5 to 40 amino acid residues, preferably 5 to 25 amino acids, still preferably 10 to 25 amino acids.

[0223] The linker is usually rich in glycine for flexibility, as well as serine or threonine for solubility, but linkers comprising amino acids randomly selected from the group consisting of valine, leucine, isoleucine, serine, threonine, lysine, arginine, histidine, aspartate, glutamate, asparagine, glutamine, glycine, and proline may also be suitable. A well-suited linker L2 according to the present disclosure contains or consists of glycine and serine residues and is for example of the format (GGGGS)p, wherein p is an integer from 1 to 8, notably from 1 to 4, advantageously 2, 3 or 4. In some embodiments, the linker L2 is (GGGGS)3(SEQ ID NO: 119). In some embodiments, the linker is selected from the group consisting of the amino acid sequences GGGGGSGGSGGSGGSGGSGGSGGSGGSGGG (SEQ ID NO: 35), SGGGGSGGGGS (SEQ ID NO: 120), SGGGGSGGGGSAP (SEQ ID NO: 121), NFSQP (SEQ ID NO: 122), KRTVA (SEQ ID NO: 123), GGGSGGGG (SEQ ID NO: 124), GGGGSGGGGS (SEQ ID NO: 125), GGGGSGGGGSGGGGS (SEQ ID NO: 119), THTCPPCPEPKSSDK (SEQ ID NO: 126), GGGS (SEQ ID NO: 127), EAAKEAAKGGGGS (SEQ ID NO: 128), EAAKEAAK (SEQ ID NO: 129), GGGGS (SEQ ID NO: 82), (SG)m where m is an integer comprised between 1 and 7, GGSSGSGSGSTGTSSSGTGTSAGTTGTSASTSGSGSGGGGGSGGGGSAGG (SEQ ID NO: 130), GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 131), GGGGGSGGGGSGGGGS (SEQ ID NO: 136), GGGGSGGGGSGGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 279), EAAAK (SEQ ID NO: 247), or (EAAAK; SEQ ID NO: 247)nwith n being a integer from 2 to 10, preferably from 3 to 6, still preferably n = 6 (sequence SEQ ID NO: 248).

[0224] In some embodiments the linker L1 comprises or consist of sequence SEQ ID NO: 35, and the linker L2 comprises or consist of sequence SEQ ID NO: 136.

[0225] In some embodiments the linker L1 comprises or consist of sequence SEQ ID NO: 82, and the linker L2 comprises or consist of sequence SEQ ID NO: 35, or SEQ ID NO: 136.

[0226] In some embodiments the linker L1 comprises or consist of sequence SEQ ID NO: 35, and the linker L2 comprises or consist of sequence SEQ ID NO: 136, SEQ ID NO: 247, or SEQ ID NO: 248.In some embodiments the linker L1 comprises or consist of sequence SEQ ID NO: 248, and the linker L2 comprises or consist of sequence SEQ ID NO: 35, SEQ ID NO: 82, SEQ ID NO: 247, or SEQ ID NO: 248.

[0227] In some embodiments, the protein comprises SEQ ID NO: 87 (as in e.g. ORK1_P_404, format ORK1 -Fc-05) or SEQ ID NO: 88 (as in e.g. ORK1_P_405, format ORK1-Fc-06), SEQ ID NO: 168 (as in e.g. ORK1_P_516, format ORK1-Fc-09), or SEQ ID NO: 161 ( format ORK1-Fc-07, ORK1-509, or ORK1-Fc-08, ORK1-510) or SEQ ID NO: 159(ORK1 -Fc-09, ORK1-507), or SEQ ID NO: 164 (ORKI-Fc-11, ORK1-512), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains IL-10 and CSF1 activities.

[0228] In some embodiments, the protein comprises a Fc1 domain comprising or consisting of SEQ ID NO: 149, SEQ ID NO: 20, SEQ ID NO: 137, SEQ ID NO: 140, SEQ ID NO: 141, SEQ ID NO: 142, or SEQ ID NO: 143.

[0229] In some embodiments, the protein comprises a Fc2 domain comprising or consisting of SEQ ID NO: 109, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: 148 or SEQ ID NO: 149.

[0230] In some embodiments, the protein comprises a combination of Fc1 domain and Fc2 domain as disclosed for any of the Protein 1 and Protein 2 of Table 1 that are structurally organised according to format ORK1_Fc_05, format ORK1_Fc_06, or format ORK1_Fc_09. For instance, Fc1 domain comprises or consists of SEQ ID NO: 20, and Fc2 domain comprises or consists of SEQ ID NO: 109; or Fc1 domain comprises or consists of SEQ ID NO: 139, and Fc2 domain comprises or consists of SEQ ID NO: 144; or Fc1 domain comprises or consists of SEQ ID NO: 140, and Fc2 domain comprises or consists of SEQ ID NO: 145.

[0231] In some embodiments, the protein comprises:

[0232] a) a first polypeptide chain comprising any one of SEQ ID NO: 87 to SEQ ID NO:

[0233] 104, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto, and

[0234] b) a second polypeptide chain comprising SEQ ID NO: 109, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto,

[0235] wherein the first and second polypeptides dimerise, and

[0236] wherein said protein retains IL-10 and CSF1 activities.

[0237] In some embodiments, the protein comprises:a) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 88 (as in e.g. QRK1_P_405), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto;

[0238] b) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 101 (as in e.g. ORK1_P_494), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto;

[0239] c) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 161 (as in e.g. ORK1_P_510), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto;

[0240] d) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 164 (as in e.g. ORK1_P_512), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto;

[0241] e) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 165 (as in e.g. ORK1_P_513), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto;

[0242] f) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 167 (as in e.g. ORK1_P_515), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto;

[0243] g) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 184 (as in e.g. ORK1_P_532), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto;

[0244] h) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 187 (as in e.g. ORK1_P_535), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto;

[0245] i) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 207 (as in e.g. ORK1_P_555), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto;

[0246] j) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 208 (as in e.g. ORK1_P_556), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto;

[0247] k) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 215 (as in e.g. ORK1_P_563), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto;

[0248] l) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 216 (as in e.g. ORK1_P_564), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto;m) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 218 (as in e.g. ORK1_P_566), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto;

[0249] n) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 228 (as in e.g. ORK1_P_576), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto;

[0250] o) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 230 (as in e.g. ORK1_P_578), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto;

[0251] p) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 231 (as in e.g. ORK1_P_579), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto;

[0252] q) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 236 (as in e.g. ORK1_P_587), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto;

[0253] r) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 239 (as in e.g. ORK1_P_590), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto;

[0254] s) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 240 (as in e.g. ORK1_P_591), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto; or

[0255] t) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 241 (as in e.g. ORK1_P_592), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto.

[0256] Advantageously, the first polypeptide retains IL-10 and CSF1 activities.

[0257] In some embodiments, the second polypeptide comprises or consists of SEQ ID NO: 109. For instance, SEQ ID NO 165, 167, 184, 187, 207, 208, 215, 216, 218, 228, 230, 231, 236, corresponding to the knob domain of ORK1_P_513, 515, 532, 535, 555, 556, 563, 564, 566, 576, 578, 579, 587 respectively, pair with SEQ ID NO: 109, corresponding to the hole Fc domain, to form the complete molecule.

[0258] In some embodiments, the protein comprises:

[0259] a) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 88, and a second polypeptide comprising or consisting of sequence SEQ ID NO: 109 (as in e.g. ORK1_P_405);b) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 101, and a second polypeptide comprising or consisting of sequence SEQ ID NO: 109 (as in e.g. ORK1_P_494);

[0260] c) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 161, and a second polypeptide comprising or consisting of sequence SEQ ID NO: 243 (as in e.g. ORK1_P_510);

[0261] d) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 164, and a second polypeptide comprising or consisting of sequence SEQ ID NO: 109 (as in e.g. ORK1_P_512),

[0262] e) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 165, and a second polypeptide comprising or consisting of sequence SEQ ID NO: 109 (as in e.g. ORK1_P_513);

[0263] f) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 167, and a second polypeptide comprising or consisting of sequence SEQ ID NO: 109 (as in e.g. ORK1_P_515);

[0264] g) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 184, and a second polypeptide comprising or consisting of sequence SEQ ID NO: 109 (as in e.g. ORK1_P_532);

[0265] h) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 187, and a second polypeptide comprising or consisting of sequence SEQ ID NO: 109 (as in e.g. ORK1_P_535);

[0266] i) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 207, and a second polypeptide comprising or consisting of sequence SEQ ID NO: 109 (as in e.g. ORK1_P_555);

[0267] j) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 208, and a second polypeptide comprising or consisting of sequence SEQ ID NO: 109 (as in e.g. ORK1_P_556);

[0268] k) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 215, and a second polypeptide comprising or consisting of sequence SEQ ID NO: 109 (as in e.g. ORK1_P_563);

[0269] l) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 216, and a second polypeptide comprising or consisting of sequence SEQ ID NO: 109 (as in e.g. ORK1_P_564);

[0270] m) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 218, and a second polypeptide comprising or consisting of sequence SEQ ID NO: 109 (as in e.g. ORK1_P_566);n) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 228, and a second polypeptide comprising or consisting of sequence SEQ ID NO: 109 (as in e.g. ORK1_P_576);

[0271] o) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 230, and a second polypeptide comprising or consisting of sequence SEQ ID NO: 109 (as in e.g. ORK1_P_578);

[0272] p) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 231, and a second polypeptide comprising or consisting of sequence SEQ ID NO: 109 (as in e.g. ORK1_P_579); or

[0273] q) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 236, and a second polypeptide comprising or consisting of sequence SEQ ID NO: 109 (as in e.g. ORK1_P_587);

[0274] r) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 239, and a second polypeptide comprising or consisting of sequence SEQ ID NO: 109 (as in e.g. ORK1_P_590);

[0275] s) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 240, and a second polypeptide comprising or consisting of sequence SEQ ID NO: 109 (as in e.g. ORK1_P_591); or

[0276] t) a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 241, and a second polypeptide comprising or consisting of sequence SEQ ID NO: 109 (as in e.g. ORK1_P_592);

[0277] wherein the first and second polypeptides dimerise and the protein retains IL-10 and CSF1 activities.

[0278] More generally, according to some embodiments, the protein comprises a combination of first (‘Protein 1’) and second (‘Protein 2’) polypeptides as disclosed for any one of ORK1 constructs of Table 1 that are structurally organised according to format ORK1_Fc_05, format ORK1_Fc_06, or format ORK1_Fc_09.

[0279] Protein comprising two single chain polypeptides having IL-10 and CSF1 activities and one immunoglobulin Fc fragment

[0280] In some embodiments, the protein comprises two SC polypeptides having IL-10 and CSF1 activities and one immunoglobulin Fc fragment. In this aspect, the protein comprises two polypeptide chains comprising the SC polypeptide having dual IL-10 and CSF1 activities fused to a Fc fragment through a peptide linker L2.

[0281] Preferably, the two SC polypeptides having IL-10 and CSF1 activities are identical.In some embodiments, the protein comprises two polypeptide chains that each comprise, in the N-terminus to C-terminus direction, a CSF1 monomer, a peptide linker L1, an IL-10 domain which is an IL-10 monomer or a SC dimeric IL-10 monomer, a peptide linker L2, and a Fc domain, wherein the two polypeptide chains dimerise through their Fc domains, thereby forming a Fc fragment. In some embodiments the peptide linker L2 connects the C-terminus of the IL-10 domain with the hinge of the Fc domain; the protein then comprises two polypeptide chains comprising or consisting of formula (III) (format ORK1_Fc_02 or ORK1_Fc_04 as shown in Figure 2):

[0282] CSF1 monomer-L1-IL-10 domain-L2-Fc1 wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and

[0283] CSF1 monomer-L1-IL-10 domain-L2-Fc2 wherein Fc2 is a Fc domain comprising hinge-CH2-CH3,

[0284] wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains.

[0285] In some embodiments, the protein comprises two polypeptide chains that each comprise, in the N-terminus to C-terminus direction, a Fc domain, a peptide linker L2, a CSF1 monomer, a peptide linker L1, and an IL-10 domain which is an IL-10 monomer or a SC dimeric IL-10 monomer, wherein the two polypeptide chains dimerise through their Fc domains, thereby forming a Fc fragment. In some embodiments the peptide linker L2 connects the C-terminus of the Fc fragment with the N-terminus of the CSF1 monomer; the protein then comprises two polypeptide chains comprising or consisting of formula (IV) (format ORK1_Fc_01 or ORK1_Fc_03 as shown in Figure 2):

[0286] Fc1-L2-CSF1 monomer-L1-IL-10 domain, wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and

[0287] Fc2-L2-CSF1 monomer-L1-IL-10 domain, wherein Fc2 is a Fc domain comprising hinge-CH2-CH3

[0288] wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains.

[0289] In some embodiments, the linker L2 connecting the single chain polypeptide having IL-10 and CSF1 activities and the immunoglobulin Fc fragment is preferably a peptide sequence that comprises from, or consists of, 3 to 45 amino acid residues, preferably 5 to 40 amino acid residues, preferably 5 to 25 amino acids, still preferably 10 to 25 amino acids.

[0290] The linker is usually rich in glycine for flexibility, as well as serine or threonine for solubility, but linkers comprising amino acids randomly selected from the group consisting of valine, leucine, isoleucine, serine, threonine, lysine, arginine, histidine, aspartate, glutamate, asparagine, glutamine, glycine, and proline may also be suitable. A well-suitedlinker L2 according to the present disclosure contains or consists of glycine and serine residues and is for example of the format (GGGGS)p, wherein p is an integer from 1 to 8, notably from 1 to 4, advantageously 2, 3 or 4. In some embodiments, the linker L2 is (GGGGS)3(SEQ ID NO: 119). In some embodiments, the linker is selected from the group consisting of the amino acid sequences SGGGGSGGGGS (SEQ ID NO: 120), SGGGGSGGGGSAP (SEQ ID NO: 121), NFSQP (SEQ ID NO: 122), KRTVA (SEQ ID NO: 123), GGGSGGGG (SEQ ID NO: 124), GGGGSGGGGS (SEQ ID NO: 125), GGGGSGGGGSGGGGS (SEQ ID NO: 119), THTCPPCPEPKSSDK (SEQ ID NO: 126), GGGS (SEQ ID NO: 127), EAAKEAAKGGGGS (SEQ ID NO: 128), EAAKEAAK (SEQ ID NO: 129), GGGGS (SEQ ID NO: 82), (SG)m where m is an integer comprised between 1 and 7, GGSSGSGSGSTGTSSSGTGTSAGTTGTSASTSGSGSGGGGGSGGGGSAGG (SEQ ID NO: 130), GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 131) GGGGGSGGGGSGGGGS (SEQ ID NO: 136), EAAAK(SEQ ID NO: 247), or(EAAAK; SEQ ID NO: 247)nwith n being a integer from 2 to 10, preferably from 3 to 6, still preferably n = 6 (sequence SEQ ID NO: 248).

[0291] In some embodiments the linker L1 comprises or consist of sequence SEQ ID NO: 35, and the linker L2 comprises or consist of sequence SEQ ID NO: 136.

[0292] In some embodiments the linker L1 comprises or consist of sequence SEQ ID NO: 82, and the linker L2 comprises or consist of sequence SEQ ID NO: 35, or SEQ ID NO: 136.

[0293] In some embodiments the linker L1 comprises or consist of sequence SEQ ID NO: 35, and the linker L2 comprises or consist of sequence SEQ ID NO: 136, SEQ ID NO: 247, or SEQ ID NO: 248.

[0294] In some embodiments the linker L1 comprises or consist of sequence SEQ ID NO: 248, and the linker L2 comprises or consist of sequence SEQ ID NO: 35, SEQ ID NO: 82, SEQ ID NO: 247, or SEQ ID NO: 248.

[0295] In some embodiments, in the protein, the Fc1 and Fc2 domains are identical.

[0296] In some embodiments, the protein comprises two identical polypeptide chains.

[0297] In some embodiments, the protein comprises sequence on the protein formats SEQ ID NO: 83 (ORK1-P-400), SEQ ID NO: 84 (ORK1-P-401), SEQ ID NO: 85 (ORK1-P-402), SEQ ID NO: 86 (ORK1-P-403), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains IL-10 and CSF1 activities.Fc fragment or Fc domain

[0298] In some embodiments, the immunoglobulin Fc fragment or Fc domain is of the isotype IgG, such as lgG1, lgG2, lgG3, and lgG4, preferably lgG1 or lgG4. In some embodiments, the immunoglobulin Fc fragment or Fc domain is an engineered Fc fragment or Fc domain.

[0299] Accordingly, in some embodiments the protein comprises a Fc fragment that comprises two polypeptides that comprise a hinge region, an IgG CH2 domain and an IgG CH3 domain. Accordingly, in some embodiments the protein comprises two polypeptide chains that each comprise a Fc domain that comprise a hinge region, an IgG CH2 domain and an IgG CH3 domain.

[0300] Preferably the Fc fragment is silenced, i.e. the Fc fragment is mutated to reduce or eliminate binding to Fc gamma receptors and / or complement protein C1 q and thereby reduce or abolish immune effector functions.

[0301] In certain embodiments, a human IgG heavy chain Fc fragment or domain extends from Cys226 to the carboxyl-terminus of the heavy chain. However, the C-terminal lysine (Lys447) of the Fc fragment or domain may or may not be present, without affecting the structure or stability of the Fc fragment. Unless otherwise specified herein, numbering of amino acid residues in the IgG or Fc region is according to the EU numbering system for antibodies, also called the EU index, as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991.

[0302] In certain embodiments, Fc fragment or domain refers to an immunoglobulin IgG heavy chain constant region comprising a hinge region (starting at Cys226), an IgG CH2 domain and CH3 domain. The term “hinge region” or “hinge sequence” as used herein refers to the amino acid sequence that is, in some instance, located between the linker L2 and the CH2 domain. In certain embodiments, the hinge region comprises the amino acid sequence CPPCP (SEQ ID NO: 106), a sequence found in the native lgG1 or lgG2 hinge region at positions 239-243, sequence CPRCP (SEQ ID NO: 107), a sequence found in the native lgG3 hinge region at positions 239-241 B, sequence CPSCP (SEQ ID NO: 108), a sequence found in the native lgG4 hinge region at positions 239-243, to facilitate dimerization. In certain other embodiments, the Fc fragment or domain starts at the hinge region and extends to the C-terminus of the IgG heavy chain. In certain particular embodiments, the Fc fragment or domain comprises the Fc fragment or domain of human lgG1, lgG2, lgG3 or lgG4. In certain particular embodiments, the Fc fragment or domain comprises the CH2 and CH3 domain of lgG4. In certain other particular embodiments, the Fc fragment or domain comprises the CH2 and CH3 domain of lgG1. In certainembodiments, the IgG CH2 domain starts at Ala 231. In certain other embodiments, the CH3 domain starts at Gly 341. It is understood that the C-terminus Lys residue of human IgG can be optionally absent. In some embodiments, the Fc fragment or domain is from human lgG1 heavy chain as shown in SEQ ID NO: 249. The lgG1 Fc fragment typically comprises or consists of sequence SEQ ID NO: 250, or amino acids 1-226 or 1-225 of SEQ ID NO: 250 (Fc with absent C-terminal Lys, or Gly-Lys), a silenced variant thereof.

[0303] In certain embodiments, the Fc fragment is a human lgG4 Fc comprising substitution S228P in the hinge region and R409K in the CH3 region.

[0304] In certain embodiments, the Fc fragment is engineered to modulate Fc mediated antibody functions. In some embodiments, the Fc fragment of the present disclosure does not induce antibody dependent cellular cytotoxicity (ADCC) and / or is an “Fc silent” antibody.

[0305] Examples of silent lgG4 Fc fragments comprise mutations at positions 234, 235, 236, 237, 238, 265 and 329 in the lgG4 Fc amino acid sequence (Ell numbering). For example, lgG4 Fc silent fragment may include any one of the following mutations or combinations of mutations: F234A / L235A (FALA); L235E; F234A / L235A / G237A / P238S; and F234S / L235T / G236R (STR).

[0306] Examples of silent IgG1 Fc fragments comprise mutations at positions 234, 235, 236, 239, 265, 297, 329 and / or 331 in the IgG1 Fc amino acid sequence (EU numbering), corresponding to positions 14, 15, 16, 19, 45, 77, 109 and / or 112 of SEQ ID NO: 250. Another silent lgG1 Fc fragment comprises the N297A mutation, which results in aglycosylated or non-glycosylated Fc fragments. For example, lgG1 Fc silent fragment may include any one of the following mutations or combinations of mutations: D265A / P329A; L234F / L235Q / K322Q (FQQ); L234A / L235A (LALA); L234A / L235A / K322A (LALAKA); N297Q (aglycosyl); L234F / L235E / P331S (FES); L234A / L235A / G237A (LALAGA); L234A / L235E; L234A / L235A / G237A / P238S / H268A / A330S / P331 S; L234A / L235A / P329G (LALAPG), e.g. an lgG1 Fc comprising or consisting of SEQ ID NO: 139 or SEQ ID NO: 144; F234S / L235T / G236R (STR), e.g. an lgG1 Fc comprising or consisting of SEQ ID NO: 142, SEQ ID NO: 147, or SEQ ID NO: 252; L234A / L235A / P329S (LALAPS), e.g. an lgG1 Fc comprising or consisting of SEQ ID NO: 140 or SEQ ID NO: 145; L234A / L235A / D265 (LALADS), e.g. an lgG1 Fc comprising or consisting of SEQ ID NO: 141 or SEQ ID NO: 146; and G236R / L328R.

[0307] For certain proteins whose Fc Region-containing first and second polypeptide chains are not identical, it is desirable to reduce or prevent homodimerization from occurring between the CH2-CH3 domains of two first polypeptide chains or between the CH2-CH3 Domains of two third polypeptide chains. The CH2 and / or CH3 domains of such polypeptidechains need not to be identical in sequence, and advantageously are modified to foster complexing between the two polypeptide chains. For example, an amino acid substitution (preferably a substitution with an amino acid comprising a bulky side group forming a "knob", e.g., tryptophan) can be introduced into the CH2 or CH3 Domain such that steric interference will prevent interaction with a similarly mutated domain and will obligate the mutated domain to pair with a domain into which a complementary, or accommodating mutation has been engineered, i.e., "the hole" (e.g., a substitution with serine, alanine or valine). Such sets of mutations can be engineered into any pair of polypeptides comprising CH2-CH3 domains that forms a Fc Region to foster heterodimerization (also called knobs-in-holes (KIH) Fc heterodimerization). Methods of protein engineering to favor heterodimerization over homodimerization are well known in the art, in particular with respect to the engineering of immunoglobulin-like molecules, and are encompassed herein (see e.g., Ridgway et al. (1996), Protein Engr. 9:617-621, Atwell et al. (1997), J. Mol. Biol.

[0308] 270: 26-35, and Xie etal. (2005), J. Immunol. Methods 296:95-101; each of which is hereby incorporated herein by reference in its entirety).

[0309] A preferred knob is created by modifying an IgG Fc Region to contain the modification T366W. A preferred hole may also be created by modifying an IgG Fc Region to contain the modification T366S, L368A and Y407V.

[0310] According to some embodiments of the invention, the first Fc fragment comprises an S354C or Y349C mutation and the second heavy chain comprises a Y349C or S354C mutation.

[0311] In some embodiments, the Fc fragment or Fc pair comprises or consist of sequence SEQ ID NO: 20 (lgG4 Fc FALA (knob)) and / or SEQ ID NO: 109 (lgG4 Fc FALA (hole)), or SEQ ID NO: 143 (lgG4 silent FALA (R409K mutation, knob) and / or SEQ ID NO: 148 (lgG4 silent FALA (R409K mutation, hole).

[0312] In some embodiments, the Fc fragment or Fc pair comprises or consist of sequence SEQ ID NO: 137 (lgG1 murine silent (knob)) and / or SEQ ID NO: 138 (lgG1 murine silent (hole)).

[0313] In some embodiments, the lgG1 Fc fragment or Fc pair comprises or consist of sequence SEQ ID NO 142 (lgG1 silent STR (knob)) and / or SEQ ID NO 147 (lgG1 silent STR (hole)); SEQ ID NO 139 (lgG1 silent LALAPG (knob)) and / or SEQ ID NO 144 (lgG1 silent LALAPG (hole)); SEQ ID NO 140 (lgG1 silent LALAPS (knob)) and / or SEQ ID NO 145 (lgG1 silent LALAPS (hole)); or SEQ ID NO 141 (lgG1 silent LALADS (knob)) and / or SEQ ID NO 146 (lgG1 silent LALADS (hole)).Protein having dual IL-10 and CSF1 activities and a moiety increasing half-life (format ORK1 Fc 08)

[0314] The invention also relates to a protein having dual IL-10 and CSF1 activities and a moiety increasing half-life.

[0315] In this embodiment, a protein is provided which comprises two polypeptide chains wherein a first polypeptide chain comprises, in the N-terminus to C-terminus direction, a CSF1 monomer fused to an immunoglobulin Fc fragment, and second polypeptide chain comprises, in the N-terminus to C-terminus direction, an immunoglobulin Fc fragment fused to an IL-10 domain, wherein the IL-10 domain is an IL-10 monomer ora single chain dimeric IL-10, and wherein the two polypeptide chains dimerise through their Fc domains. In some aspects the IL-10 domain is a single chain dimeric IL-10.

[0316] In some embodiments, the protein comprises two polypeptide chains comprising or consisting of:

[0317] CSF1 monomer-L2-Fc1, wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and

[0318] Fc2-L2-IL-10 domain, wherein Fc2 is a Fc domain comprising hinge-CH2-CH3, wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains. In some aspects the IL-10 domain is a single chain dimeric IL-10.

[0319] The CSF1 monomer, IL-10 domain, linker L2, and immunoglobulin Fc fragments Fc1 and Fc2 are as defined above.

[0320] In some embodiments, the protein comprises a first polypeptide chain comprising or consisting of sequence SEQ ID NO: 161, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto, and a second polypeptide comprising or consisting of sequence SEQ ID NO: 243 (as in e.g.

[0321] or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto, wherein the two polypeptide chains dimerise and the protein retains IL- 10 and CSF1 activities

[0322] In some embodiments, the protein comprises a first polypeptide chain comprising or consisting of sequence SEQ I D NO: 161, and a second polypeptide comprising or consisting of sequence SEQ ID NO: 243 (as in e.g. wherein the two polypeptide chains dimerisePharmaceutical composition

[0323] The invention further relates to a pharmaceutical composition comprising the protein comprising the single chain polypeptide having IL-10 and CSF1 activities and an immunoglobulin Fc fragment, and a pharmaceutically acceptable carrier.

[0324] A suitable pharmaceutically acceptable ‘carrier’ (such as diluents, adjuvants, excipients or vehicles) is described in " Remington's Pharmaceutical Sciences" by E. W. Martin. Pharmaceutical compositions of the invention are formulated to conform to regulatory standards and can be administered orally, intravenously, topically, or subcutaneously or via other standard routes. As used herein, the term ‘carrier’ includes any and all solvents, dispersion media, vehicles, coatings, diluents, antibacterial and antifungal agents, isotonic and absorption delaying agents, buffers, carrier solutions, suspensions, colloids, and the like. The use of such media and agents for pharmaceutical active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions. The phrase ‘pharmaceutically acceptable’ or ‘pharmacologically-acceptable’ refers to molecular entities and compositions that do not produce an allergic or similar untoward reaction when administered to a human or a non-human mammal.

[0325] Acceptable pharmaceutical vehicles can be liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. The pharmaceutical vehicles can be saline, gum acacia, gelatin, starch paste, talc, keratin, colloidal silica, urea, and the like. In addition, auxiliary, stabilising, thickening, lubricating and colouring agents may be used. When administered to a subject, the pharmaceutically acceptable vehicles are preferably sterile. Water is a suitable vehicle particularly when the compound of the invention is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid vehicles, particularly for injectable solutions. Suitable pharmaceutical vehicles also include excipients such as starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like. The present compositions, if desired, can also contain minor amounts of wetting or emulsifying agents, or buffering agents.

[0326] The medicaments and pharmaceutical compositions of the invention can take the form of liquids, solutions, suspensions, lotions, gels, tablets, pills, pellets, powders, modified-release formulations (such as slow or sustained-release), suppositories, emulsions, aerosols, sprays, capsules (for example, capsules containing liquids or powders),liposomes, microparticles or any other suitable formulations known in the art. Other examples of suitable pharmaceutical vehicles are described in Remington's Pharmaceutical Sciences, Alfonso R. Gennaro ed., Mack Publishing Co. Easton, Pa., 19th ed., 1995, see for example pages 1447-1676.

[0327] In some embodiments the therapeutic compositions or medicaments of the invention are formulated in accordance with routine procedures as a pharmaceutical composition adapted for oral administration (more suitably for human beings). Compositions for oral delivery may be in the form of tablets, lozenges, aqueous or oily suspensions, granules, powders, emulsions, capsules, syrups, or elixirs, for example. Thus, in various embodiments, the pharmaceutically acceptable vehicle may be a capsule, tablet or pill.

[0328] Orally administered compositions may contain one or more agents, for example, sweetening agents such as fructose, aspartame or saccharin; flavouring agents such as peppermint, oil of Wintergreen, or cherry; colouring agents; and preserving agents, to provide a pharmaceutically palatable preparation. When the composition is in the form of a tablet or pill, the compositions may be coated to delay disintegration and absorption in the gastrointestinal tract, so as to provide a sustained release of active agent over an extended period of time. Selectively permeable membranes surrounding an osmotically active driving compound are also suitable for orally administered compositions. In these dosage forms, fluid from the environment surrounding the capsule is imbibed by the driving compound, which swells to displace the agent or agent composition through an aperture. These dosage forms can provide an essentially zero order delivery profile as opposed to the spiked profiles of immediate release formulations. A time delay material such as glycerol monostearate or glycerol stearate may also be used. Oral compositions can include standard vehicles such as mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, etc. Such vehicles are preferably of pharmaceutical grade. For oral formulations, the location of release may be the stomach, the small intestine (the duodenum, the jejunem, or the ileum), or the large intestine. The person skilled in the art is able to prepare formulations that will not dissolve in the stomach, yet will release the material in the duodenum or elsewhere in the intestine. Suitably, the release will avoid the deleterious effects of the stomach environment, either by protection of the peptide (or derivative) or by release of the peptide (or derivative) beyond the stomach environment, such as in the intestine. To ensure full gastric resistance a coating impermeable to at least pH 5.0 would be essential. Examples of the more common inert ingredients that are used as enteric coatings are cellulose acetate trimellitate (CAT), hydroxypropylmethylcellulose phthalate (HPMCP), HPMCP 50, HPMCP 55, polyvinyl acetate phthalate (PVAP), EudragitL30D, Aquateric, cellulose acetate phthalate (CAP), Eudragit L, Eudragit S, and Shellac, which may be used as mixed films.

[0329] To aid dissolution of the therapeutic agent(s) into the aqueous environment a surfactant might be added as a wetting agent. Surfactants may include anionic detergents such as sodium lauryl sulfate, dioctyl sodium sulfosuccinate and dioctyl sodium sulfonate. Cationic detergents might be used and could include benzalkonium chloride or benzethomium chloride. Potential nonionic detergents that could be included in the formulation as surfactants include: lauromacrogol 400, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil 10, 50 and 60, glycerol monostearate, polysorbate 20, 40, 60, 65 and 80, sucrose fatty acid ester, methyl cellulose and carboxymethyl cellulose. These surfactants, when used, could be present in the formulation of the peptide or nucleic acid or derivative either alone or as a mixture in different ratios.

[0330] Typically, compositions for intravenous administration comprise sterile isotonic aqueous buffer. Where necessary, the compositions may also include a solubilising agent.

[0331] Mixtures of the polypeptides as described herein may be prepared in water suitably mixed with one or more excipients, carriers, or diluents. Dispersions may also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations may contain a preservative to prevent the growth of microorganisms. The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases the form may be sterile and may be sufficiently fluid to enable injection by an appropriate syringe. Suitably, the composition is stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and / or vegetable oils. Proper fluidity may be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of a dispersion and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.For parenteral administration in an aqueous solution, for example, the solution may be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose. These particular aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous, intratumoral and intraperitoneal administration. In this connection, sterile aqueous media that can be employed will be known to those of skill in the art.

[0332] Another suitable route of administration for the pharmaceutical compositions of the invention is via pulmonary or nasal delivery.

[0333] Additives may be included to enhance cellular uptake of a therapeutic agent of the invention, such as the fatty acids, oleic acid, linoleic acid and linolenic acid.

[0334] Therapeutic applications

[0335] The protein comprising the single chain polypeptide having IL-10 and CSF1 activities and immunoglobulin Fc fragment as disclosed herein, or pharmaceutical composition comprising it, is for use as a medicament.

[0336] Accordingly, the invention further relates to a method of treatment which comprises administering the protein comprising the single chain polypeptide having IL-10 and CSF1 activities and immunoglobulin Fc fragment to a subject in need thereof.

[0337] The subject may be a human or non-human mammal such as a rodent (mouse or rat), a primate (e.g. a monkey), a pig, etc.

[0338] The subject in need thereof is in particular a subject suffering from an inflammatory disease. The single chain polypeptide having IL-10 and CSF1 activities is designed to preferentially activate myeloid cells (monocytes, macrophages, dendritic cells) that coexpress CSF1R and IL-10R - and that induce anti-inflammatory responses - over CD8 T cells and B cells expressing IL-10R that could induce cytotoxic responses and autoantibodies when activated.

[0339] Accordingly, the invention further relates to the protein comprising the single chain polypeptide and immunoglobulin Fc fragment for use for treating an inflammatory disease. Also provided is a method of treating an inflammatory disease which comprises administering the protein comprising the single chain polypeptide having IL-10 and CSF1 activities of the invention to a subject in need thereof.

[0340] In some embodiments, the inflammatory disease is an inflammatory and / or autoimmune disease of the gut such as inflammatory bowel disease, including Crohn’s disease and ulcerative colitis, or Behget Disease, Rheumatoid Arthritis, Type I diabetes, Psoriasis, ARDS (Acute Respiratory Distress Syndrome), fibrosis, allergy or asthma.Treatment regimens may vary, and often depend on the type and / or location of the disease, the stage / progression of the disease, and / or the health and age of the patient. The skilled clinician will be able to determine a suitable therapeutic treatment regime under each circumstance.

[0341] The invention will be further illustrated in view of the following figures and examples.

[0342] FIGURES

[0343] Figure 1. Diagram displaying possible fusion proteins of IL-10 and CSF1 (A), as well as of FoldikinelO and CSF1 (B). (C): display of how the IL-10 WT monomer fused to CSF1 might aggregate or generate concatemers.

[0344] Figure 2. Structural organisation of different proteins comprising a single chain polypeptide having IL-10 and CSF1 activities and an immunoglobulin Fc fragment.

[0345] Symetric molecules comprising two single chain polypeptides having IL-10 and CSF1 activities and one immunoglobulin Fc fragment (ORK1_Fc_01, ORK1_Fc_02, ORK1_Fc_03, ORK1_Fc_04) and asymmetric molecules comprising one single chain polypeptide having IL-10 and CSF1 activities and one immunoglobulin Fc fragment (ORK1_Fc_05, ORK1_Fc_06). Fc fragment is from lgG4. In asymmetric molecules, Fc fragment comprises stabilization mutation S228P and silent FALA.

[0346] Figure 3. ORK1-405 shows selectivity towards myeloid cells compared to lymphoid cells. (A) PBMC from healthy donors were plated on in 96-well plates and stimulated next day with IL10, ORK1-068 or ORK1-405 for 20 min at 37°C. Cells were stained with CD14 Alexa647 (Biolegend) and CD3 APC / Cy7 (Biolegend), fixed with 4% PFA and permeabilized for intracellular staining with 80% MetOH. Cells were stained with AlexaFluor488 pSTAT3 (pY705) (BD) for 1h. (B) Samples were analyzed with LSRII cytometer and MFI values obtained from STAT3 activation were plotted using GraphPad Prism software. One representative result with two technical replicates is shown as mean with error bar depicting the SEM, similar results were obtained from 8 different donors. Each biological replicate was normalized by assigning the highest IL10 MFI value of the top concentration as 100% and the lowest MFI value of an untreated control as 0%. The MFI from the samples treated with ORK1 was normalized accordingly.

[0347] Figure 4. ORK1-405 inhibits monocyte activation. (A) CD14+ cells were isolated from total human PBMCs. Purified monocytes were stimulated with LPS in the presence or absence of different concentrations of IL-10, ORK1-068 or ORK1-405. (B) TNFa levels in the supernatants were determined by ELISA 24h after stimulation. Similar results obtained with IL-6.Figure 5. ORK1-405 is less potent than IL-10 at activating CD8 T cells. (A) CD8 T cells were isolated from human PBMCs using positive microbeads selection (Miltenyi). The lymphocytes were stimulated with anti-CD3 and anti-CD28 for 72h. Then, cells were counted, replated, and treated with IL10, ORK1-068 or ORK1-405 with IL-2 for 48h. (B) Cell supernatants were obtained and GzmB was detected by ELISA. Independent data of 4 experiments are shown in the graph.

[0348] Figure 6. ORK1-405 is less potent than IL-10 at activating B cells. (A) B cells were isolated from human PBMCs using positive microbeads selection (Miltenyi). The isolated lymphocytes were stimulated with CpG (T-independent conditions) and IL10, ORK1-068 or ORK1-405 at dO and d2. After 7 days cell supernatants were obtained for the determination of IgG levels by ELISA (B) and analysis of plasma cell differentiations (C) by flow cytometry. Similar results were obtained when stimulating with CD40L (T-dependent conditions). Independent data of 4 independent experiment are shown in the graph.

[0349] Figure 7. ORK1-405 binds preferentially to monocytes. (A) PBMC from healthy donors were plated on in 96-well plates and treated when specified with saturating concentrations of rCSF1, rlL-10 or both (5 min, 37°C), after that cells were washed and resuspended in buffer containing sodium azide 0.1%. Then, ORK1-405 is added and incubated for 20 min on ice. (B) Cells were washed and resuspended in stain buffer containing the mix of antibodies: CD14 Alexa 647, CD3 APC / Cy7 and anti-human lgG4-FC PE. Finally, cells were resuspended in stain buffer. Samples were analyzed with LSRII cytometer and MFI values obtained from anti-human lgG4-Fc PE were plotted using GraphPad Prism software.

[0350] Figure 8. ORK1-405 induces the differentiation of monocyte-derived macrophages. (A) PBMCs were isolated from human buffy coats and CD14+ cells were purified by positive microbead selection (Miltenyi). Monocytes were cultured for 7 days in the presence or absence of: CSF1, ORK1-068 or ORK1-405. Cytokines were added every 2 days. (B) At day 7 cells were counted and characterized by FACS (CD163, CD86).

[0351] Figure 9. ORK1-405 induces regulatory macrophages. (A) PBMCs were isolated from human buffy coats and CD14+ cells were purified by positive selection (Miltenyi). Monocytes were cultured for 7 days in the presence or absence of: CSF1, ORK1-068 or ORK1-405. Cytokines were added every 2 days. (B) At day 7 cells were stimulated with LPS±IFNg and cytokines levels in the supernatants were determined by ELISA 17h after stimulation. Mean +SEM of 4-6 independent experiments is shown in the graph.

[0352] Figure 10. Influence of Fc formats. (A) PBMC from healthy donors were plated o / n in 96-well plates and stimulated next day with ORK1-405 (format ORK1_Fc_06) and ORK1-400 (format ORK1_Fc_01) for 20 min at 37°C. Cells were stained with CD14 Alexa647(Biolegend) and CD3 APC / Cy7 (Biolegend), fixed with 4% PFA and permeabilized for intracellular staining with 80% MetOH. Cells were stained with AlexaFluor488 pSTAT3 (pY705) (BD) for 1h. (B) Samples were analyzed with LSRII cytometer and MFI values obtained from STAT3 activation were plotted using GraphPad Prism software. One representative result with two technical replicates is shown as mean with error bar depicting the SEM. Each biological replicate was normalized by assigning the highest IL10 MFI value of the top concentration as 100% and the lowest MFI value of an untreated control as 0%. The MFI from the samples treated with both formats was normalized accordingly.

[0353] Figure 11. HEK-Blue 10 activity of ORK1-405. (A) IL10 reporter HEK cells were seeded in 96 well plate at 0.4M / ml. Serial dilutions of IL10, ORK1-068 and ORK1-405 were added to the cells o / n. The day after 20ul of supernatant was transferred to a new plate and 180ul of Quanti-Blue Solution was added. Plate was incubated between 15 min-1h and the OD at 620nm was measured with a Tekan plate reader (B).

[0354] Figure 12. ORK1-Fc protects mice from chronic TNBS-induced colitis.

[0355] (A) Evaluation of therapeutic efficacy of ORK1-405, compared to ORK1-068, (in 2 doses / week intraperitoneally at 5pg per mice) in an experimental model of chronic IBD induced by TNBS.

[0356] TNBS was intrarectally infused in male Balb / c mice at two increasing doses (0.8 mg / mouse at day 0 and 1.0 mg / mouse at day 7). The experimental Groups included 10 mice per group and were as follows:

[0357] 1. Intrarectal TNBS / 50% ethanol+treatment with PBS

[0358] 2. Intrarectal TNBS / 50% ethanol+treatment with ORK1-068 (dose 5 pg per mice) 3. Intrarectal TNBS / 50% ethanol+treatment with ORK1-405(dose 5 pg per mice). Mice treated with ORK1-405 had less body weight loss compared to both ORK1-068-treated or vehicle treated after 12 days.

[0359] Figure 13. ORK1 sequence variants. (A) PBMC from healthy donors were plated o / n in 96-well plates and stimulated next day with IL10, ORK1 variants or ORK1-405 for 20 min at 37°C. Cells were stained with CD14 Alexa647 (Biolegend) and CD3 APC / Cy7 (Biolegend), fixed with 4% PFA and permeabilized for intracellular staining with 80% MetOH. Cells were stained with AlexaFluor488 pSTAT3 (pY705) (BD) for 1h. (B)(C) Samples were analyzed with LSRII cytometer and MFI values obtained from STAT3 activation were plotted using GraphPad Prism software. One representative result with two technical replicates is shown as mean with error bar depicting the SEM, similar results were obtained from 4 different donors. Each biological replicate was normalized by assigning the highest IL10 MFI value of the top concentration as 100% and the lowest MFI value of anuntreated control as 0%. The MFI from the samples treated with ORK1-variants and ORK1-405 as a control were normalized accordingly.

[0360] Figure 14. ORK1 single chain IL-10 sequence. (A) PBMC from healthy donors were plated o / n in 96-well plates and stimulated next day with IL10, ORK1-068 or ORK1-455 (sc) for 20 min at 37°C. Cells were stained with CD14 Alexa647 (Biolegend) and CD3 APC / Cy7 (Biolegend), fixed with 4% PFA and permeabilized for intracellular staining with 80% MetOH. Cells were stained with AlexaFluor488 pSTAT3 (pY705) (BD) for 1h. (B) Samples were analyzed with LSRII cytometer and MFI values obtained from STAT3 activation were plotted using GraphPad Prism software. One representative result with two technical replicates is shown as mean with error bar depicting the SEM, similar results were obtained from 3 different donors. Each biological replicate was normalized by assigning the highest IL10 MFI value of the top concentration as 100% and the lowest MFI value of an untreated control as 0%. The MFI from the samples treated with ORK1-068 and ORK1-455 (sc) were normalized accordingly.

[0361] Figure 15. ORK1-405 extended CSF1 version. (A) PBMC from healthy donors were plated o / n in 96-well plates and stimulated next day with IL10, ORK1-405 or ORK1-497 for 20 min at 37°C. Cells were stained with CD14 Alexa647 (Biolegend) and CD3 APC / Cy7 (Biolegend), fixed with 4% PFA and permeabilized for intracellular staining with 80% MetOH. Cells were stained with AlexaFluor488 pSTAT3 (pY705) (BD) for 1h. (B) Samples were analyzed with LSRII cytometer and MFI values obtained from STAT3 activation were plotted using GraphPad Prism software. One representative result with two technical replicates is shown as mean with error bar depicting the SEM, similar results were obtained from 3 different donors. Each biological replicate was normalized by assigning the highest IL10 MFI value of the top concentration as 100% and the lowest MFI value of an untreated control as 0%. The MFI from the samples treated with ORK1-405 and ORK1-497 were normalized accordingly.

[0362] Figure 16. Structure-Activity Relationship: IL-10 Signaling in Monocytes is CSF1-Dependent. (A) PBMC from healthy donors were plated o / n in 96-well plates and stimulated next day with IL-10 (not shown), ORK1-068 or ORK1-341 for 20 min at 37°C. Cells were stained with CD14 Alexa647 (Biolegend) and CD3 APC / Cy7 (Biolegend), fixed with 4% PFA and permeabilized for intracellular staining with 80% MetOH. Cells were stained with AlexaFluor488 pSTAT3 (pY705) (BD) for 1h. (B) PBMC from healthy donors were plated o / n in 96-well plates and treated next day when specified with saturating concentrations of CSF1 (37°C, 5 min) then stimulated with IL-10 (not shown) or ORK1-068 for 20 min at 37°C. Cells were stained with CD14 Alexa647 (Biolegend) and CD3 APC / Cy7 (Biolegend), fixed with 4% PFA and permeabilized for intracellular staining with 80%MetOH. Cells were stained with AlexaFluor488 pSTAT3 (pY705) (BD) for 1 h. Samples were analyzed with LSRII cytometer and MFI values obtained from STAT3 activation were plotted using GraphPad Prism software. One representative result with two technical replicates is shown as mean with error bar depicting the SEM, similar results were obtained from 2 different donors. Each biological replicate was normalized by assigning the highest IL-10 MFI value of the top concentration as 100% and the lowest MFI value of an untreated control as 0%. The MFI from the samples treated with ORK1-068 and ORK1-341 were normalized accordingly.

[0363] Figure 17. ORK1-494 ratio between T cell and monocyte in STAT3 activation, CD8 T cell activation (GzmB) and TNFa production. Sequence optimization in ORK1-494 maintains selectivity towards myeloid cells compared to lymphoid cells. (A) Total PBMC from healthy donors were treated with IL-10, ORK1-068, ORK1-405 and ORK1-494. Cells were stained with CD14 and CD3 antibodies and intracellular staining for pSTAT3. Samples were analyzed with LSRII cytometer and MFI values obtained from STAT3 activation were plotted using GraphPad Prism software. Ratio between EC50 from T cells vs EC50 from monocytes is shown in the graph. (B) CD8 T cells were activated with a-CD3 / a-CD28, activated cells were treated with IL-10, ORK1-405 and ORK1-494 and after 2 days supernatants were collected. GzmB was determined by ELISA. (C) Macrophage differentiated after 7 days from monocytes with GM-CSF, CSF1, ORK1-405 or ORK1-494 were stimulated overnight with LPS or LPS+IFN-y. TNFa was measured by ELISA.

[0364] Figure 18. ORK1-494 Binding to Monocytes is CSF1-Dependent. (A) Total PBMC from healthy donors were treated when specified with saturating concentrations of rCSF1, rlL-10 or both together. After 5 min at 37°C, cells were washed and resuspend in buffer containing sodium azide 0,1%. After that, cells were treated with ORK1-494 for 20 min on ice. Cells were washed and resuspended in stain buffer containing the mix of antibodies: CD14 Alexa 647, CD3 APC / Cy7 and anti-human lgG4-FC PE. (B) Samples were analysed with LSRII cytometer and MFI values obtained from anti-human lgG4-FC PE were plotted on graphs in comparison with control sample with no blocking conditions.

[0365] Figure 19. ORK1-494 improves body weight recovery and reduces colitis scores in mice exposed to chronic TNBS. Evaluation of the therapeutic efficacy of ORK1-494 was performed in an experimental model of chronic IBD induced by TNBS (Trinitrobenzene sulfonic acid (TNBS)-induced colitis mouse model). TNBS was intrarectally infused in male Balb / c mice at four increasing doses (0.8 mg / mouse at day 0; 1.0 mg / mouse at day 7; 1.2 mg / mouse at day 14; and 1.5 mg / mouse at day 21).

[0366] The experimental Groups included 10 mice per group and were as follows:

[0367] 1. Healthy control mice with no TNBS or ORK1 treated2. Intrarectal TNBS / 50% ethanol+treatment with PBS

[0368] 3. Intrarectal TNBS / 50% ethanol+treatment with ORK1-494 (dose 1 g per mice) 4. Intrarectal TNBS / 50% ethanol+treatment with ORK1-494 (dose 5 pg per mice) 5. Intrarectal TNBS / 50% ethanol+treatment with ORK1-494 (dose 10 pg per mice) ORK1-494 was administered intraperitoneally, at defined dose, 2 times per week during 4 weeks. Graphs show data related to body weight loss (A), colitis score (B) and survival rates (C). Protective effects were dose dependent.

[0369] Figure 20. ORK1-494 protects mice from chronic TNBS-induced colitis overtime with no treatment repetition after 2 weeks. Evaluation of the therapeutic efficacy of ORK1-494 was performed in an experimental model of chronic IBD induced by TNBS (Tri nitrobenzene sulfonic acid (TNBS)-induced colitis mouse model). TNBS was intrarectally infused in male Balb / c mice at four increasing doses (0.8 mg / mouse at day 0; 1.0 mg / mouse at day 7; 1.2 mg / mouse at day 14; and 1.5 mg / mouse at day 21).

[0370] The experimental Groups included 10 mice per group and were as follows:

[0371] 1. Healthy control mice with no TNBS or ORK1 treated

[0372] 2. Intrarectal TNBS / 50% ethanol+treatment with PBS

[0373] 3. Intrarectal TNBS / 50% ethanol+treatment with ORK1-494 (dose 5 pg per mice)

[0374] 4. Intrarectal TNBS / 50% ethanol+treatment with ORK1-494 (dose 10 pg per mice) ORK1-494 were administered intraperitoneally, at defined dose, 2 times per week during only the first two weeks, after 4 doses, mice were not treated anymore with ORK1-494 but continued to inject TNBS for two weeks. Graphs show data related to body weight loss (A), colitis score (B) and survival rates (C).

[0375] Figure 21. Spatial Orientation and Structure to Optimize Function. Tested N-to-C and C-to-N orientations of CSF1 and IL- 10 together with two CSF1 molecules linked and hybrid human-viral IL-10 to assess impact on monocyte binding and T cell activity. Asymmetric formats (Fc-05 to Fc-12) were evaluated to assess how the positioning of IL-10, CSF-1, and the Fc region affects monocyte binding and T-cell activity. Linker 1 (L1) is defined as the linker connecting the IL-10 domain to the CSF-1 domain. Linker 1.1 (L1.1) is defined as intrapolypeptide linker connecting one CSF-1 domain to another CSF-1 domain. Linker 2 (L2) is defined as the linker connecting the cytokine domain (either IL-10 or CSF-1) to the FC domain.

[0376] Figure 22. Some optimized formats show improved selectivity towards myeloid cells compared to lymphoid cells. (A) Total PBMC from healthy donors were treated with IL-10, ORK1-494, ORK1-507, ORK1-509, ORK1-510, ORK-512, ORK1-513 ORK1-525. Cells were stained with CD14 and CD3 antibodies and intracellular staining for pSTAT3.

[0377] (B) Samples were analyzed with LSRII cytometer and MFI values obtained from STAT3activation were plotted using GraphPad Prism software. Each biological replicate was normalized by assigning the highest IL-10 % of positive cells value of the top concentration as 100% and the lowest value of an untreated control as 0%.

[0378] Figure 23. Optimized formats bind preferentially to monocytes. (A) Total PBMC from healthy donors were treated with ORK1-494, ORK1-510 and ORK1-513. Cells were washed and resuspended in stain buffer containing the mix of antibodies: CD14 Alexa 647, CD3 APC / Cy7 and anti-human lgG4-FC PE. Finally, cells were resuspended in stain buffer.

[0379] (B) Samples were analyzed with LSRII cytometer and MFI values obtained from anti-lgG4 were plotted using GraphPad Prism software.

[0380] Figure 24. Optimized formats (ORK1-510 and ORK1-513) show increased ratio between T cells and monocytes in STAT3 activation. (A) Total PBMC from healthy donors were treated with IL-10, ORK1-494, ORK1-510 and ORK1-513. Cells were stained with CD14 and CD3 antibodies and intracellular staining for pSTAT3. Samples were analyzed with LSRII cytometer and values obtained from % of positive cells from STAT3 activation were plotted using GraphPad Prism software. Each biological replicate was normalized by assigning the highest IL-10 % of positive cells value of the top concentration as 100% and the lowest value of an untreated control as 0%. The samples treated with ORK1 variants were normalized accordingly. Data was interpolated to 50% of maximum stimulation. Ratio between values from T cells vs monocytes is shown in the graph. (B) Dose-response curves % of positive cells from STAT3 activation in monocytes and T cells.

[0381] Figure 25. Optimized formats induce regulatory macrophages. (A) CD14+ cells were purified from total PBMC of healthy donors. Monocytes were cultured for 7 days in the presence or absence of: CSF1, ORK1-494, ORK1-510 and ORK1-513 at 1.08nM. Cytokines were added every 2 days. At day 7, cells were counted and characterized by LSRII cytometer (CD163, CD206, CD86 and HLA-DR). (B) CD14+ cells were purified from total PBMC of healthy donors. Monocytes were cultured for 7 days in the presence or absence of: CSF1, ORK1-494, ORK1-510 and ORK1-513 at 1.08nM. Cytokines were added every 2 days. At day 7 media was refreshed, and cells were stimulated overnight with LPS and IFN-y. After harvesting supernatants, TNFa production was determined by ELISA.

[0382] Figure 26. ORK1-513 is less potent than previous formats activating CD8 T cells and B cells. (A) B cells were isolated from human PBMCs using positive microbeads selection (Miltenyi). The isolated lymphocytes were stimulated with CD40L (T-dependent conditions) and IL10, ORK1-494, ORK1-510 and ORK1-513 variants at dO and d2. After 7 days plasma cell differentiation was analyzed by flow cytometry and cell supernatants were obtained for the determination of IgG levels by ELISA (data not shown). Similar results wereobtained when stimulating with CpG (T-independent conditions). (B) CD8 T cells were isolated from human PBMCs using positive microbeads selection (Miltenyi). Cells were activated for 3 days with α-CD3 and α-CD28 and treated with IL-10, ORK1-494, ORK1-510 and ORK1-513 with IL-2 and after 2 days supernatants were collected. GzmB was determined by ELISA.

[0383] Figure 27. Regulatory macrophages induced by optimized variants reduce T cell activation. (A) CD14+ cells were purified from total PBMC of healthy donors. Monocytes were cultured for 5 days in the presence or absence of: CSF1, ORK1-494, ORK1-510 and ORK1-513 at 1.08nM. Cytokines were added every 2 days. At day 5, cells were counted and replated to a 96 round well plate (40.000 cells / well) and last dose of cytokines was added. After 2 days CD3 T cells from another donor were purified and added to macrophages after refreshing media. 120.000 CD3 T cells were added per well in a final ratio of 1:3. Macrophages were activated with LPS ± NOD2 agonist and after 2 days supernatants were collected, and T cells were counted. (B) IFN-y, GzmB, TNFa and IL-23 production in co-cultures was determined by ELISA.

[0384] Figure 28. ORK1-513 shows limited efficacy in chronic TNBS-induced colitis compared to ORK1-494. Evaluation of the therapeutic efficacy of ORK1-513 was performed in an experimental model of chronic IBD induced by TNBS (Trinitrobenzene sulfonic acid (TNBS)-induced colitis mouse model), with comparison with ORK1-494.(A) TNBS was intrarectally infused in male Balb / c mice at three increasing doses (0.8 mg / mouse at day 0; 1.0 mg / mouse at day 7; and 1.2 mg / mouse at day 14).

[0385] The experimental groups included 15 mice per group and were as follows:

[0386] 1. Intrarectal TNBS / 50% ethanol+treatment with PBS

[0387] 2. Intrarectal TNBS / 50% ethanol+treatment with ORK1-513 (dose 1 pg per mice) 3. Intrarectal TNBS / 50% ethanol+treatment with ORK1-513 (dose 5 pg per mice) 4. Intrarectal TNBS / 50% ethanol+treatment with ORK1-513 (dose 10 pg per mice) 5. Intrarectal TNBS / 50% ethanol+treatment with ORK1-494 (dose 5 pg per mice) ORK1-513 and ORK1-494 were administered intraperitoneally, at defined dose, 2 times per week for 3 weeks.

[0388] Graphs shown data related to body weight loss, (B), colitis score, (C) and survival rates (D).

[0389] Figure 29. PK studies ORK1-068 and ORK1-513. (A) ORK1-068 was administered IV at a 0.5mg / kg into C57BL / 6J female mice. Blood samples were collected at several timepoints 5min,1h, 3h, 7h, 11 h, 24h. Serum ORK1-513 concentration was measured using IL-10 and CSF1 ELISA methods kit. ORK1-068 terminal half-life (T1 / 2) was between 0.995-1.01h while rlL-10 was 0.710h. The ORK1-068 Cmax and exposure (AUCO-tau) after0.5mg / kg IV injection was higher than obtained with IL-10. (B) ORK1-513 was administered in 3 different groups, in single or repeated dosing, injected IV into C57BL / 6J female mice aged 9 weeks at indicated doses (according to the table 1). Blood samples were collected at several timepoints 5min, 30min, 1 h, 3h, 6h, 24h, 48h, 72h,120h, 168h, 240 and 336h. Serum ORK1-513 concentration was measured using IL-10 and CSF1 ELISA methods kit.

[0390] (C) ORK1-513 improves terminal half-life (T1 / 2) in comparison with ORK1-068. Data was evaluated using GraphPad Prism software.

[0391] Figure 30. ORK1 biases IL-10 signaling toward monocytes via CSF1-CSF1R. (A) Total PBMC from healthy donors were treated when specified with saturating concentrations of rCSF1. After that, samples were stimulated with ORK1-494 or IL-10 as a control. Cells were stained with CD14 and CD3 antibodies and intracellular staining for pSTAT3. (B) Samples were analyzed with LSRII cytometer and values obtained from % of positive cells from STAT3 activation were plotted using GraphPad Prism software.

[0392] Figure 31. Binding affinity of mutations for CSF1. (A)Total PBMC from healthy donors were treated with different ORK1 variants with single mutations designed for improving binding affinity to CSF1 (Table). (B) Cells were washed and stained with CD14 and lgG4. Samples were analyzed with LSRII cytometer and MFI values obtained from anti-lgG4 were plotted using GraphPad Prism software.

[0393] Figure 32. pSTAT3 activation by affinity mutations for CSF1. (A) Total PBMC from healthy donors were treated with IL-10 and ORK1 variants with selected mutations to increase binding affinity to CSF1. Cells were stained with antibodies specific for CD14 and CD3 (data not shown), followed by intracellular staining for pSTAT3. (B) Samples were analyzed with LSRII cytometer and values corresponding to STAT3 activation in monocytes (% of positive cells) were quantified using GraphPad Prism software.

[0394] Figure 33. CSF1 affinity mutation (ORK1-532) increases function and outperforms previous formats in selectivity and function. (A) Total PBMC from healthy donors were treated with IL-10, ORK1-494, ORK1-513 and ORK1-532. Cells were stained with CD14 and CD3 antibodies and intracellular staining for pSTAT3. (B) Samples were analyzed with LSRII cytometer and values obtained from % of positive cells from STAT3 activation were plotted using GraphPad Prism software. Each biological replicate was normalized by assigning the highest IL-10 % of positive cells value of the top concentration as 100% and the lowest value of an untreated control as 0%. The samples treated with ORK1 variants were normalized accordingly. Data was interpolated to 50% of maximum stimulation. Ratio between values from T cells vs monocytes is shown in the graph. (C) Dose-response curves of % of positive cells from STAT3 activation in monocytes and T cells from samples treated with ORK1-513 and ORK1-532.Figure 34. ORK1-532 improves selective binding to monocytes and increases IL-10 signaling. (A) Total PBMC from healthy donors were treated with or without saturating concentrations of rCSF1. After that, samples were stimulated with ORK1-532 or IL-10 as a control. Cells were stained with CD14 and CD3 antibodies and intracellular staining for pSTAT3. (B) Samples were analyzed with LSRII cytometer and values obtained from % of positive cells from STAT3 activation were plotted using GraphPad Prism software.

[0395] Figure 35. ORK1-532 variant induces regulatory macrophages without activating T cells. (A) CD14+ cells were purified from total PBMC of healthy donors. Monocytes were cultured for 7 days in the presence or absence of: CSF1, ORK1-513 or ORK1-532. Cytokines were added every 2 days. Macrophages were stimulated overnight with LPS+IFN-y. TNFa was measured by ELISA. Data from 4 different donors is shown. (B) CD8 T cells were isolated from human PBMCs using positive microbeads selection (Miltenyi). Cells were activated for 3 days with a-CD3 and a-CD28 and treated with IL-10, ORK1-532 or ORK1-513 with IL-2 and after 2 days supernatants were collected. GzmB was determined by ELISA. Data from 4-8 different donors is shown.

[0396] Figure 36. ORK1-532 alleviates clinical symptoms in a TNBS mouse model of colitis and other inflammatory diseases compared to ORK1-494. Evaluation of the therapeutic efficacy of ORK1-532 was performed in an experimental model of chronic IBD induced by TNBS (Tri nitrobenzene sulfonic acid (TNBS)-induced colitis mouse model), with comparison with ORK1-494. (A) TNBS was intrarectally infused in male Balb / c mice at three increasing doses (0.8 mg / mouse at day 0; 1.0 mg / mouse at day 7; and 1.2 mg / mouse at day 14).

[0397] The experimental Groups included 15 mice per group and were as follows:

[0398] 1. Intrarectal TNBS / 50% ethanol+treatment with PBS

[0399] 2. Intrarectal TNBS / 50% ethanol+treatment with ORK1-532 (dose 5 g per mice) 3. Intrarectal TNBS / 50% ethanol+treatment with ORK1-494 (dose 5 pg per mice) ORK1-532 and ORK1-494 were administered intraperitoneally, at defined dose, 2 times per week for 2 weeks, TNBS was injected for an additional week, for a total duration of 3 weeks.

[0400] The results show body weight loss (B), colitis score (C) and survival (D) compared to both ORK1-494 treated or vehicle-treated mice, after 21 days treatment.

[0401] Figure 37. Linkers Design. (A) In ORK1-513 scaffold L1 binds CSF1 to IL10 and L2 IL10 to Fc. Types of linkers and combinations to generate different variants. FL consists of SEQ I D NO: 35; FS consists of SEQ I D NO: 82; RL consists of SEQ I D NO: 248; RS consists of SEQ ID NO: 247 (B)(C)

[0402] Figure 38. Short L1 Linkers Improve Monocyte Potency and Restore IL-10-Like Vmax. Total PBMC from healthy donors were treated with IL-10, ORK1-513 and linkervariants designed. Cells were stained with antibodies specific for CD14 and CD3, followed by intracellular staining for pSTAT3. Samples were analyzed with LSRII cytometer and mean fluorescence intensity (MFI) values corresponding to STAT3 activation were quantified using GraphPad Prism software. (A) Graph to illustrate Vmax from ORK1-555 compared to ORK1-513 and IL- 10 as a control. (B) Graph correlation between Vmax values (relative to IL-10) and EC50 in monocytes from pSTAT3 activation, all linker variants were included and ORK1-513 was used as a control.

[0403] Figure 39. Short L1 Linkers Drive Optimal Monocyte / T Cell Selectivity. Total PBMC from healthy donors were treated with IL-10, ORK1-513 or ORK1-551-566. Cells were stained with CD14 and CD3 antibodies and intracellular staining for pSTAT3. Samples were analyzed with LSRII cytometer and values obtained from % of positive cells from STAT3 activation were plotted using GraphPad Prism software. (A) Graph correlation between ratio of T cell vs monocyte in pSTAT3 activation and EC50 on monocytes in pSTAT3 activation, all linker variants were included and ORK1-513 was used as a control.

[0404] (B) Ratio between (50% maximum stimulation in pSTAT3 activation relative to IL-10) values from T cells vs monocytes is shown in the graph. Data from linker variants ORK1-551 to ORK1-566 are shown in relative to ORK1-513 as a control.

[0405] Figure 40. Synergistic effects on single mutations and linker modification. (A) Total PBMC from healthy donors were treated with IL-10, ORK1-513, ORK1-532, ORK1-555 or ORK1-587. Cells were stained with CD14 and CD3 antibodies and intracellular staining for pSTAT3. Samples were analyzed with LSRII cytometer and values obtained from % of positive cells from STAT3 activation were plotted using GraphPad Prism software.

[0406] (B) Each biological replicate was normalized by assigning the highest IL-10 MFI value of the top concentration as 100% and the lowest value of an untreated control as 0%. The samples treated with IL-10 or ORK1 variants were normalized accordingly. Data is shown in a dose-response graph and interpolated to 50% of maximum stimulation.

[0407] Figure 41. ORK1 optimization. Description of molecule structure, number of the molecules and description of changes applied to improve function. ORK1 Fc was optimized followed a systematic, multi-parametric approach. Initial format ORK1-068, then ORK1-405 adding lgG4 and later sequence optimization in ORK1-494. Screening was performed to determine the optimal spatial orientation of the cytokine domains (CSF1 and IL-10) and the Fc subunit within the molecular construct, ORK1-513. In parallel, multiple CSF1 affinity variants were evaluated (ORK1-532), together with systematic optimization of linker length and rigidity between the individual domains (ORK1-555, ORK1-563, ORK1-564 and ORK1-566). Later, affinity mutation was synergized with linker modification (ORK1-587).EXAMPLES

[0408] Example 1: Production of CSF1-IL10 combined bioactivity in a single-chain protein

[0409] Our goal is to create a dual molecule that can activate IL- 10 signaling exclusively in cells expressing CSF1R, which are predominantly myeloid cells and no other cells such as CD8+ T cells. To achieve this, we have engineered variants of IL-10 and / or Foldikine-10 with reduced affinity to IL10RA and IL10RB linked to a CSF1 monomer. The CSF1 component acts as a targeting tool to circulating monocytes, facilitating preferential binding of IL- 10 to these cells.

[0410] Example 2: Generation of single-chain (SC) molecules fused to Fc fragment The principles of protein expression are as follows.

[0411] In bold is highlighted the signal peptide (SP). Signal peptide is NOT included in final ORF as it is processed.

[0412] • In italic is included the tag used for purification (TEV-his-hibit). Tag is included in the ORF.

[0413] • Some proteins are the result of the co-transfection of two vectors. The two vectors are displayed. Knob chain or hole chain is specified.

[0414] • For context. Specified origin of the signal peptide at the end of the document

[0415] > ORK1_P_068 with signal peptide and tag MTAPGAAGRCPPTTWLGSLLLLVCLLASRSITEEVSEYCSHMIGSGHLQSLQRLIDSQM ETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLK SCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGMTQSENSCTHFPGNLPNMLRDLR DAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDI KAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFI NYI EAYMTM N NGGLDYLPNM LRDLRDAFSRVKTFFQM KDQLDN LLLKESLLEDFKGYLG CQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKA VEQVKNAFN KLQEKGIYKAMSEFDI Fl NYI EAYMTM KI RN GENL YFQSGGHHHHHHGGGV SGWRLFKKIS*

[0416] (SEQ ID NO: 132)

[0417] > ORK1_P_455 with signal peptide and tag MTAPGAAGRCPPTTWLGSLLLLVCLLASRSITEEVSEYCSHMIGSGHLQSLQRLIDSQM ETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLK SCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNML RDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAEN QDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSE FDIFINYI EAYMTM KIRNGGGSGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSR VKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVN SLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAY MTM KI RN GENL YFQSGGHHHHHHGGG VSG WRLFKKIS*(SEQ ID NO: 133)

[0418] Signal peptide context:

[0419] MEWSWVFLFFLSVTTGVHS (SEQ ID NO: NO: 110) [Taken from antibodies. Used as universal in different constructs];

[0420] MTAPGAAGRCPPTTWLGSLLLLVCLLASRSIT (SEQ ID NO: 112) [From human CSF1],

[0421] Table 1 discloses the different polypeptides constituting the protein(s) comprising single chain polypeptide having IL- 10 and CSF1 activities fused to a Fc fragment.Table 1: Description of CSF1-IL10 proteins: description of Polypeptides for Protein 1 and Protein 2

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[0614] ::OPlasmid generation

[0615] All plasmids generated in this work were assembled following the Gibson method (Gibson DG et al. Nat Methods. 2009 May;6(5):343-5). When required, Integrated DNA Technologies (IDT) Corporation performed gene synthesis (gBIock double-stranded fragments) and oligonucleotides synthesis. Gene amplification was carried out with Phusion DNA polymerase (Thermo Fisher Scientific) and transformed in Escherichia coli DH5-alpha competent cells (NEB).

[0616] For mammalian protein expression in ExpiCHO cells, the vector used was pcDNA3.1 (V790-20, Invitrogen). All the plasmids were verified by Sanger sequencing (Eurofins Genomics).

[0617] Expression in CHO cells

[0618] ExpiCHO protein expression kit cells were purchased from Thermofisher (A29133). For a low-scale production (2.5 mL), 24 deep well plates were used (AXYPDW10ML24CS, Merck) and covered by gas-permeable film (ThermoFisher). The day -1 of production, ExpiCHO-cells were split to a final density of 3 x 106viable cells / mL. Grown at 37°C, 8% CO2 and shaking at 110rpm. The Day 0 of production, ExpiCHO cells were split to a final density of 3 x 106viable cells / mL per sample aliquot 2.5mL of the cell suspension into one well of the plate. Per sample, 2ug of DNA to 200ul of cold OptiPRO SFM & 9.0 pL ExpiFectamine CHO Reagent was added. Then, 200 pL of the complexation mixture was added to each relevant well. All the plate was covered with a gas permeable seal and placed at 37°C, 8% CO2 and 225rpm. The day 1 after 18-22 hours post-transfection, ExpiFectamine CHO Enhancer and ExpiCHO Feed was added to each well. For an entire plate of 24 wells, 400 pL ExpiFectamine CHO Enhancer and 16 mL ExpiCHO Feed in a conical tube were mix. From this mixture, 600 pLwas added to each well of the plate. Place at 37C, 8% CO2 and 225rpm. After 4 days the samples were harvested. Plates were centrifuged 5 minutes at 4600 rpm at room temperature. Supernatants were aliquoted, snap frozen liquid nitrogen and stored at -80°C.

[0619] Protein purification

[0620] Affinity purification of Fc proteins was performed with HiTrap MabSelect PrismA. Binding buffer was PBS, 850 mM NaCI. Elution was performed with the following buffer: 0.1 M Na-citrate pH 3.3. Samples were neutralized right after elution with 1 M Tris-HCI, pH 9 (10% v / v). Purity, as defined by the fraction of Fc proteins, was determined by HPLC-SEC. SDS-PAGE of purified products was performed to verify the fragmentation and / oraggregation status of the final material. When necessary, a size exclusion chromatography was performed.

[0621] Colorimetric analysis in HEK-Blue™ Cells for IL-10 activity

[0622] Human IL-10 Reporter HEK-Blue™ cell line carrying a SEAP reporter construct was purchased from InvivoGen (InvivoGen, San Diego, CA, USA). Cell line was grown in DMEM (Lonza, BE12-604F) supplemented with 10% FBS, 2 mM L-glutamine and specified selection antibiotics by the manufacturer in each of the reporter cells. Cells were passed when 70% confluence was reached, following the manufacturer’s recommendation.

[0623] After supernatant quantification, samples were adjusted to fit into the dynamic range of the HEK reporter cells using DMEM media for performing the different dilutions. Each of the top concentration was set for each reporter cell to match saturation, and 11 serial dilutions were performed (1 / 3) for each sample. Then, HEK reporter cells were prepared according to manufacturer instructions. Cells were incubated 60 min at 37°C, and absorbance (630 nm) was measured in the spectrophotometer Tecan i-control, 2.0.10.0.

[0624] Example 3: in v / Yro functional evaluation of the dual CSF1-IL10 cytokine PBMC (150,000 cells / well) from healthy donors were plated on in 96-well plates and stimulated next day with rl L10, ORK1-068 or ORK1-405 (8 dilutions 1 / 3) for 20 min at 37°C. Cells were stained with CD14 Alexa647 (Biolegend) and CD3 APC / Cy7 (Biolegend), fixed with 4% PFA and permeabilized for intracellular staining with 80% MetOH. Cells were stained with AlexaFluor488 pSTAT3 (pY705) (BD) for 1 h (Figure3). Samples were analyzed with LSRII cytometer and MFI values obtained from STAT3 activation were plotted using GraphPad Prism software. One representative result with two technical replicates is shown as mean with error bar depicting the SEM. Similar results were obtained in 10 different donors. Each biological replicate was normalized by assigning the highest IL10 MFI value of the top concentration as 100% and the lowest MFI value of an untreated control as 0%. The MFI from the samples treated with ORK1 was normalized accordingly. In contrast, the ORK1-068 and ORK1-405 preferentially activates monocytes when compared to T cells.

[0625] These results show that ORK1-405 shows selectivity towards myeloid cells compared to lymphoid cells, in contrast with IL-10 and more selectivity compared to ORK1-068.

[0626] Monocytes (CD14+) (150,000 cells / well) purified from PBMCs from healthy donors were stimulated with 10 ng / ml of LPS (Sigma) in the presence or absence of different concentrations of rlL-10 or ORK1-068 (0.00001 to 1 nM) or of ORK1-405 (Figure 4). The TN Fa production measured in the presence of rlL-10, ORK1-068 or ORK1-405, show thatORK1-405 inhibits monocytes activation similarly to IL- 10 (dotted line indicates the levels of LPS alone, as a control) (Figure 4 B).

[0627] Similar results obtained with IL-6 or when using total PBMCs (data not shown).

[0628] ORK1-405, ORK1-068 and IL-10, respective potencies at activating CD8 T and B cells were further characterized.

[0629] CD8 T cells were isolated from human PBMCs using positive microbeads selection (Miltenyi). The lymphocytes (1.5 million / ml) were stimulated with anti-CD3 (ThermoFisher) (1 pg / ml) and anti-CD28 (ThermoFisher) (3 pg / ml) for 72h. Then, cells were splited, replated and treated with IL10, ORK1-068 or ORK1-405 (10 nM per treatment, 6 dilutions 1 / 4) with IL-2 (5 ng / ml) for 48h (Figure 5 A). Cell supernatants were harvested, and Granzyme B (GzmB) was detected by ELISA (Figure 5 B).

[0630] The results show that ORK1-405 (Figure 5 C) is less potent than IL-10 and ORK1-068 at activating CD8 T cells.

[0631] B cells were isolated from human PBMCs using positive microbeads selection (Miltenyi). The isolated lymphocytes (50,000 cells / well) were stimulated with CpG (InvivoGene) (1 pg / ml) and IL10, ORK1-068 orORK1-405 (10 nM per treatment, 8 dilutions 1 / 3) at dO and d2. After 7 days cell supernatants were obtained for the determination of IgG levels by ELISA (Figure 6 B) and cells were analyzed by flow cytometry to determine plasma cell differentiation (PCs) (Figure 6C). Similar results were obtained when stimulating with CD40L. The results show that ORK1-405 (Figure. 6 B) (Figure. 6 C) is less potent than ORK1-068 and IL-10 at activating B cells.

[0632] PBMC from healthy donors were plated o / n in 96-well plates and treated when specified with saturating concentrations of rCSF1, rlL-10 or both (5 min, 37°C), after that cells were washed and resuspend in buffer containing sodium azide 0,1%. Then, ORK1-405 (15nM per treatment, 8 dilutions 1 / 3) is added and incubated 20 min on ice. Cells were washed and resuspend in stain buffer containing the mix of antibodies: CD14 Alexa 647, CD3 APC / Cy7 and anti-human lgG4-FC PE (Fig 7A). Finally, cells were resuspended in stain buffer. Samples were analysed with LSRII cytometer and MFI values obtained from anti-human lgG4-FC PE were plotted using GraphPad Prism software (Fig 7B). The results show a preferential binding of ORK1 to monocytes (CD14+) driven by CSF1 binding to CSF1R.

[0633] Additionally, PBMCs were isolated from human buffy coats and CD14+ cells were purified by positive microbead selection (Miltenyi). Monocytes (0.5 million / ml) were culturedfor 7 days in the presence or absence of: rCSF1, ORK1-068 or ORK1-405 (0.56nM per treatment). Cytokines were added every 2 days. At day 7, alive cells were counted and characterized by FACS (CD163, CD86) (Figure 8A).

[0634] The results show that ORK1-405, similarly to ORK1-068, induces the differentiation of regulatory monocyte-derived macrophages, as identified by the CD163 expression (Figure 8B).

[0635] Further characterization was performed to determine the phenotype of the induced monocyte-derived macrophages.

[0636] PBMCs were isolated from human buffy coats and CD14+ cells were purified by positive selection (Miltenyi). Monocytes (0.5 million / ml) were cultured for 7 days in the presence or absence of: rCSF1, ORK1-068 or ORK1-405 (0.56nM). Cytokines were added every 2 days. At day 7, cells were stimulated with LPS, with or without IFNy (Figure 9 A). TNF-a levels in the supernatants were determined by ELISA, 17h after stimulation. CSF1-differentiated macrophages produce TNF-a, which is further enhanced by the addition of IFNy. In contrast, ORK1-068 and ORK1-405-differentiated macrophages do not produce TNF-a, even when stimulated with LPS+IFN-y.

[0637] These results show that ORK1-405-induced macrophages have a more regulatory phenotype compared to CSF1 and ORK1-068 (Figure 9B).

[0638] Example 4: in vivo, ORK1-405 alleviates clinical symptoms in a mouse model of colitis

[0639] Evaluation of the therapeutic efficacy of ORK1-405 was performed in an experimental model of chronic IBD induced by TNBS (Trinitrobenzene sulfonic acid (TNBS)-induced colitis mouse model), with comparison with ORK1-068.

[0640] TNBS was intrarectally infused in male Balb / c mice at two increasing doses (0.8 mg / mouse at day 0 and 1.0 mg / mouse at day 7).

[0641] The experimental Groups included 10 mice per group and were as follows:

[0642] 1. Intrarectal TNBS / 50% ethanol+treatment with vehicle

[0643] 2. Intrarectal TNBS / 50% ethanol+treatment with ORK1-405 (dose 5 pg per mice) 3. Intrarectal TNBS / 50% ethanol+treatment with ORK1-068 (dose 5 pg per mice) ORK1-405 and ORK1-068 were administered intraperitoneally, at defined dose, 2 times per week.

[0644] The results show that mice treated with ORK1-405 had less body weight loss compared to both ORK1-068-treated or vehicle-treated mice, after 12 days. (Figure 12).Example 5: Optimization of the dual CSF1 -IL-10 cytokine

[0645] PBMC (150,000 cells / well) from healthy donors were plated on in 96-well plates and stimulated next day with saturating concentrations of rCSF1 for 5 min when indicated. Then samples were treated with ORK1-068, ORK1-341 and rlL-10 as a control (data not shown) (10 nM per treatment, 8 dilutions 1 / 3) for 20 min at 37°C. Cells were stained with CD14 and CD3, fixed with 4% PFA and permeabilized for intracellular staining with 80% MetOH. Cells were stained with pSTAT3 (pY705) (BD) for 1h (Figure 16). Each biological replicate was normalized by assigning the highest IL10 MFI value of the top concentration as 100% and the lowest MFI value of an untreated control as 0%. The MFI from the samples treated with ORK1 variants was normalized accordingly. In Figure 16A, ORK1-341 contain a mutation in CSF1 that is unable bind to CSF1R. In Figure 16B, an excess of soluble rCSF1 was added to saturate CSF1 receptors. In both examples ORK1-068 activates STAT3 preferentially in monocytes but this difference is lost when ORK1 is unable to bind to CSF1R.

[0646] Different sequence modifications were added to improve ORK1-405. The new ORK1-494 was compared to previous formats. PBMC (150,000 cells / well) from healthy donors were plated on in 96-well plates. Cells were treated next day with ORK1-068, ORK1-405, ORK1-494 or rlL-10 as a control (10 nM per treatment, 8 dilutions 1 / 3) for 20 min at 37°C. Cells were stained with CD14 and CD3, fixed with 4% PFA and permeabilized for intracellular staining with 80% MetOH. Cells were stained with pSTAT3 (pY705) (BD) for 1h. Each biological replicate was normalized by assigning the highest IL10 MFI value of the top concentration as 100% and the lowest MFI value of an untreated control as 0%. The MFI from the samples treated with ORK1 variants was normalized accordingly. EC50 from dose-response for each donor was represented and fold change between T cell values vs monocytes values was indicated in the graph. (Figure 17 A).

[0647] CD8 T cells were isolated from human PBMCs using positive microbeads selection (Miltenyi). The lymphocytes (1.5 million / ml) were stimulated with anti-CD3 (ThermoFisher) (1 pg / ml) and anti-CD28 (ThermoFisher) (3 pg / ml) for 72h. Then, cells were splited, replated and treated with IL10, ORK1-405 or ORK1-494 (10 nM per treatment, 6 dilutions 1 / 4) with IL-2 (5 ng / ml) for48h. Cell supernatants were harvested and GzmB was detected by ELISA. (Figure 17 B)

[0648] The results show that ORK1-405 and ORK1-494 are similar but less potent than IL-10 at activating CD8 T cells.

[0649] PBMCs were isolated from human buffy coats and CD14+ cells were purified by positive selection (Miltenyi). Monocytes (0.5 million / ml) were cultured for 7 days in thepresence or absence of: GM-CSF, CSF1, ORK1-405 or ORK1-494 (0.56nM). Cytokines were added every 2 days. At day 7, cells were stimulated with LPS, with or without IFNy (Figure 17C). TNF-a levels in the supernatants were determined by ELISA, 17h after stimulation. GM-CSF and CSF-1 -differentiated macrophages produce large amounts TNF-a. In contrast, ORK1-068 and ORK1-405-differentiated macrophages do not produce TNF-a, even when stimulated with LPS+IFNy.

[0650] ORK1-494 selectively target monocytes via CSF1R. PBMC from healthy donors were plated o / n in 96-well plates and treated when specified with saturating concentrations of rCSF1, rlL-10 or both (5 min, 37°C), after that cells were washed and resuspend in buffer containing sodium azide 0,1%. Then, ORK1-494 (15nM per treatment, 7 dilutions 1 / 3) is added and incubated 20’ on ice. Cells were washed and resuspended in stain buffer containing the mix of antibodies: CD14, CD3 and anti-human lgG4-FC. Finally, cells were resuspended in stain buffer. Samples were analysed with LSRII cytometer and MFI values obtained from anti-human lgG4-FC were plotted using GraphPad Prism software (Fig 18B). The results show a preferential binding to monocytes (CD14+). Binding of ORK1-494 to human PBMCs is predominantly restricted to CD14+monocytes and is inhibited by CSF1R blockade, by not by IL-10R blockade, confirming CSF1R-dependent targeting. Minimal binding to CD3+T cells suggest low off-target activity.

[0651] Example 6: In vivo, ORK1-494 protects mice from chronic TNBS-induced colitis.

[0652] ORK1-494 protects mice from chronic TNBS-induced colitis. TNBS was intrarectally infused in male Balb / c mice at four increasing doses (0.8 mg / mouse at day 0; 1.0 mg / mouse at day 7; 1.2 mg / mouse at day 14; and 1.5 mg / mouse at day 21). ORK1-494 was administered intraperitoneally, at defined dose, 2 times per week for 4 weeks.

[0653] The results show that ORK1-494 improves body weight recovery and reduces colitis scores in mice exposed to chronic TNBS compared to TNBS untreated mice, after 28 days of treatment. Protective effects observed were dose-dependent, also in survival rates. (Figure 19)

[0654] The results show that mice treated with ORK1-494 had less weight loss (Figure 19A), improved colitis score (Figure 19B) and survival (Figure 19C) compared to TNBS untreated mice, after 28 days of treatment. ORK1-494 dose-dependently improves survival rates in TNBS-induced chronic colitis.

[0655] The experimental Groups included 10 mice per group and were as follows:

[0656] 1. Healthy control mice with no TNBS or ORK1 treated

[0657] 2. Intrarectal TNBS / 50% ethanol+treatment with PBS3. I ntrarectal TNBS / 50% ethanol+treatment with ORK1-494 (dose 1 g per mice) 4. Intrarectal TNBS / 50% ethanol+treatment with ORK1-494 (dose 5 pg per mice) 5. Intrarectal TNBS / 50% ethanol+treatment with ORK1-494 (dose 10 pg per mice)

[0658] In Figure 20, similar experimental model of chronic IBD induced by TNBS was performed. TNBS was intrarectally infused in male Balb / c mice at four increasing doses (0.8 mg / mouse at day 0; 1.0 mg / mouse at day 7; 1.2 mg / mouse at day 14; and 1.5 mg / mouse at day 21). Same mice groups (10 mice / group) were included except for “Intrarectal TNBS / 50% ethanol+treatment with ORK1-494 (dose 1 pg per mice)”. ORK1-494 were administered for 2 weeks intraperitoneally, at defined dose, 2 times per week, after 4 doses mice was not treated anymore with ORK1-494 but continued to inject TNBS for two more weeks. ORK1-494 protects mice from chronic TNBS-induced colitis over time with no treatment repetition after 2 weeks, indicating a durable effect on inflammation and tissue repair.

[0659] Example 7: Spatial domain orientation affects binding and signaling Different spatial domain orientations were tested to optimize the molecule. In figure 21 all these formats designed were represented indicating position of CSF1, sclL-10, asymmetric Fc IgG and linkers.

[0660] In figure 22, PBMC (150,000 cells / well) from healthy donors were plated on in 96-well plates. Next day samples were treated with different ORK1 variants or rlL-10 as a control (15 nM per treatment, 8 dilutions 1 / 3) for 20 min at 37°C. Cells were stained with CD14 and CD3, fixed with 4% PFA and permeabilized for intracellular staining with 80% MetOH. Cells were stained with pSTAT3 (pY705) (BD) for 1h. Each biological replicate was normalized by assigning the highest IL10 MFI value of the top concentration as 100% and the lowest MFI value of an untreated control as 0%. The MFI from the samples treated with ORK1 variants was normalized accordingly. STAT3 activation assays revealed that CSF1 position in N-terminal was the preferred orientation, because of IL-10 signaling was reduced on T cells, maintaining preferential signaling for monocytes.

[0661] In figure 24, selected variants of interest were tested in a higher number of donors confirming that in monocytes, ORK1-513, ORK1-494 and ORK1-510 maintain full IL-10 antiinflammatory activity. ORK1-510 induces stronger pSTAT3 responses in monocytes without affecting T cells, albeit with reduced maximal activation compared to ORK1-494. In contrast, ORK1-513 maintains monocyte pSTAT3 responses while significantly reducing T cell activation, supporting partial agonism with selective immunomodulation. 50% maximumstimulation values from dose-response for each donor was represented and fold change between the median of T cell values vs monocytes values was indicated.

[0662] Comparing variants of interest on affinity to CSF1, ORK1-510 and ORK1-513 display higher mean fluorescence intensity (MFI) on monocytes compared to ORK1-494, while maintaining minimal interaction with T cells indicating higher binding affinity to CSF1R. (Figure 23).

[0663] Further characterization was performed to characterize the phenotype of the induced monocyte-derived macrophages. PBMCs were isolated from human buffy coats and CD14+ cells were purified by positive microbead selection (Miltenyi). Monocytes (0.5 million / ml) were cultured for 7 days in the presence or absence of: rCSF1, ORK1-494, ORK1-510 or ORK1-513 (1.08 nM per treatment). Cytokines were added every 2 days. At day 7, alive cells were counted and characterized by FACS (CD163, CD206, CD86 and HLA-DR). ORK1-510 and ORK1-513 promote differentiation of monocytes into macrophages expressing regulatory markers (CD163, CD206) with downregulation of inflammatory makers (CD86, HLA-DR) comparable to ORK1-494 (Figure 25A).

[0664] Similar experimental design was performed to assess the potency of IL-10 inhibiting inflammatory responses, after ORK1 treatment for 7 days cells were stimulated with LPS+IFN-y, supernatants were harvested and TNFa production was measured by ELISA (Figure 25B). In monocyte-derived macrophages, ORK1-510 and ORK1-513 retain the capacity to suppress TNFa production at levels comparable to ORK1-494.

[0665] ORK1-510 and ORK1-513 respective potencies activating CD8 T cells and B cells. B cells were isolated from human PBMCs using positive microbeads selection (Miltenyi). The isolated lymphocytes were stimulated with CD40L (T-dependent conditions) and IL10, ORK1-494, ORK1-510 and ORK1-513 variants at dO and d2. After 7 days plasma cell differentiation was analyzed by flow cytometry and cell supernatants were obtained for the determination of IgG levels by ELISA (data not shown) (Figure 26A). CD8 T cells were isolated from human PBMCs using positive microbeads selection (Miltenyi). Cells were activated for 3 days with α-CD3 and α-CD28 and treated with IL-10, ORK1-494, ORK1-510 and ORK1-513 with IL-2 and after 2 days supernatants were collected. GzmB was determined by ELISA (Figure 26B). Compared to ORK1-494 and ORK1-510, ORK1-513 achieves >8-fold reduction in CD8+T cell Granzyme B expression and >5-fold reduction in PC differentiation without loss of IL-10 function in monocytes. This selectivity suggests an improved safety profile through decoupling of proinflammatory lymphocyte responses.Co-cultures with CD14+ and CD3+ cells purified from total PBMC of healthy donors were performed to confirm the anti-inflammatory response of ORK1 -differentiated macrophages. Monocytes were cultured for 5 days in the presence or absence of: CSF1, ORK1-494, ORK1-510 and ORK1-513 at 1.08nM. Cytokines were added every 2 days. At day 5, cells were counted and replated to a 96 round well plate (40,000 cells / well) and last dose of cytokines was added. After 2 days CD3 T cells from another donor were purified and added to macrophages after refreshing media. 120.000 CD3 T cells were added per well in a final ratio of 1:3. Macrophages were activated with LPS ± NOD2 agonist and after 2 days supernatants were collected, and T cells were counted. ORK1-494, ORK1-510 and ORK1-513 suppress IFN-y, TNFa, IL-23 and GzmB expression, and T cell expansion in macrophage-T cell co-cultures, with no clear differences compared to each other. These results confirm that optimized variants retain anti-inflammatory control over lymphocyte responses without increasing off-target activity (Figure 27).

[0666] Example 8. In vivo, ORK1-513 shows limited efficacy in chronic TNBS model.

[0667] Evaluation of the therapeutic efficacy of ORK1 -513 was performed in an experimental model of chronic IBD induced by TNBS (Trinitrobenzene sulfonic acid (TNBS)-induced colitis mouse model), with comparison with ORK1-494.

[0668] TNBS was intrarectally infused in male Balb / c mice at three increasing doses (0.8 mg / mouse at day 0; 1.0 mg / mouse at day 7 and 1.2 mg / mouse at day 14).

[0669] The experimental groups included 15 mice per group and were as follows

[0670] 1. Intrarectal TNBS / 50% ethanol+treatment with PBS

[0671] 2. Intrarectal TNBS / 50% ethanol+treatment with ORK1-513 (dose 1 pg per mice) 3. Intrarectal TNBS / 50% ethanol+treatment with ORK1-513 (dose 5 pg per mice) 4. Intrarectal TNBS / 50% ethanol+treatment with ORK1-513 (dose 10 pg per mice) 5. Intrarectal TNBS / 50% ethanol+treatment with ORK1-494 (dose 5 pg per mice) ORK1-513 and ORK1-494 were administered intraperitoneally, at defined dose, 2 times per week for 3 weeks.

[0672] The results show that mice treated with ORK1-494 had less body weight loss (Figure 28B), improved colitis score (Figure 28C) and survival rates (Figure 28D) compared to both ORK1-513 treated or vehicle-treated mice. Altogether, ORK1-513 shows reduced efficacy in chronic TNBS-induced colitis compared to ORK1-494.

[0673] Example 9: PK studies ORK1-068 and ORK1-513

[0674] ORK1-068 and rlL-10 were administered IV at 0.5mg / kg into C57BL / 6J female mice at a n=3 per group. Blood samples were collected at several timepoints 5min,1h, 3h, 7h,11h, 24h. ORK1-068 serum concentration was measured using IL-10 and CSF1 ELISA methods kit(Human IL-10 Uncoated ELISA Kit, Thermo Catalog Number 88-7106; Human M-CSF antibody Abeam Catalog Number ab256655) and rlL-10 using Human IL-10 Uncoated ELISA Kit, Thermo Catalog Number 88-7106. ORK1-068 terminal half-life (T1 / 2) was between 0.995-1.01h while rlL-10 was 0.710h. The ORK1-068 Cmax and exposure (AUCO-tau) after 0.5mg / kg IV injection was higher than obtained with IL-10. Therefore, the dual CSF1-IL10 without IgG improved the half-life in respect to rl L10 but not dramatically (Figure 29A). The graph shows plasma drug concentration ORK1-068 and rlL10 versus time following administration. For clarity, graph A from Figure 29 shows molecule concentration data only data until 9h timepoint.

[0675] ORK1-513 was administered in 3 different groups at n=9 per group IV at 0.1mg / kg as a single dose, 0.5mg / kg as a single dose and 0.5mg / kg every 3 days (DO, D3, D6, D9, D12 and D15) for two weeks into C57BL / 6J female mice (according to the table of Figure 29). Each single dose group was divided into 3 subgroups and blood samples were collected at several timepoints 5min, 3h, 48h, 168h; 30 min, 6h, 72h, 240h; 1 h, 24h, 120h and 336h. In the repeated dosing group, blood samples were collected at DO, 5 min, 3h, 24h, pre-dose D9, D15 + 5 min, 3h, 24h, 168h; DO, 30min, 6h, 48h; D9 + 5min; D15 + 30min, 6h, 48h; DO, 1h, 12h, 72h (predose D6); pre-dose D15; D15, 1h, 12h, 72h.

[0676] Serum ORK1-513 concentration was measured using IL-10 and CSF1 ELISA methods kit. (Human IL-10 Uncoated ELISA Kit, Thermo Catalog Number 88-7106; Human M-CSF antibody Abeam Catalog Number ab256655). ORK1-513 in the Cmax and exposure (AUCO-tau) after 0.5mg / kg IV injection was 5,260 ng / ml and 17,221 h*ng / ml, respectively; 353ng / mL and 509h*ng / mL after 0.1mg / kg IV injection and 6,753ng / ml and 32,886 h*ng / ml in the repeated dose group at IV 0.5mg / kg. ORK1-513 terminal half-life (T1 / 2) was 5.34h in the single dose IV 0.5 mg / kg group, and 1.19 h in the single dose 0.1 mg / kg IV group and 4.92h in the group administered for 2 weeks every 3 days at a dose IV 0.5 mg / kg group. Therefore, the dual CSF1-IL10 with Fc improved the half-life in respect to non-Fc fused protein ORK1-068. ORK1-513 had a better half-life compared to native ORK1-068 non-Fc fused, 4.92-5.34h vs 1.01 h as shown in the graph in Figure 29. Figure 29B shows plasma drug concentration ORK1-513 in indicated dose and administration regimes versus time following administration. Figure 29C compares plasma drug concentration ORK1-513 in indicated dose vs ORK1-068. For clarity, Figure 29 graph B and C shows only molecule concentration until 12h timepoint.Example 10: Optimization through binding affinity mutations to CSF1.

[0677] Previous data reported demonstrated that increase binding translates in increased IL-10 signaling in monocytes compared to T cells. The increased IL-10 signaling could be mediated by CSF1-CSF1R interaction (Figure 30). The optimization strategy in this case was based on increase selectivity towards monocytes by increasing CSF1 affinity to CSF1 R based on a high-throughput mutational screening.

[0678] In Figure 31, data in Table represent all the single mutations selected from the screening. Binding assays were performed to identify variants with highest binding to CSF1R indicating most of the variants increasing binding in comparison to ORK1-513 (Figure 31 B).

[0679] Total PBMC from healthy donors were treated with IL-10, or ORK1-513 compare to different ORK1 candidates (ORK1-527 to ORK1-550) which include different single CSF1 affinity mutations (described in table of Figure 31). Cells were stained with CD14 and CD3 (data not shown) antibodies and intracellular staining for pSTAT3. In figure 32, all selected affinity mutations were tested in pSTAT3 in monocytes (CD14+). ORK1-532 was included as one of the best performing variants lowering the 50% maximum stimulation in pSTAT3 in monocytes

[0680] Ratio between values of 50% maximum stimulation from T cells vs monocytes is shown in figure 33, where ORK1-532 outperforms previous variants as ORK-513 or ORK1-494 and IL-10 as a control. Dose-response curves from % of positive cells from STAT3 activation in monocytes and T cells from samples treated with ORK1-513 and ORK1-532 (Figure 33C) show that ORK1-532 behave similar as ORK1-513 in T cells but in contrast, ORK1-532 monocyte pSTAT3 shows an improved profile compared to ORK1-513, making ORK1-532 an improved variant. Data show that CSF1 affinity mutation ORK1-532 increase selectivity to monocytes and outperforms previous formats in selectivity and function.

[0681] In ORK1-532 variant, IL-10 signaling is mainly driven by CSF1-CSF1R interaction. Total PBMC from healthy donors were treated when specified withsaturating concentrations of rCSF1. After that, samples were stimulated with ORK1-532 or IL-10 as a control. Cells were stained with CD14 and CD3 antibodies and intracellular staining for pSTAT3. (Figure 34) Increase binding translates in increased IL-10 signaling in monocytes compared to T cells mediated by CSF1-CSF1R interaction.

[0682] ORK1-532 variant induces regulatory macrophages without activating T cells.CD14+ cells were purified from total PBMC of healthy donors. Monocytes were cultured for 7 days in the presence or absence of: CSF1, ORK1-513 or ORK1-532. Cytokines were added every 2 days. Macrophages were stimulated overnight with LPS+IFN-y. TNFa was measured by ELISA. In monocyte-derived macrophages, ORK1-532 outperforms previous formats and maintain the highest IL- 10 anti-inflammatory activity (Figure 35A)

[0683] CD8 T cells were isolated from human PBMCs using positive microbeads selection (Miltenyi). Cells were activated for 3 days with α-CD3 and α-CD28 and treated with IL-10, ORK1-532 or ORK1-513 with IL-2 and after 2 days supernatants were collected. GzmB was determined by ELISA. ORK1-532 is similar at ORK1-513, both reducing activation of CD8 T cells (Figure 35B).

[0684] All data generated from experiments performed with single affinity mutations to CSF1 are reported in Table 2.Table 2: Evaluation of variants including single affinity mutations to CSF1 in induction of regulatory macrophages without T cell activation

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[0732] Following the optimization process, 12 most promising single affinity mutations to CSF1 were selected and all possible combinations were generated for testing in vitro (table 3).

[0733] Table 3: Variants with combination of affinity mutations to CSF1

[0734] Variant Mutations

[0735] ORK1-567 S18I, T64H, M65F, R66V, L85l, R86Y, V120F

[0736] ORK1-568 E5R, S18I, T64H, R66L, L85V, R86Y, V120F

[0737] ORK1-569 S18I, T64H, R66L, V78Y, L85I, R86Y, V120F

[0738] ORK1-570 S18I, T64H, R66L, L85l, R86Y, V120F

[0739] ORK1-571 S18I, T64H, R66Y, L85l, R86Y, V120F

[0740] ORK1-572 S18I, T64H, A74D, L85l, R86Y, V120F

[0741] ORK1-573 S18I, T64H, R66I, L85I, R86Y

[0742] ORK1-574 S18I, T64H, R66V, A74D, L85I

[0743] ORK1-575 E5R, T64H. R66L. L85I, R86Y

[0744] ORK1-576 S18I, T64H, R66V, A74D, L85V, R86Y, V120F

[0745] ORK1-577 S18I, T64H, R66I, A74D, L85V, R86Y. V120F

[0746] ORK1-578 S18I, T64H, R66V, A74D, L85I, R86Y. V120F

[0747] ORK1-579 S18I, T64H, R66I, A74D, L85I, R86Y. V120F

[0748] ORK1-580 S18I, T64H, R66V, A74D, R86Y, V120F

[0749] ORK1-581 S18I, T64H, R66V, A74D, L85Y, R86Y. V120F

[0750]

[0751] All data generated from experiments performed were reported in table 4. ORK1-576, ORK1-578 and ORK1-579 were selected as best performing variants with multiple mutations but none of them outperformed ORK1-532.Table 4: Evaluation of variants including multiple affinity mutations to CSF1 in induction of regulatory macrophages without T cell activation

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[0792] o n oExample 11: In vivo optimization through binding affinity mutations to CSF1.

[0793] Evaluation of the therapeutic efficacy of ORK1-532 was performed in an experimental model of chronic IBD induced by TNBS (Trinitrobenzene sulfonic acid (TNBS)-induced colitis mouse model), with comparison with ORK1-494 at one dose.

[0794] TNBS was intrarectally infused in male Balb / c mice at three increasing doses (0.8 mg / mouse at day 0; 1.0 mg / mouse at day 7; and 1.2 mg / mouse at day 14).

[0795] The experimental Groups included 15 mice per group and were as follows:

[0796] 1. Intrarectal TNBS / 50% ethanol+treatment with PBS

[0797] 2. Intrarectal TNBS / 50% ethanol+treatment with ORK1-532 (dose 5 pg per mice) 3. Intrarectal TNBS / 50% ethanol+treatment with ORK1-494 (dose 5 pg per mice)

[0798] ORK1-532 and ORK1-494 were administered intraperitoneally, at defined dose, 2 times per week for 2 weeks. The results show that mice treated with ORK1-532 had less body weight loss (Figure 36B), improved colitis score (Figure 36C) and survival (Figure 36D) compared to both ORK1-494 treated or vehicle-treated mice, after 21 days treatment. ORK1-532 alleviates clinical symptoms in a TNBS mouse model of colitis and other inflammatory diseases compared to ORK1-494.

[0799] Example 12: Optimization through linker length and composition

[0800] ORK1-513 was selected as scaffold for linker modification. In Figure 37B, several linker types were described combining short / long and / or rigid / flexible. In Figure 37C, different combinations of L1 and L2 in type of linker and the variants produced were described.

[0801] Total PBMC from healthy donors were treated with IL-10, ORK1-513 and linker variants designed. Cells were stained with antibodies specific for CD14 and CD3, followed by intracellular staining for pSTAT3. MFI values corresponding to STAT3 activation were indicated in Figure 38 A. Graph to illustrate Vmax restore in ORK1-555 compared to ORK1-513 and IL-10 as a control.

[0802] In Figure 38B correlation between Vmax values (relative to IL- 10) and EC50 in monocytes from pSTAT3 activation, all linker variants were included and ORK1-513 was used as a control. 5 variants were selected as best performers with higher Vmax and low EC50 in monocytes (ORK1-564, ORK1-563, ORK1-555, ORK1-556 and ORK1-566), all of them with short L1 linkers. Short L1 linkers improve monocyte potency and restore IL-10-like Vmax.

[0803] The correlation between ratio of T cell vs monocyte pSTAT3 activation and EC50 on monocytes in pSTAT3 activation identify 4 of the 5 previously selected variants in Figure 38(ORK1-564, ORK1-566, ORK1-563 and ORK1-555). Figure 39A illustrates that Short L1 Linkers Drive Optimal Monocyte / T Cell Selectivity. In Figure 39B, ratio between (50% maximum stimulation in pSTAT3 activation relative to IL-10) values from T cells vs monocytes is shown in the graph. Data from linker variants ORK1-551 to ORK1-566 is shown in relative to ORK1-513 as a control, the selected variants of interest are indicated in darker color. All data generated from experiments performed with linker combinations were reported in table 4.

[0804] ORK1-555 selected as candidate for linker modification, together with variant selected from single affinity mutations (ORK1-532), synergies the effects observed in both variants.

[0805] Total PBMC from healthy donors were treated with IL-10, ORK1-513, ORK1-532, ORK1-555 or ORK1-587. Cells were stained with CD14 and CD3 antibodies and intracellular staining for pSTAT3. Data is shown in a dose-response graph and interpolated to 50% of maximum stimulation. ORK1-587 has improved IL-10 signaling due to single mutation from ORK1-532 and increased Vmax due to linker modification from ORK1-555 (Figure 40).

[0806] SEQUENCES

[0807] Mutations are in bold. Signal peptides used to secrete the proteins which are cleaved upon secretion are not showed. We used for proteins starting by IL10 and FoldikinelO the signal sequence: MHSSALLCCLVLLTGVRA (SEQ ID NO: 69). In the case of proteins starting by CSF1 we used MTAPGAAGRCPPTTWLGSLLLLVCLLASRSIT (SEQ ID NO:70). In Fc fusion we used MEWSWVFLFFLSVTTGVHS (SEQ ID NO:110).

[0808] SEQ ID NO: 1 foldikinelO (with mutations D84E ( first IL-10 domain), and C108N in second IL-10 domain, short Nter, X3-L1.1-Z sequence NNGGLDYL (SEQ ID NO: 71 ) ):

[0809] TQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQF YLEEVMPQAENQEPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIY KAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKG YLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQV KNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 2 QRK10-002 (short Nter, X3-L1.1-Z sequence NFGGLDYL (SEQ ID NO: 45 ) ) TQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQF YLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIY KAMSEFDIFINYIEAYMTMNFGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKG YLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQV KNAFNKLQEKGIYKAMSEFDIFINYIEAYMTM SEQ ID NO: 3 QRK10-003 ( short Nter, X3-L1. 1-Z sequence LYKTITPEFA (SEQ ID NO: 46 )TQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQF YLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIY KAMSEFDIFINYIEAYLYKTITPEFANMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYL GCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKN AFNKLQEKGIYKAMSEFDIFINYIEAYMTM SEQ ID NO: 4 ORK10-005 (short Nter, X3-L1.1-Z sequence KDKDIRDGDELA (SEQ ID NO: 47 ) ) TQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQF YLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIY KAMSEFDIFINYIEAKDKDIRDGDELANMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGY LGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVK NAFNKLQEKGIYKAMSEFDIFINYIEAYMTM SEQ ID NO: 5 Foldikine 10 (without Methionine, only C108N in second IL-10 domain, and N-terminal completed, X3-L1.1-Z sequence MTMNNGGLDYLP ( SEQ ID NO: 263 ) ) SPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALS EMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQ EKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLL EDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSK AVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 6 Long CSFl-FoldikinelO EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVVTKPDCNCLYPKAIPSSDPASVSPHQPLAPSMAPVAGLTWEDSEGTE GSSLLPGEQPLHTVDPGSAKQRPPRGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGTQSENSCTHFP GNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAEN QDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFIN YIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMI QFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKG IYKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 7 short CSFl-FoldikinelO EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGN LPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQD PDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYI EAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQF YLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIY KAMSEFDIFINYIEAYMTMKIRN

[0810] SEQ ID NO: 8 long CSFl-scILlO ( from SEQ ID 104 ) EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVVTKPDCNCLYPKAIPSSDPASVSPHQPLAPSMAPVAGLTWEDSEGTE GSSLLPGEQPLHTVDPGSAKQRPPRGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENS CTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVM PQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEF DIFINYIEAYMTMKIRNGGGSGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMK DQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLR RCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNSEQ ID NO: 9 QRKMCSF_013 KACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFNECEDQDRRQPGTKTTGKVSEY CSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTP NAIAIVQLQELSLRLKSCFTKDY SEQ ID NO: 10 ORKMCSF_014 KACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECGKPPQYIDRHTQPGQPNAT SEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRD NTPNAIAIVQLQELSLRLKSCFTKDY SEQ ID NO: 11 IL10:

[0811] SPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALS EMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQ EKGIYKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 12 CSF1 short version without valine EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQD SEQ ID NO: 13 IL-10-CSF1 SPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALS EMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQ EKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGEEVSEYCS HMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNA IAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNI FSKNCN NSFAECSSQD SEQ ID NO: 14 FoldikinelO-CSFl TQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQF YLEEVMPQAENQEPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIY KAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKG YLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQV KNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGSGGSGGSGGSGGSGGSGGSGGG EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQD SEQ ID NO: 17 CSF1 long version EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDWTKPDCNCLYPKAIPSSDPASVSPHQPLAPSMAPVAGLTWEDSEGTE GSSLLPGEQPLHTVDPGSAKQRPPR SEQ ID NO: 19 ORK1_068_P EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGMTQSENSCTHFPGNLPNM LRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIK AHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYM TMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEE VMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMS EFDIFINYIEAYMTMKIRNSEQ ID NO: 20 engineered IgG4 Fc FALA ( knob) ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 21 short CSF1 ( including valine) -FoldikinelO (from SEQ ID 5 ) EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 22 single chain dimeric IL-10 domain in ORK1-455 and ORKl-p-482 SPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALS EMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQ EKGIYKAMSEFDIFINYIEAYMTMKIRNGGGSGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDA FSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSE EDI FINY IEAYMTMKIRN SEQ ID NO: 25 CSFl-30aa IL10 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGN LPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQD PDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYI EAYMTMKIRN SEQ ID NO: 30 Nt Ct linker

[0812] NGGLDY SEQ ID NO: 31 Nt Ct linker

[0813] YKTIT SEQ ID NO: 32 Nt Ct linker

[0814] DKDIRDGD SEQ ID NO: 33 linker LI 15

[0815] GGSGGSGGSGGSGGG SEQ ID NO: 34 linker LI 25

[0816] GGGSGGSGGSGGSGGSGGSGGSGGG SEQ ID NO: 35 linker LI 30

[0817] GGGGGSGGSGGSGGSGGSGGSGGSGGSGGG SEQ ID NO: 36 linker LI 35 GGGGGSGGGGSGGSGGSGGSGGSGGSGGSGGSGGG SEQ ID NO: 45NFGGLDYL

[0818] SEQ ID NO: 46

[0819] LYKTITPEFA SEQ ID NO: 47

[0820] KDKDIRDGDELA SEQ ID NO: 48

[0821] FGGLDY SEQ ID NO: 65 N-terminal part of SC dimeric IL-10 TQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQF YLEEVMPQAENQ SEQ ID NO: 66 C-terminal part of SC dimeric IL-10 NMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPD IKAHVNSLGENLKTLRLRLRR SEQ ID NO: 67

[0822] MTMN SEQ ID NO: 68

[0823] PE FA SEQ ID NO: 71

[0824] NNGGLDYL SEQ ID NO: 76

[0825] MTMKIRN SEQ ID NO: 77

[0826] MTMK I RNNNGGL DY L P SEQ ID NO: 79 (CSF1 Short version ending in QD, sc IL-10 with D84E, C108N (2nd IL10 domain) short N term) EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGTQSENSCTHFPGNLPNML RDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIKA HVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMT MNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEV MPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSE FDIFINYIEAYMTMKIRN SEQ ID NO: 80 (CSF1 long version ( SEQ ID NO: 17 ), sc IL-10 with D84E, C108N (2nd IL10 domain) short N term) ORK1 P 497 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVVTKPDCNCLYPKAIPSSDPASVSPHQPLAPSMAPVAGLTWEDSEGTE GSSLLPGEQPLHTVDPGSAKQRPPRGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGTQSENSCTHFP GNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAEN QEPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFIN YIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKG IYKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 81

[0827] SPGQG SEQ ID NO: 82

[0828] GGGGS SEQ ID NO: 83 QRKl-Fc-01 (Protein 1 of QRKl_P_400 ) ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQR LIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRL KSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGGG GGSGGSGGSGGSGGSGGSGGSGGSGGGMTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQL DNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTLRLRLRRCH RFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAF SRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGE NLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINYIEAYMTMKIRN SEQ ID NO: 84 QRKl-Fc-02 ( Protein 1 of QRK1_P_4O1 ) EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGMTQSENSCTHFPGNLPNM LRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIK AHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYM TMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEE VMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMS EFDIFINYIEAYMTMKIRNGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDT LMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNG KEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWE SNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 85 QRKl-Fc-03 ( Protein 1 of ORK1_P_402 ) ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSEEVSEY CSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTP NAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNI FSKN CNNSFAECSSQDGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGMTQSENSCTHFPGNLPNMLRDLRD AFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSL GENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINYIEAYMTMNNGG LDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAE NQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FI NYIEAYMTMKIRN SEQ ID NO: 86 QRKl-Fc-04 ( Protein 1 of ORK1_P_403 )EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGMTQSENSCTHFPGNLPNM LRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIK AHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYM TMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEE VMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMS EFDIFINYIEAYMTMKIRNGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYR VVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTC LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALH NHYTQKSLSLSLG SEQ ID NO: 87 ORKl-Fc-05 ( Protein 1 of ORK1_P_404 ) EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGMTQSENSCTHFPGNLPNM LRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIK AHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYM TMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEE VMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMS EFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKD TLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEW ESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 88 ORKl-Fc-06 ( Protein 1 of ORK1_P_405 ) ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQ RLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLR LKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGG GGGSGGSGGSGGSGGSGGSGGSGGSGGGMTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDA FSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 89 Protein 1 of ORK1_P_438 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQ RLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLR LKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGG GGGSGGSGGSGGSGGSGGSGGSGGSGGGMTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDA FSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNSEQ ID NO: 90 Protein 1 of ORK1_P_439 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQ RLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLR LKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGG GGGSGGSGGSGGSGGSGGSGGSGGSGGGMTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDA FSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 91 Protein 1 of ORK1_P_440 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQ RLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLR LKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGG GGGSGGSGGSGGSGGSGGSGGSGGSGGGMTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDA FSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRAHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 92 Protein 1 of ORK1_P_441 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQ RLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLR LKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGG GGGSGGSGGSGGSGGSGGSGGSGGSGGGMTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDA FSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 93 Protein 1 of ORK1_P_473 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQ RLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLR LKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGG GGGSGGSGGSGGSGGSGGSGGSGGSGGGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQL DNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCH RFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAF SRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGE NLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINYIEAYMTMKIRNSEQ ID NO: 94 Protein 1 of ORK1_P_474 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQ RLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLR LKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGG GGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQ MKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLR LRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRD LRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHV NSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMK IRN SEQ ID NO: 95 Protein 1 of ORK1_P_475 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQ RLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLR LKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGG GGGSGGSGGSGGSGGSGGSGGSGGSGGGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQL DNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCH RFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAF SRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGE NLKTLRLRLRRAHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINYIEAYMTMKIRN SEQ ID NO: 96 Protein 1 of ORK1_P_476 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQ RLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLR LKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGG GGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQ MKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLR LRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRD LRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHV NSLGENLKTLRLRLRRAHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMK IRN SEQ ID NO: 97 Protein 1 of ORK1_P_482 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQ RLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLR LKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGG GGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQ MKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLR LRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGSGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSE MIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQE KGIYKAMSEFDI FINYIEAYMTMKIRN SEQ ID NO: 98 Protein 1 of ORK1_P_485 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQ RLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLR LKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGG GGGSGGSGGSGGSGGSGGSGGSGGSGGGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQL DNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTLRLRLRRCH RFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAF SRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGE NLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINYIEAYMTMKIRN SEQ ID NO: 99 Protein 1 of ORK1_P_486 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQ RLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLR LKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGG GGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQ MKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTLRLR LRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRD LRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHV NSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMK IRN SEQ ID NO: 100 Protein 1 of ORK1_P_493 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQ RLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLR LKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDVG GGGGSGGSGGSGGSGGSGGSGGSGGSGGGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDA FSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSE EDI FINYIEAYMTMKIRN SEQ ID NO: 101 Protein 1 of ORK1_P_494 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQ RLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLR LKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDVG GGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRL RLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINYIEAYMTMNNGGLDYLPNMLR DLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAH VNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTM KIRN SEQ ID NO: 102 Protein 1 of ORK1_P_495 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQ RLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLR LKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDVG GGGGSGGSGGSGGSGGSGGSGGSGGSGGGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDA FSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 103 Protein 1 of ORK1_P_496 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQ RLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLR LKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDVG GGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTLRL RLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINYIEAYMTMNNGGLDYLPNMLR DLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAH VNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTM KIRN SEQ ID NO: 104 Protein 1 of ORK1-497_P ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQ RLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLR LKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDVV TKPDCNCLYPKAIPSSDPASVSPHQPLAPSMAPVAGLTWEDSEGTEGSSLLPGEQPLHTVDPGSAK QRPPRGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTF FQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLR LRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNML RDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKA HVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMT MKIRN*

[0829] SEQ ID NO: 105 ORK1_455_P EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQD PDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYI EAYMTMKIRNGGGSGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLL LKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLP CENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 106

[0830] CPPCP SEQ ID NO: 107

[0831] CPRCP SEQ ID NO: 108

[0832] CPSCP SEQ ID NO: 109 engineered IgG4 Fc FALA (hole ) ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVCT LPPSQEEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: NO: 110 signal peptide

[0833] MEWSWVFLFFLSVTTGVHS SEQ ID NO: NO: 111 signal peptide

[0834] MPRGFTWLRYLGIFLGVALG SEQ ID NO: NO: 112 signal peptide MTAPGAAGRCPPTTWLGSLLLLVCLLASRSIT SEQ ID NO: 119 (GGGGS)3

[0835] GGGGSGGGGSGGGGS SEQ ID NO: 120

[0836] SGGGGSGGGGS SEQ ID NO: 121

[0837] SGGGGSGGGGSAP SEQ ID NO: 122

[0838] NFSQP SEQ ID NO: 123

[0839] KRTVA SEQ ID NO: 124

[0840] GGGSGGGG SEQ ID NO: 125

[0841] GGGGSGGGGS SEQ ID NO: 126

[0842] THTCPPCPEPKSSDK SEQ ID NO: 127GGGS

[0843] SEQ ID NO: 128

[0844] EAAKEAAKGGGGS SEQ ID NO: 129

[0845] EAAKEAAK SEQ ID NO: 130 GGSSGSGSGSTGTSSSGTGTSAGTTGTSASTSGSGSGGGGGSGGGGSAGG SEQ ID NO: 131

[0846] GGGGSGGGGSGGGGSGGGGS SEQ ID NO: 132 ORK1 P 068 with signal peptide and tag MTAPGAAGRCPPTTWLGSLLLLVCLLASRSITEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQIT FEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDK ACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSG GSGGSGGSGGSGGGMTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDF KGYLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVE QVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQL DNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNH RFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGWYFQSGGHHHHH HGGG VSGWRL FKKIS SEQ ID NO: 133 ORK1-455 P with signal peptide and HiBit tag MTAPGAAGRCPPTTWLGSLLLLVCLLASRSITEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQIT FEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDK ACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSG GSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESL LEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKS KAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGSGGGGSSPGQGTQSENSCTHF PGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAE NQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FI NY I E AYMTMK I RNGW YFQSGGHHHHHHGGGVSGWRL FKKIS SEQ ID NO: 136 linker L2

[0847] GGGGGSGGGGSGGGGS SEQ ID NO 137: engineered IgGl murine Fc ( knob) VPRDCGCKPCICTVKEVSKVFIFPPKPKDVLTITLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHT AQTQPREEQFNSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISKTKGRPKAPQVCTIP PPKEQMAKDKVSLWCMITDFFPEDITVEWQWNGQPAENYKNTQPIMNTNGSYFVYSKLNVQKSNWE AGNTFTCSVLHEGLHNHHTEKSLSHSPGK SEQ ID NO 138: engineered IgGl murine Fc (hole) VPRDCGCKPCICTVKEVSKVFIFPPKPKDVLTITLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHT AQTQPREEQFNSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISKTKGRPKAPQVYTIP PPKCQMAKDKVSLSCAITDFFPEDITVEWQWNGQPAENYKNTQPIMNTNGSYFVVSKLNVQKSNWE AGNTFTCSVLHEGLHNHHTEKSLSHSPGK SEQ ID NO 139: engineered IgGl Fc LALAPG ( knob) DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPG SEQ ID NO 140: engineered IgGl Fc LALAPS ( knob) DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALSAPIEKTISKAKGQPREPQVYTLP PCREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPG SEQ ID NO 141: engineered IgGl Fc ( knob) DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALSAPIEKTISKAKGQPREPQVYTLP PCREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPG SEQ ID NO 142: engineered IgGl Fc STR ( knob) DKTHTCPPCPAPESTRGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALSAPIEKTISKAKGQPREPQVYTLP PCREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPG SEQ ID NO 143: engineered IgG4 Fc IgG4 FALA (R409K mutation, knob ) ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO 144: engineered IgGl Fc LALAPG (hole) DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVCTLP PSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPG SEQ ID NO 145: engineered IgGl Fc LALAPS (hole) DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALSAPIEKTISKAKGQPREPQVCTLP PSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPG SEQ ID NO 146: engineered IgGl Fc LALADS (hole) DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALSAPIEKTISKAKGQPREPQVCTLP PSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPG SEQ ID NO 147: engineered IgGl Fc STR (hole ) DKTHTCPPCPAPESTRGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALSAPIEKTISKAKGQPREPQVCTLP PSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPG SEQ ID NO 148: engineered IgG4 Fc FALA (R409K mutation, hole)ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVCT LPPSQEEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO 149: IgG4 Fc FALA ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 150: Protein 1 of ORK1_P _498 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSKEVSEHCSHMIGNGHLKVLQ QLIDSQMETSCQIAFEFVDQEQLDDPVCYLKKAFFLVQDIIDETMRFKDNTPNANATERLQELSNN LNSCFTKDYEEQNKACVRTFHETPLQLLEKIKNFFNETKNLLEKDWNIFTKNCNNSFAKCSSRDVG GGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRL RLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINYIEAYMTMNNGGLDYLPNMLR DLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAH VNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTM KIRN SEQ ID NO: 151: Protein 1 of ORK1_P _499 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSKEVSEHCSHMIGNGHLKVLQ QLIDSQMETSCQIAFEFVDQEQLDDPVCYLKKAFFLVQDIIDETMRFKDNTPNANATERLQELSNN LNSCFTKDYEEQNKACVRTFHETPLQLLEKIKNFFNETKNLLEKDWNIFTKNCNNSFAKCSSRDVG GGGGSGGSGGSGGSGGSGGSGGSGGSGGGSRGQYSREDNNCTHFPVGQSHMLLELRTAFSQVKTFF QTKDQLDNILLTDSLMQDFKGYLGCQALSEMIQFYLVEVMPQAEKHGPEIKEHLNSLGEKLKTLRM RLRRCHRFLPCENKSKAVEQVKSDFNKLQDQGVYKAMNEFDI FINCIEAYMMIKMKSGGGSGGGGS SRGQYSREDNNCTHFPVGQSHMLLELRTAFSQVKTFFQTKDQLDNILLTDSLMQDFKGYLGCQALS EMIQFYLVEVMPQAEKHGPEIKEHLNSLGEKLKTLRMRLRRCHRFLPCENKSKAVEQVKSDFNKLQ DQGVYKAMNEFDIFINCIEAYMMIKMKS SEQ ID NO: 152: Protein 1 of ORK1_P _500 VPRDCGCKPCICTVKEVSKVFIFPPKPKDVLTITLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHT AQTQPREEQFNSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISKTKGRPKAPQVCTIP PPKEQMAKDKVSLWCMITDFFPEDITVEWQWNGQPAENYKNTQPIMNTNGSYFVYSKLNVQKSNWE AGNTFTCSVLHEGLHNHHTEKSLSHSPGKGGGGGSGGGGSGGGGSKEVSEHCSHMIGNGHLKVLQQ LIDSQMETSCQIAFEFVDQEQLDDPVCYLKKAFFLVQDIIDETMRFKDNTPNANATERLQELSNNL NSCFTKDYEEQNKACVRTFHETPLQLLEKIKNFFNETKNLLEKDWNIFTKNCNNSFAKCSSRDVGG GGGSGGSGGSGGSGGSGGSGGSGGSGGGSRGQYSREDNNCTHFPVGQSHMLLELRTAFSQVKTFFQ TKDQLDNILLTDSLMQDFKGYLGCQALSEMIQFYLVEVMPQAEKHGPEIKEHLNSLGEKLKTLRMR LRRCHRFLPCENKSKAVEQVKSDFNKLQDQGVYKAMNEFDIFINCIEAYMMIKMKSGGGSGGGGSS RGQYSREDNNCTHFPVGQSHMLLELRTAFSQVKTFFQTKDQLDNILLTDSLMQDFKGYLGCQALSE MIQFYLVEVMPQAEKHGPEIKEHLNSLGEKLKTLRMRLRRCHRFLPCENKSKAVEQVKSDFNKLQD QGVYKAMNEFDI FINCIEAYMMIKMKS SEQ ID NO: 153: Protein 1 of ORK1_P_501ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQ RLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLR LKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDVG GGGGSGGSGGSGGSGGSGGSGGSGGSGGGTDQCDNFPQMLRDLRDAFSRVKTFFQTKDEVDNLLLK ESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPEAKDHVNSLGENLKTLRLRLRRCHRFLPCE NKSKAVEQIKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTIKARGGGSGGGGSTDQCDNFPQMLRD LRDAFSRVKTFFQTKDEVDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPEAKDHV NSLGENLKTLRLRLRRCHRFLPCENKSKAVEQIKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTIK AR SEQ ID NO: 154: Protein 1 of ORK1_P_502 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQ RLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLR LKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDVG GGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRL RLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINYIEAYMTMKIRNGGGSGGGGS TDQCDNFPQMLRDLRDAFSRVKTFFQTKDEVDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMP QAENQDPEAKDHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQIKNAFNKLQEKGIYKAMSEFD IFINYIEAYMTIKAR SEQ ID NO: 155: Protein 1 of ORK1 P 503 (Hole plasmid is ORK1_V_534, SEQ ID NO: 144 ) DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLP PCREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQR LIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRL KSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDVGG GGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQ MKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLR LRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRD LRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHV NSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMK IRN SEQ ID NO: 156: Protein 1 of ORK1_P_504 DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALSAPIEKTISKAKGQPREPQVYTLP PCREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQR LIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRL KSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDVGG GGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQ MKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLR LRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRD LRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHV NSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMK IRNSEQ ID NO: 157: Protein 1 of ORK1_P_505 DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALSAPIEKTISKAKGQPREPQVYTLP PCREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQR LIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRL KSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDVGG GGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQ MKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLR LRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRD LRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHV NSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMK IRN SEQ ID NO: 158: Protein 1 of ORK1_P_506 DKTHTCPPCPAPESTRGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALSAPIEKTISKAKGQPREPQVYTLP PCREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQR LIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRL KSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDVGG GGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQ MKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLR LRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRD LRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHV NSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMK IRN SEQ ID NO: 159: Protein 1 of ORK1_P_507 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSSPGQGTQSENSCTHFPGNLP NMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPD IKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEA YMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYL EEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKA MSEFDIFINYIEAYMTMKIRNGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGEEVSEYCSHMIGSGH LQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQ ELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECS SQDV SEQ ID NO: 160: Protein 1 of ORK1_P_508 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSTDQCDNFPQMLRDLRDAFSR VKTFFQTKDEVDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPEAKDHVNSLGENL KTLRLRLRRCHRFLPCENKSKAVEQIKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTIKARGGGSG GGGSTDQCDNFPQMLRDLRDAFSRVKTFFQTKDEVDNLLLKESLLEDFKGYLGCQALSEMIQFYLE EVMPQAENQDPEAKDHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQIKNAFNKLQEKGIYKAM SEFDIFINYIEAYMTIKARGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGEEVSEYCSHMIGSGHLQ SLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQ DV SEQ ID NO: 161: Protein 1 of ORK1_P_509 and ORK1_P_510 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGESKYGPPCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYR VVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWC LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALH NHYTQKSLSLSLG SEQ ID NO: 163: Protein 1 of ORK1_P_511 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGEEVSEYCSHMIG SGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIV QLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFA ECSSQDVGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKG LPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 164: Protein 1 of ORK1_P_512 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGGGSGGSGGSGGSGGSGGSGGSGGSGGGHDKACVRTFYET PLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFNECEDQDRRQPGTKTTGKVSEYCSHMIGSGHL QSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQE LSLRLKSCFTKDYEEGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPN MLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDI KAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAY MTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLE EVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAM SEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 165: Protein 1 of ORK1_P_513 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 166: Protein 1 of ORK1_P_514 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQ RLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLR LKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDVG GGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRL RLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINYIEAYMTMNNGGLDYLPNMLR DLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAH VNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTM KIRN SEQ ID NO: 167: Protein 1 of ORK1_P_515 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMKIRNGGGSGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNL LLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFL PCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSES KYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHN AKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLP PCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQ EGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 168: Protein 1 of ORK1_P_516 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSSPGQGTQSENSCTHFPGNLP NMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPD IKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEA YMTMKIRNGGGSGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLK ESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCE NKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGSGGSGGSGGSGGS GGSGGSGGGEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLV QDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNE TKNLLDKDWNIFSKNCNNS F AE C S S Q D V SEQ ID NO: 177: Protein 1 of ORK1_P_525 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMKIRNGGGSGGGGSTDQCDNFPQMLRDLRDAFSRVKTFFQTKDEVDNLLLKESLLEDFKG YLGCQALSEMIQFYLEEVMPQAENQDPEAKDHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQI KNAFNKLQEKGIYKAMSEFDIFINYIEAYMTIKARGGGGGSGGGGSGGGGSESKYGPPCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSL WCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEA LHNHYTQKSLSLSLG SEQ ID NO: 178: Protein 1 of ORK1_P_526 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSSPGQGTQSENSCTHFPGNLP NMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPD IKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEA YMTMKIRNGGGSGGGGSTDQCDNFPQMLRDLRDAFSRVKTFFQTKDEVDNLLLKESLLEDFKGYLG CQALSEMIQFYLEEVMPQAENQDPEAKDHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQIKNA FNKLQEKGIYKAMSEFDIFINYIEAYMTIKARGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGEEVS EYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDN TPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFS KNCNNSFAECSSQDV SEQ ID NO: 179: Protein 1 of ORK1_P_527 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 180: Protein 1 of ORK1_P_528 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAGLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 181: Protein 1 of ORK1_P_529 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSIRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 182: Protein 1 of ORK1_P_530 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVEDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 183: Protein 1 of ORK1_P_531 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 184: Protein 1 of ORK1_P_532 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTYR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 185: Protein 1 of ORK1_P_533 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMI FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 186: Protein 1 of ORK1_P_534 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNDIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 187: Protein 1 of ORK1_P_535 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIYQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 188: Protein 1 of ORK1_P_536 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLYELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 189: Protein 1 of ORK1_P_537 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFQETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 190: Protein 1 of ORK1_P_538 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCQNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 191: Protein 1 of ORK1_P_539 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLYVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 192: Protein 1 of ORK1_P_540 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 193: Protein 1 of ORK1_P_541 EEVSRYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 194: Protein 1 of ORK1_P_542 EEVSEYCSQMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 195: Protein 1 of ORK1_P_543 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG

[0848] SEQ ID NO: 196: Protein 1 of ORK1_P_544 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTEDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 197: Protein 1 of ORK1_P_545 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDYACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 198: Protein 1 of ORK1_P_546 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDN NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 199: Protein 1 of ORK1_P_547 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLQKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 200: Protein 1 of ORK1_P_548 EEVSEYCSHMIGSGHLQLLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 201: Protein 1 of ORK1_P_549 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKANLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 202: Protein 1 of ORK1_P_550 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVFDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 203: Protein 1 of ORK1_P_551 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGESKYGPPCPPCP APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQF NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQ VSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVM HEALHNHYTQKSLSLSLG SEQ ID NO: 204: Protein 1 of ORK1_P_552 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKC KVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 205: Protein 1 of ORK1_P_553 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNEAAAKEAAAKEAAAKEAAAKEAAAKEAAAKESKYGPPCPPCP APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQF NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQ VSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVM HEALHNHYTQKSLSLSLG SEQ ID NO: 206: Protein 1 of ORK1_P_554 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNEAAAKESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKC KVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 207: Protein 1 of ORK1_P_555 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDA FSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSE EDI FINY IEAYMTMKIRNG GGGGSGGSGGSGGSGGSGGSGGSGGSGGGESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKC KVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 208: Protein 1 of ORK1_P_556 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDA FSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSE EDI FINY IEAYMTMKIRNG GGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVD GVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREP QVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTV DKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 209: Protein 1 of ORK1_P_557 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDA FSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNE AAAKEAAAKEAAAKEAAAKEAAAKEAAAKESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKC KVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 210: Protein 1 of ORK1_P_558 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDA FSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNE AAAKESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVD GVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREP QVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTV DKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 211: Protein 1 of ORK1_P_559 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVEAAAKEAAAKEAAAKEAAAKEAAAKEAAAKSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGESKYGPPCPPCP APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQF NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQ VSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVM HEALHNHYTQKSLSLSLG SEQ ID NO: 212: Protein 1 of ORK1_P_560 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVEAAAKEAAAKEAAAKEAAAKEAAAKEAAAKSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKC KVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 213: Protein 1 of ORK1_P_561 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVEAAAKEAAAKEAAAKEAAAKEAAAKEAAAKSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNEAAAKEAAAKEAAAKEAAAKEAAAKEAAAKESKYGPPCPPCP APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQF NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQ VSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVM HEALHNHYTQKSLSLSLG SEQ ID NO: 214: Protein 1 of ORK1_P_562 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVEAAAKEAAAKEAAAKEAAAKEAAAKEAAAKSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNEAAAKESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKC KVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 215: Protein 1 of ORK1_P_563 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVEAAAKSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDA FSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSE EDI FINY IEAYMTMKIRNG GGGGSGGSGGSGGSGGSGGSGGSGGSGGGESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKC KVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 216: Protein 1 of ORK1_P_564 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVEAAAKSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDA FSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSE EDI FINY IEAYMTMKIRNG GGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVD GVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREP QVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTV DKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG

[0849] SEQ ID NO: 217: Protein 1 of ORK1_P_565 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVEAAAKSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDA FSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSE EDI FINY IEAYMTMKIRNEAAAKEAAAKEAAAKEAAAKEAAAKEAAAKESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKC KVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG

[0850] SEQ ID NO: 218: Protein 1 of ORK1_P_566 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVEAAAKSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDA FSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNE AAAKESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVD GVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREP QVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTV DKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 219: Protein 1 of ORK1_P_567 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHFV FRDNTPNAIAIVQLQELSIYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 220: Protein 1 of ORK1_P_568 EEVSRYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHML FRDNTPNAIAIVQLQELSVYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 221: Protein 1 of ORK1_P_569 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHML FRDNTPNAIAIYQLQELSIYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 222: Protein 1 of ORK1_P_570 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHML FRDNTPNAIAIVQLQELSIYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 223: Protein 1 of ORK1_P_571 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMY FRDNTPNAIAIVQLQELSIYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 224: Protein 1 of ORK1_P_572 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMR FRDNTPNDIAIVQLQELSIYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 225: Protein 1 of ORK1_P_573 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMI FRDNTPNAIAIVQLQELSIYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 226: Protein 1 of ORK1_P_574 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMV FRDNTPNDIAIVQLQELSIRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 227: Protein 1 of ORK1_P_575 EEVSRYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHML FRDNTPNAIAIVQLQELSIYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 228: Protein 1 of ORK1_P_576 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMV FRDNTPNDIAIVQLQELSVYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 229: Protein 1 of ORK1_P_577 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMI FRDNTPNDIAIVQLQELSVYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 230: Protein 1 of ORK1_P_578 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMV FRDNTPNDIAIVQLQELSIYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 231: Protein 1 of ORK1_P_579 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMI FRDNTPNDIAIVQLQELSIYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 232: Protein 1 of ORK1_P_580 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMV FRDNTPNDIAIVQLQELSLYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 233: Protein 1 of ORK1_P_581 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMV FRDNTPNDIAIVQLQELSYYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 234: Protein 1 of ORK1_P_582 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGESKYGPPCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYR VVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWC LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALH NHYTQKSLSLSLGGGGGGSGGGGSGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRL RLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINYIEAYMTMNNGGLDYLPNMLR DLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAH VNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTM KIRN SEQ ID NO: 235: Protein 1 of ORK1_P_583 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG

[0851] SEQ ID NO: 236: Protein 1 of ORK1_P_587 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTYR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDA FSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSE EDI FINY IEAYMTMKIRNG GGGGSGGSGGSGGSGGSGGSGGSGGSGGGESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKC KVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 237: Protein 1 of ORK1_P_588 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTYR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDA FSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSE EDI FINY IEAYMTMKIRNG GGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQE DPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKT ISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 238: Protein 1 of ORK1_P_589 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTYR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLG SEQ ID NO: 239: Protein 1 of ORK1_P_590 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTYR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVEAAAKSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDA FSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSE EDI FINY IEAYMTMKIRNG GGGGSGGSGGSGGSGGSGGSGGSGGSGGGESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKC KVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 240: Protein 1 of ORK1_P_591 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTYR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVEAAAKSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDA FSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSE EDI FINY IEAYMTMKIRNG GGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVD GVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREP QVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTV DKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 241: Protein 1 of ORK1_P_592 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTYR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVEAAAKSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNE AAAKESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVD GVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREP QVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTV DKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO 242: Protein 2 of ORK1_P_509 SPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALS EMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQ EKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLL EDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSK AVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCP PCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPRE EQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVCTLPPSQEEMT KNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSRLTVDKSRWQEGNVFSC SVMHEALHNHYTQKSLSLSLG SEQ ID NO 243: Protein 2 of ORK1_P_510 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVCT LPPSQEEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSSPGQGTQSENSCTHFPGNLP NMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPD IKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEA YMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYL EEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKA MSEFDIFINYIEAYMTMKIRN SEQ ID NO: 244 ORK1-341_P EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQALSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGMTQSENSCTHFPGNLPNM LRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIK AHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYM TMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEE VMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMS EFDIFINYIEAYMTMKIRN SEQ ID NO: 245 CSF1 ( (amino acids E1-V151 of mature human CSF1 protein) EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDV SEQ ID NO: 255 ( scIL-10 with D84E, C108N ( 2nd IL10 domain), Metl, short Nter) MTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQEPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFK GYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQ VKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNSEQ ID NO: 256 ( scIL-10 with Metl, short Nter) MTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFK GYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQ VKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 257 ( scIL-10 with C108N ( 2nd IL10 domain), Metl, short Nter ) MTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFK GYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQ VKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 258 ( scIL-10 with D84E, Metl, short Nter) MTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQEPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFK GYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQ VKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 259 ( scIL-10 with C108N ( 2nd IL10 domain), short N term) TQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQF YLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIY KAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKG YLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQV KNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 261 ( scIL-10 with C108A ( 2nd IL10 domain), short N term) SPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALS EMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQ EKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLL EDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRAHRFLPCENKSK AVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 262 ( scIL-10 with C108A ( 2nd IL10 domain) SPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALS EMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQ EKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLL EDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRAHRFLPCENKSK AVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 263

[0852] MTMNNGGLDYLP SEQ ID NO: 264 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 265 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDA FSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 266 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTYR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 267 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTYR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDA FSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSE EDI FINY IEAYMTMKIRN SEQ ID NO: 268 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMKIRNGGGSGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNL LLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFL PCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRN

[0853] SEQ ID NO: 269 CSFl-cpCSFl - scIL-10 C108N ( 2nd IL10 domain) of ORK1_P_512 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGGGSGGSGGSGGSGGSGGSGGSGGSGGGHDKACVRTFYET PLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFNECEDQDRRQPGTKTTGKVSEYCSHMIGSGHL QSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQE LSLRLKSCFTKDYEEGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPN MLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDI KAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAY MTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLE EVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAM SEFDIFINYIEAYMTMKIRNSEQ ID NO: 270 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVEAAAKSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDA FSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLG ENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 273 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMV FRDNTPNDIAIVQLQELSVYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 274 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMV FRDNTPNDIAIVQLQELSIYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 275 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMI FRDNTPNDIAIVQLQELSIYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPG NLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDI FINY IEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQ FYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGI YKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 276 circular permutant CSF1 HDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFNECEDQDRRQPGTKTTGKVS EYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDN TPNAIAIVQLQELSLRLKSCFTKDYEE SEQ ID NO: 277 single chain dimeric CSF1 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDVGGGGGSGGGGSGGSGGSGGSGGSGGSGGSGGSGGGHDKACVRTFYET PLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFNECEDQDRRQPGTKTTGKVSEYCSHMIGSGHL QSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQE LSLRLKSCFTKDYEESEQ ID NO: 278 CSF1 (amino acids E1-V151 of mature human CSF1 protein with M65Y mutation) EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTYR FRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDW NIFSKNCNNSFAECSSQDV SEQ ID NO: 279

[0854] GGGGSGGGGSGGGGSGGGGSGGGGSGGGGS

Claims

CLAIMS1. A protein comprising a single chain polypeptide having IL-10 and CSF1 activities modified to increase its half-life.

2. The protein according to claim 1, which comprises a single chain polypeptide having IL-10 and CSF1 activities fused to an immunoglobulin Fc fragment or to albumin, or albumin fragment, or to an anti-albumin antibody, or anti-albumin antibody fragment, or conjugated with a poly(ethylene glycol) (PEG) molecule.

3. The protein according to claim 1 or 2, wherein the single chain polypeptide having IL-10 and CSF1 activities comprises in the N-terminus to C-terminus direction, a CSF1 domain, a peptide linker L1, and an IL-10 domain, wherein the IL-10 domain is an IL-10 monomer or a single chain dimeric IL-10, and wherein the CSF1 domain is a CSF1 monomer or a single chain dimeric CSF1.

4. The protein according to any one of claims 1 to 3, which comprises one single chain polypeptide having IL-10 and CSF1 activities and one immunoglobulin Fc fragment, wherein the single chain polypeptide having IL-10 and CSF1 activities is fused to the immunoglobulin Fc fragment through a peptide linker L2 connecting (i) the C-terminus of the IL-10 domain with the N-terminus of one hinge region of the Fc fragment, or (ii) the C-terminus of the one of the CH3 domains of the Fc fragment with the N-terminus of the CSF1 monomer.

5. The protein according to any one of claims 1 to 3, which comprises two single chain polypeptides having IL-10 and CSF1 activities and one immunoglobulin Fc fragment.

6. The protein according to claim 5, which comprises two polypeptide chains that each comprise, in the N-terminus to C-terminus direction, a CSF1 domain which is a CSF1 monomer or a single chain dimeric CSF1, a peptide linker L1, an IL-10 domain which is an IL-10 monomer or a single chain dimeric IL-10 monomer, a peptide linker L2, and a Fc domain, wherein the two polypeptide chains dimerise through their Fc domains.

7. The protein according to claim 5, which comprises two polypeptide chains that each comprise, in the N-terminus to C-terminus direction, a Fc domain, a peptide linker L2, a CSF1 domain which is a CSF1 monomer or a single chain dimeric CSF1, a peptide linker L1, and IL-10 domain which is an IL-10 monomer or a single chain dimeric IL-10 monomer, wherein the two polypeptide chains dimerise through their Fc domains.

8. The protein according to any one of claims 1 to 7, wherein the CSF1 monomer is a CSF1 monomer that comprises sequence SEQ ID NO:12, SEQ ID NO: 17,or SEQ ID NO: 245, or from a single chain dimeric CSF1 comprising sequence SEQ ID NO: 277, or a sequence at least 80% identical thereto and that retains CSF1 activity.

9. The protein according to any one of claims 1 to 7, wherein the CSF1 domain is a polypeptide that comprises a sequence differing from SEQ ID NO: 12, SEQ ID NO: 245, SEQ ID NO: 17, or SEQ ID NO: 277, by one or more mutations comprising:a) substitution M65Y;b) substitutions S18I, T64H, R66V, A74D, L85V, R86Y, and V120F;c) substitutions S18I, T64H, R66V, A74D, L85I, R86Y, and V120F; or d) substitutions S18I, T64H, R66I, A74D, L85I, R86Y, and V120F.

10. The protein according to any one of claims 1 to 9, wherein the IL- 10 domain is an IL-10 monomer and the single chain polypeptide having IL-10 and CSF1 activities comprises sequence SEQ ID NO: 25, or a sequence at least 80% identical thereto and that retains IL-10 and CSF1 activities.

11. The protein according to any one of claims 1 to 9, wherein the IL- 10 domain is a single chain dimeric IL-10 that comprises sequence SEQ ID NO: 22, SEQ ID NO: 5, or SEQ ID NO: 1, or a sequence at least 80% identical to mentioned sequences and that retains at least the same stability, and / or at least the same level of interaction with IL- 10 receptor, and the same IL- 10 activity.

12. The protein according to any one of claims 1 to 11, wherein one or more of the following conditions are met:(i) the peptide linker L1 comprises from 5 to 50 amino acid residues;(ii) the peptide linker L1 is a flexible peptide sequence composed of Gly and Ser residues, in different proportion;(iii) the peptide linker L1 comprises sequence SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, or SEQ ID NO: 36; SEQ ID NO: 82, SEQ ID NO: 247, SEQ ID NO: 248,(iv) the peptide linker L1 comprises sequence (SEQ ID NO:82)n, n being an integer from 1 to 10.

13. The protein according to any one of claims 1-12, wherein the single chain polypeptide having IL- 10 and CSF1 activities comprises sequence SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 105, or any one of sequence SEQ ID NO: 264 to SEQ ID NO: 269, and SEQ ID NO: 273 to SEQ ID NO: 275, or a sequence at least 80% identical thereto and that retains IL-10 and CSF1 activities.

14. The protein according to any one of claims 1-13, wherein one or more of the following conditions are met:(i) the peptide linker L2 comprises from 3 to 45 amino acid residues;(ii) the peptide linker L2 is a flexible peptide sequence composed of Gly and Ser residues;(iii) the peptide linker L2 comprises sequence (GGGGS)p, wherein p is an integer from 1 to 8;(iv) the peptide linker L2 comprises any one of sequence SEQ ID NO: 82, SEQ ID NO: 136, SEQ ID NO: 35, SEQ ID NO: 247, SEQ ID NO: 248, SEQ ID NO: 279, and SEQ ID NO: 119 to SEQ ID NO: 131.

15. The protein according to any one of claims 1-14, wherein the immunoglobulin Fc fragment or Fc domain is of the isotype IgG, an is optionally engineered.

16. The protein according to any one of claims 1 to 15, which comprises a first polypeptide comprising:i) sequence SEQ ID NO: 88, SEQ ID NO: 101, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 167, SEQ ID NO: 184, SEQ ID NO: 187, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 218, SEQ ID NO: 228, SEQ ID NO: 230, SEQ ID NO: 231, SEQ ID NO: 236, SEQ ID NO:239, SEQ ID NO: 240, SEQ ID NO: 241, or a sequence at least 80% identical thereto and that retains IL- 10 and CSF1 activities, and a second polypeptide comprising SEQ ID NO: 109, orii) a first polypeptide comprising sequence SEQ ID NO: 155, or a sequence at least 80% identical thereto and that retains IL-10 and CSF1 activities and a second polypeptide comprising SEQ ID NO: 144.

17. A protein having dual IL-10 and CSF1 activities that comprises two polypeptide chains comprising, in the N-terminus to C-terminus direction, a CSF1 monomer fused to an immunoglobulin Fc fragment, and second polypeptide chain comprises, in the N-terminus to C-terminus direction, an immunoglobulin Fc fragment fused to an IL-10 domain, wherein the IL-10 domain is an IL-10 monomer or a single chain dimeric IL-10, and wherein the two polypeptide chains dimerise through their Fc domains.

18. The protein according to claim 17, which comprises two polypeptide chains comprising:(i) CSF1 monomer-linker L2-Fc1, wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and(ii) Fc2- linker L2-IL-10 domain,wherein Fc2 is a Fc domain comprising hinge-CH2-CH3,wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains, and wherein CSF1 monomer, IL-10 domain, linker L2, Fc1 and Fc2 are as defined in any preceding claims.

19. The protein according to claim 17 or 18, wherein the protein comprises a first polypeptide chain comprising sequence SEQ ID NO: 161, and a second polypeptide comprising sequence SEQ ID NO: 24320. A pharmaceutical composition comprising the protein according to any one of claims 1 to 19 and a pharmaceutically acceptable carrier.

21. A protein according to any one of claims 1 to 19, or a pharmaceutical composition according to claim 20, for use as a medicament.

22. A protein according to any one of claims 1 to 19, or a pharmaceutical composition according to claim 20, for use for treating an inflammatory disease.

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