Anti-sting antibodies and methods of use thereof

Abstract

The present disclosure relates to novel antibodies and antigen-binding fragments thereof that specifically bind to human stimulator of interferon genes (STING), and uses of these antibodies and antigen-binding fragments thereof for treating diseases and disorders (e.g., inflammatory diseases or an autoimmune diseases).

Description

ANTI-STING ANTIBODIES AND METHODS OF USE THEREOF CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to United States Provisional Application No. 63 / 692,993 filed September 10, 2024, the content of which is incorporated by reference in its entirety herein. SEQUENCE LISTING

[0002] This application contains an electronic Sequence Listing which has been submitted in XML file format with this application, the entire content of which is incorporated by reference herein in its entirety. The Sequence Listing XML file submitted with this application is entitled “14727-035-228_SEQLISTING.xml”, was created on September 5, 2025, and is 143,089 bytes in size.1. FIELD

[0003] The present disclosure relates to novel antibodies and antigen-binding fragments thereof that specifically bind to human stimulator of interferon genes (STING), and uses of these antibodies and antigen-binding fragments thereof for treating diseases and disorders (e.g., inflammatory diseases or autoimmune diseases).2. BACKGROUND

[0004] The human immune system is a multi-faceted network of functionally diverse cells expressing a broad array of receptors that collectively function to respond to infection, eliminate pre-cancerous cells, and maintain metabolic health. Breakdown of this delicately poised immune response is typically life limiting; however, even subtle changes in its ability to distinguish an invading pathogen from the host can give rise to a spectrum of autoimmune diseases. Autoimmune diseases are caused by immune system disorders as well as self-attack and are a serious risk to human health. Indeed, there are over 80 different types of autoimmune diseases which affect approximately 5%–8% of the world population and cause tremendous suffering to patients while also representing a major global socioeconomic issue.

[0005] Most autoimmune diseases remain incurable due to the causative factors that often require long-term or lifelong medication. Presently, synthetic drugs approved for theNAI-5002871589v11treatment of autoimmune diseases are dominated by Janus kinase (JAK) inhibitors. JAK1, JAK2, JAK3, and the non-receptor tyrosine-protein kinase TYK2 are intracellular tyrosine kinases that act in concert with seven different signal transducer and activator of transcription (STAT) transcription factors in mediating the signaling of more than 50 distinct cytokines, as well as hormones and growth factors. Pharmacological inhibition of cytokine signaling by blocking the JAK-STAT pathway is increasingly used to treat autoimmune diseases. Examples of synthetic and biologic drugs for treating autoimmune diseases are reviewed for example in Fugger et al., Cell 181 (2020), 63-80.

[0006] However, current immune-modulatory drugs used in the treatment of autoimmune diseases are broadly acting and associated with side effects such as infection and malignant disease. Thus, it is urgent to explore novel therapeutical targets more closely implicated in autoimmune diseases and to develop corresponding specific molecule drugs.3. SUMMARY

[0007] The present disclosure provides the embodiments as characterized in the claims, disclosed in the description and illustrated in the Examples and Figures further below. Thus, the present disclosure provides antibodies being specific for human STING, which preferably recognize the C-terminal domain of STING and interfere with STING signaling resulting in reduction of inflammatory cytokine production. Furthermore, the present disclosure generally relates to antagonistic / inhibitory anti-STING antibodies for use as a medicament, for example in the treatment of autoimmune and (auto)inflammatory diseases.

[0008] In accordance with the present disclosure anti-STING antibodies have been isolated from blood samples of healthy human volunteers and cloned, that could be demonstrated to reduce STING-mediated inflammatory cytokine production in both human and mouse cells, and to attenuate the pathological features of autoinflammatory disease, i.e., excessive cytokine production in mice. Using the well-known potent, selective antagonist of STING, H- 151, a small covalent inhibitor of STING preventing the dimerization (multimerization) required for the activation of STING that has noteworthy inhibitory activity both in cells and in vivo towards autoinflammatory disease as a positive control, experiments performed within the scope of the present disclosure surprisingly revealed that the presently disclosed anti-STING antibodies inhibited the release of IFN- and prevented the phosphorylation ofTANK-binding kinase 1 (TBK1) in mouse splenocytes and in differentiated THP-1 cells, which had been stimulated with cyclic GAMP or CMA to at least the same extent as H-151;NAI-5002871589v12see Examples 7 to 11. The presently disclosed anti-STING antibodies also inhibited therelease of pro-inflammatory cytokines such as IL-6, IP-10 and IFN- in differentiated THP-1cells at a dose-dependent manner, where the STING pathway of the THP-1 cells was activated with HT DNA (deoxyribonucleic acid from herring testes) (see Example 17). Importantly, the in vivo therapeutic effects of the presently disclosed anti-STING antibodies were demonstrated in two lupus mouse models, Trex1- / - mice (see Example 12) and NZB / W mice (see Example 18).

[0009] In general, the release of pro-inflammatory cytokines, like type I interferons (IFNs) is the first-line defense to protect the body from potential threats and is precisely controlled by a complex biological regulation process. In particular, pro-inflammatory cytokines continuously activate several inflammation-related signaling pathways to ignite the immune system and execute elimination programs. However, it happens that the body loses its ability to distinguish an invading pathogen from the host leading to the production of too many inflammatory signals (also referred to as cytokine storm), which can lead to excess inflammation and conditions like autoimmune diseases. Thus, compounds inhibiting the release of cytokines are promising candidates for the treatment of autoimmune diseases.

[0010] The complex biological regulation process behind the release of pro-inflammatory cytokines involves STING that acts as a sensor of cytosolic DNA, including exogenous and endogenous DNA. The activation of the STING pathway by recognition of cytosolic DNA can induce the production of type I interferons (IFNs) and other cytokines, and modulation of STING activity has recently become an attractive therapeutic strategy in the field of immunology for the treatment of cancers, autoimmune diseases, and inflammatory diseases.

[0011] In more detail, double-stranded DNA (dsDNA) in the cytoplasm, which can be derived from pathogens (e.g., bacteria or viruses) and “self” cells (e.g., cancers, dead cells, or damaged mitochondria), binds to and activates cyclic guanosine monophosphate-adenosine monophosphate synthase (cGAS). The activated DNA-cGAS then catalyzes the synthesis of 2’,3’-cyclic GMP-AMP (cGAMP) using cytosolic GTP and ATP as substrates. After the binding of cGAMP to STING, STING is activated and the complex translocates from the endoplasmic reticulum (ER) to the Golgi apparatus, where STING recruits and activates TANK-binding kinase 1 (TBK1). The activated TBK1 in turn phosphorylates STING, which results in the recruitment and phosphorylation of interferon regulatory factor 3 (IRF3). The phosphorylated IRF3 is dimerized and enters the nucleus, resulting in the expression of type I IFNs and inflammatory cytokines and chemokines. The IFNs and cytokines have significant immune-stimulatory functions by promoting the priming and activation of T cells, dendriticNAI-5002871589v13cells (DCs), and natural killer (NK) cells. In addition, bacteria-derived cyclic dinucleotides (CDNs), including cyclic dimeric GMP (c-di-GMP or CDG), cyclic dimeric AMP (c-di-AMP or CDA), and 3’,3’-cGAMP, can also directly bind to and activate STING like cGAMP. Thus, the persistent or chronic inflammatory signaling that links to the activation of cGAS- STING signaling is prone to developing the autoimmune disease including autoinflammatory syndromes like Aicardi-Goutières syndrome (AGS), STING-associated vasculopathy of infancy (SAVI), systemic lupus erythematosus (SLE), and COPA syndrome, autoimmune neurodegenerative diseases like amyotrophic lateral sclerosis (ALS), Niemann-Pick disease type C (NPC), and multiple sclerosis, other cGAS-STING related autoimmune diseases like familial chilblain lupus (FCL), rheumatoid arthritis, and Sjögren’s syndrome, etc. cGAS- STING pathway inhibitors such as H-151 have been shown to effectively improve the symptoms of the autoimmune diseases ALS and psoriasis by decreasing the inflammatory signaling in animal models, thereby becoming a promising therapeutic agent for autoimmune diseases. Details about the cGAS–STING pathway are subject to several reviews, see for example Kong et al., Journal of Advanced Research 44 (2023), 119-133, Zhang et al., Front Immunol. 13 (2022), 954129, Decout et al., Nat Rev Immunol 21 (2021), 548-569, Skopelja- Gardner et al., Nature Reviews Nephrology 18 (2022), 558–572, and Hopfner and Hornung, Nature Reviews Molecular Cell Biology 21 (2020), 501–521.

[0012] Thus, since the exemplary anti-STING antibody of the present disclosure has beenshown to inhibit the release of IFN- - an inflammatory cytokine - and thus, inhibits cytokinesignalling, and has been shown to prevent phosphorylation of TBK1 and thus acts as cGAS- STING pathway inhibitor similar to the well-established STING antagonist H-151. It is prudent to expect that antagonistic anti-STING antibodies are suitable for the treatment of diseases related to excessive STING mediated cytokine production like in autoimmune and (auto)inflammatory diseases. This has been confirmed by in vivo experiments showing that treatment of mice having an increased amount of relevant inflammatory cytokines (inflammatory mouse model) with an anti-STING antibody of the present disclosure reducedthe level of IFN- and IL-6 similar to H-151; see Example 12.

[0013] As mentioned above, several treatment strategies for autoimmune diseases are known, for example inhibiting cytokine signaling by blocking the JAK-STAT pathway and inhibiting the cGAS-STING pathway, for which usually small molecules are employed. Such small molecule inhibitors are for example described in Zhang et al., Front Immunol. 13 (2022), 954129, Kong et al., Journal of Advanced Research 44 (2023), 119-133, and Decout et al., Nat Rev Immunol 21 (2021), 548-569. However anti-STING antibodies have not beenNAI-5002871589v14reported as suitable candidates for the treatment of autoimmune diseases and thanks to the results obtained during the experiments performed within the scope of the present disclosure and corresponding in vivo data, for the first time it has been credibly shown that anti-STING antibodies are a promising treatment option. It is prudent to expect that anti-STING antibodies act more specific and have less severe side effects than other broadly acting immune-modulatory drugs used in the treatment of autoimmune diseases since they target one of the essential proteins, i.e., STING, mediating autoimmune process in innate immune system.

[0014] Thus, the present disclosure relates in general to antagonistic anti-STING antibodies, in particular anti-STING antibodies, for use in the treatment of an an autoimmune or (auto)inflammatory disease, for example chronic and acute inflammatory diseases.

[0015] Furthermore, it has been shown that the anti-STING antibodies of the present disclosure bind to the C-terminal domain of STING. In addition, antibody Ab1 has been shown to bind to a C-terminus short peptide (see Example 2) and antibodies Ab2 and Ab3 each bind to an epitope comprising the amino acid residues 300 to 308 of human STING and to an epitope comprising the amino acid residues 245-255 of human STING, respectively. Example 13 further shows that Ab1 bound to an epitope located within the C-terminal domain of STING, where the epitope comprises amino acid residues S358, G367, M368, E369, and P371 of human STING protein (SEQ ID NO:34).

[0016] Human STING (UniProt ID: Q86WV6 · STING_HUMAN) is a transmembrane protein located on the endoplasmic reticulum (ER) and consists of an N-terminal transmembrane structural domain (NTD) containing four transmembrane helices TM1 (residues 18-34), TM2 (residues 45-69), TM3 (residues 92-106), TM4 (residues 117-134) and a globular C-terminal structural domain (CTD, residues 135-379), wherein the latter one comprises a ligand-binding domain (LBD), an IRF3-binding domain, and a TBK1-binding motif.

[0017] Accordingly, an antagonistic anti-STING antibody for use in accordance with the present disclosure preferably binds to the C-terminal domain of the STING protein and an epitope located within said C-terminal domain, respectively. In certain embodiments, the epitope comprises amino acid residues S358, G367, M368, E369, and P371 of human STING protein (SEQ ID NO:34). In certain embodiments, the anti-STING antibody for use in accordance with the present disclosure is the antibody of the present disclosure as characterized further below.NAI-5002871589v15

[0018] As further illustrated in the Examples, human monoclonal anti-STING antibodies have been cloned and identified that bind to STING, in particular to the C-terminal domain of the STING protein. The binding specificity of anti-STING antibodies was determined by ELISA assays, Western Blot analysis, immunohistochemistry, and immunofluorescence analysis.

[0019] ELISA assays and Western Blot analysis have shown strong and selective binding of the anti-STING antibodies on both recombinant human and mouse STING protein as well as on the C-terminal portion of the human, cyno and rat (Example 2). Furthermore, Western Blot analysis as performed in Example 3 showed concentration dependent binding of anti- STING antibodies to the STING protein. Immunocytochemistry analysis revealed specific binding of anti-STING antibodies in human PBMCs and human derived splenocytes demonstrating specific target binding of anti-STING antibodies to human cells; see Example 4. Immunofluorescence analysis showed specific binding of anti-STING antibodies to STING in human lung adenocarcinoma tissue as well as in colon cancer tissue (see Example 5).

[0020] Thus, the experiments described in the appended Examples demonstrate that the anti- STING antibodies identified in accordance with the present disclosure specifically bind human STING under different conditions and are thus also useful in laboratory research. For example, anti-STING antibodies recognized mouse STING protein with substantially the same affinity and thus can be used in mouse disease models without the need of genetically engineering the animal to express human STING while the endogenous copy of STING must be knocked out.

[0021] In certain embodiments, the antibody of the present disclosure may be characterized by the complementarity determining regions (CDRs) or hypervariable regions of the variable heavy (VH) and variable light (VL) chain comprising the amino acid sequence of SEQ ID: 2 and SEQ ID NO:7 as shown in FIG. 1A. In another embodiment, the antibody of the present disclosure may be characterized by the CDRs or hypervariable regions of the VH and VL chain comprising the amino acid sequence of SEQ ID NO:12 and SEQ ID NO:17 as shown in FIG. 1B. In a further embodiment, the antibody of the present disclosure may be characterized by the CDRs or hypervariable regions of the VH and VL chain comprising the amino acid sequence of SEQ ID NO:22 and SEQ ID NO:27 as shown in FIG. 1C.

[0022] As mentioned above, the anti-STING antibodies of the present disclosure binds to the C-terminal domain of the STING protein and an epitope located within said C-terminal domain, respectively. As further referred to above, the anti-STING antibodies of the presentdisclosure have also been shown to inhibit the release of IFN- in mouse splenocytesNAI-5002871589v16(Example 7) and in differentiated THP-1 cells (Example 8), and prevent phosphorylation of TBK1 in differentiated THP-1 cells (Example 9), and in mouse splenocytes (Example 10). The STING antagonistic properties of the antibody which match those of STING antagonistH-151 and its ability to inhibit the release of IFN- , as one of the major inflammatorycytokine involved in autoimmune and inflammatory disease, as exemplary shown for antibody Ab1 in in vivo mouse studies (see Example 12), makes it is prudent to expect that the anti-STING antibodies of the present disclosure and equivalent anti-STING antagonist antibodies are useful for immunotherapy of autoimmune and (auto)inflammatory diseases.

[0023] As pointed out in the Examples, in certain embodiments, the anti-STING antibody of the present disclosure prevents phosphorylation of TANK-binding kinase 1 (TBK1). In addition, or alternatively, the antibody of the present disclosure is capable of reducing thelevel of human IFN- in differentiated human THP-1 derived macrophages, in particular inphorbol 12-myristate 13-acetate (PMA)-differentiated human THP-1-derived macrophages, which have been stimulated with the STING agonist cGAMP.

[0024] The present disclosure further relates to an antibody having the same binding specificities as any one of antibodies Ab1, Ab2, and Ab3, i.e., any antibody which has the immunological characteristics of (i) binding to the C-terminal domain of STING; and (ii) being capable of preventing phosphorylation of TBK1; and / or (iii) being capable of reducing the level of hIFN- in PMA-differentiated humanTHP-1-derived macrophages stimulated with cGAMP.

[0025] The mentioned features can be easily determined in accordance with the experiments and assays disclosed in the appended Examples, wherein antibodies Ab1, Ab2, and Ab3, respectively. Typically, an antagonistic anti-STING antibody will compete with the corresponding reference antibody for binding to the C-terminal domain of STING.

[0026] As further shown in Example 2, antibody Ab1 binds with high affinity (EC50value of 1 nM, for example 224 pM as determined in Example 2) to the C-terminus STING short peptide within the C-terminal domain of STING. The C-terminal domain of STING is comprised of a ligand-binding domain (LBD, residues 157-335), an IRF3-binding domain (residues 362-366), and a TBK1-binding motif (TBM, residues 369-377). Thus, in certain embodiments, the anti-STING antibody of the present disclosure binds to the C-terminal tail of STING protein.

[0027] While the invention is illustrated and described by way of reference to the antibody originally obtained in the experiments performed in accordance with the present disclosureNAI-5002871589v17and described in the Examples it is to be understood that the antibody of the present disclosure includes synthetic and biotechnological derivatives of an antibody which means any engineered antibody or antibody-like STING binding molecule, synthesized by chemical or recombinant techniques, which retain one or more of the functional properties of the subject antibody illustrated in the Examples, in particular preventing phosphorylation ofTBK1 and / or reducing the level of hIFN- in differentiated human THP-1-derivedmacrophages, preferably both, and as a structural feature preferably recognizing and binding the C-terminal domain of STING. Hence, since proof-of-concept for using antagonistic anti- STING antibodies in the treatment of conditions caused by inflammatory cytokine production is provided for the first time, the present disclosure paves the way for a novel immunotherapy for the treatment of autoimmune and (auto)inflammatory diseases and thus generally relates to a method of treating an inflammatory or autoimmune disease in a subject, wherein the method comprises administering an antagonistic anti-STING antibody, preferably wherein the antibody specifically binds to the C-terminal domain of STING to a subject in need thereof.

[0028] In certain embodiments, the antibody or antigen-binding fragment disclosed herein is internalized into monocytes as measured by Western blot and immunocytochemistry (ICC). In certain embodiments, the antibody or antigen-binding fragment disclosed herein is internalized into monocytes as measured by fluorescent labeling. In certain embodiments, the antibody or antigen-binding fragment disclosed herein is co-localized with STING in cells as measured by confocal microscopy. In certain embodiments, the internalization and / or colocalization occur in vitro, ex vivo or in vivo.

[0029] In one aspect, the present disclosure provides an antibody or an antigen-binding fragment thereof that can specifically bind human stimulator of interferon genes (STING), wherein the antibody or antigen-binding fragment thereof comprises: (i) a heavy chain variable region (VH) comprising a heavy chain variable region complementarity determining region 1 (VH-CDR1), a heavy chain variable region complementarity determining region 2 (VH-CDR2), and a heavy chain variable region complementarity determining region 3 (VH-CDR3) as set forth in a VH comprising the amino acid sequence of SEQ ID NO:2; and a light chain variable region (VL) comprising a light chain variable region complementarity determining region 1 (VL-CDR1), a light chain variable region complementarity determining region 2 (VL-CDR2), and a light chainNAI-5002871589v18variable region complementarity determining region 3 (VL-CDR3) as set forth in a VL comprising the amino acid sequence of SEQ ID NO:7; (ii) a VH comprising a VH-CDR1, a VH-CDR2, and a VH-CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO:12; and a VL comprising a VL-CDR1, a VL-CDR2, and a VL-CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO:17; or (iii) a VH comprising a VH-CDR1, a VH-CDR2, and a VH-CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO:22; and a VL comprising a VL-CDR1, a VL-CDR2, and a VL-CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO:27.

[0030] In certain embodiments, the VH-CDR1 comprises the amino acid sequence of SEQ ID NO:3, the VH-CDR2 comprises the amino acid sequence of SEQ ID NO:4, the VH-CDR3 comprises the amino acid sequence of SEQ ID NO:5, the VL-CDR1 comprises the amino acid sequence of SEQ ID NO:8, the VL-CDR2 comprises the amino acid sequence of SEQ ID NO:9, and the VL-CDR3 comprises the amino acid sequence of SEQ ID NO:10, and wherein the amino acid sequences of the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL- CDR2, and VL-CDR3 are determined by Kabat definition.

[0031] In certain embodiments, (i) the VH comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:2, or the VL comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:7; (ii) the VH comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:2, and the VL comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:7; or (iii) the VH comprises the amino acid sequence of SEQ ID NO:2 or the VL comprises the amino acid sequence of SEQ ID NO:7.

[0032] In certain embodiments, the VH comprises the amino acid sequence of SEQ ID NO:2 and the VL comprises the amino acid sequence of SEQ ID NO:7.

[0033] In certain embodiments, the VH-CDR1 comprises the amino acid sequence of SEQ ID NO:13, the VH-CDR2 comprises the amino acid sequence of SEQ ID NO:14, the VH- CDR3 comprises the amino acid sequence of SEQ ID NO:15, the VL-CDR1 comprises the amino acid sequence of SEQ ID NO:18, the VL-CDR2 comprises the amino acid sequence ofNAI-5002871589v19SEQ ID NO:19, and the VL-CDR3 comprises the amino acid sequence of SEQ ID NO:20, wherein the amino acid sequences of the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL- CDR2, and VL-CDR3 are determined by Kabat definition.

[0034] In certain embodiments, (i) the VH comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:12, or the VL comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:17; (ii) the VH comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:12, and the VL comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:17; or (iii) the VH comprises the amino acid sequence of SEQ ID NO:12 or the VL comprises the amino acid sequence of SEQ ID NO:17.

[0035] In certain embodiments, the VH comprises the amino acid sequence of SEQ ID NO:12 and the VL comprises the amino acid sequence of SEQ ID NO:17.

[0036] In certain embodiments, the VH-CDR1 comprises the amino acid sequence of SEQ ID NO:23, the VH-CDR2 comprises the amino acid sequence of SEQ ID NO:24, the VH- CDR3 comprises the amino acid sequence of SEQ ID NO:25, the VL-CDR1 comprises the amino acid sequence of SEQ ID NO:28, the VL-CDR2 comprises the amino acid sequence of SEQ ID NO:29, and the VL-CDR3 comprises the amino acid sequence of SEQ ID NO:30, wherein the amino acid sequences of the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL- CDR2, and VL-CDR3 are determined by Kabat definition.

[0037] In certain embodiments, (i) the VH comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:22, or the VL comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:27; (ii) the VH comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:22, and the VL comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:27; or (iii) the VH comprises the amino acid sequence of SEQ ID NO:22 or the VL comprises the amino acid sequence of SEQ ID NO:27.NAI-5002871589v110

[0038] In certain embodiments, the VH comprises the amino acid sequence of SEQ ID NO:22 and the VL comprises the amino acid sequence of SEQ ID NO:27.

[0039] In another aspect, the present disclosure provides an antibody or an antigen-binding fragment thereof that competes with an anti-STING antibody or an antigen-binding fragment thereof disclosed herein for specifically binding to an epitope located within the C-terminal domain of STING.

[0040] In certain embodiments, the C-terminal domain of STING comprises the amino acid sequence of SEQ ID NO:31. In certain embodiments, the epitope is a conformational epitope or a linear epitope. In certain embodiments, the epitope comprises amino acid residues S358, G367, M368, E369, and P371 of human STING, wherein the amino acid positions are numbered according to the positions in the amino acid sequence of SEQ ID NO:34.

[0041] In certain embodiments, (i) the antibody or antigen-binding fragment thereof can inhibit the phosphorylation of TANK binding kinase 1 (TBK1) in cells with activated STING pathway in vitro, ex vivo or in vivo; and / or (ii) the antibody or antigen-binding fragment thereof can inhibit the release of an inflammatory cytokine in cells with activated STING pathway in vitro, ex vivo or in vivo; (iii) the antibody or antigen-binding fragment thereof can inhibit the release of an inflammatory cytokine in cells of a Trex1 null mouse or reduce the level of an inflammatory cytokine of a NZB / W mouse.

[0042] In another aspect, the present disclosure provides an antagonistic antibody or an antigen-binding fragment thereof that can specifically bind human STING, wherein: (i) the antibody or antigen-binding fragment thereof can specifically bind an epitope located within the C-terminal domain of STING; (ii) the antibody or antigen-binding fragment thereof can inhibit the phosphorylation of TBK1 in cells with activated STING pathway in vitro, ex vivo or in vivo; and / or (iii) the antibody or antigen-binding fragment thereof can inhibit the release of an inflammatory cytokine in cells with activated STING pathway in vitro, ex vivo or in vivo; and / or (iv) the antibody or antigen-binding fragment thereof can inhibit the release of an inflammatory cytokine in cells of a Trex1 null mouse or reduce the level of an inflammatory cytokine of a NZB / W mouse.NAI-5002871589v111

[0043] In certain embodiments, the C-terminal domain of STING comprises the amino acid sequence of SEQ ID NO:31. In certain embodiments, the epitope is a conformational epitope or a linear epitope. In certain embodiments, the epitope comprises amino acid residues S358, G367, M368, E369, and P371 of human STING, wherein the amino acid positions are numbered according to the positions in the amino acid sequence of SEQ ID NO:34.

[0044] In certain embodiments, the inhibition of the phosphorylation of TBK1 in the cells with activated STING pathway is indicated by a reduced pTBK1 level in the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof as compared to cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof. In certain embodiments, the level of pTBK1 is measured by an immunohistochemistry assay. In certain embodiments, the level of pTBK1 is measured by Western Blot. In certain embodiments, the level of pTBK1 is measured by immunocytochemistry (ICC). In certain embodiments, the level of pTBK1 is measured as the ratio between pTBK1 and total TBK1 levels, or the ratio between pTBK1 and total GAPDH levels.

[0045] In certain embodiments, the inhibition of the release of the inflammatory cytokine is indicated by a reduced level of the inflammatory cytokine released by the cells with activated STING pathway or the cells of the Trex1 null mouse and being contacted or administered with the antibody or antigen-binding fragment thereof as compared to cells with activated STING pathway or cells of a Trex1 null mouse but without being contacted or administered with the antibody or antigen-binding fragment thereof. In certain embodiments, the level of the inflammatory cytokine is measured by an immunoassay or RT-qPCR. In certain embodiments, the immunoassay is ELISA or HTRF (homogeneous time resolvedfluorescence). In certain embodiments, the inflammatory cytokine is IL-6, IFN- , and / orISG-15.

[0046] In certain embodiments, the cells are immune cells. In certain embodiments, the cells are differentiated THP-1 cells or splenocytes. In certain embodiments, the differentiated THP-1 cells are macrophages. In certain embodiments, the THP-1 cells are human THP-1 cells. In certain embodiments, the differentiated THP-1 cells are PMA-differentiated human THP-1-derived macrophages. In certain embodiments, the cells are heart tissue cells of the Trex1 null mouse. In certain embodiments, the splenocytes are mouse splenocytes. In certain embodiments, the STING pathway is activated by 10-carboxymethyl-9-acridanone (CMA) or cGAMP.NAI-5002871589v112

[0047] In certain embodiments, the pTBK1 level in the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less of the pTBK1 level in the cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof, as measured by Western Blot or ICC, and the cells are PMA-differentiated human THP-1- derived macrophages or mouse splenocytes with STING pathway activated by cGAMP or CMA.

[0048] In certain embodiments, the level of the inflammatory cytokine released by the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less of cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof, as measured by ELISA or HTRF, and the cells are PMA-differentiated human THP- 1-derived macrophages or mouse splenocytes with STING pathway activated by cGAMP,and the inflammatory cytokine is IFN- .

[0049] In certain embodiments, the level of the inflammatory cytokine released by the cells from the Trex1 null mouse administered with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less of cells from a Trex1 null mouse without being administered with the antibody or antigen-binding fragment thereof, as measured by RT-qPCR, and the cells are heart tissue cells, and the inflammatory cytokine isIFN- , IL-6, and / or ISG-15.

[0050] In certain embodiments, the antibody or antigen-binding fragment thereof is an antagonistic antibody or antigen-binding fragment thereof. In certain embodiments, the antibody is a recombinant antibody and / or a monoclonal antibody.

[0051] In certain embodiments, the antibody comprises a heavy chain constant region and / or a light chain constant region. In certain embodiments, the heavy chain constant region is of human IgG type. In certain embodiments, the heavy chain constant region is of IgG1 type or human IgG4 type. In certain embodiments, the light chain constant region comprises a human kappa light chain constant region or a human lambda light chain constant region.

[0052] In certain embodiments, the heavy chain constant region comprises the amino acid sequence of SEQ ID NO:129 or SEQ ID NO:132, and / or the light chain constant region comprises the amino acid sequence of SEQ ID NO:130 or SEQ ID NO:131.NAI-5002871589v113

[0053] In certain embodiments, the antibody comprises an Fc region comprising one or more mutations. In certain embodiments, the one or more mutations decrease immune effector function, increase immune effector function, or increase half-life of the antibody.

[0054] In certain embodiments, the antigen-binding fragment a single chain Fv fragment (scFv), an F(ab'), an F(ab), an F(ab')2, an Fd, an Fv, a single-chain antibody, a disulfide- linked Fv (sdFv), a VHH, or a nanobody.

[0055] In certain embodiments, the antibody or antigen-binding fragment thereof is attached to a detectable label, wherein the detectable label is an enzyme, a radioisotope, a fluorescent compound, a chemiluminescent compound, a bioluminescent compound, a tag, a flag or a heavy metal.

[0056] In another aspect, the present disclosure provides a pharmaceutical composition comprising an anti-STING antibody or an antigen-binding fragment thereof disclosed herein and a pharmaceutically acceptable carrier.

[0057] In another aspect, the present disclosure provides a formulation suitable for in vivo administration comprising an anti-STING antibody or an antigen-binding fragment thereof disclosed herein. In certain embodiments, the formulation is suitable for in vivo intravenous, intraperitoneal, subcutaneous, or intramuscular injection.

[0058] In another aspect, the present disclosure provides an isolated polynucleotide or a set of polynucleotides encoding an anti-STING antibody or an antigen-binding fragment thereof disclosed herein.

[0059] In another aspect, the present disclosure provides a vector or a set of vectors comprising a polynucleotide or a set of polynucleotides disclosed herein.

[0060] In another aspect, the present disclosure provides an isolated cell comprising a polynucleotide or a set of polynucleotides or a vector or a set of vectors disclosed herein.

[0061] In another aspect, the present disclosure provides a method of making an antibody or an antigen-binding fragment thereof, comprising: (i) culturing a cell disclosed herein, and (ii) isolating the antibody or antigen-binding fragment thereof.

[0062] In another aspect, the present disclosure provides an antibody or an antigen-binding fragment thereof produced by a method disclosed herein.

[0063] In another aspect, the present disclosure provides a method of detecting STING in a biological sample, comprising: (i) contacting the biological sample with an anti-STING antibody or an antigen- binding fragment thereof disclosed herein; andNAI-5002871589v114(ii) detecting the binding between the antibody or antigen-binding fragment thereof and STING in the biological sample, optionally wherein the detecting comprises using flow cytometry, immunohistochemistry (IHC), Western Blot analysis, ELISA, or mass spectrometry.

[0064] In another aspect, the present disclosure provides a method for screening for an antibody or an antigen-binding fragment thereof that can specifically bind human STING, comprising measuring the binding of the antibody or antigen-binding fragment thereof to an epitope located within the C-terminal domain of STING, and selecting the antibody if the binding affinity is higher than an isotype control antibody. In certain embodiments, the binding is measured by ELISA. In certain embodiments, the C-terminal domain of STING comprises the amino acid sequence of SEQ ID NO:31. In certain embodiments, the epitope is a conformational epitope or a linear epitope. In certain embodiments, the epitope comprises amino acid residues S358, G367, M368, E369, and P371 of human STING, wherein the amino acid positions are numbered according to the positions in the amino acid sequence of SEQ ID NO:34.

[0065] In another aspect, the present disclosure provides a method of inhibiting the phosphorylation of TBK1 in cells with activated STING pathway, inhibiting the release of an inflammatory cytokine in cells with activated STING pathway, and / or inhibiting the release of an inflammatory cytokine in cells of a Trex1 null mouse, comprising contacting the cell or administering to the mouse with an anti-STING antibody or an antigen-binding fragment thereof disclosed herein, wherein the method is an in vitro, ex vivo or in vivo method.

[0066] In certain embodiments of the antibody or antigen-binding fragment thereof or method disclosed herein, the inhibition of the phosphorylation of TBK1 in the cells with activated STING pathway is indicated by a reduced pTBK1 level in the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof as compared to cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof. In certain embodiments, the level of pTBK1 is measured by an immunohistochemistry assay. In certain embodiments, the immunohistochemistry assay is Western Blot or immunocytochemistry (ICC). In certain embodiments, the level of pTBK1 is measured as the ratio between pTBK1 and total TBK1 levels, or the ratio between pTBK1 and total GAPDH levels.

[0067] In certain embodiments of the antibody or antigen-binding fragment thereof or method disclosed herein, the inhibition of the release of the inflammatory cytokine is indicated by a reduced level of the inflammatory cytokine released by the cells with activatedNAI-5002871589v115STING pathway or the cells of the Trex1 null mouse and being contacted or administered with the antibody or antigen-binding fragment thereof as compared to cells with activated STING pathway or cells of a Trex1 null mouse but without being contacted or administered with the antibody or antigen-binding fragment thereof. In certain embodiments, the level of inflammatory cytokine is measured by an immunoassay or RT-qPCR. In certain embodiments, the immunoassay is ELISA, HTRF (homogeneous time resolved fluorescence), or MSD (Meso Scale Discovery) multiplex assay.

[0068] In certain embodiments of the antibody or antigen-binding fragment thereof or method disclosed herein, reducing of the level the inflammatory cytokine is indicated by a reduced level of the inflammatory cytokine in serum of the NZB / W mouse being administered with the antibody or antigen-binding fragment thereof as compared to the level of the inflammatory cytokine in serum of a NZB / W mouse but without being administered with the antibody or antigen-binding fragment thereof. In certain embodiments, the level of the inflammatory cytokine is measured by an immunoassay. In certain embodiments, the immunoassay is MSD multiplex assay.

[0069] In certain embodiments of the antibody or antigen-binding fragment thereof ormethod disclosed herein, the inflammatory cytokine is IL-6, IP-10, IFN- , and / or ISG-15.

[0070]

[0063] In certain embodiments, the cells are immune cells. In certain embodiments, the cells are differentiated THP-1 cells or splenocytes. In certain embodiments, the differentiated THP-1 cells are macrophages. In certain embodiments, the THP-1 cells are human THP-1 cells. In certain embodiments, the differentiated THP-1 cells are PMA- differentiated human THP-1-derived macrophages. In certain embodiments, the splenocytes are mouse splenocytes. In certain embodiments, the cells are heart tissue cells from the Trex1 null mouse. In certain embodiments, the STING pathway is activated by 10-carboxymethyl- 9-acridanone (CMA), cyclic GMP-AMP (cGAMP), or HT DNA (deoxyribonucleic acid from herring testes).

[0071] In certain embodiments of the antibody or antigen-binding fragment thereof or method disclosed herein, the pTBK1 level in the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less of the pTBK1 level in the cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof, as measured by Western Blot or ICC, and the cells are PMA-differentiated human THP-1-derived macrophages or mouse splenocytes with STING pathway activated by cGAMP or CMA.NAI-5002871589v116

[0072] In certain embodiments of the antibody or antigen-binding fragment thereof or method disclosed herein, the level of the inflammatory cytokine released by the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less of cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof, as measured by ELISA or HTRF, and the cells are PMA-differentiated human THP-1-derived macrophages or mouse splenocytes with STING pathway activated by cGAMP, and theinflammatory cytokine is IFN- .

[0073] In certain embodiments of the antibody or antigen-binding fragment thereof or method disclosed herein, the level of the inflammatory cytokine released by the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less of cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof, as measured by MSD multiplex assay, and the cells are PMA-differentiated human THP-1- derived macrophages with STING pathway activated by HT DNA, and the inflammatorycytokine is IFN- , IL-6, and / or IP-10.

[0074] In certain embodiments of the antibody or antigen-binding fragment thereof or method disclosed herein, the level of the inflammatory cytokine released by the cells from the Trex1 null mouse administered with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less of cells from a Trex1 null mouse without being administered with the antibody or antigen-binding fragment thereof, as measured by RT-qPCR, and the cells are heart tissue cells, and the inflammatory cytokine is IFN- , IL-6,and / or ISG-15.

[0075] In certain embodiments of the antibody or antigen-binding fragment thereof or method disclosed herein, the level of the inflammatory cytokine in serum of the NZB / W mouse administered with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, about 80% or less, or about 90% or less of serum from a NZB / W mouse without being administered with the antibody or antigen-binding fragment thereof, as measured by MSD multiplex assay, and the inflammatory cytokine is IL-6.NAI-5002871589v117

[0076] In another aspect, the present disclosure provides a method of treating a disease or a disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an antibody or an antigen-binding fragment thereof disclosed herein, a pharmaceutical composition disclosed herein, or a formulation disclosed herein. In one aspect, the present disclosure provides an anti-STING antibody or an antigen-binding fragment thereof disclosed herein, a pharmaceutical composition disclosed herein, or a formulation disclosed herein, for use in treating a disease or a disorder in a subject in need thereof. In one aspect, the present disclosure provides use of an anti-STING antibody or an antigen-binding fragment thereof disclosed herein, a pharmaceutical composition disclosed herein, or a formulation disclosed herein in the manufacture of a medicament for treating a disease or a disorder in a subject in need thereof. In certain embodiments, the disease or disorder is an inflammatory disease or an autoimmune disease. In certain embodiments, the inflammatory disease is an autoinflammatory disease. In certain embodiments, the inflammatory diseases is an acute inflammatory disease. In certain embodiments, the inflammatory diseases is a chronic inflammatory disease. In certain embodiments, the disease or disorder is Aicardi-Goutières syndrome (AGS), STING-associated vasculopathy of infancy (SAVI), lupus, systemic lupus erythematosus (SLE), COPA syndrome, amyotrophic lateral sclerosis (ALS), Niemann-Pick disease type C (NPC), multiple sclerosis, familial chilblain lupus (FCL), rheumatoid arthritis, Sjögren’s syndrome, secondary Sjögren’s syndrome, Alzheimer disease, ischemic brain injury, Parkinson disease, neurodegeneration, Huntington disease, frontotemporal dementia, age-dependent macular degeneration, traumatic brain injury, nonalcoholic steatohepatitis, alcoholic liver disease, acute pancreatitis, silica- induced fibrosis, sepsis, myocardial infarction, chronic heart failure, or colorectal cancer.

[0077] In another aspect, the present disclosure provides methods of inhibiting STING pathway, inhibiting the phosphorylation of TBK1, and / or inhibiting the release of an inflammatory cytokine in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an anti-STING antibody or an antigen-binding fragment thereof disclosed herein, a pharmaceutical composition disclosed herein, or a formulation disclosed herein.

[0078] In certain embodiments, inhibiting STING pathway is indicated by a reduced level of pTBK1 and / or a reduced level of an inflammatory cytokine in a sample of the subject. In certain embodiments, inhibiting the phosphorylation of TBK1 is indicated by a reduced level of pTBK1 in a sample of the subject. In certain embodiments, inhibiting the release of an inflammatory cytokine is indicated by a reduced level of the inflammatory cytokine in aNAI-5002871589v118sample of the subject. In certain embodiments, the inflammatory cytokine is IL-6, IP-10,IFN- , and / or ISG-15. In certain embodiments, the sample is a blood sample, a serumsample, a tissue sample, or a cell sample. In certain embodiments, the reduced level of pTBK1 and / or reduced level of the inflammatory cytokine is reduced as compared to the level of pTBK1 and / or the level of the inflammatory cytokine measured before the administration of the anti-STING antibody or antigen-binding fragment thereof or to a reference level. In certain embodiments, the level of pTBK1 is measured as the ratio between pTBK1 and total TBK1 levels, or the ratio between pTBK1 and total GAPDH levels.

[0079] In another aspect, the present disclosure provides an immunogen comprising a stabilized epitope comprising or consisting of amino acid residues S358, G367, M368, E369, and P371 of human STING, wherein the amino acid positions are numbered according to the positions in the amino acid sequence of SEQ ID NO:34. In certain embodiments, the epitope is a conformational epitope or a linear epitope. In certain embodiments, the immunogen is stabilized by epitope scaffolding or structural modifications.

[0080] In another aspect, the present disclosure provides a method of generating an anti- STING antibody, comprising administering an immunogen disclosed herein to a subject, and isolating the anti-STING antibody from the subject. 3.1 Illustrative Embodiments

[0081] The present disclosure provides the following non-limiting embodiments: 1. An antibody or an antigen-binding fragment thereof that can specifically bind human stimulator of interferon genes (STING), wherein the antibody or antigen-binding fragment thereof comprises: (i) a heavy chain variable region (VH) comprising a heavy chain variable region complementarity determining region 1 (VH-CDR1), a heavy chain variable region complementarity determining region 2 (VH-CDR2), and a heavy chain variable region complementarity determining region 3 (VH-CDR3) as set forth in a VH comprising the amino acid sequence of SEQ ID NO:2; and a light chain variable region (VL) comprising a light chain variable region complementarity determining region 1 (VL-CDR1), a light chain variable region complementarity determining region 2 (VL-CDR2), and a light chain variable region complementarity determining region 3 (VL-CDR3) as set forth in a VL comprising the amino acid sequence of SEQ ID NO:7; (ii) a VH comprising a VH-CDR1, a VH-CDR2, and a VH-CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO:12; and a VLNAI-5002871589v119comprising a VL-CDR1, a VL-CDR2, and a VL-CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO:17; or (iii) a VH comprising a VH-CDR1, a VH-CDR2, and a VH-CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO:22; and a VL comprising a VL-CDR1, a VL-CDR2, and a VL-CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO:27. 2. The antibody or antigen-binding fragment thereof of embodiment 1, wherein the VH- CDR1 comprises the amino acid sequence of SEQ ID NO:3, the VH-CDR2 comprises the amino acid sequence of SEQ ID NO:4, the VH-CDR3 comprises the amino acid sequence of SEQ ID NO:5, the VL-CDR1 comprises the amino acid sequence of SEQ ID NO:8, the VL- CDR2 comprises the amino acid sequence of SEQ ID NO:9, and the VL-CDR3 comprises the amino acid sequence of SEQ ID NO:10, and wherein the amino acid sequences of the VH-CDR1, VH-CDR2, VH-CDR3, VL- CDR1, VL-CDR2, and VL-CDR3 are determined by Kabat definition. 3. The antibody or antigen-binding fragment thereof of embodiment 2, wherein (i) the VH comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:2, or the VL comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:7; (ii) the VH comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:2, and the VL comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:7; or (iii) the VH comprises the amino acid sequence of SEQ ID NO:2 or the VL comprises the amino acid sequence of SEQ ID NO:7. 4. The antibody or antigen-binding fragment thereof of embodiment 2 or 3, wherein the VH comprises the amino acid sequence of SEQ ID NO:2 and the VL comprises the amino acid sequence of SEQ ID NO:7. 5. The antibody or antigen-binding fragment thereof of embodiment 1, wherein the VH- CDR1 comprises the amino acid sequence of SEQ ID NO:13, the VH-CDR2 comprises the amino acid sequence of SEQ ID NO:14, the VH-CDR3 comprises the amino acid sequence of SEQ ID NO:15, the VL-CDR1 comprises the amino acid sequence of SEQ ID NO:18, the VL-CDR2 comprises the amino acid sequence of SEQ ID NO:19, and the VL-CDR3 comprises the amino acid sequence of SEQ ID NO:20, andNAI-5002871589v120wherein the amino acid sequences of the VH-CDR1, VH-CDR2, VH-CDR3, VL- CDR1, VL-CDR2, and VL-CDR3 are determined by Kabat definition. 6. The antibody or antigen-binding fragment thereof of embodiment 5, wherein (i) the VH comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:12, or the VL comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:17; (ii) the VH comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:12, and the VL comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:17; or (iii) the VH comprises the amino acid sequence of SEQ ID NO:12 or the VL comprises the amino acid sequence of SEQ ID NO:17. 7. The antibody or antigen-binding fragment thereof of embodiment 5 or 6, wherein the VH comprises the amino acid sequence of SEQ ID NO:12 and the VL comprises the amino acid sequence of SEQ ID NO:17. 8. The antibody or antigen-binding fragment thereof of embodiment 1, wherein the VH- CDR1 comprises the amino acid sequence of SEQ ID NO:23, the VH-CDR2 comprises the amino acid sequence of SEQ ID NO:24, the VH-CDR3 comprises the amino acid sequence of SEQ ID NO:25, the VL-CDR1 comprises the amino acid sequence of SEQ ID NO:28, the VL-CDR2 comprises the amino acid sequence of SEQ ID NO:29, and the VL-CDR3 comprises the amino acid sequence of SEQ ID NO:30, and wherein the amino acid sequences of the VH-CDR1, VH-CDR2, VH-CDR3, VL- CDR1, VL-CDR2, and VL-CDR3 are determined by Kabat definition. 9. The antibody or antigen-binding fragment thereof of embodiment 8, wherein (i) the VH comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:22, or the VL comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:27; (ii) the VH comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:22, and the VL comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:27; orNAI-5002871589v121(iii) the VH comprises the amino acid sequence of SEQ ID NO:22 or the VL comprises the amino acid sequence of SEQ ID NO:27. 10. The antibody or antigen-binding fragment thereof of embodiment 8 or 9, wherein the VH comprises the amino acid sequence of SEQ ID NO:22 and the VL comprises the amino acid sequence of SEQ ID NO:27. 11. An antibody or an antigen-binding fragment thereof that competes with the antibody or antigen-binding fragment thereof of any one of embodiments 1-10 for specifically binding to an epitope located within the C-terminal domain of STING. 12. The antibody or antigen-binding fragment thereof of embodiment 11, wherein the C- terminal domain of STING comprises the amino acid sequence of SEQ ID NO:31. 13. The antibody or antigen-binding fragment thereof of embodiment 11 or 12, wherein the epitope is a conformational epitope or a linear epitope. 14. The antibody or antigen-binding fragment thereof of any one of embodiment 11-13, wherein the epitope comprises amino acid residues S358, G367, M368, E369, and P371 of human STING, wherein the amino acid positions are numbered according to the positions in the amino acid sequence of SEQ ID NO:34. 15. The antibody or antigen-binding fragment thereof of any one of embodiments 11-14, wherein the (i) the antibody or antigen-binding fragment thereof can inhibit the phosphorylation of TANK binding kinase 1 (TBK1) in cells with activated STING pathway in vitro, ex vivo or in vivo; and / or (ii) the antibody or antigen-binding fragment thereof can inhibit the release of an inflammatory cytokine in cells with activated STING pathway in vitro, ex vivo or in vivo; (iii) the antibody or antigen-binding fragment thereof can inhibit the release of an inflammatory cytokine in cells of a Trex1 null mouse or reduce the level of an inflammatory cytokine of a NZB / W mouse. 16. An antagonistic antibody or an antigen-binding fragment thereof that can specifically bind human STING, wherein: (i) the antibody or antigen-binding fragment thereof can specifically bind an epitope located within the C-terminal domain of STING; (ii) the antibody or antigen-binding fragment thereof can inhibit the phosphorylation of TBK1 in cells with activated STING pathway in vitro, ex vivo or in vivo;NAI-5002871589v122(iii) the antibody or antigen-binding fragment thereof can inhibit the release of an inflammatory cytokine in cells with activated STING pathway in vitro, ex vivo or in vivo; and / or (iv) the antibody or antigen-binding fragment thereof can inhibit the release of an inflammatory cytokine in cells of a Trex1 null mouse or reduce the level of an inflammatory cytokine of a NZB / W mouse. 17. The antibody or antigen-binding fragment thereof of embodiment 16, wherein the C- terminal domain of STING comprises the amino acid sequence of SEQ ID NO:31. 18. The antibody or antigen-binding fragment thereof of embodiment 16 or 17, wherein the epitope is a conformational epitope or a linear epitope. 19. The antibody or antigen-binding fragment thereof of any one of embodiments 15-18, wherein the epitope comprises amino acid residues S358, G367, M368, E369, and P371 of human STING, wherein the amino acid positions are numbered according to the positions in the amino acid sequence of SEQ ID NO:34. 20. The antibody or antigen-binding fragment thereof of any one of embodiments 15-19, wherein the inhibition of the phosphorylation of TBK1 in the cells with activated STING pathway is indicated by a reduced pTBK1 level in the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof as compared to cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof, optionally wherein the level of pTBK1 is measured by an immunohistochemistry assay, further optionally wherein the immunohistochemistry assay is Western Blot or immunocytochemistry (ICC). 21. The antibody or antigen-binding fragment thereof of any one of embodiments 15- 19, wherein the inhibition of the release of the inflammatory cytokine is indicated by a reduced level of the inflammatory cytokine released by the cells with activated STING pathway or the cells of the Trex1 null mouse and contacted or administered with the antibody or antigen-binding fragment thereof as compared to cells with activated STING pathway or cells of a Trex1 null mouse but without being contacted or administered with the antibody or antigen-binding fragment thereof, optionally wherein the level of the inflammatory cytokine is measured by an immunoassay or RT-qPCR, optionally wherein the immunoassay is ELISA, HTRF (homogeneous time resolved fluorescence), or MSD (Meso Scale Discovery) multiplex assay. 22. The antibody or antigen-binding fragment thereof of any one of embodiments 15- 19, wherein reducing the level of the inflammatory cytokine is indicated by a reduced level ofNAI-5002871589v123the inflammatory cytokine in serum of the NZB / W mouse being administered with the antibody or antigen-binding fragment thereof as compared to the level of the inflammatory cytokine in serum of a NZB / W mouse but without being administered with the antibody or antigen-binding fragment thereof, further optionally wherein the level of the inflammatory cytokine is measured by an immunoassay, further optionally wherein the immunoassay is MSD multiplex assay. 23. The antibody or antigen-binding fragment thereof of any one of embodiments 15-22,wherein the inflammatory cytokine is IL-6, IP-10, IFN- , and / or ISG-15.24. The antibody or antigen-binding fragment thereof of any one of embodiments 15-23, wherein the cells are immune cells, heart tissue cells of the Trex1 null mouse, differentiated THP-1 cells, or mouse splenocytes. 25. The antibody or antigen-binding fragment thereof of any one of embodiments 15-24, wherein the STING pathway is activated by 10-carboxymethyl-9-acridanone (CMA), cyclic GMP-AMP (cGAMP), or HT DNA (deoxyribonucleic acid from herring testes). 26. The antibody or antigen-binding fragment thereof of any one of embodiments 20-25, wherein the pTBK1 level in the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less of the pTBK1 level in the cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof, as measured by Western Blot or ICC, and the cells are PMA-differentiated human THP-1-derived macrophages or mouse splenocytes with STING pathway activated by cGAMP or CMA. 27. The antibody or antigen-binding fragment thereof of any one of embodiments 21-25, wherein the level of the inflammatory cytokine released by the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less of cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof, as measured by ELISA or HTRF, and the cells are PMA-differentiated human THP-1-derived macrophages or mouse splenocytes with STING pathway activated by cGAMP, and the inflammatorycytokine is IFN- .28. The antibody or antigen-binding fragment thereof of any one of embodiments 21-25, wherein the level of the inflammatory cytokine released by the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof is aboutNAI-5002871589v12420% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less of cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof, as measured by MSD multiplex assay, and the cells are PMA-differentiated human THP-1-derived macrophageswith STING pathway activated by HT DNA, and the inflammatory cytokine is IFN- , IL-6,and / or IP-10. 29. The antibody or antigen-binding fragment thereof of any one of embodiments 21-25, wherein the level of the inflammatory cytokine released by the cells from the Trex1 null mouse administered with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less of cells from a Trex1 null mouse without being administered with the antibody or antigen-binding fragment thereof, as measured by RT-qPCR, and thecells are heart tissue cells, and the inflammatory cytokine is IFN- , IL-6, and / or ISG-15.30. The antibody or antigen-binding fragment thereof of any one of embodiments 22-25, wherein the level of the inflammatory cytokine in serum of the NZB / W mouse administered with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, about 80% or less, or about 90% or less of serum from a NZB / W mouse without being administered with the antibody or antigen-binding fragment thereof, as measured by MSD multiplex assay, and the inflammatory cytokine is IL-6. 31. The antibody or antigen-binding fragment thereof of any one of embodiments 1-30, which is an antagonistic antibody or antigen-binding fragment thereof. 32. The antibody or antigen-binding fragment thereof of any one of embodiments 1-31, wherein the antibody is a recombinant antibody and / or a monoclonal antibody. 33. The antibody or antigen-binding fragment thereof of any one of embodiments 1-32, wherein the antibody comprises a heavy chain constant region and / or a light chain constant region. 34. The antibody or antigen-binding fragment thereof of embodiment 33, wherein the heavy chain constant region is of human IgG type, optionally wherein the heavy chain constant region is of IgG1 type or human IgG4 type. 35. The antibody or antigen-binding fragment thereof of embodiment 33 or 34, wherein the light chain constant region comprises a human kappa light chain constant region or a human lambda light chain constant region.NAI-5002871589v12536. The antibody or antigen-binding fragment thereof of any one of embodiments 33-35, wherein the heavy chain constant region comprises the amino acid sequence of SEQ ID NO:129 or SEQ ID NO:132, and / or the light chain constant region comprises the amino acid sequence of SEQ ID NO:130 or SEQ ID NO:131. 37. The antibody or antigen-binding fragment thereof of any one of embodiments 33-36, wherein the antibody comprises an Fc region comprising one or more mutations, optionally wherein the one or more mutations decrease immune effector function, increase immune effector function, or increase half-life of the antibody. 38. The antibody or antigen-binding fragment thereof of any one of embodiments 1-37, wherein the antigen-binding fragment a single chain Fv fragment (scFv), an F(ab’), an F(ab), an F(ab’)2, an Fd, an Fv, a single-chain antibody, a disulfide-linked Fv (sdFv), a VHH, or a nanobody. 39. The antibody or antigen-binding fragment thereof of any one of embodiments 1-37, wherein the antibody or antigen-binding fragment thereof is attached to a detectable label, wherein the detectable label is an enzyme, a radioisotope, a fluorescent compound, a chemiluminescent compound, a bioluminescent compound, a tag, a flag or a heavy metal. 40. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof of any one of embodiments 1-39 and 46 and a pharmaceutically acceptable carrier. 41. A formulation suitable for in vivo administration comprising the antibody or antigen- binding fragment thereof of any one of embodiments 1-39 and 46, optionally wherein the formulation is suitable for in vivo intravenous, intraperitoneal, subcutaneous, or intramuscular injection. 42. An isolated polynucleotide or a set of polynucleotides encoding the antibody or antigen-binding fragment thereof of any one of embodiments 1-39 and 46. 43. A vector or a set of vectors comprising the polynucleotide or set of polynucleotides of embodiment 42. 44. An isolated cell comprising the polynucleotide or set of polynucleotides of embodiment 42 or the vector or set of vectors of embodiment 43. 45. A method of making an antibody or an antigen-binding fragment thereof, comprising: (i) culturing the cell of embodiment 44, and (ii) isolating the antibody or antigen-binding fragment thereof. 46. An antibody or an antigen-binding fragment thereof produced by the method of embodiment 45. 47. A method of detecting STING in a biological sample, comprising:NAI-5002871589v126(i) contacting the biological sample with the antibody or antigen-binding fragment thereof of any one of embodiments 1-39 and 46; and (ii) detecting the binding between the antibody or antigen-binding fragment thereof and STING in the biological sample, optionally wherein the detecting comprises using flow cytometry, immunohistochemistry (IHC), Western Blot analysis, ELISA, or mass spectrometry. 48. A method for screening for an antibody or an antigen-binding fragment thereof that can specifically bind human STING, comprising measuring the binding of the antibody or antigen-binding fragment thereof to an epitope located within the C-terminal domain of STING, and selecting the antibody if the binding affinity is higher than an isotype control antibody, optionally wherein the binding is measured by ELISA. 49. The method of embodiment 48, wherein the C-terminal domain of STING comprises the amino acid sequence of SEQ ID NO:31. 50. The method of embodiment 48 or 49, wherein the epitope is a conformational epitope or a linear epitope. 51. The method of any one of embodiment 48-50, wherein the epitope comprises amino acid residues S358, G367, M368, E369, and P371 of human STING, wherein the amino acid positions are numbered according to the positions in the amino acid sequence of SEQ ID NO:34. 52. A method of inhibiting the phosphorylation of TBK1 in cells with activated STING pathway, inhibiting the release of an inflammatory cytokine in cells with activated STING pathway, inhibiting the release of an inflammatory cytokine in cells of a Trex1 null mouse, and / or reducing the level of an inflammatory cytokine of a NZB / W mouse comprising contacting the cells or administering to the mouse with the antibody or antigen-binding fragment thereof of any one of embodiments 1-39 and 46, wherein the method is an in vitro, ex vivo or in vivo method. 53. The method of embodiment 52, wherein the inhibition of the phosphorylation of TBK1 in the cells with activated STING pathway is indicated by a reduced pTBK1 level in the cells with activated STING pathway and being contacted with the antibody or antigen- binding fragment thereof as compared to cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof, optionally wherein the level of pTBK1 is measured by an immunohistochemistry assay, optionally wherein the level of pTBK1 is measured by an immunohistochemistry assay, further optionally, the immunohistochemistry assay is Western Blot or immunocytochemistry (ICC).NAI-5002871589v12754. The method of embodiment 52, wherein the inhibition of the release of the inflammatory cytokine is indicated by a reduced level of the inflammatory cytokine released by the cells with activated STING pathway or the cells of the Trex1 null mouse and being contacted or administered with the antibody or antigen-binding fragment thereof as compared to cells with activated STING pathway or the cells of the Trex1 null mouse but without being contacted or administered with the antibody or antigen-binding fragment thereof, optionally wherein the level of the inflammatory cytokine is measured by an immunoassay or RT-qPCR, optionally wherein the immunoassay is ELISA or HTRF. 55. The method of embodiment 52, wherein reducing the level of the inflammatory cytokine is indicated by a reduced level of the inflammatory cytokine in serum of the NZB / W mouse being administered with the antibody or antigen-binding fragment thereof as compared to the level of the inflammatory cytokine in serum of a NZB / W mouse but without being administered with the antibody or antigen-binding fragment thereof, optionally wherein the level of the inflammatory cytokine is measured by an immunoassay, further optionally wherein the immunoassay is MSD multiplex assay. 56. The method of any one of embodiments 52-55, wherein the inflammatory cytokine isIL-6, IFN- , IP-10, and / or ISG-15.57. The method of any one of embodiments 52-56, wherein the cells are immune cells, heart tissue cells from the Trex1 null mouse, differentiated THP-1 cells, or mouse splenocytes. 58. The method of any one of embodiments 52-57, wherein the STING pathway is activated by CMA, cGAMP, or HT DNA. 59. The method of any one of embodiments 53-58, wherein the pTBK1 level in the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less of the pTBK1 level in the cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof, as measured by Western Blot or ICC, and the cells are PMA-differentiated human THP-1-derived macrophages or mouse splenocytes with STING pathway activated by cGAMP or CMA. 60. The method of any one of embodiments 54-58, wherein the level of the inflammatory cytokine released by the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less ofNAI-5002871589v128cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof, as measured by ELISA or HTRF, and the cells are PMA- differentiated human THP-1-derived macrophages or mouse splenocytes with STINGpathway activated by cGAMP, and the inflammatory cytokine is IFN- .61. The method of any one of embodiments 52-56, wherein the level of the inflammatory cytokine released by the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less of cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof, as measured by MSD multiplex assay, and the cells are PMA-differentiated human THP-1-derived macrophages with STING pathway activated byHT DNA, and the inflammatory cytokine is IFN- , IL-6, and / or IP-10.62. The method of any one of embodiments 54-58, wherein the level of the inflammatory cytokine released by the cells from the Trex1 null mouse administered with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less of cells from a Trex1 null mouse without being contacted with the antibody or antigen-binding fragment thereof, as measured by RT-qPCR, and the cells are heart tissue cells, and the inflammatorycytokine is IFN- , IL-6, and / or ISG-15.63. The method of any one of embodiments 55-58, wherein the level of the inflammatory cytokine in serum of the NZB / W mouse administered with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, about 80% or less, or about 90% or less of serum from a NZB / W mouse without being administered with the antibody or antigen-binding fragment thereof, as measured by MSD multiplex assay, and the inflammatory cytokine is IL- 6. 64. A method of treating a disease or a disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the antibody or antigen- binding fragment thereof of any one of embodiments 1-39 and 46, the pharmaceutical composition of embodiment 40, or the formulation of embodiment 41. 65. The antibody or antigen-binding fragment thereof of any one of embodiments 1-39 and 46, the pharmaceutical composition of embodiment 40, or the formulation of embodiment 41 for use in treating a disease or a disorder in a subject in need thereofNAI-5002871589v12966. Use of the antibody or antigen-binding fragment thereof of any one of embodiments 1-39 and 46, the pharmaceutical composition of embodiment 40, or the formulation of embodiment 41 in the manufacture of a medicament for treating a disease or a disorder in a subject in need thereof. 67. The method of embodiment 64, antibody or antigen-binding fragment thereof for use in accordance with embodiment 65, or use of embodiment 66, wherein the disease or disorder is an autoimmune disease or an inflammatory disease. 68. The method of embodiment 64 or 67, the antibody or antigen-binding fragment thereof for use in accordance with embodiment 65 or 67, or the use of embodiment 66 or 67, wherein the disease or disorder is Aicardi-Goutières syndrome (AGS), STING-associated vasculopathy of infancy (SAVI), lupus, systemic lupus erythematosus (SLE), COPA syndrome, amyotrophic lateral sclerosis (ALS), Niemann-Pick disease type C (NPC), multiple sclerosis, familial chilblain lupus (FCL), rheumatoid arthritis, Sjögren’s syndrome, secondary Sjögren’s syndrome, Alzheimer disease, ischemic brain injury, Parkinson disease, neurodegeneration, Huntington disease, frontotemporal dementia, age-dependent macular degeneration, traumatic brain injury, nonalcoholic steatohepatitis, alcoholic liver disease, acute pancreatitis, silica- induced fibrosis, sepsis, myocardial infarction, chronic heart failure, or colorectal cancer. 69. An immunogen comprising a stabilized epitope, wherein the epitope comprises or consists of amino acid residues S358, G367, M368, E369, and P371 of human STING, wherein the amino acid positions are numbered according to the positions in the amino acid sequence of SEQ ID NO:34. 70. The immunogen of embodiment 69, wherein the immunogen is stabilized by epitope scaffolding or structural modifications. 71. A method of generating an anti-STING antibody, comprising administering the immunogen of embodiment 69 or 70 to a subject, and isolating the anti-STING antibody from the subject, wherein in one embodiment the subject in not a human subject. 72. A method of inhibiting STING pathway, inhibiting the phosphorylation of TBK1, and / or inhibiting the release of an inflammatory cytokine in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the antibody or antigen-binding fragment thereof of any one of embodiments 1-39 and 46, the pharmaceutical composition of embodiment 40, or the formulation of embodiment 41. 73. The method of embodiment 72, wherein (i) inhibiting STING pathway is indicated by a reduced level of pTBK1 and / or a reduced level of an inflammatory cytokine in a sample ofNAI-5002871589v130the subject; (ii) inhibiting the phosphorylation of TBK1 is indicated by a reduced level of pTBK1 in a sample of the subject; and / or (iii) inhibiting the release of an inflammatory cytokine is indicated by a reduced level of the inflammatory cytokine in a sample of the subject, optionally wherein the reduced level of pTBK1 and / or reduced level of the inflammatory cytokine is reduced as compared to the level of pTBK1 and / or the level of the inflammatory cytokine measured before the administration of the anti-STING antibody or antigen-binding fragment thereof or to a reference level. 74. The method of embodiment 72 or 73, wherein the inflammatory cytokine is IL-6, IP-10, IFN- , and / or ISG-15.75. The method of embodiment 73 or 74, wherein the sample is a blood sample, a serum sample, a tissue sample, or a cell sample. 4. BRIEF DESCRIPTION OF THE FIGURES

[0082] FIGS. 1A-1C provide amino acid sequences of the variable regions, i.e., heavy chain and kappa light chain (VH, VL) of the anti-STING specific antibody Ab1 (FIG. 1A), Ab2 (FIG. 1B), and Ab3 (FIG. 1C) of the present disclosure. Framework (FR) and complementarity determining regions (CDRs) are indicated with the CDRs being underlined. The Kabat numbering scheme was used. Depicted are the variable heavy chain VH and light chain VL sequence of antibody Ab1 as set forth in SEQ ID NOs: 2 and 7, of antibody Ab2 as set forth in SEQ ID NOs: 12 and 17, and of antibody Ab3 as set forth in SEQ ID NOs: 22 and 27. As further explained in the description, within CDRs and / or framework region conservative amino acid substitutions are preferred which take into account the physicochemical properties of the original amino acid either alone or with an adjacent amino acid as illustrated in Mirsky et al., Mol. Biol. Evol. 32 (2014) 806-819 at page 813, Figure 6 in particular the AB or LG model, for example such that the position of two amino acids is exchanged.

[0083] FIG. 2 Anti-STING antibody Ab1 of the present disclosure showed specific binding to human and mouse STING, as well as to human, cyno and rat C-terminus STING peptides. Binding activity of Ab1 was assessed on recombinant human STING protein, recombinant mouse STING protein, C-terminus human STING peptide (Peptide sequence: H-Val-Thr- Val-Gly-Ser-Leu-Lys-Thr-Ser-Ala-Val-Pro-Ser-Thr-Ser-Thr-Met-Ser-Gln-Glu-Pro-Glu-Leu- Leu-Ile-Ser-Gly-Met-Glu-Lys-Pro-Leu-Pro-Leu-Arg-Thr-Asp-Phe-Ser-OH; SEQ ID NO:31), C-terminus cyno STING peptide (Peptide sequence: H-Arg-Gln-Glu-Lys-Lys-Glu-Glu-Val-NAI-5002871589v131Thr-Val-Gly-Ser-Leu-Lys-Asn-Ser-Ala-Val-Pro-Ser-Thr-Ser-Thr-Met-Ser-Gln-Glu-Pro-Glu- Leu-Leu-Ile-Ser-Gly-Met-Glu-Lys-Pro-Leu-Pro-Leu-Arg-Thr-Asp-Phe-Ser-OH; SEQ ID NO:32), C-terminus rat STING peptide (Peptide sequence: H-Lys-Glu-Glu-Val-Thr-Met-Ser- Gly-Pro-Pro-Thr-Ser-Val-Ala-Pro-Arg-Pro-Ser-Leu-Leu-Ser-Gln-Glu-Pro-Arg-Leu-Leu-Ile- Ser-Gly-Met-Glu-Gln-Pro-Leu-Pro-Leu-Arg-Thr-Asp-Leu-Ile-OH; SEQ ID NO:33), BSA and E. coli lysates by ELISA. Line depicts sigmoidal curve fit. Data is displayed as mean ± SEM.

[0084] FIGS. 3A-3C Anti-STING antibody Ab1 of the present disclosure showed specific binding to human and mouse STING ((FIG. 3A) and (FIG. 3C) in a concentration-dependent manner, but no binding to E. coli cell lysates and BSA (FIG. 3B). Binding specificity of Ab1 (10 g / ml) was assessed on different amounts of recombinant human STING protein (Alexotech), E. coli lysates, BSA, recombinant mouse STING by Western Blot. Numbers indicate molecular weight in kilo Dalton (kDa).

[0085] FIGS. 4A & 4B Anti-STING antibody Ab1 of the present disclosure showed specific binding to STING in human cells. Binding of Ab1 (1 g / ml) to human PBMNCs (FIG. 4A) and human splenocytes (FIG. 4B) was assessed by Immunocytochemistry. Positive Cy3 signal was detected in cells stained with Ab1. No signal was observed with human IgG control (isotype control antibody, 5 g / ml) and with an anti-human control (secondary antibody control, diluted 1:250). Nuclei were visualized with DAPI. Images were acquired with a 20× objective. Scale bar: 5 μm.

[0086] FIG. 5 Anti-STING antibody Ab1 of the present disclosure showed specific binding to STING in human tissue. Binding of Ab1 to human lung adenocarcinoma was assessed by immunofluorescence. Positive fluorescent signal was detected in tissues stained with Ab1 (50 g / ml). Tissue stained with a commercial anti-STING antibody (Invitrogen D2P2F, diluted 1:100) was used as positive control. No signal was observed with anti-human and anti-rabbit controls (secondary antibody controls, diluted 1:400). Nuclei were visualized with DAPI. Images were acquired with a 20× objective. Scale bar: 20 μM.

[0087] FIGS. 6A-6D Anti-STING antibody Ab1 of the present disclosure showed specific binding to STING in human tissue. Binding of Ab1 to non demented control human brain tissue was assessed by immunohistochemistry. Positive signal was observed in tissues stained with Ab1 (10 g / ml) (FIG. 6B). Tissue stained with a commercial anti-STING antibody (Proteintech 66680-1 Ig, diluted 1:1000) was used as positive control (FIG. 6A). No signal was observed with human IgG control (isotype control antibody, 10 g / ml) (FIG. 6C) andNAI-5002871589v132with anti-human and anti-mouse controls (secondary antibody controls, diluted 1:350) (FIG. 6D). Scale bar: 50 μM.

[0088] FIG. 7 Anti-STING antibody Ab1 of the present disclosure inhibited IFN- release bymouse splenocytes in vitro. IFN- levels in cell medium were assessed by ELISA. Incubationwith cGAMP (23.6 g / ml) for 24 hours was used to activate STING pathway, which triggersIFN- release. Reduction in IFN- concentration in cell medium was observed when cellswere treated for 2 hours with Ab1 (5 g / ml). Human IgG control (isotype control antibody, 5 g / ml) did not affect IFN- release. Data is displayed as IFN- concentration in pg / ml.

[0089] FIG. 8 Anti-STING antibody Ab1 of the present disclosure inhibited IFN- release bydifferentiated THP-1 cells in vitro, in a dose-dependent manner. IFN- levels in cell mediumwere assessed by HTRF. Incubation with cGAMP (23.6 g / ml) for 3 hours was used toactivate STING pathway, which triggers IFN- release. Progressive reduction in IFN- levelsin cell medium was observed when cells were treated overnight with increasing concentrations of Ab1 (0.5, 1 and 5 μg / ml). Human IgG control (isotype control antibody, 10 g / ml) did not affect IFN- release. Data is displayed as Delta ratio.

[0090] FIGS. 9A & 9B Anti-STING antibody Ab1 of the present disclosure led to a concentration-dependent decrease in pTBK1 levels in differentiated THP-1 cells. (FIG. 9A) Reduction in phosphorylation of TBK1 (pTBK1) was assessed in THP-1 cell lysates by Western Blot. Numbers indicate molecular weight in kilo Dalton (kDa). Overnight incubation with PMA (500 ng / ml) was used to differentiate the cells, while 3 hours of incubation with cGAMP (750 ng / ml) allowed activation of STING pathway, which led to phosphorylation of TBK1. A small molecule STING inhibitor (H-151, 0.5 M for 2 hour) was included as positive control. Progressive reduction in pTBK1 levels was observed when cells are treated overnight with increasing concentrations of Ab1. Human IgG control (isotype controlantibody) did not affect IFN- release. Total TBK1 levels were used for normalization andshowed no variation among different treatments. (FIG.9B) Quantification of the ratio between pTBK1 and total TBK1 Western Blot bands. Data is displayed as mean + SD.

[0091] FIGS. 10A-10C Anti-STING antibody Ab1 of the present invention led to a decrease in pTBK1 levels in PBMCs. (FIG. 10A) Reduction in phosphorylation of TBK1 (pTBK1) was assessed in PBMCs lysates by Western Blot. Numbers indicate molecular weight in kilo Dalton (kDa). 4 hours of incubation with cGAMP (2.5μg / ml) was used to activate the STING pathway, which leads to phosphorylation of TBK1. A small molecule STING inhibitor (H151, 0.5 M for 2 hours) was included as positive control. Reduction in pTBK1 levels was observed after 4h therapeutic treatment with Ab1. Human IgG control (isotype controlNAI-5002871589v133antibody) did not affect pTBK1 levels. Total TBK1 and GAPDH levels are used for normalization and showed no variation among different treatments. Quantification of the ratio between pTBK1 and total TBK1 (FIG. 10B) or GAPDH (FIG. 10C) Western Blot bands. Data is displayed as mean + SD.

[0092] FIGS. 11A-11F Anti-STING antibody Ab1 of the present disclosure led to a reduction in pTBK1 levels in mouse splenocytes, upon ex vivo CMA-mediated STING pathway activation. Reduction in phosphorylation of TBK1 was assessed in mouse splenocytes by immunocytochemistry. (FIG. 11B) Positive Cy5 signal was detected in cells treated for 3 hours with CMA (250 g / ml) in contrast to (FIG. 11A) untreated cells. (FIG. 11E) Reduction in signal was observed upon overnight treatment with Ab1 (10 g / ml), (FIG. 11C) as well as with the small molecule STING inhibitor (H-151, 0.5 M, 2 hours). (FIG. 11D) Human IgG control (isotype control antibody, 10 g / ml) did not affect pTBK1 levels. No signal was detected with anti-human control (secondary antibody control, diluted 1:250). Nuclei were visualized with DAPI. Images were acquired with a 20x objective. Scale bar: 5 μm. (FIG. 11F) Quantification of percentage of Cy5 (pTBK1) positive cells, normalized by DAPI signal.

[0093] FIGS. 12A-12F Anti-STING antibody Ab1 of the present disclosure led to a reduction in pTBK1 levels in mouse splenocytes, upon ex vivo cGAMP-mediated STING pathway activation. Reduction in phosphorylation of TBK1 was assessed in mouse splenocytes by immunocytochemistry. (FIG. 12B) Positive Cy5 signal was detected in cells treated for 3 hours with cGAMP (750 ng / ml) in contrast to (FIG. 12A) untreated cells. (FIG. 12E) Reduction in signal was observed upon overnight treatment with Ab1 (10 g / ml), (FIG. 12C) as well as with the small molecule STING inhibitor (H-151, 0.5 M, 2 hours). (FIG. 12D) Human IgG control (isotype control antibody, 10 g / ml) did not affect pTBK1 levels. No signal was detected with anti-human control (secondary antibody control, diluted 1:250). Nuclei were visualized with DAPI. Images were acquired with a 20x objective. Scale bar: 5 μm. (FIG. 12F) Quantification of percentage of Cy5 (pTBK1) positive cells, normalized by DAPI signal.

[0094] FIG. 13 Schematic representation of the study design employed for the in vivo study to assess anti-STING antibody Ab1 efficacy in Trex1- / - mice. Twelve 4-5 week-old mice were assigned to each of the following groups: untreated, IgG2a murine control (50 mg / kg), Ab1 (0.5 mg / kg), Ab1 (5 mg / kg), Ab1 (50 mg / kg), positive control (250 ug / mouse).

[0095] FIG. 14 shows the body weight change rate in Trex1 - / - mice receiving the indicated treatments. A concentration-dependent correlation was observed upon administration of anti-NAI-5002871589v134STING antibody lead candidate Ab1, with 5 mg / kg dose exhibiting comparable levels of body weight change as the positive control treatment groups, whereas the 50 mg / kg dose of Ab1 showed the most pronounced beneficial increase. Data are shown as mean ± SEM. Normality - Lognormality test applied. Statistical analysis by Two Way-ANOVA.

[0096] FIGS. 15A-15C show the mRNA levels of key STING-related cytokines (IFN-(FIG. 15A), ISG-15 (FIG. 15B) and IL-6 (FIG. 15C) measured by RT-qPCR in heart tissue of Trex1 - / - mice receiving the indicated treatments (at day 42). Data are shown as mean ± SEM. Normality - Lognormality test applied. Statistical analysis by One Way- ANOVA or Kruskal-Wallis. Multiple comparisons to untreated or isotype group.

[0097] FIGS. 16A & 16B show the results of epitope mapping using overlapping peptide array representing human STING protein. FIG.16A shows binding of the anti-STING antibody lead candidate Ab1 (at 1 g / ml) of the present disclosure the indicated peptides. FIG. 16B shows the absence of binding of the anti-human secondary antibody negative control (at 1:20,000 dilution).

[0098] FIG. 17 shows the peptides amino acid sequences, the sequences of the peptides to which Ab1 antibody bound are in bold.

[0099] FIGS. 18A-18B show the results of enzyme-linked immunosorbent assay (ELISA) assay, measuring the binding of the anti-STING antibody lead candidate Ab1 at four dilutions (at 50 nM, 10 nM, 2 nM, and 0 nM, columns from left to right for each peptide group were 50 nM, 10 nM, 2 nM, and 0 nM) to the indicated peptides (FIG. 18A, peptides 42-65; FIG. 18B, peptides 66-91) in the epitope mapping using alanine replacement peptides and shortening of peptides representing C-terminal human STING protein. BSA and E. coli lysate were used as negative controls.

[0100] FIG. 19A shows the peptides amino acid sequences, the sequences of the peptides to which Ab1 antibody bound are in bold.

[0101] FIG. 19B illustrates confirmed epitope (greyed out amino acid residues) for Ab1 binding on human STING peptide .

[0102] FIG. 20 is a schematic representation of the study design used to assess in vivo target engagement of the anti-STING antibody Ab1 in C57BL / 6J mice. Male mice (11– 12 weeks old) were divided into three groups: Ab1 at 50 mg / kg (n=4), isotype-matched negative control at 50 mg / kg (n=4), and no injection (n=3). Twenty-four hours after a single intravenous injection, the mice were sacrificed, and organs were harvested. CD11b+ monocytes from the blood and spleen were isolated using Magnetic-activated cell sorting (MACS) for subsequent analysis by Western blotting (WB) and immunocytochemistry (ICC).NAI-5002871589v135

[0103] FIG. 21 shows that the anti-STING antibody Ab1 of the present disclosure was able to bind its target in CD11b+ monocytes isolated from the blood of mice injected with the antibody. The presence of the human antibody was assessed by Western blotting in both CD11b+ cell lysates and flow-through fractions. A lower signal was observed in cells isolated from mice injected with the isotype control antibody, reflecting the absence of the target. Molecular weight markers (in kilodaltons, kDa) are indicated.

[0104] FIG. 22 shows that the anti-STING antibody Ab1 of the present disclosure was able to bind its target in CD11b+ monocytes isolated from the spleen of mice injected with the antibody. The presence of the human antibody was assessed by Western blotting in both CD11b+ cell lysates and flow-through fractions. A lower signal was observed in cells isolated from mice injected with the isotype control antibody, indicating the absence of the target. Molecular weight markers (in kilodaltons, kDa) are indicated.

[0105] FIG. 23 is a schematic representation of the protocol used for the sedimentation and immunocytochemistry staining of CD11b+ monocytes isolated from mouse blood and spleen. Anti-human Cy3-conjugated antibody was used for staining Ab1 and isotype control antibodies, while DAPI was used for nuclear staining.

[0106] FIGS. 24A-24F show that the anti-STING antibody Ab1 of the present disclosure was able to bind its target in CD11b+ monocytes isolated from the blood of mice injected with the antibody. Binding of Ab1 to its target was assessed in CD11b+ monocytes isolated from pooled blood of mice injected with the antibody (FIGS. 24A & 24B), with the isotype control (FIG.24C), or not injected (FIG. 24D) using immunocytochemistry. Nuclei were visualized with DAPI. Images were acquired with a 20x objective, and the scale bar represents 5 μm. A positive Cy3 signal was detected in cells derived from mice injected with Ab1, while lower to no signal was observed in cells from isotype and non-injected controls. The results were confirmed by quantification of the Cy3-to-DAPI signal ratio (FIG. 24E) and the percentage of covered area (FIG.24F).

[0107] FIGS. 25A-25F show that the anti-STING antibody Ab1 of the present disclosure was able to bind its target in CD11b+ monocytes isolated from the spleens of mice injected with the antibody. Binding of Ab1 to its target was assessed in CD11b+ monocytes isolated from pooled spleens of mice injected with the antibody (FIGS. 25A & 25B), with the isotype control (FIG. 25C), or not injected (FIG. 25D) using immunocytochemistry. Nuclei were visualized with DAPI. Images were acquired with a 20x objective, and the scale bar represents 5 μm. A positive Cy3 signal was detected in cells derived from mice injected with Ab1, while lower to no signal was observed in cells from isotype and non-injectedNAI-5002871589v136controls. The results were confirmed by quantification of the Cy3-to-DAPI signal ratio (FIG. 25E) and the percentage of covered area (FIG. 25F).

[0108] FIGS. 26A & 26B are schematic representation of the experiment performed to assess antibody internalization in THP-1 cells. Live cell imaging was performed following a 24h pre-incubation with the antibody (FIG. 26A) or directly after antibody addition to the cells (FIG.26B)

[0109] FIG. 27 shows that THP-1 cells incubated with Ab1 labeled with Alexa Fluor 488 (permanent fluorescence) or pHrodo Red (fluorescent in acidic environments with a gradient from early endosome to lysosome). Positive A488 signal visible few hours after antibody incubation indicated vesicular uptake of the STING antibody followed by a progressive uptake in the endosomes characterized by increasing pHrodo signal over time. Diffuse signal observed after 23h suggested possible endosomal escape of the antibody to the cytoplasm.

[0110] FIG. 28 shows that THP-1 cells incubated with Ab1 labeled with Alexa Fluor 488 (permanent fluorescence) or pHrodo Red (fluorescent in acidic environments with a gradient from early endosome to lysosome). Increasing pHrodo Red signal and merged A488 and pHrodo Red signal over time indicated continued endosomal uptake.

[0111] FIG. 29 shows that THP-1 cells incubated with an isotype control antibody labeled with Alexa Fluor 488 (permanent fluorescence) or pHrodo Deep Red (fluorescent in acidic environments with a gradient from late endosome to lysosome). Positive dot-like A488 signal indicated vesicular uptake of the antibody and diffuse signal suggested possible endosomal escape of the antibody to the cytoplasm. Detection of pHrodo Deep Red signal after longer incubation times revealed localization of the antibody within late endosomes or lysosomes.

[0112] FIGS. 30A & 30B are representative high-resolution images of THP-1 cells incubated with 200 nM STING antibody labeled with Alexa Fluor 488 for (FIG. 30A) 48 hours or (FIG. 30B) 72 hours. Dot-like structures (see arrowheads) indicated vesicular uptake of the antibody and diffuse signal suggested endosomal escape of the antibody to the cytoplasm.

[0113] FIGS. 31A & 31B are representative high-resolution images of THP-1 cells incubated with 200 nM isotype control antibody labeled with Alexa Fluor 488 for (FIG. 31A) 48 hours or (FIG. 31B) 72 hours. Dot-like structures (see arrowheads) indicated vesicular uptake of the antibody and diffuse signal suggested endosomal escape of the antibody to the cytoplasm.NAI-5002871589v137

[0114] FIGS. 32A & 32B are representative high-resolution images of HeLa cells incubated with 200 nM STING antibody labeled with Alexa Fluor 488 for (FIG. 32A) 48 hours or (FIG. 32B) 72 hours. Dot-like structures indicated vesicular uptake of the antibody and diffuse signal suggested endosomal escape of the antibody to the cytoplasm.

[0115] FIGS. 33A & 33B is a Western Blot analysis of THP-1 cells incubated with STING antibody labeled with Alexa Fluor 488 for 6h, 24h, 48h or 72h. Numbers indicate molecular weight in kilo Dalton (kDa). GAPDH was used as loading control. Positive A488 signal indicated intracellular uptake of Ab1-A488 (FIG. 33A). Quantification of the ratio between A488 and GAPDH bands revealed time-dependent internalization pattern of Ab1- A488 in PMA-differentiated THP-1 cells (FIG. 33B).

[0116] FIGS. 34 shows EGFP signal detected in HeLa cells transduced with lentiviruses encoding the following constructs: EGFP only (positive control for successful transduction), STING-EGFP, EGFP-STING, GAPDH-EGFP and EGFP-GAPDH. Positive EGFP signal detected in the cells confirmed successful transduction with all constructs and generation of stable cell lines.

[0117] FIG. 35 is a schematic representation of the experiment performed to assess Ab1-A647 internalization and co-localization with STING-EGFP.

[0118] FIGS. 36A-36G are representative confocal images of HeLa cells expressing STING-EGFP and incubated for 48h with Ab1-A647 or Isotype-A647. Actin staining allowed visualization of the actin filaments and confirmed antibody internalization (FIG. 36A). 3D representation of a HeLa cell expressing STING-EGFP showing minimal overlap between Isotype-A647 and STING-EGFP signal (FIG. 36B). 3D representation of a HeLa cell expressing STING-EGFP showing a pronounced overlap between Ab1-A647 and STING-EGFP signal (FIG. 36C). Comparative quantification (two-tailed t-test, p<0.05) of the number of pixels (FIG. 36D) and Comparative quantification (two-tailed t-test, p<0.05) of the signal intensity indicated significant overlap of Ab1-A647 with the STING-EGFP signal (FIG. 36E). Correlation analysis between the number of STING-EGFP-positive pixels and Ab1-positive pixels (FIG. 36F). Correlation analysis between the number of STING- EGFP-positive pixels and Isotype-positive pixels (FIG. 36G).

[0119] FIGS. 37A-37C depict levels of cytokine IL-6 (FIG. 37A), IP-10 (FIG. 37B),and IFN- (FIG. 37C) assessed by MSD multiplex assay as demonstrated in Example 17.

[0120] FIGS 38A-38E depict in vivo study to assess anti-STING antibody Ab1 efficacy in NZB / W mice as demonstrated in Example 18. FIG. 38A is a schematic showing of the study design. FIG. 38B shows the percentage of surviving mice upon weeklyNAI-5002871589v138assessment in NZB / W mice. FIG. 38C shows the blood urea nitrogen (BUN) levels assessed in NZB / W mice after study endpoint. FIG. 38D shows the amelioration of kidney lesion score assessed in NZB / W mice after study endpoint. FIG. 38E shows the IL-6 levels assessed by MSD multiplex assay in NZB / W mice serum after study endpoint. 5. DETAILED DESCRIPTION

[0121] The present disclosure provides novel anti-STING antibodies and antigen- binding fragments thereof (Section 5.2). The present disclosure also provides polynucleotides encoding the presently disclosed anti-STING antibodies and antigen-binding fragments thereof, vectors comprising the presently disclosed polynucleotides, and cells comprising the presently disclosed polynucleotides and vectors (Section 5.3). The present disclosure further provides pharmaceutical compositions comprising the presently disclosed anti-STING antibodies and antigen-binding fragments thereof, polynucleotides, vectors, and cells (Section 5.4). The present disclosure further provides methods of making the anti-STING antibodies and antigen-binding fragments thereof (Section 5.5). The present disclosure also provides methods of using the anti-STING antibodies and antigen-binding fragments thereof for detecting STING in biological samples and for treating diseases and disorders (Section 5.5).

[0122] Since the exemplary anti-STING antibody of the present disclosure has beenshown to reduce the level of IFN- in differentiated human THP-1 cells stimulated withcGAMP, wherein the ability to reduce the level of IFN- has been confirmed in in vivomouse studies, and has been shown to prevent phosphorylation of TBK1, similar to the well- established STING antagonist H-151, and since the antibody is of human origin, i.e., maturation of the original antibody in the human body, it is prudent to expect that antagonistic anti-STING antibodies are useful in therapy of autoimmune and (auto)inflammatory diseases. More specifically, the present disclosure provides the embodiments as characterized in the claims, disclosed in the description and illustrated in the Examples and Figures further below. 5.1 Definitions

[0123] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, the exemplary methods and materials are described below. All publications, patent applications, patents, and otherNAI-5002871589v139references mentioned herein are incorporated by reference in their entirety. The materials, methods, and examples are illustrative only and not intended to be limiting. Unless otherwise stated, a term as used herein is given the definition as provided in the Oxford Dictionary of Biochemistry and Molecular Biology, Oxford University Press, 1997, revised 2000 and reprinted 2003, ISBN 0198506732; Second edition published 2006, ISBN 0-19-852917-1 978-0-19852917-0.

[0124] For the avoidance of any doubt it is emphasized that the expressions “in some embodiments”, “in certain embodiments”, “in certain instances”, “in some instances”, “in a further embodiment”, “in certain embodiments” and the like are used and meant such that any of the embodiments described therein are to be read with a mind to combine each of the features of those embodiments and that the disclosure has to be treated in the same way as if the combination of the features of those embodiments would be spelled out in one embodiment. The same is true for any combination of embodiments and features of the appended claims and illustrated in the Examples, which are also intended to be combined with features from corresponding embodiments disclosed in the description, wherein only for the sake of consistency and conciseness the embodiments are characterized by dependencies while in fact each embodiment and combination of features, which could be construed due to the (multiple) dependencies must be seen to be literally disclosed and not considered as a selection among different choices.

[0125] The term “antagonist” refers to an agent that significantly inhibits (either partially or completely) the biological activity of a target molecule, in the present case STING. Unless specifically indicated otherwise, the terms (i) “antagonistic” and “antagonist” and (ii) “inhibitory” and “inhibitory” may be used interchangeably herein.

[0126] An “antagonistic anti-STING antibody” and “anti-STING antagonist antibody”, respectively, refers to an anti-STING antibody (including any STING binding antibody format, fragment and derivative thereof) that effects decreased STING signalling. In certain embodiments, decreasing STING signalling comprises reducing or preventingphosphorylation of TBK1 and / or reducing the level of human IFN- in differentiated humanTHP-1 derived macrophages, in particular in PMA-differentiated human THP-1-derived macrophages, which have been stimulated with the STING agonist cGAMP as illustrated in the appended Examples.

[0127] While the present disclosure may be described for the sake of conciseness by way of reference to an antibody or antibodies, unless stated otherwise synthetic and biotechnological derivatives thereof as well as equivalent STING binding molecules areNAI-5002871589v140meant and included within the meaning of the term “antibody”. Accordingly, the term “antibody” as referred to herein includes whole antibodies and any antigen binding fragment (i.e., “antigen-binding portion”) or single chain versions thereof including synthetic derivatives, or biotechnological derivative thereof. An “antibody” refers, in certain embodiments, to a glycoprotein comprising at least two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds, or an antigen binding portion thereof. Each heavy chain is comprised of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region (CH) is comprised of three domains, CH1, CH2 and CH3. Each light chain is comprised of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region is comprised of one domain, CL. The VH and VL regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDRs), interspersed with regions that are more conserved, termed framework regions (FR). Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy- terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen. The constant regions of the antibodies may mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and various complement proteins.

[0128] “Antibody” as used herein also includes antibody fragments (and / or polypeptides that comprise antibody fragments) that are derived from the full-length antibody and retain antigen (e.g., STING) binding characteristics of the antibody from which the respective fragment was derived. Non-limiting examples of antibody fragments include Fab, Fab’, F(ab’)2, Fv, scFv, (scFv)2, single chain antibody molecule, dual variable domain antibody, single variable domain, linear antibody, V region, multispecific antibodies formed from antibody fragments, F(ab)2, Fd, Fc, diabody, disulfide-linked Fvs (dsFv), and single- domain antibody (e.g., nanobody). As used herein, the term “anti-STING antibody” also includes antigen-binding (i.e., STING-binding) fragments of the full-length anti-STING antibody.

[0129] The exact boundaries of CDRs can be defined differently according to different methods. In some embodiments, the positions of the CDRs or framework regions within a light or heavy chain variable domain can be as defined by Kabat et al., NIH Publication 91 (1991), 3242. In such cases, the CDRs can be referred to as “Kabat CDRs” (e.g., “Kabat LCDR2” or “Kabat HCDR1”). In certain embodiments, the positions of theNAI-5002871589v141CDRs of a light or heavy chain variable region can be as defined by Chothia et al., Nature 342 (1989), 877- 883). Accordingly, these regions can be referred to as “Chothia CDRs” (e.g., “Chothia LCDR2” or “Chothia HCDR3”). In certain embodiments, the positions of the CDRs of the light and heavy chain variable regions can be as defined by a Kabat-Chothia combined definition. In such embodiments, these regions can be referred to as “combined Kabat-Chothia CDRs”. Thomas et al., Mol Immunol 33 (1996), 1389-1401 exemplifies the identification of CDR boundaries according to Kabat and Chothia definitions. In certain embodiments, the positions of the CDRs or framework regions within a light or heavy chain variable domain can be as defined by the International Immunogenetics database (IMGT) standard. Marie-Paule Lefranc et al., Developmental & Comparative Immunology 27 (2003), 55-77 exemplify the identification of and CDR boundaries according to IMGT standard. Accordingly, these regions can be referred to as “IMGT CDRs” (e.g., “IMGT-LCDR2” or “IMGT-HCDR3”).

[0130] As the location of the CDRs within the structure of the immunoglobulin variable domain is conserved between species and present in structures called loops, by using numbering systems that align variable domain sequences according to structural features, CDR and framework residues and are readily identified by one skilled in the art. Various systems known in the art or disclosed herein represent different ways of delineating CDRs, and when they are used to define the same antibody, they are considered equivalent. The residues from each of these CDRs are exemplified in the table below. Exemplary CDRs According to Various Numbering Systems IMGT Kabat AbM Chothia Contact VH-CDR1 27-38 31-35 26-35 26-32 30-35 VH-CDR2 56-65 50-65 50-58 53-55 47-58 VH-CDR3 105-117 95-102 95-102 96-101 93-101 VL-CDR1 27-38 24-34 24-34 26-32 30-36 VL-CDR2 56-65 50-56 50-56 50-52 46-55 VL-CDR3 105-117 89-97 89-97 91-96 89-96

[0131] The specific CDR sequences defined in the anti-STING antibodies herein (e.g., Ab1, Ab2, Ab3) are based on Kabat definition. However, it will be understood that reference to a heavy chain variable region CDR or CDRs and / or a light chain variable region CDR or CDRs of a specific antibody will encompass all CDR definitions as known to those of skill in the art. For example, depending on the position of the first CDR the following CDRs might be shifted in either direction. Accordingly, in case of any inadvertent errors or inconsistencies regarding indication of CDRs in FIGS. 1A-1C and / or the sequence listing, theNAI-5002871589v142person skilled in the art on the basis of the disclosure content of the present application is well in the position to determine the correct CDR sequences in accordance with various definitions known in the art.

[0132] The term “antigen-binding fragment” of an antibody (or simply “antibody fragment”), as used herein, refers to one or more fragments or portions of an antibody that retain the ability to specifically bind to an antigen. Such “fragments” are, for example between about 8 and about 1500 amino acids in length, suitably between about 8 and about 745 amino acids in length, suitably about 8 to about 300, for example about 8 to about 200 amino acids, or about 10 to about 50 or 100 amino acids in length. It has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments encompassed within the term “antigen-binding fragment” of an antibody include (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CH1 domains; (ii) a F(ab’)2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) an Fd fragment consisting of the VH and CH1 domains; (iv) an Fv fragment consisting of the VL and VH domains of a single arm of an antibody, (v) an dAb fragment (Ward et al., Nature 341 (1989), 544-546), which consists of a VH domain; and (vi) an isolated complementarity determining region (CDR) or (vii) a combination of two or more isolated CDRs which may optionally be joined by a synthetic linker, for example, antibody fragments comprising variable heavy chain (VH) CDRs and / or variable light chain (VL) CDRs, e.g., VHCDR1-3 and / or VLCDR1-3. Furthermore, although the two domains of the Fv fragment, VL and VH, are coded for by separate genes, they can be joined, using recombinant methods, by a synthetic linker that enables them to be made as a single protein chain in which the VL and VH regions pair to form monovalent molecules (known as single chain Fv (scFv)); see e.g., Bird et al., Science 242 (1988), 423-426; and Huston et al. Proc. Natl. Acad. Sci. USA 85 (1988), 5879-5883. Such single chain antibodies are also intended to be encompassed within the term “antigen- binding fragment” of an antibody. These antibody fragments are obtained using conventional techniques known to those with skill in the art, and the fragments are screened for utility in the same manner as are intact antibodies. Antigen-binding portions can be produced by recombinant DNA techniques, or by enzymatic or chemical cleavage of intact immunoglobulins.

[0133] Furthermore, regarding STING protein and anti-STING antibodies, their recombinant production in a host cell, purification, modification, formulation in a pharmaceutical composition and therapeutic use as well as terms and feature common in theNAI-5002871589v143art can be relied upon by the person skilled in art when carrying out the present disclosure as claimed; see, e.g., Antibodies A Laboratory Manual 2ndedition, 2014 by Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA, wherein also antibody purification and storage; engineering antibodies, including use of degenerate oligonucleotides, 5’-RACE, phage display, and mutagenesis, immunoblotting protocols and the latest screening and labeling techniques are described.

[0134] As used herein, reference to “about” of a given value includes (and describes) embodiments that are directed to the given value and embodiments with 10% or less variation of the given value. 5.2 Anti-STING Antibodies and Antigen-Binding Fragments Thereof

[0135] The present disclosure provides novel anti-STING antibodies and antigen- binding fragments thereof that bind to certain epitopes of human STING. The present disclosure also provides novel anti-STING antibodies and antigen-binding fragments thereof that have certain antagonistic activity on STING signaling pathway. The present disclosure further provides novel anti-STING antibodies and antigen-binding fragments thereof comprising specific CDR and / or VH / VL sequences (e.g., sequences of Tables 1-3).

[0136] As shown by the experiments performed within the scope of the presentdisclosure, antibody Ab1 inhibits the release of IFN- in cGAMP stimulated mousesplenocytes in vitro and inhibits the release of IFN- in cGAMP stimulated differentiatedTHP-1 cells, incubation of cGAMP or CMA stimulated mouse splenocytes with antibody Ab1 leads to a decrease in phosphorylated TBK1 levels, and incubation of cGAMP stimulated differentiated THP-1 cells with antibodies Ab1, Ab2 and Ab3 leads to a decrease in phosphorylated TBK1 levels.

[0137] Thus, the anti-STING antibody of the present disclosure can inhibit the releaseof IFN- in human and mouse cells, in particular in cGAMP stimulated mouse splenocytesand in cGAMP stimulated differentiated THP-1 cells. In addition, or alternatively, the anti- STING antibody of the present disclosure can prevent phosphorylation of TBK1 in human and mouse cells, in particular in cGAMP / CMA stimulated mouse splenocytes and in cGAMP stimulated differentiated THP-1 cells.

[0138] In certain embodiments, the anti-STING antibody or antigen-binding thereof can inhibit the phosphorylation of TBK1 in cells with activated STING pathway. In certain embodiments, the inhibition is indicated by a reduced pTBK1 level in the cells with activated STING pathway and being contacted with the anti-STING antibody or antigen-binding fragment thereof as compared to cells with activated STING pathway but without beingNAI-5002871589v144contacted with the anti-STING antibody or antigen-binding fragment thereof. In certain embodiments, the level of pTBK1 is measured by an immunohistochemistry assay, for example, a Western Blot or an immunocytochemistry (ICC). In certain embodiments, the cells are immune cells. In certain embodiments, the cells are lymphoid cells. In certain embodiments, the cells are myeloid cells. In certain embodiments, the cells are differentiated THP-1 cells. In certain embodiments, the cells are splenocytes. In certain embodiments, the differentiated THP-1 cells are macrophages. In certain embodiments, the THP-1 cells are human THP-1 cells. In certain embodiments, the differentiated THP-1 cells are PMA- differentiated human THP-1-derived macrophages. In certain embodiments, the cells are peripheral blood mononuclear cells (PBMCs). In certain embodiments, the splenocytes are mouse splenocytes. In certain embodiments, the STING pathway is activated by CMA, cGAMP, or HT DNA. In certain embodiments, the anti-STING antibody or antigen-binding thereof can inhibit the phosphorylation of TBK1 in cells with activated STING pathway in vitro, ex vivo or in vivo.

[0139] In certain embodiments, the pTBK1 level in the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less of the pTBK1 level in the cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof, as measured by Western Blot or ICC, and the cells are PMA-differentiated human THP-1- derived macrophages or mouse splenocytes with STING pathway activated by cGAMP or CMA. In certain embodiments, the pTBK1 level in the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof is about 40% or less of the pTBK1 level in the cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof as measured by ICC, wherein the cells are mouse splenocytes with STING pathway activated by CMA. In certain embodiments, the pTBK1 level in the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof is about 50% or less of the pTBK1 level in the cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof as measured by ICC, wherein the cells are mouse splenocytes with STING pathway activated by cGAMP. In certain embodiments, the pTBK1 level in the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof is about 60% or less of the pTBK1 level in the cells with activated STING pathway but without being contacted with the antibody orNAI-5002871589v145antigen-binding fragment thereof as measured by Western Blot, wherein the cells are PMA- differentiated human THP-1-derived macrophages with STING pathway activated by cGAMP. In certain embodiments, the pTBK1 level in the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof is about 40% or less or 60% or less of the pTBK1 level in the cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof as measured by Western Blot, wherein the cells are PBMCs with STING pathway activated by cGAMP.

[0140] In certain embodiments, the anti-STING antibody or antigen-binding thereof can inhibit the release of an inflammatory cytokine in cells with activated STING pathway. In certain embodiments, the anti-STING antibody or antigen-binding thereof can inhibit the release of an inflammatory cytokine in cells of a Trex1 null mouse. In certain embodiments, the anti-STING antibody or antigen-binding thereof can reduce the level of an inflammatory cytokine of an NZB / W mouse. In certain embodiments, the inhibition of the release of the inflammatory cytokine is indicated by a reduced level of the inflammatory cytokine released by the cells with activated STING pathway or cells of the Trex1 null mouse and being contacted or administered with the antibody or antigen-binding fragment thereof as compared to cells with activated STING pathway or cells of a Trex1 null mouse but without being contacted or administered with the antibody or antigen-binding fragment thereof. In certain embodiments, the reduction of the level of the inflammatory cytokine is indicated by the reduced level of the inflammatory cytokine in serum of the NZB / W mouse being administered with the antibody or antigen-binding fragment thereof as compared to a NZB / W mouse but without being administered with the antibody or antigen-binding fragment thereof. In certain embodiments, the level of the inflammatory cytokine is measured by an immunoassay, for example, ELISA, HTRF, or MSD multiple assay. In certain embodiments, the level of the inflammatory cytokine is measured by RT-qPCR. In certain embodiments,the inflammatory cytokine is IFN- . In certain embodiments, the inflammatory cytokine isIL-6. In certain embodiments, the inflammatory cytokine is ISG-15. In certain embodiments, the inflammatory cytokine is IP-10. In certain embodiments, the cells are immune cells. In certain embodiments, the cells are lymphoid cells. In certain embodiments, the cells are myeloid cells. In certain embodiments, the cells are differentiated THP-1 cells. In certain embodiments, the cells are splenocytes. In certain embodiments, the differentiated THP-1 cells are macrophages. In certain embodiments, the THP-1 cells are human THP-1 cells. In certain embodiments, the differentiated THP-1 cells are PMA-differentiated human THP-1-derived macrophages. In certain embodiments, the splenocytes are mouseNAI-5002871589v146splenocytes. In certain embodiments, the cells of the Trex1 null mouse are tissue cells (e.g., heart tissue cells). In certain embodiments, the STING pathway is activated by CMA, cGAMP, or HT DNA. In certain embodiments, the anti-STING antibody or antigen-binding thereof can inhibit the release of an inflammatory cytokine in cells with activated STING pathway in vitro, ex vivo or in vivo.

[0141] In certain embodiments, the level of the inflammatory cytokine released by the cells with activated STING pathway and being contacted with the antibody or antigen- binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less of cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof, as measured by ELISA or HTRF, and the cells are PMA-differentiated human THP-1-derived macrophages or mouse splenocytes with STING pathway activated bycGAMP, and the inflammatory cytokine is IFN- . In certain embodiments, the IFN- levelreleased by the cells with activated STING pathway and being contacted with the antibody orantigen-binding fragment thereof is about 20% or less of the IFN- level released by the cellswith activated STING pathway but without being contacted with the antibody or antigen- binding fragment thereof as measured by ELISA, wherein the cells are mouse splenocyteswith STING pathway activated by cGAMP. In certain embodiments, the IFN- levelreleased by the cells with activated STING pathway and being contacted with the antibody orantigen-binding fragment thereof is about 50% or less or about 80% or less of the IFN- levelreleased by the cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof as measured by HTRF, wherein the cells are PMA-differentiated human THP-1-derived macrophages with STING pathway activated by cGAMP.

[0142] In certain embodiments, the level of the inflammatory cytokine released by the cells with activated STING pathway and being contacted with the antibody or antigen- binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less of cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof, as measured by MSD multiplex assay, and the cells are PMA-differentiated human THP-1-derived macrophages with STING pathway activated by HT DNA, and theinflammatory cytokine is IFN- , IL-6, and / or IP-10.

[0143] In certain embodiments, the IFN- mRNA level in a tissue sample (e.g., hearttissue) of a Trex1 null mouse being administered with the antibody or antigen-bindingNAI-5002871589v147fragment thereof is about 20% or less or about 80% or less of the IFN- mRNA level in atissue sample (e.g., heart tissue) of a Trex1 null mouse but without being administered with the antibody or antigen-binding fragment thereof as measured by RT-qPCR. In certain embodiments, the IL-6 mRNA level in a tissue sample (e.g., heart tissue) of a Trex1 null mouse being administered with the antibody or antigen-binding fragment thereof is about 60% or less or about 80% or less of the IL-6 mRNA level in a tissue sample (e.g., heart tissue) of a Trex1 null mouse but without being administered with the antibody or antigen- binding fragment thereof as measured by RT-qPCR. In certain embodiments, the ISG-15 mRNA level in a tissue sample (e.g., heart tissue) of a Trex1 null mouse being administered with the antibody or antigen-binding fragment thereof is about 80% or less of the ISG-15 mRNA level in a tissue sample (e.g., heart tissue) of a Trex1 null mouse without being administered with the antibody or antigen-binding fragment thereof as measured by RT- qPCR.

[0144] In certain embodiments, the level of the inflammatory cytokine in serum of the NZB / W mouse administered with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, about 80% or less, or about 90% or less of serum from a NZB / W mouse without being administered with the antibody or antigen-binding fragment thereof. In certain embodiments, the inflammatory cytokine is measured by MSD multiplex assay. In certain embodiments, the inflammatory cytokine is IL-6.

[0145] The antibody or antigen-binding fragment thereof of the present disclosure can be further characterized by binding to the C-terminal domain of the STING protein and to an epitope located within said C-terminal domain of the STING protein, respectively. In addition, antibody Ab1 has been shown to bind to a C-terminus short peptide (see Example 2) and antibodies Ab2 and Ab3 each bind to an epitope comprising the amino acid residues 300 to 308 of human STING and to an epitope comprising the amino acid residues 245-255 of human STING, respectively.

[0146] In certain embodiments, the anti-STING antibody or antigen-binding thereof can specifically bind to the C-terminal domain of the STING protein. In certain embodiments, the C-terminal domain comprises the amino acid sequence of SEQ ID NO:31 (setting forth the amino acid sequence of C-terminus human STING peptide). In certain embodiments, the anti-STING antibody or antigen-binding thereof can specifically bind to an epitope within the C-terminal domain of the STING protein. In certain embodiments, the anti-STING antibody or antigen-binding thereof can specifically bind to an epitopeNAI-5002871589v148comprising amino acid residues S358, G367, M368, E369, and P371 of human STING protein, where the amino acid positions are numbered according to the positions in the amino acid sequence of SEQ ID NO:34 (setting forth the amino acid sequence of human STING protein isoform 1 (NCBI Reference No. NP_938023.1)). In certain embodiments, the epitope is a conformational epitope. In certain embodiments, the epitope is a linear epitope.

[0147] The present disclosure also provides an immunogen comprising an epitope within the C-terminal domain of the human STING protein. In certain embodiments, the immunogen comprises the amino acid sequence of SEQ ID NO:31. In certain embodiments, the immunogen comprises an epitope of the human STING protein. In certain embodiments, the immunogen comprises a stabilized epitope, wherein the epitope comprises or consisting of amino acid residues S358, G367, M368, E369, and P371 of human STING, wherein the amino acid positions are numbered according to the positions in the amino acid sequence of SEQ ID NO:34. In certain embodiments, the epitope is a conformational epitope. In certain embodiments, the epitope is a linear epitope.

[0148] In certain embodiments, the anti-STING antibody or antigen-binding thereof can specifically bind an epitope comprising the amino acid residues 300 to 308 of human STING protein. In certain embodiments, the anti-STING antibody or antigen-binding thereof can specifically bind an epitope comprising the amino acid residues 245-255 of human STING protein. In certain embodiments, the amino acid positions are numbered according to the positions in the amino acid sequence of SEQ ID NO:34.

[0149] As illustrated in the Examples and in the Figures, the antibodies of the present disclosure have been originally isolated from human donors and are shown to specifically recognize human STING. Antibody Ab1 has been also shown to recognize mouse STING, which makes it particularly suitable for preclinical mouse studies. By “specifically recognizing STING”, “binding STING”, “antibody specific to / for STING” and “anti-STING antibody” is meant specifically, generally, and collectively, antibodies to the native form of STING or peptides derived from STING, in particular short peptides of the C-terminus of STING. Provided herein are antibodies selective for STING protein. Binding specificity of an anti-STING antibody may be measured, for example, by determination of binding affinity to recombinant full-length STING or to STING peptides as for examples measured by the assay in Example 2 herein, or by equivalent assays.

[0150] In this context, in order to obtain a measure of the binding affinity, the EC50of the antibodies of the invention in the ELISA performed in Example 2 was determined. As demonstrated, the antibodies of the present disclosure display a particularly high apparentNAI-5002871589v149binding affinity as determined by the EC50 value. The term “EC50”, in the context of an in vitro, ex vivo or in vivo assay using an antibody or an antigen-binding fragment thereof, refers to the concentration of an antibody or an antigen-binding fragment thereof that induces a response that is 50% of the maximal response, i.e., halfway between the maximal response and the baseline.

[0151] In certain embodiments, an anti-STING antibody or an antigen-binding fragment thereof disclosed herein has an EC50of about 1 nM or less in binding to human STING as measured by ELISA. In certain embodiments, an anti-STING antibody or an antigen-binding fragment thereof disclosed herein has an EC50of about 900 nM or less, or about 850 nM or less in binding to human STING as measured by ELISA. In certain embodiments, an anti-STING antibody or an antigen-binding fragment thereof disclosed herein has an EC50 of about 20 nM or less, about 18 nM or less, or about 15 nM or less in binding to human STING as measured by ELISA. In certain embodiments, an anti-STING antibody or an antigen-binding fragment thereof disclosed herein has an EC50 of about 900 pM in binding to human STING as measured by ELISA. In certain embodiments, an anti- STING antibody or an antigen-binding fragment thereof disclosed herein has an EC50of about 850 pM in binding to human STING as measured by ELISA. In certain embodiments, an anti-STING antibody or an antigen-binding fragment thereof disclosed herein has an EC50 of 844 pM in binding to human STING as measured by ELISA. In certain embodiments, an anti-STING antibody or an antigen-binding fragment thereof disclosed herein has an EC50 of about 18 nM or about 15 nM in binding to human STING as measured by ELISA.

[0152] In certain embodiments, an anti-STING antibody or an antigen-binding fragment thereof disclosed herein has an EC50of about 600 pM or less, about 500 pM or less, or about 475 pM or less in binding to mouse STING as measured by ELISA. In certain embodiments, an anti-STING antibody or an antigen-binding fragment thereof disclosed herein has an EC50of about 475 pM in binding to mouse STING as measured by ELISA.

[0153] In certain embodiments, an anti-STING antibody or an antigen-binding fragment thereof disclosed herein has an EC50 of about 300 pM or less, about 280 pM or less, or about 260 pM or less in binding to a C-terminus human STING measured by ELISA. In certain embodiments, an anti-STING antibody or an antigen-binding fragment thereof disclosed herein has an EC50 of about 260 pM in binding to a C-terminus human STING measured by ELISA. In certain embodiments, an anti-STING antibody or an antigen-binding fragment thereof disclosed herein has an EC50 of 257 pM in binding to a C-terminus humanNAI-5002871589v150STING measured by ELISA. In certain embodiments, the C-terminus human STING consists of the amino acid sequence of SEQ ID NO:31.

[0154] In certain embodiments, an anti-STING antibody or an antigen-binding fragment thereof disclosed herein has an EC50 of about 300 pM or less, about 280 pM or less, or about 250 pM or less in binding to a C-terminus cynomolgus monkey STING measured by ELISA. In certain embodiments, an anti-STING antibody or an antigen-binding fragment thereof disclosed herein has an EC50of about 250 pM in binding to a C-terminus cynomolgus monkey STING measured by ELISA. In certain embodiments, an anti-STING antibody or an antigen-binding fragment thereof disclosed herein has an EC50of 233 pM in binding to a C- terminus cynomolgus monkey STING measured by ELISA. In certain embodiments, the C- terminus cynomolgus monkey STING consists of the amino acid sequence of SEQ ID NO:32.

[0155] In certain embodiments, an anti-STING antibody or an antigen-binding fragment thereof disclosed herein has an EC50of about 700 pM or less, about 680 pM or less, or about 650 pM or less in binding to a C-terminus rat STING measured by ELISA. In certain embodiments, an anti-STING antibody or an antigen-binding fragment thereof disclosed herein has an EC50of about 650 pM in binding to a C-terminus rat STING measured by ELISA. In certain embodiments, an anti-STING antibody or an antigen-binding fragment thereof disclosed herein has an EC50 of 648 pM in binding to a C-terminus rat STING measured by ELISA. In certain embodiments, the C-terminus rat STING consists of the amino acid sequence of SEQ ID NO:33.

[0156] However, also depending on the antibody format, for example whether an IgG1, IgG4 or antibody fragments are used, like Fab fragments, the EC50values may deviate and may be for example higher than the values mentioned above and in the Examples. Accordingly, in this context, the term “about” means within one order of magnitude and / or within the same order of magnitude variation of a given value, for example, the EC50may be ±10 nM of a given value.

[0157] In certain embodiments, the anti-STING antibody of the present disclosure recognizes human STING in various environments, for example it binds to STING in human cells (see Example 4) and human tissue (see Examples 5 and 6), which can be determined by immunocytochemistry and immunofluorescence.

[0158] In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof has at least one of: (i) the binding specificity disclosed herein (e.g., specifically binding to an epitope comprising amino acid residues S358, G367, M368, E369, and P371 of human STING protein); (ii) the antagonistic properties disclosed herein (e.g., inhibitoryNAI-5002871589v151effects on the phosphorylation of TBK1 and / or the release of of IFN- in cells with activatedSTING pathway); and (iii) the binding affinity disclosed herein (e.g., an EC50 for human STING of about 1 nM as measured by ELISA). In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof further comprises a VH comprising VH-CDRs 1-3, and a VL chain comprising VL-CDRs 1-3 as disclosed herein (e.g., CDRs disclosed in Tables 1-3).

[0159] The present disclosure also provides anti-STING antibodies or an antigen- binding fragments thereof that competes with a presently disclosed anti-STING antibody or antigen-binding fragment thereof for specifically binding to an epitope located within the C- terminal domain of STING. In certain embodiments, the epitope comprises amino acid residues S358, G367, M368, E369, and P371 of human STING protein, where the amino acid positions are numbered according to the positions in the amino acid sequence of SEQ ID NO:34. In certain embodiments, the epitope is a conformational epitope. In certain embodiments, the epitope is a linear epitope.

[0160] In certain embodiments, the competing antibody or antigen-binding fragment thereof has (i) the antagonistic properties disclosed herein (e.g., inhibitory effects on thephosphorylation of TBK1 and / or the release of of IFN- in cells with activated STINGpathway); and / or (ii) the binding affinity disclosed herein (e.g., an EC50 for human STING of about 1 nM as measured by ELISA).

[0161] The present disclosure is illustrated with anti-STING antibodies which are characterized by comprising in their variable region, i.e., binding domain, the variable heavy (VH) and variable light (VL) chain having the amino acid sequences depicted in FIGS. 1A-1C, respectively. The corresponding nucleotide and amino acid sequences are set forth in Tables 1-3 below.

[0162] As always, the variable domains of each chain contain three hypervariable loops named complementarity determining regions (CDRs, CDR-1,-2, and -3). The CDRs are separated by structurally conserved regions called framework regions (FR-1,-2,-3, and -4) that form a “core” ß-sheet structure displaying these loops on the surface of the variable domain. The length and composition of the CDR sequences are highly variable, especially in the CDR3. The CDRs are approximated to the paratope of the antibody that interacts with the antigen and therefore contains the antigen-binding residues. Accordingly, it is common to define an antibody by its six CDRs. Exemplary sets of CDRs in the above amino acid sequences of the VH and VL chains are depicted in FIGS. 1A-1C. However, as discussed in the following the person skilled in the art is well aware of the fact that in addition orNAI-5002871589v152alternatively CDRs may be used, which differ in their amino acid sequence from those set forth in any one of FIGS. 1A-1C by one, two, three or even more amino acids, especially in case of CDR2 and CDR3. As mentioned in the Figure legend of FIGS. 1A-1C, the person skilled in the art can easily identify the CDRs according to common principles, for example as summarized in www.bioinf.org.uk / abs. In this context, while the CDRs of the antibodies depicted in FIGS. 1A-1C are indicated according to Kabat et al. the person skilled in the art knows that a number of definitions of the CDRs are commonly in use, i.e., the (i) Kabat definition based on sequence variability, which is the most commonly used; (ii) Chothia definition based on the location of the structural loop regions; (iii)AbM definition as a compromise between the two used by Oxford Molecular’s AbM antibody modelling software; and (iv) Contact definition that has been recently introduced by and is based on an analysis of the available complex crystal structures. This definition is likely to be the most useful for performing mutagenesis to modify the affinity of an antibody since these are residues which take part in interactions with the antigen. For lists of CDR residues making contact in each antibody with summary data for each CDR see, e.g., www.bioinf.org.uk / abs which also refers to antibody modelling software such as abYmod available at abymod.abysis.org.

[0163] In certain embodiments, the present disclosure provides an anti-STING antibody or an antigen-binding fragment thereof, a synthetic derivative, or a biotechnological derivative thereof, wherein the anti-STING antibody, antigen-binding fragment thereof, or derivative thereof comprises a variable heavy (VH) chain comprising VH complementarity determining regions (CDRs) 1, 2, and 3, and a variable light (VL) chain comprising VL CDRs 1, 2, and 3 as defined by Kabat, wherein: (i) (a) the VH-CDR1 comprises the amino acid sequence of SEQ ID NO:3 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions, preferably one or two amino acid substitutions and / or deletion, more preferably one or two amino acid substitutions, (b) the VH-CDR2 comprises the amino acid sequence of SEQ ID NO:4 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions, preferably one or two amino acid substitutions and / or deletion, more preferably one or two amino acid substitutions,NAI-5002871589v153(c) the VH-CDR3 comprises the amino acid sequence of SEQ ID NO:5 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions, preferably one or two amino acid substitutions and / or deletion, more preferably one or two amino acid substitutions, (d) the VL-CDR1 comprises the amino acid sequence of SEQ ID NO:8 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions, preferably one or two amino acid substitutions and / or deletion, more preferably one or two amino acid substitutions, (e) the VL-CDR2 comprises the amino acid sequence of SEQ ID NO:9 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions, preferably one or two amino acid substitutions and / or deletion, more preferably one or two amino acid substitutions, and (f) the VL-CDR3 comprises the amino acid sequence of SEQ ID NO:10 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions, preferably one or two amino acid substitutions and / or deletion, more preferably one or two amino acid substitutions; or (g) the VH-CDR1 comprises the amino acid sequence of SEQ ID NO:13 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions, preferably one or two amino acid substitutions and / or deletion, more preferably one or two amino acid substitutions, (h) the VH-CDR2 comprises the amino acid sequence of SEQ ID NO:14 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions, preferably one or two amino acid substitutions and / or deletion, more preferably one or two amino acid substitutions, (i) the VH-CDR3 comprises the amino acid sequence of SEQ ID NO:15 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions, preferably one or two amino acid substitutions and / or deletion, more preferably one or two amino acid substitutions, (j) the VL-CDR1 comprises the amino acid sequence of SEQ ID NO:18 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions, preferably one or two amino acid substitutions and / or deletion, more preferably one or two amino acid substitutions, (k) the VL-CDR2 comprises the amino acid sequence of SEQ ID NO:19 or a variant thereof, wherein the variant comprises one or more amino acidNAI-5002871589v154substitutions and / or deletions, preferably one or two amino acid substitutions and / or deletion, more preferably one or two amino acid substitutions, and (l) the VL-CDR3 comprises the amino acid sequence of SEQ ID NO:20 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions, preferably one or two amino acid substitutions and / or deletion, more preferably one or two amino acid substitutions; or (m) the VH-CDR1 comprises the amino acid sequence of SEQ ID NO:23 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions, preferably one or two amino acid substitutions and / or deletion, more preferably one or two amino acid substitutions, (n) the VH-CDR2 comprises the amino acid sequence of SEQ ID NO:24 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions, preferably one or two amino acid substitutions and / or deletion, more preferably one or two amino acid substitutions, (o) the VH-CDR3 comprises the amino acid sequence of SEQ ID NO:25 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions, preferably one or two amino acid substitutions and / or deletion, more preferably one or two amino acid substitutions, (p) the VL-CDR1 comprises the amino acid sequence of SEQ ID NO:28 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions, preferably one or two amino acid substitutions and / or deletion, more preferably one or two amino acid substitutions, (q) the VL-CDR2 comprises the amino acid sequence of SEQ ID NO:29 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions, preferably one or two amino acid substitutions and / or deletion, more preferably one or two amino acid substitutions, and (r) the VL-CDR3 comprises the amino acid sequence of SEQ ID NO:30 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions, preferably one or two amino acid substitutions and / or deletion, more preferably one or two amino acid substitutions.

[0164] In certain embodiments, the anti-STING antibody, antigen-binding fragment thereof, or derivative thereof comprises:NAI-5002871589v155(i) the VH chain comprising the amino acid sequence depicted in SEQ ID NO:2 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions; and / or (ii) the VL comprising the amino acid sequence depicted in SEQ ID NO:7, or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions; or (iii) the VH comprising the amino acid sequence depicted in SEQ ID NO:12 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions; and / or (iv) the VL comprising the amino acid sequence depicted in SEQ ID NO:17, or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions; or (v) the VH comprising the amino acid sequence depicted in SEQ ID NO:22 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions; and / or (vi) the VL comprising the amino acid sequence depicted in SEQ ID NO:27, or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions.

[0165] In certain embodiments, the VH and VL chain amino acid sequences are at least 90% identical to SEQ ID NO:2 and 7, respectively, to SEQ ID NO:12 and 17, respectively or SEQ ID NO:22 and 27, respectively.

[0166] In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises six CDRs of an antibody designated as Ab1, Ab2, or Ab3 based on the Kabat definition as set forth in Tables 1-3 and FIGS. 1A-1C.NAI-5002871589v156Table 1: Antibody Ab1 VH-CDR1 VH-CDR2 VH-CDR3 SYWMS SIKQDGSQKPYVDSVKG VGYPHAFDI (SEQ ID NO:3) (SEQ ID NO:4) (SEQ ID NO:5) Heavy chain variable region (VH) EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMSWVRQAPGKGLEWVASIKQDGSQKPY VDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARVGYPHAFDIWGQGTMVTVSS (SEQ ID NO:2) Exemplary nucleotide sequence encoding VH GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTC TCATGTGCAGCCTCTGGATTCACCTTTAGTAGTTATTGGATGAGCTGGGTCCGCCAGGCT CCAGGGAAGGGACTGGAGTGGGTGGCCAGCATAAAGCAAGATGGAAGTCAGAAACCCTAT GTGGACTCTGTGAAGGGGCGATTCACCATCTCCAGAGACAACGCCAAGAACTCACTGTAT CTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGTCGGC TACCCCCATGCTTTTGATATCTGGGGCCAGGGGACAATGGTCACCGTCTCCTCA (SEQ ID NO:1) Exemplary heavy chain constant region (CH) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGG PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO:129) VL-CDR1 VL-CDR2 VL-CDR3 TGTSSDIGNYNVVS EVTKRPS FSYAGSSTWV (SEQ ID NO:8) (SEQ ID NO:9) (SEQ ID NO:10) Light chain variable region (VL) QSALTQPASVSGSPGQSITISCTGTSSDIGNYNVVSWYQHHPGKAPKLMIYEVTKRPSGV SSRFSGSKSGDTASLTISGLQAEDEADYYCFSYAGSSTWVFGGGTKLTVL (SEQ ID NO:7) Exemplary nucleotide sequence encoding VL CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATC TCCTGCACTGGAACCAGCAGTGATATTGGGAATTATAATGTTGTCTCCTGGTACCAACAC CACCCAGGCAAAGCCCCCAAGCTCATGATTTATGAGGTCACTAAGCGGCCCTCAGGGGTA TCTAGTCGCTTCTCTGGCTCCAAGTCTGGCGACACGGCCTCCCTGACAATCTCTGGGCTC CAGGCCGAGGACGAGGCTGATTATTACTGCTTCTCATATGCAGGTAGTAGCACTTGGGTG TTCGGCGGAGGGACCAAGCTGACCGTCCTA (SEQ ID NO:6) Exemplary light chain constant region (CL) GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSK QSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:130) - CDR sequences are determined based on Kabat definition - Underlined, nucleotides or amino acids indicate the CDR coding regions in the variable chain sequenceNAI-5002871589v157Table 2: Antibody Ab2 VH-CDR1 VH-CDR2 VH-CDR3 NYGMH VTRYDGTNKKYGDSVKG GDLWSAYYYSGMDV (SEQ ID NO:13) (SEQ ID NO:14) (SEQ ID NO:15) Heavy chain variable region (VH) EVQLVESGGGVVQPGGSLRLSCAASGFTFSNYGMHWVRQAPGKGLEWVAVTRYDGTNKKY GDSVKGRFTISRDNSKNTLYLQMNSLRPEDTAVYYCAKGDLWSAYYYSGMDVWGQGTTVT VSS (SEQ ID NO:12) Exemplary nucleotide sequence encoding VH GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTAGTCCAGCCTGGGGGGTCCCTGAGACTC TCCTGTGCAGCGTCTGGATTCACGTTCAGCAATTATGGCATGCACTGGGTCCGCCAGGCT CCAGGCAAGGGGCTGGAGTGGGTGGCAGTTACACGGTATGATGGAACTAATAAAAAATAT GGAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAACTCCAAGAACACGCTGTAT CTGCAAATGAACAGCCTGAGACCTGAGGACACGGCCGTGTATTACTGTGCGAAAGGGGAT CTTTGGAGTGCATACTACTACTCCGGAATGGACGTCTGGGGCCAAGGGACCACGGTCACC GTCTCCTCA (SEQ ID NO:11) Exemplary heavy chain constant region (CH) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGG PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO:129) VL-CDR1 VL-CDR2 VL-CDR3 RASQSISTYLN AASSLQS QQSYSTPFT (SEQ ID NO:18) (SEQ ID NO:19) (SEQ ID NO:20) Light chain variable region (VL) DIQMTQSPSSLSASVGDRVTITCRASQSISTYLNWYQQTPGKAPKLLIYAASSLQSGVPS RFSGSGSGTDFTLTISSLQAEDFATYYCQQSYSTPFTFGPGTKVEIK (SEQ ID NO:17) Exemplary nucleotide sequence encoding VL GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTCGGAGACAGAGTCACC ATCACTTGCCGGGCAAGTCAGAGCATTAGCACCTATTTAAATTGGTATCAACAGACGCCC GGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCA AGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAAGCT GAAGATTTTGCAACTTATTACTGTCAACAGAGTTACAGTACCCCATTCACTTTCGGCCCT GGGACCAAGGTGGAGATCAAA (SEQ ID NO:16) Exemplary light chain constant region (CL) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQD SKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO:131) - CDR sequences are determined based on Kabat definition - Underlined, nucleotides or amino acids indicate the CDR coding regions in the variable chain sequenceNAI-5002871589v158Table 3: Antibody Ab3 VH-CDR1 VH-CDR2 VH-CDR3 NYWIS RIDPTDSYTNYSPSFQG QPRNGDLTFDY (SEQ ID NO:23) (SEQ ID NO:24) (SEQ ID NO:25) Heavy chain variable region (VH) QVQLVQSGAEVKKPGESLRISCQGSGYSFSNYWISWVRQMPGKGLEWMGRIDPTDSYTNY SPSFQGHVTISVDKSISTASLQWNSLKASDTAMYYCARQPRNGDLTFDYWGQGSLVTVSS (SEQ ID NO:22) Exemplary nucleotide sequence encoding VH CAGGTGCAGCTGGTGCAGTCCGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATC TCCTGTCAGGGTTCTGGATACAGCTTTAGCAACTACTGGATCAGCTGGGTGCGCCAGATG CCCGGGAAAGGCCTGGAGTGGATGGGGAGGATTGATCCTACTGACTCTTATACCAACTAC AGCCCGTCCTTCCAAGGCCACGTCACCATCTCAGTTGACAAGTCCATCAGCACTGCCTCC CTGCAGTGGAACAGCCTGAAGGCCTCGGACACCGCCATGTATTACTGTGCGAGACAGCCC CGGAACGGTGACTTAACCTTTGACTATTGGGGCCAGGGATCCCTGGTCACCGTCTCCTCA (SEQ ID NO:21) Exemplary heavy chain constant region (CH) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGG PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO:129) VL-CDR1 VL-CDR2 VL-CDR3 SGSSSNIGYNAVS DNYKRPP GTWDSSLSAEV (SEQ ID NO:28) (SEQ ID NO:29) (SEQ ID NO:30) Light chain variable region (VL) QAVVTQPPSVSAAPGQKVTISCSGSSSNIGYNAVSWYQQFPGTAPKLLINDNYKRPPGIP DRFSGSKSGTSATLGITGLQTGDEADYYCGTWDSSLSAEVFGGGTKLTVL (SEQ ID NO:27) Exemplary nucleotide sequence encoding VL CAGGCTGTGGTGACTCAGCCGCCCTCAGTGTCTGCGGCCCCAGGACAGAAGGTCACCATC TCCTGCTCTGGAAGCAGCTCCAACATTGGATATAATGCTGTATCCTGGTACCAGCAGTTC CCAGGAACAGCCCCCAAACTCCTCATTAATGACAATTATAAGCGACCCCCAGGGATTCCT GACCGATTCTCTGGCTCCAAGTCTGGCACGTCAGCCACCCTGGGCATCACCGGACTCCAG ACTGGGGACGAGGCCGATTATTACTGCGGAACATGGGATAGCAGCCTGAGTGCTGAGGTA TTCGGCGGAGGGACCAAGCTGACCGTCCTA (SEQ ID NO:26) Exemplary light chain constant region (CL) GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSK QSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:130) - CDR sequences are determined based on Kabat definition - Underlined, nucleotides or amino acids indicate the CDR coding regions in the variable chain sequenceNAI-5002871589v159

[0167] In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH-CDR1, a VH-CDR2, and a VH-CDR3 from antibody Ab1 (Table 1); or (b) a VL-CDR1, a VL-CDR2, and a VL-CDR3 from antibody Ab1 (Table 1). In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH-CDR1, a VH-CDR2, and a VH-CDR3 from antibody Ab1 (Table 1); and (b) a VL-CDR1, a VL-CDR2, and a VL-CDR3 from antibody Ab1 (Table 1).

[0168] In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH-CDR1, a VH-CDR2, and a VH-CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO:2; or (b) a VL-CDR1, a VL-CDR2, and a VL-CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO:7. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH-CDR1, a VH-CDR2, and a VH-CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO:2; and (b) a VL-CDR1, a VL-CDR2, and a VL- CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO:7.

[0169] In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO:3, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO:4, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO:5; or (b) a VL-CDR1 comprising the amino acid sequence of SEQ ID NO:8, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO:9, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO:10. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO:3, a VH- CDR2 comprising the amino acid sequence of SEQ ID NO:4, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO:5; and (b) a VL-CDR1 comprising the amino acid sequence of SEQ ID NO:8, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO:9, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO:10.

[0170] In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH comprising a VH-CDR1, a VH-CDR2, and a VH-CDR3 disclosed herein, or (b) a VL comprising a VL-CDR1, a VL-CDR2, and a VL-CDR3 disclosed herein. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH comprising a VH-CDR1, a VH-CDR2, and a VH-CDR3 disclosed herein, and (b) a VL comprising a VL-CDR1, a VL-CDR2, and a VL-CDR3 disclosed herein. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises a VH at least 75%, at least 80%, at least 85%, at least 90%, atNAI-5002871589v160least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO:2; and / or a VL at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO:7. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:2, or a VL comprising the amino acid sequence of SEQ ID NO:7. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:2, and a VL comprising the amino acid sequence of SEQ ID NO:7.

[0171] In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises a VH at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO:1; and / or a VL at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO:6. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO:1, or a VL comprising the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO:6. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO:1, and a VL comprising the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO:6.

[0172] In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH-CDR1, a VH-CDR2, and a VH-CDR3 from antibody Ab2 (Table 2); or (b) a VL-CDR1, a VL-CDR2, and a VL-CDR3 from antibody Ab2 (Table 2). In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH-CDR1, a VH-CDR2, and a VH-CDR3 from antibody Ab2 (Table 2); and (b) a VL-CDR1, a VL-CDR2, and a VL-CDR3 from antibody Ab2 (Table 2).

[0173] In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH-CDR1, a VH-CDR2, and a VH-CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO:12; or (b) a VL-CDR1, a VL-CDR2, and a VL-CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO:17. InNAI-5002871589v161certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH-CDR1, a VH-CDR2, and a VH-CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO:12; and (b) a VL-CDR1, a VL-CDR2, and a VL- CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO:17.

[0174] In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO:13, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO:14, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO:15; or (b) a VL-CDR1 comprising the amino acid sequence of SEQ ID NO:18, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO:19, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO:20. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO:13, a VH- CDR2 comprising the amino acid sequence of SEQ ID NO:14, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO:15; and (b) a VL-CDR1 comprising the amino acid sequence of SEQ ID NO:18, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO:19, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO:20.

[0175] In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH comprising a VH-CDR1, a VH-CDR2, and a VH-CDR3 disclosed herein, or (b) a VL comprising a VL-CDR1, a VL-CDR2, and a VL-CDR3 disclosed herein. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH comprising a VH-CDR1, a VH-CDR2, and a VH-CDR3 disclosed herein, and (b) a VL comprising a VL-CDR1, a VL-CDR2, and a VL-CDR3 disclosed herein. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises a VH at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO:12; and / or a VL at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO:17. In certain embodiments, the anti- STING antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:12, or a VL comprising the amino acid sequence of SEQ ID NO:17. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:12, and a VL comprising the amino acid sequence of SEQ ID NO:17.NAI-5002871589v162

[0176] In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises a VH at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO:11; and / or a VL at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO:16. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO:11, or a VL comprising the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO:16. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO:11, and a VL comprising the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO:16.

[0177] In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH-CDR1, a VH-CDR2, and a VH-CDR3 from antibody Ab3 (Table 3); or (b) a VL-CDR1, a VL-CDR2, and a VL-CDR3 from antibody Ab3 (Table 3). In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH-CDR1, a VH-CDR2, and a VH-CDR3 from antibody Ab3 (Table 3); and (b) a VL-CDR1, a VL-CDR2, and a VL-CDR3 from antibody Ab3 (Table 3).

[0178] In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH-CDR1, a VH-CDR2, and a VH-CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO:22; or (b) a VL-CDR1, a VL-CDR2, and a VL-CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO:27. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH-CDR1, a VH-CDR2, and a VH-CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO:22; and (b) a VL-CDR1, a VL-CDR2, and a VL- CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO:27.

[0179] In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO:23, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO:24, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO:25; or (b) a VL-CDR1 comprising the amino acid sequence of SEQ ID NO:28, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO:29, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO:30. InNAI-5002871589v163certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO:23, a VH- CDR2 comprising the amino acid sequence of SEQ ID NO:24, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO:25; and (b) a VL-CDR1 comprising the amino acid sequence of SEQ ID NO:28, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO:29, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO:30.

[0180] In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH comprising a VH-CDR1, a VH-CDR2, and a VH-CDR3 disclosed herein, or (b) a VL comprising a VL-CDR1, a VL-CDR2, and a VL-CDR3 disclosed herein. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises (a) a VH comprising a VH-CDR1, a VH-CDR2, and a VH-CDR3 disclosed herein, and (b) a VL comprising a VL-CDR1, a VL-CDR2, and a VL-CDR3 disclosed herein. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises a VH at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO:22; and / or a VL at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO:27. In certain embodiments, the anti- STING antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:22, or a VL comprising the amino acid sequence of SEQ ID NO:27. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:22, and a VL comprising the amino acid sequence of SEQ ID NO:27.

[0181] In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises a VH at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO:21; and / or a VL at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO:26. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO:21, or a VL comprising the amino acid sequence encoded by theNAI-5002871589v164nucleotide sequence of SEQ ID NO:26. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO:21, and a VL comprising the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO:26.

[0182] In certain embodiments, the anti-STING antibody comprises a heavy chain constant region at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO:129; and / or a light chain constant region at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO:130. In certain embodiments, the anti-STING antibody comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO:129, or a light chain constant region comprising the amino acid sequence of SEQ ID NO:130. In certain embodiments, the anti-STING antibody comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO:129 and a light chain constant region comprising the amino acid sequence of SEQ ID NO:130.

[0183] In certain embodiments, the anti-STING antibody comprises a heavy chain constant region at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO:129; and / or a light chain constant region at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO:131. In certain embodiments, the anti-STING antibody comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO:129, or a light chain constant region comprising the amino acid sequence of SEQ ID NO:131. In certain embodiments, the anti-STING antibody comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO:129 and a light chain constant region comprising the amino acid sequence of SEQ ID NO:131.

[0184] In certain embodiments, the anti-STING antibody comprises a heavy chain constant region at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO:132; and / or a light chain constant region at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or atNAI-5002871589v165least 99% identical to the amino acid sequence of SEQ ID NO:130. In certain embodiments, the anti-STING antibody comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO:132, or a light chain constant region comprising the amino acid sequence of SEQ ID NO:130. In certain embodiments, the anti-STING antibody comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO:132 and a light chain constant region comprising the amino acid sequence of SEQ ID NO:130.

[0185] In certain embodiments, the anti-STING antibody comprises a heavy chain constant region at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO:132; and / or a light chain constant region at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO:131. In certain embodiments, the anti-STING antibody comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO:132, or a light chain constant region comprising the amino acid sequence of SEQ ID NO:131. In certain embodiments, the anti-STING antibody comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO:132 and a light chain constant region comprising the amino acid sequence of SEQ ID NO:131.

[0186] In certain embodiments, preferably one or more of the CDRs according to the Kabat definition are maintained substantially unchanged. However, under the simplified assumption that the paratope corresponds to the CDRs, the Chothia definition of the CDRs may be used in addition or alternatively as they correlate very well with the structural loops present in the variable regions. Thus, in order to provide anti-STING antibodies equivalent to subject antibodies Ab1, Ab2, and Ab3, preferably at least one or two of said one or more, preferably not more than two amino acid substitutions if made in the CDRs as defined according to Kabat are made outside the CDRs as defined by Chothia and / or IMGT and most preferably outside the overlap of the CDRs as defined according to Kabat and Chothia.

[0187] For example, regarding amino acid substitutions within the CDRs, variable heavy and light chain and framework amino acid sequences, respectively, preferably conservative amino acid substitutions are performed in accordance with the most frequently exchanged amino acids as analyzed and described by Mirsky et al., Mol. Biol. Evol. 32 (2014), 806-819; see Figure 6 at page 813 of Mirsky et al. In particular, within VH-CDR1, S may be substituted with T; within VH-CDR3, V may be substituted with E, T may be substituted with S and / or M may be substituted with V; within VL-CDR1, R may beNAI-5002871589v166substituted with K, R may be substituted with E, and / or T may be substituted; within VL- CDR2, S may be substituted with A and / or A may be substituted with G; and in VL-CDR3, P may be substituted with S. As mentioned, preferably amino acid substitutions are selected which belong to the same category in either or preferably both models LG and AB shown in Figure 6 of Mirsky et al. (2014), supra, with the LG model being preferred for the tendency to keep amino acid properties, and wherein the amino acid substitutions are selected preferably such that the physiochemical properties of the original amino acid is substantially maintained, i.e., hydrophobic, polar or charged property or for example that in case two or more amino acid substitutions are performed, they compensate each other so as to provide the physicochemical property of the surface all together. In certain embodiments, the antibody of the invention comprises a variant of the amino acid sequence of the VH and / or VL region which is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the VH and VL regions depicted in FIGS. 1A-1C, respectively.

[0188] Of course, besides theoretical considerations also experimental approaches exist for identifying CDR variants within a reasonable time and undue burden. For example, Tiller et al., in Front Immunol. 8 (2017), 986 describe facile affinity maturation of antibody variable domains using natural diversity mutagenesis. Indeed, already a few years earlier Rajpal et al., in PNAS 102 (2005), 8466–8471 reported a general method for greatly improving the affinity of antibodies by using combinatorial libraries and illustrated theirmethod with anti-TNF- antibody D2E7 (HUMIRA©) identifying 38 substitutions in 21CDR positions that resulted in higher affinity binding to TNF- . More recently, Cannon etal., in PLOS Computational Biology, https: / / doi.org / 10.1371 / journal.pcbi.1006980 May 1, 2019 described experimentally guided computational antibody affinity maturation with de novo docking, modelling and rational design in silico affinity maturation, together with alanine scanning, that allowed fine-tuning the protein-protein docking model to subsequently enable the identification of two single-point mutations that increase the affinity of a hybridoma-derived antibody, AB1 for its antigen murine CCL20.

[0189] Accordingly, though each antibody is unique and may have distinct features, nevertheless once a lead candidate has been provided the person skilled in the art in consideration of the teaching of the present disclosure as disclosed in the present application, as well as in view of the computational design and experimental approaches developed so far is able to arrive at equivalent anti-STING antibodies which keep the desired features of the antibody such as those described for the anti-STING antibodies illustrated in the Examples and specifically defined in the claims. In this context, it is well understood that the variantNAI-5002871589v167antibody substantially maintains the binding specificity of the parent antibody, for example recognizing and binding human STING, in particular the C-terminal domain of humanSTING, and preventing phosphorylation of TBK1 or reducing the level of hIFN- in PMA-differentiated human THP-1-derived macrophages stimulated with STING agonist cGAMP, or for example competing with the parent antibody, i.e., with any one of antibodies Ab1, Ab2, and Ab3 for binding to the C-terminal domain of STING. Preferably however, the antibody of the present disclosure comprises in one or both of its immunoglobulin chains one, two or all three CDRs of the variable regions as set forth in FIGS. 1A-1C or one, two or all three CDRs which are 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the CDRs of the variable regions as set forth in FIGS. 1A-1C. In addition, or alternatively, one or more framework regions (FRs) from the FRs are 80% identical to the corresponding FRs depicted in FIGS. 1A-1C, respectively, preferably 85%, 90%, 95%, 96, 97%, 98%, 99% or 100% identical to the FRs depicted in FIGS. 1A, 1B and 1C, respectively. In some embodiments, 1, 2, 3, or all 4 FRs (each being at least 90%, 90-95%, and / or 95-99% identical to the FRs shown in FIGS. 1A-1C, respectively is / are present.

[0190] As known in the art, CDR3 of the variable heavy chain (VH-CDR3) seems to mainly determine antigen specificity; see, e.g., Xu and Davis, Immunity 13 (2000), 37-45. In this context, it was noted that it is the diversity of heavy-chain CDR3s that drive specificity, whereas VH-CDR1 and VH-CDR2 residues are broadly cross-reactive and subject to improvement by somatic hypermutation; see Davis, Semin. Immunol. 16 (2004), 239-243. Accordingly, in certain embodiments the antibody of the present disclosure, which has the immunological characteristics of any of the reference antibodies and being capable of competing with their binding to STING in the C-terminal domain comprise in their variable region at least VH-CDR3 of the corresponding reference antibody or a VH-CDR3 which amino acid sequence is at least 90% identical to the reference VH-CDR3, preferably 95% identical and more preferably 96%, 97%, 98%, 99% or 100% identical. For example, a variant antibody of a reference antibody may retain VH-CDR3 of the reference (parent) antibody while VH-CDR1 and / or VH-CDR2 may contain one or more amino acid substitutions; see supra.

[0191] In a further additional or alternative embodiment of the present disclosure the anti-STING antibody, antigen-binding fragment, synthetic or biotechnological variant thereof can be optimized to have appropriate binding affinity to the target and stability properties. Therefore, at least one amino acid in the CDR or variable region, which is prone to modifications selected from the group consisting of glycosylation, oxidation, deamination,NAI-5002871589v168peptide bond cleavage, iso-aspartate formation and / or unpaired cysteine is substituted by a mutated amino acid that lacks such alteration or wherein at least one carbohydrate moiety is deleted or added chemically or enzymatically to the antibody, see, e.g. Liu et al., J. Pharm. Sci. 97 (2008), 2426-2447; Beck et al., Nat. Rev. Immunol.10 (2010), 345-352; Haberger et al., MAbs. 6 (2014), 327-339.

[0192] An immunoglobulin or its encoding cDNA may be further modified. Thus, in a further embodiment, the method of the present disclosure comprises any one of the step(s) of producing a chimeric antibody, murinized antibody, single-chain antibody, Fab-fragment, bi-specific antibody, fusion antibody, labeled antibody or an analog of any one of those. Corresponding methods are known to the person skilled in the art and are described, e.g., in Harlow and Lane “Antibodies, A Laboratory Manual”, CSH Press, Cold Spring Harbor (1988) First edition; Second edition by Edward A. Greenfield, Dana-Farber Cancer Institute © 2014, ISBN 978-1-936113-81-1. For example, Fab and F(ab’)2 fragments may be produced recombinantly or by proteolytic cleavage of immunoglobulin molecules, using enzymes such as papain (to produce Fab fragments) or pepsin (to produce F(ab’)2 fragments). F(ab’)2 fragments contain the variable region, the light chain constant region (CL) and the CH1 domain of the heavy chain. Such fragments are sufficient for use, for example, in immunodiagnostic procedures involving coupling the immunospecific portions of immunoglobulins to detecting reagents such as radioisotopes.

[0193] In certain embodiments, the antibody of the present disclosure may thus be provided in a format selected from the group consisting of a single chain Fv fragment (scFv), an F(ab’) fragment, an F(ab) fragment, and an F(ab’)2fragment, an Fd, an Fv, a single-chain antibody, a disulfide-linked Fv (sdFv), a VHH, and a nanobody, and / or which is a chimeric murine-human or a murinized antibody.

[0194] The perceived advantages of using small Fab and scFv engineered antibody formats which lack the effector function include minimizing the risk of triggering inflammatory side reactions. Furthermore, besides scFv and single-domain antibodies retain the binding specificity of full-length antibodies, they can be expressed as single genes and intracellularly in mammalian cells as intrabodies, with the potential for alteration of the folding, interactions, modifications, or subcellular localization of their targets; see for review, e.g., Miller and Messer, Molecular Therapy 12 (2005), 394-401.

[0195] The five primary classes of immunoglobulins are IgG, IgM, IgA, IgD and IgE. These are distinguished by the type of heavy chain found in the molecule. IgG molecules have heavy chains known as gamma-chains; IgMs have mu-chains; IgAs have alpha-chains;NAI-5002871589v169IgEs have epsilon-chains; and IgDs have delta-chains; see for review, e.g., Schroeder et al.,Structure and function of immunoglobulins. J. Allergy Clin. Immunol. 125 (2010), S41 S52.In principle, the antibodies of the present disclosure may be of any kind of class and antibody fragment as long as the binding specificity towards STING as and illustrated in the appended Examples for the corresponding reference antibody remains unaffected in kind. However, preferably complete IgG antibodies are used, wherein the antibody comprises a constant domain. The constant domain may be native, i.e., originally cloned together with the variable domain or heterologous, for example, a murine constant in case animal studies are envisaged. Preferably, the constant domain is of human origin with a different IgG subtype, e.g., IgG1 versus IgG4 or a different allotype and allele, respectively, compared to the constant domain of the antibody as naturally occurred in human. The definition of “allotypes” requires that antibody reagents are available to determine the allotypes serologically. If the determination is only done at the sequence level, the polymorphisms have to be described as “alleles”. This does not hinder to establish a correspondence with allotypes if the correspondence allele- allotype has been experimentally proven, or if the individual sequence is identical to a sequence for which it has been demonstrated.

[0196] In certain embodiments of the present disclosure, the constant domain is heterologous to at least one of the CDRs and the VH and VL chains, respectively, e.g., an immunoglobulin heavy chain constant domain and / or immunoglobulin light chain constant domain, preferably of the IgG type. In addition, or alternatively, the heterologous part of the antibody may be a mammalian secretory signal peptide. Put in other words, in certain embodiments the anti-STING antibody of the present disclosure is a (i) fusion protein comprising a polypeptide sequence which is heterologous to the VH region and / or VL region, or at least one CDR; and / or (ii) a non-natural variant of a polypeptide derived from an immunoglobulin, said non-natural variant comprising a heavy chain constant region that comprises one or more amino acid deletions, substitutions, and / or additions relative to a wild type polypeptide. For example, the human constant domain of the recombinant antibody of the present disclosure may be of a different IgG isotype than the constant domain of the parent antibody as naturally produced by the memory B cell or of a different allotype, for example to avoid or reduce immunogenicity which can happen as a result of allo- immunization; see, e.g., for review Jefferis and Lefranc, MAbs 1 (2009), 332–338.

[0197] In certain embodiments, the antibody disclosed herein is of IgA type. In certain embodiments, the antibody disclosed herein is not IgA type. In certain embodiments, the antibody disclosed herein is of IgG type. In certain embodiments, the antibody of theNAI-5002871589v170present disclosure comprises an immunoglobulin heavy chain constant region (also referred to as “heavy chain constant region” herein) and / or an immunoglobulin light chain constant region (also referred to as “light chain constant region” herein).

[0198] In certain embodiments, the immunoglobulin heavy constant region is of the IgG type. In certain embodiments, the immunoglobulin heavy constant region is of the IgG1 type. In certain embodiments, the immunoglobulin heavy constant region is of the IgG1 type and G1m3 allotype. In certain embodiments, the immunoglobulin heavy constant region is of the IgG4 type. In certain embodiments, the heavy chain constant region is not IgA type.

[0199] In certain embodiments, the heavy chain constant region is of human IgG type. In certain embodiments, the heavy chain constant region is of human IgG1 type. In certain embodiments, the heavy chain constant region is of human IgG1 type and G1m3 allotype. In certain embodiments, the heavy chain constant region is of human IgG4 type.

[0200] In certain embodiments, the light chain constant region comprises a human kappa light chain constant region. In certain embodiments, the light chain constant region comprises a human lambda light chain constant region.

[0201] In certain embodiments, the heavy chain constant region comprises an amino acid sequence at least 80%, at least 90%, at least 95%, or at least 99% identical to the amino acid sequence of SEQ ID NO:129. In certain embodiments, the heavy chain constant region comprises the amino acid sequence of SEQ ID NO:129. In certain embodiments, the heavy chain constant region comprises an amino acid sequence at least 80%, at least 90%, at least 95%, or at least 99% identical to the amino acid sequence of SEQ ID NO:132. In certain embodiments, the heavy chain constant region comprises the amino acid sequence of SEQ ID NO:132. In certain embodiments, the light chain constant region comprises an amino acid sequence at least 80%, at least 90%, at least 95%, or at least 99% identical to the amino acid sequence of SEQ ID NO:130. In certain embodiments, the light chain constant region comprises the amino acid sequence of SEQ ID NO:130. In certain embodiments, the light chain constant region comprises an amino acid sequence at least 80%, at least 90%, at least 95%, or at least 99% identical to the amino acid sequence of SEQ ID NO:131. In certain embodiments, the light chain constant region comprises the amino acid sequence of SEQ ID NO:131.

[0202] The four subclasses, IgG1, IgG2, IgG3, and IgG4, which are highly conserved, differ in their constant region, particularly in their hinges and upper CH2 domains. These regions are involved in binding to both IgG-Fc receptors (FcgR) and C1q. As a result, the different subclasses have different effector functions, both in terms of triggering FcgR-NAI-5002871589v171expressing cells, resulting in phagocytosis or antibody-dependent cell-mediated cytotoxicity, and activating complement. The Fc regions also contain a binding epitope for the neonatal Fc receptor (FcRn), responsible for the extended half-life, placental transport, and bidirectional transport of IgG through mucosal surfaces. However, FcRn is also expressed in myeloid cells, where it participates in both phagocytosis and antigen presentation together with classical FcgR and complement. How these properties, IgG-polymorphisms and post-translational modification of the antibodies in the form of glycosylation, affect IgG-function is described in Vidarsson et al., (2014) IgG subclasses and allotypes: from structure to effector function. Front. Immunol. 5:520. doi:10.3389 / fimmu.2014.00520 and de Taeye et al., Antibodies 2019, 8, 30; doi:10.3390 / antib8020030. Preferably, the immunoglobulin heavy and / or light chain constant domain present in the antibody of the present disclosure is of the IgG type.

[0203] Accordingly, in certain embodiments of the present disclosure a specific IgG type is preferred, for example the IgG4 or IgG1 isotype and / or the constant region of the antibody, or antigen-binding fragment, variant, or derivative thereof has been altered so as to provide desired biochemical characteristics. In particular, in certain embodiments the Fc portion of the antibody may be mutated to alter, i.e., to decrease or increase immune effector function or to increase its half-life using techniques known in the art. Thus, in certain embodiments the Fc portion of the antibody is mutated to decrease immune effector function and in another embodiment the Fc portion of the antibody is mutated to increase immune effector function. In another embodiment, the antibody is mutated to increase its half-life. In certain embodiments, the antibody comprises an Fc region comprising one or more mutations. In certain embodiments, the one or more mutations decrease the immune effector function of the antibody. In certain embodiments, the one or more mutations increase the immune effector function of the antibody. In certain embodiments, the one or more mutations increase the half-life of the antibody.

[0204] For example, it may be that constant region modifications consistent with the instant invention moderate complement binding and thus reduce the serum half-life and nonspecific association of a conjugated cytotoxin. Other modifications of the constant region may be used to modify disulfide linkages or oligosaccharide moieties that allow for enhanced tissue antigen interaction due to increased antigen specificity or antibody flexibility. Furthermore, mutations in the Fc region can be made that lead to enhanced antibody dependent cell-mediated cytotoxicity (ADCC) or antibody-dependent cellular phagocytosis(ADCP) via increasing Fc RIIIa binding and / or decreasing Fc RIIIb binding and viaincreasing Fc RIIa binding and / or Fc RIIIa binding, respectively. For example, theNAI-5002871589v172GASDALIE Fc mutant (G236A / S239D / A330L / I332E) exhibits a higher affinity for Fc RIIIa.Another possibility is the enhancement of complement-dependent cytotoxicity (CDC) via increasing C1q binding and / or hexamerization.

[0205] In certain embodiments, certain antibodies for use in the diagnostic and treatment methods described herein have a constant region, e.g., an IgG heavy chain constant region, which is altered to eliminate glycosylation, referred to elsewhere herein as aglycosylated or “agly” antibodies. Such “agly” antibodies may be prepared enzymatically as well as by engineering the consensus glycosylation site(s) in the constant region. It is believed that “agly” antibodies have a reduced effector function and thus an improved safety and stability profile in vivo. Methods of producing aglycosylated antibodies, having desired effector function are found for example in international application WO 2005 / 018572, which is incorporated by reference in its entirety. A further approach to reduce the effector functionof antibodies is the reduction of Fc R and C1q binding by mutations in the Fc region.

[0206] A summary is for example given in the review of Wang et al., Protein Cell 9 (2018), 63-73, wherein Table 1 provides examples of modifications to modulate antibody effector function and the half-life of an antibody and which mutations described therein are herein incorporated by reference. The resulting physiological profile, bioavailability and other biochemical effects of the modifications, such as recognizing and binding STING may easily be measured and quantified using well know immunological techniques without undue experimentation.

[0207] As mentioned, in some instances inflammatory responses should be avoided for which reason effector functions of the constant domain of the antibody may be attenuated or eliminated altogether. For example, recombinant human IgG antibodies (hIgGs)completely devoid of binding to Fc receptors (Fc Rs) and complement protein C1q, and thuswith abolished immune effector functions, are of use for various therapeutic applications. It was found that the combination of Leu234Ala and Leu235Ala (commonly called LALAmutations) or the SPLE mutation eliminated Fc RIIa binding and were shown to eliminatedetectable binding to Fc RI, IIa, and IIIa for both IgG1 and IgG4 and that the LALA-PGmutation was an improvement over LALA mutations alone in that they nullified Fc function in mouse and human IgG; for corresponding review see, e.g., Saunders (2019) Conceptual Approaches to Modulating Antibody Effector Functions and Circulation Half-Life. Front. Immunol. 10:1296.doi: 10.3389 / fimmu.2019.01296 and Schlothauer et al., Protein Engineering, Design and Selection 29 (2016), 457-466.NAI-5002871589v173

[0208] Another early approach to reduce effector function is to mutate the glycosylation site at N297 with mutations such as N297A, N297Q, and N297G. The half-life of an antibody can be increased via introducing the following mutations M252Y / S254T / T256E or M428L / N434S; see Wang et al. 2018.

[0209] In this context, antibody effector function does not seem to be necessary for antibody binding to STING and preventing TBK1 phosphorylation and reducing the level ofhIFN- since the small compound H-151 has also been shown to prevent TBK1phosphorylation; see Example 9.

[0210] Thus, in certain embodiments the antibody is of the IgG1 class or isotype preferably, wherein the antibody is an IgG1 variant comprising the amino acid substitutions L234A, L235A (LALA) and preferably the amino acid substitutions L234A, L235A, P329G(LALA-PG). For example, in order to avoid recruitment of immune cells through Fc -receptors and enable a short systemic half-life in circulation, Fc R binding can be abolishedby introduction of P329G LALA mutations, see Schlothauer et al. (2016), supra, while FcRn binding and recycling can be abolished by introduction of Triple A (I253A, H310A, H435A) mutations; see, e.g., Regula et al., EMBO Mol. Med., 8 (2016), 1265-1288.

[0211] Furthermore, human immunoglobulin G isotype 4 (IgG4) antibodies are potential candidates for antibody therapy when reduced immune effector functions are desirable. Thus, in another embodiment, the antibody is of the IgG4 class or isotype. IgG4 antibodies are dynamic molecules able to undergo a process known as Fab arm exchange (FAE). This results in functionally monovalent, bispecific antibodies (bsAbs) with unknown specificity and hence, potentially, reduced therapeutic efficacy. Thus, the antibody of the present disclosure is of the IgG4 class or isotype including the S228P mutation. The S228P mutation prevents in vivo and in vitro IgG4 Fab-arm exchange as demonstrated using a combination of novel quantitative immunoassays and physiological matrix preparation; see Silva et al., J. Biol. Chem. 290 (2015), 5462–5469.

[0212] In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof of the present disclosure (e.g., an anti-STING antibody or an antigen-binding fragment thereof of Section 5.2) is a recombinant antibody or an antigen-binding fragment thereof. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof of the present disclosure (e.g., an anti-STING antibody or an antigen-binding fragment thereof of Section 5.2) is a monoclonal antibody or an antigen-binding fragment thereof. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof of the present disclosure (e.g., an anti-STING antibody or an antigen-bindingNAI-5002871589v174fragment thereof of Section 5.2) is an isolated antibody or an antigen-binding fragment thereof. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof of the present disclosure (e.g., an anti-STING antibody or an antigen-binding fragment thereof of Section 5.2) is an antibody having one or both of its C-terminal lysine residues removed from its heavy chains or an antigen-binding fragment thereof. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof of the present disclosure (e.g., an anti-STING antibody or an antigen-binding fragment thereof of Section 5.2) is an antibody having a non-human glycosylation pattern or an antigen-binding fragment thereof. In certain embodiments, the antibody having a non-human glycosylation pattern is produced in a cell (e.g., CHO cell) cultured in vitro (see Section 5.3). In certain embodiments, the non-human glycosylation pattern is conferred by CHO cells. In certain embodiments, the anti-STING antibody or antigen-binding fragment thereof of the present disclosure (e.g., an anti-STING antibody or an antigen-binding fragment thereof of Section 5.2) is in a lyophilized format. 5.3 Polynucleotides, Vectors, and Cells

[0213] The present disclosure also provides one or more polynucleotide(s) or a set of polynucleotides encoding the anti-STING antibody or antigen-binding fragment thereof of the present disclosure (e.g., an anti-STING antibody or an antigen-binding fragment thereof of Section 5.2) or an immunoglobulin VH and VL thereof or a heavy chain or a light chain thereof, or a polynucleotide or a set of polynucleotides complementary thereto, preferably wherein the polynucleotide(s) or set of polynucleotides is (are) cDNA.

[0214] Polynucleotides disclosed herein can be RNA or DNA (e.g., cDNA, genomic DNA, or synthetic DNA), and can be double-stranded or single-stranded, and if single stranded can be the coding strand or non-coding (anti-sense) strand. In certain embodiments, polynucleotides disclosed herein are cDNA.

[0215] In certain embodiments of the present disclosure, the polynucleotide comprises, consists essentially of, or consists of a nucleic acid having a polynucleotide sequence encoding the VH or VL chain of an anti-STING antibody as depicted in Tables 1-3. In this respect, the person skilled in the art will readily appreciate that the polynucleotides encoding the light and / or heavy chain may be encoded by one or more polynucleotides. In certain embodiments therefore, the polynucleotide comprises, consists essentially of, or consists of a nucleic acid having a polynucleotide sequence of the VHand the VLchain of an anti-STING antibody as depicted in Tables 1-3.NAI-5002871589v175

[0216] In certain embodiments, the polynucleotide(s) of the present disclosure comprise, consist essentially of, or consist of a nucleotide sequence of the VH and / or VL chain as set forth in SEQ ID NO:1 and / or SEQ ID NO:6, or codon degenerated version(s) thereof; as set forth in SEQ ID NO:11 and / or SEQ ID NO:16, or codon degenerated version(s) thereof; as set forth in SEQ ID NO:21 and / or SEQ ID NO:26, or codon degenerated version(s) thereof.

[0217] In certain embodiments, the present disclosure provides a polynucleotide encoding both VH and VL of an anti-STING antibody or an antigen-binding fragment thereof disclosed herein (e.g., an anti-STING antibody or an antigen-binding fragment thereof of Section 5.2). In certain embodiments, the present disclosure provides a polynucleotide encoding both heavy chain and light of an anti-STING antibody or an antigen-binding fragment thereof disclosed herein (e.g., an anti-STING antibody or an antigen-binding fragment thereof of Section 5.2). In certain embodiments, the present disclosure provides a set of polynucleotides or a polynucleotide composition encoding an anti-STING antibody or an antigen-binding fragment thereof disclosed herein (e.g., an anti-STING antibody or an antigen-binding fragment thereof of Section 5.2). In certain embodiments, at least one polynucleotide of the set of polynucleotides or polynucleotide composition encodes the VH of the anti-STING antibody or antigen-binding fragment thereof and at least one polynucleotide of the set of polynucleotides or polynucleotide composition encodes the VL of the anti-STING antibody or antigen-binding fragment thereof. In certain embodiments, at least one polynucleotide of the set of polynucleotides or polynucleotide composition encodes the heavy chain of the anti-STING antibody or an antigen-binding fragment thereof and at least one polynucleotide of the set of polynucleotides or polynucleotide composition encodes the VL of the anti-STING antibody or antigen-binding fragment thereof.

[0218] The present disclosure further provides a vector or a set of vectors comprising a polynucleotide or a set of polynucleotides disclosed herein. Such vectors are useful, for example, for amplifying the polynucleotide or set of polynucleotides in host cells to create useful quantities thereof, and for expressing an anti-STING antibody or an antigen-binding fragment thereof disclosed herein (e.g., an anti-STING antibody or an antigen-binding fragment thereof of Section 5.2).

[0219] Any suitable vectors can be used to introduce one or more polynucleotides disclosed herein into a cell. Exemplary vectors include, but not limited to, lentivirus vectors, adeno-associated viral (AAV) vectors, adenoviral (AV), and liposomal vectors.NAI-5002871589v176

[0220] The present disclosure further provides a cell (e.g., a host cell) comprising any one or more of an anti-STING antibody or an antigen-binding fragment thereof disclosed herein (e.g., an anti-STING antibody or an antigen-binding fragment thereof of Section 5.2), a polynucleotide or a set of polynucleotides disclosed herein, or a vector or a set of vectors disclosed herein. In certain embodiments, the cell expresses an anti-STING antibody or an antigen-binding fragment thereof disclosed herein (e.g., an anti-STING antibody or an antigen-binding fragment thereof of Section 5.2).

[0221] In certain embodiments, the cell replicates the polynucleotide or et of polynucleotides disclosed herein or the vector or set of vectors disclosed herein.

[0222] In certain embodiments of the present disclosure, the polynucleotide(s) are linked to a heterologous nucleic acid, for example expression control sequences such as a promoter, transcription and / or translation enhancer sequences, internal ribosome binding sites, nucleic acids encoding a peptide leader sequence for recombinant expression in a host and the like. Accordingly, the present disclosure provides a polynucleotide encoding a recombinant anti-STING antibody or STING binding fragment, synthetic derivative, or biotechnological derivative thereof, wherein the polynucleotide encodes (i) a VH chain comprising CDRs 1, 2, and 3, and / or a VL chain comprising VL CDRs 1, 2, and 3 as defined by Kabat, wherein (a) VH-CDR1 comprises the amino acid sequence of SEQ ID NO:3 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletion, preferably one or two amino acid substitutions and / or deletions, more preferably one or two amino acid substitutions, (b) VH-CDR2 comprises the amino acid sequence of SEQ ID NO:4 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletion, preferably one or two amino acid substitutions and / or deletions, more preferably one or two amino acid substitutions, (c) VH-CDR3 comprises the amino acid sequence of SEQ ID NO:5 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletion, preferably one or two amino acid substitutions and / or deletions, more preferably one or two amino acid substitutions, (d) VL-CDR1 comprises the amino acid sequence of SEQ ID NO:8 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletion, preferably one or two amino acid substitutions and / or deletions, more preferably one or two amino acid substitutions,NAI-5002871589v177(e) VL-CDR2 comprises the amino acid sequence of SEQ ID NO:9 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletion, preferably one or two amino acid substitutions and / or deletions, more preferably one or two amino acid substitutions, and (f) VL-CDR3 comprises the amino acid sequence of SEQ ID NO:10 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletion, preferably one or two amino acid substitutions and / or deletions, more preferably one or two amino acid substitutions; and / or (ii) a VH chain and / or a VL chain, wherein (a) the VH chain comprises the amino acid sequence depicted in SEQ ID NO:2 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions; and (b) the VL comprises the amino acid sequence depicted in SEQ ID NO:7, or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions; preferably wherein the VH and VL chain amino acid sequence is at least 90% identical to SEQ ID NO:2 and 7, respectively; or (iii) a VH chain comprising CDRs 1, 2, and 3, and / or a VL chain comprising VL CDRs 1, 2, and 3 as defined by Kabat, wherein (a) VH-CDR1 comprises the amino acid sequence of SEQ ID NO:13 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletion, preferably one or two amino acid substitutions and / or deletions, more preferably one or two amino acid substitutions, (b) VH-CDR2 comprises the amino acid sequence of SEQ ID NO:14 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletion, preferably one or two amino acid substitutions and / or deletions, more preferably one or two amino acid substitutions, (c) VH-CDR3 comprises the amino acid sequence of SEQ ID NO:15 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletion, preferably one or two amino acid substitutions and / or deletions, more preferably one or two amino acid substitutions, (d) VL-CDR1 comprises the amino acid sequence of SEQ ID NO:18 or a variant thereof, wherein the variant comprises one or more amino acid substitutionsNAI-5002871589v178and / or deletion, preferably one or two amino acid substitutions and / or deletions, more preferably one or two amino acid substitutions, (e) VL-CDR2 comprises the amino acid sequence of SEQ ID NO:19 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletion, preferably one or two amino acid substitutions and / or deletions, more preferably one or two amino acid substitutions, and (f) VL-CDR3 comprises the amino acid sequence of SEQ ID NO:20 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletion, preferably one or two amino acid substitutions and / or deletions, more preferably one or two amino acid substitutions; and / or (iv) a VH chain and / or a VL chain, wherein (a) the VH chain comprises the amino acid sequence depicted in SEQ ID NO:12 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions; and (b) the VL comprises the amino acid sequence depicted in SEQ ID NO:17, or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions; preferably wherein the VH and VL chain amino acid sequence is at least 90% identical to SEQ ID NO:12 and 17, respectively; or (v) a VH chain comprising CDRs 1, 2, and 3, and / or a VL chain comprising VL CDRs 1, 2, and 3 as defined by Kabat, wherein (a) VH-CDR1 comprises the amino acid sequence of SEQ ID NO:23 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletion, preferably one or two amino acid substitutions and / or deletions, more preferably one or two amino acid substitutions, (b) VH-CDR2 comprises the amino acid sequence of SEQ ID NO:24 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletion, preferably one or two amino acid substitutions and / or deletions, more preferably one or two amino acid substitutions, (c) VH-CDR3 comprises the amino acid sequence of SEQ ID NO:25 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletion, preferably one or two amino acid substitutions and / or deletions, more preferably one or two amino acid substitutions,NAI-5002871589v179(d) VL-CDR1 comprises the amino acid sequence of SEQ ID NO:28 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletion, preferably one or two amino acid substitutions and / or deletions, more preferably one or two amino acid substitutions, (e) VL-CDR2 comprises the amino acid sequence of SEQ ID NO:29 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletion, preferably one or two amino acid substitutions and / or deletions, more preferably one or two amino acid substitutions, and (f) VL-CDR3 comprises the amino acid sequence of SEQ ID NO:30 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletion, preferably one or two amino acid substitutions and / or deletions, more preferably one or two amino acid substitutions; and / or (vi) a VH chain and / or a VL chain, wherein (a) the VH chain comprises the amino acid sequence depicted in SEQ ID NO:22 or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions; and (b) the VL comprises the amino acid sequence depicted in SEQ ID NO:27, or a variant thereof, wherein the variant comprises one or more amino acid substitutions and / or deletions; preferably wherein the VH and VL chain amino acid sequence is at least 90% identical to SEQ ID NO:22 and 27, respectively.

[0223] In addition, the present disclosure provides a polynucleotide linked to a heterologous nucleic acid, wherein the polynucleotide is selected from the group consisting of: (a) a polynucleotide encoding an immunoglobulin heavy chain or a fragment thereof comprising a heavy chain variable region (VH) comprising CDRs 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs: 3, 4, and 5, respectively, and wherein the VH when paired with a light chain variable region (VL) comprising the amino acid sequence set forth in SEQ ID NO:7 binds to STING; (b) a polynucleotide encoding an immunoglobulin light chain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively, and wherein theNAI-5002871589v180VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO:2 binds to STING; (c) a polynucleotide encoding (i) an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs: 3, 4, and 5, respectively; and (ii) an immunoglobulin light chain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; (d) a polynucleotide encoding an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising the amino acid sequence set forth in SEQ ID NO:2, wherein the VH when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO:7 binds to STING; (e) a polynucleotide encoding an immunoglobulin light chain or a fragment thereof comprising a VL comprising the amino acid sequence set forth in SEQ ID NO:7, wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO:2 binds to STING; (f) a polynucleotide encoding an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising the amino acid sequence set forth in SEQ ID NO:2 and an immunoglobulin light chain or a fragment thereof comprising a VL comprising the amino acid sequence set forth in SEQ ID NO:7; (g) a polynucleotide as in any one of (a)-(f), wherein a CDR comprises one or more, preferably no more than two amino acid substitutions and / or deletions, and / or the variable region sequence is at least 90% identical to SEQ ID NO:2 or SEQ ID NO:7.

[0224] The present disclosure further relates to a polynucleotide linked to a heterologous nucleic acid, wherein the polynucleotide is selected from the group consisting of: (a) a polynucleotide encoding an immunoglobulin heavy chain or a fragment thereof comprising a heavy chain variable region (VH) comprising CDRs 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs: 13, 14, and 15, respectively, and wherein the VH when paired with a light chain variable region (VL) comprising the amino acid sequence set forth in SEQ ID NO:17 binds to STING;NAI-5002871589v181(b) a polynucleotide encoding an immunoglobulin light chain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs: 18, 19, and 20, respectively, and wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO:12 binds to STING; (c) a polynucleotide encoding (i) an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs: 13, 14, and 15, respectively; and (ii) an immunoglobulin light chain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs: 18, 19, and 20, respectively; (d) a polynucleotide encoding an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising the amino acid sequence set forth in SEQ ID NO:12, wherein the VH when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO:17 binds to STING; (e) a polynucleotide encoding an immunoglobulin light chain or a fragment thereof comprising a VL comprising the amino acid sequence set forth in SEQ ID NO:17, wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO:12 binds to STING; (f) a polynucleotide encoding an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising the amino acid sequence set forth in SEQ ID NO:12 and an immunoglobulin light chain or a fragment thereof comprising a VL comprising the amino acid sequence set forth in SEQ ID NO:17; (g) a polynucleotide as in any one of (a)-(f), wherein a CDR comprises one or more, preferably no more than two amino acid substitutions and / or deletions, and / or the variable region sequence is at least 90% identical to SEQ ID NO:12 or SEQ ID NO:17.

[0225] Alternatively, the present disclosure provides a polynucleotide linked to a heterologous nucleic acid, wherein the polynucleotide is selected from the group consisting of: (a) a polynucleotide encoding an immunoglobulin heavy chain or a fragment thereof comprising a heavy chain variable region (VH) comprising CDRs 1, 2,NAI-5002871589v182and 3 with the amino acid sequences set forth in SEQ ID NOs: 23, 24, and 25, respectively, and wherein the VH when paired with a light chain variable region (VL) comprising the amino acid sequence set forth in SEQ ID NO:27 binds to STING; (b) a polynucleotide encoding an immunoglobulin light chain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs: 28, 29, and 30, respectively, and wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO:22 binds to STING; (c) a polynucleotide encoding (i) an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs: 23, 24, and 25, respectively; and (ii) an immunoglobulin light chain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs: 28, 29, and 30, respectively; (d) a polynucleotide encoding an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising the amino acid sequence set forth in SEQ ID NO:22, wherein the VH when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO:27 binds to STING; (e) a polynucleotide encoding an immunoglobulin light chain or a fragment thereof comprising a VL comprising the amino acid sequence set forth in SEQ ID NO:27, wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO:22 binds to STING; (f) a polynucleotide encoding an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising the amino acid sequence set forth in SEQ ID NO:22 and an immunoglobulin light chain or a fragment thereof comprising a VL comprising the amino acid sequence set forth in SEQ ID NO:27; (g) a polynucleotide as in any one of (a)-(f), wherein a CDR comprises one or more, preferably no more than two amino acid substitutions and / or deletions, and / or the variable region sequence is at least 90% identical to SEQ ID NO:22 or SEQ ID NO:27.NAI-5002871589v183

[0226] In certain embodiments, the immunoglobulin of any of the preceding paragraphs when paired as VH and VL binds to human STING, for example as measured by ELISA or an equivalent assay to the assay described in the Examples.

[0227] Furthermore, the present disclosure provides a vector and vectors comprising one or more of those polynucleotides, preferably wherein the vector is an expression vector and the one or more polynucleotide(s) are operably linked to expression control sequences.

[0228] The polynucleotides may be produced and, if desired manipulated using methods well known in the art for the manipulation of nucleotide sequences, e.g., recombinant DNA techniques, site directed mutagenesis, PCR, etc. (see, for example, the techniques described in Molecular Cloning: A Laboratory Manual (Fourth Edition): Three- volume set; Green and Sambrook (2012) ISBN 10: 1936113422 / ISBN 13: 9781936113422 Cold Spring Harbor Laboratory Press; update (2014) ISBN 978-1-936113-42-2 and Ausubel et al., eds., Current Protocols in Molecular Biology, John Wiley & Sons, NY (1998) and updates, which are both incorporated by reference herein in their entireties), to generate antibodies having a different amino acid sequence, for example to create amino acid substitutions, deletions, and / or insertions.

[0229] Once a polynucleotide encoding an antibody molecule or a heavy or light chain of an antibody, or portion thereof (preferably containing the heavy or light chain variable domain), of the invention has been obtained, the vector for the production of the antibody molecule may be produced by recombinant DNA technology using techniques well known in the art. Thus, methods for preparing a protein by expressing a polynucleotide containing an antibody encoding nucleotide sequence are described herein. Methods which are well known to those skilled in the art can be used to construct expression vectors containing antibody coding sequences and appropriate transcriptional and translational control signals. These methods include, for example, in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination. The invention, thus, provides replicable vectors comprising a nucleotide sequence encoding an antibody molecule of the invention, or a heavy or light chain thereof, or a heavy or light chain variable domain, operable linked to a promoter. Such vectors may include the nucleotide sequence encoding the constant region of the antibody molecule (see, e.g., international applications WO 86 / 05807 and WO 89 / 01036; and US patent no. 5,122,464) and the variable domain of the antibody may be cloned into such a vector for expression of the entire heavy or light chain.

[0230] The term “vector” or “expression vector” is used herein to mean vectors used in accordance with the present disclosure as a vehicle for introducing into and expressing aNAI-5002871589v184desired gene in a host cell. In general, vectors compatible with the instant invention will comprise a selection marker, appropriate restriction sites to facilitate cloning of the desired gene and the ability to enter and / or replicate in eukaryotic or prokaryotic cells. The marker may provide for prototrophy to an auxotrophic host, biocide resistance (e.g., antibiotics), or resistance to heavy metals such as copper. The selectable marker gene can either be directly linked to the DNA sequences to be expressed, or introduced into the same cell by co- transformation. Additional elements may also be needed for optimal synthesis of mRNA. These elements may include signal sequences, splice signals, as well as transcriptional promoters, enhancers, and termination signals. For the expression of double-chained antibodies, a single vector or vectors encoding both the heavy and light chains may be co- expressed in the host cell for expression of the entire immunoglobulin molecule, as detailed below.

[0231] The host cell may be co-transfected with two expression vectors of the invention, the first vector encoding a heavy chain derived polypeptide and the second vector encoding a light chain derived polypeptide. The two vectors may contain identical selectable markers which enable equal expression of heavy and light chain polypeptides. Alternatively, a single vector may be used which encodes both heavy and light chain polypeptides. In such situations, the light chain is advantageously placed before the heavy chain to avoid an excess of toxic free heavy chain; see Proudfoot, Nature 322 (1986), 52; Kohler, Proc. Natl. Acad. Sci. USA 77 (1980), 2197. The coding sequences for the heavy and light chains may comprise cDNA or genomic DNA. The expression vector(s) is (are) transferred to a host cell by conventional techniques and the transfected cells are then cultured by conventional techniques to produce an antibody for use in the methods described herein. Accordingly, the present disclosure also relates to host cells comprising one or more polynucleotides or a vector or vectors of the present disclosure.

[0232] As used herein, “host cells” refers to cells which harbor vectors constructed using recombinant DNA techniques and encoding at least one heterologous gene. In descriptions of processes for isolation of antibodies from recombinant hosts, the terms “cell” and “cell culture” are used interchangeably to denote the source of antibody unless it is clearly specified otherwise. In other words, recovery of polypeptide from the “cells” may mean either from spun down whole cells, or from the cell culture containing both the medium and the suspended cells.

[0233] Antibodies used for laboratory research / diagnosis may be expressed in any suitable host, e.g., in mammalian cells, bacterial cells, yeasts, plant cells or insect cells.NAI-5002871589v185However, currently almost all therapeutic antibodies are still produced in mammalian cell lines in order to reduce the risk of immunogenicity due to altered, non-human glycosylation patterns. However, recent developments of glycosylation-engineered yeast, insect cell lines, and transgenic plants are promising to obtain antibodies with “human-like” post-translational modifications. Furthermore, smaller antibody fragments including bispecific antibodies without any glycosylation are successfully produced in bacteria and have advanced to clinical testing. The first therapeutic antibody products from a non-mammalian source can be expected in coming next years. A review on current antibody production systems that can be applied for preparing the recombinant anti-STING antibody of the present disclosure including their usability for different applications is given in Frenzel et al., Front Immunol. 4 (2013), 217, published online on July 29, 2013 doi: 10.3389 / fimmu.2013.00217 and transient expression of human antibodies in mammalian cells is described by Vazquez-Lombardi et al., Nature protocols 13 (2018), 99-117; and Hunter et al., Optimization of protein expression in mammalian cells. Current Protocols in Protein Science 95 (2019), e77. doi: 10.1002 / cpps.77. Once an antibody molecule of the invention has been recombinantly expressed, the whole antibodies, their dimers, individual light and heavy chains, or other immunoglobulin forms of the present disclosure can be purified according to standard procedures of the art, including for example, by chromatography (e.g., ion exchange, affinity, particularly by affinity for the specific antigen after Protein A, and sizing column chromatography), centrifugation, differential solubility, e.g. ammonium sulfate precipitation, or by any other standard technique for the purification of proteins; see, e.g., Scopes, “Protein Purification”, Springer Verlag, N.Y. (1982) and Antibodies A Laboratory Manual 2nd edition, 2014 by Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA. Thus, the present disclosure also relates to a method for preparing an anti-STING antibody and / or fragments thereof or immunoglobulin chain(s) thereof, said method comprising: (a) culturing the host cell as defined hereinabove, which cell comprised the polynucleotide(s) or vector(s) as defined hereinbefore under conditions allowing for expression of the anti-STING antibody, STING-binding fragment or immunoglobulin chain(s) thereof; and (b) isolating the anti-STING antibody, STING-binding fragment or immunoglobulin chain(s) thereof from the culture.

[0234] Furthermore, the present disclosure also relates to the anti-STING antibody, STING-binding fragment and immunoglobulin chain(s) thereof encoded by a polynucleotideNAI-5002871589v186as defined hereinabove and / or obtainable by the method for their recombinant production mentioned above.

[0235] In certain embodiments, the antibody polypeptide comprises an amino acid sequence or one or more moieties not normally associated with an antibody. Thus, the present disclosure further encompasses antibodies, or antigen-binding fragments, variants, or derivatives thereof of the invention conjugated to a diagnostic or therapeutic agent, like a drug. Detection can be facilitated by coupling the antibody, or antigen-binding fragment, variant, or derivative thereof to a detectable substance. Exemplary modifications are described in more detail below. For example, the antibody or STING-binding fragment thereof such a single-chain Fv antibody fragment of the invention may comprise a flexible linker sequence, or may be modified to add a functional moiety or detectable label (e.g., PEG, a drug, a toxin, or a label such as a fluorescent, (chemo / bio)luminescent, radioactive, enzyme, nuclear magnetic, heavy metal, a tag, a flag and the like); see, e.g., Antibodies A Laboratory Manual 2nd edition, 2014 by Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA for general techniques; and Falck and Müller, Antibodies 7 (2018), 4; doi:10.3390 / antib7010004 for enzyme-based labeling strategies for antibody-drug conjugates and antibody mimetics.

[0236] The cGAS-STING pathway has been implicated in systemic disease processes, including autoimmunity, autoinflammation, ageing and cancer, as well as organ-specific diseases that target the eyes, brain, lungs, heart, kidneys, skin and intestines. cGAS–STING is also involved in the response to many viral infections, which can be systemic or local. Thus, to deliver the anti-STING antibody to the site of interest, the anti-STING antibody of the present disclosure can comprise a cell, tissue, or organ specific targeting entity and / or is contained in or conjugated to a vehicle such as an exosome or nanoparticle for delivery to the respective cells, tissue, or organs.

[0237] Neuroinflammation driven by type-I interferons in the CNS is well established to exacerbate the progression of many CNS pathologies both acute and chronic. The role of STING is increasingly appreciated to instigate type-I IFN-mediated neuroinflammation and STING signaling has been detected in the brain. Thus, in certain embodiments, the anti- STING antibody of the present disclosure comprises a brain specific targeting entity and / or is contained in or conjugated to a vehicle such as an exosome or nanoparticle for delivery to the brain.

[0238] As it is well known in the art, delivery of biotherapeutics across the blood- brain barrier (BBB) is a challenge. For example, monoclonal antibodies do not cross the BBBNAI-5002871589v187efficiently, reaching a maximum of 0.11% at 1 hour after injection (Banks et al. Peptides 23 (2022), 2223-2226). The BBB is a specialized structural, physiological and biochemical barrier and serves as the first interface between the changeable environment of blood and the extracellular fluid in the CNS. The BBB regulates the homeostasis of the nervous system by strictly controlling the movement of small molecules or macromolecules from the blood to the brain. It only permits selective transport of molecules that are essential for brain function. In detail, more than 98% of small molecule drugs and almost 100% of large molecule drugs are precluded from drug delivery to brain (Redzic (2011) Fluids Barriers CNS 8, 3; Pardridge (2005) NeuroRx, 2, 3-14). Thus, the polypeptide and the antibody, antigen-binding fragment thereof, variant or derivative thereof, respectively may be modified in order to be able to penetrate the BBB.

[0239] For example, said antibodies and binding fragments can be fused to cell- penetrating peptides (CPPs), which qualify as brain targeting entity and which are usually short cationic and / or amphipathic peptides that have the ability to transport the associated molecular cargo (e.g., peptides, proteins, antibodies, etc.) across cellular membranes. However, also anionic CCPs have been reported. Examples are given in Sharma et al. (2016) Int. J. Mol. Sci. 17, 806 and instructions how to fuse an antibody with a CPP are for example provided in Gaston et al. (2019) Sci. Rep. 9, 18688 doi:10.1038 / s41598-019-55091-0. Furthermore, polyamine modification has been shown to dramatically increase the penetration of i.a. antibodies across the BBB (Poduslo and Curran (1996) J. Neurochem. 66, 1599-1609). The most investigated method to deliver macromolecules into the brain is via receptor-mediated transcytosis (RMT) and the main RMT receptors that have been studied are the transferrin receptor (TfR) and insulin receptor (IR). Thus, RMT receptors are also brain targeting entities. For example, bispecific antibodies have emerged as promising scaffolds to deliver therapeutic antibodies to the brain via engineering the antibody to incorporate one arm with specificity against a BBB RMT receptor, which drives their transmission across the BBB, and the other arm against a CNS therapeutic agent. Essentially, bispecific antibodies can be generated by fusion of antibody fragments such as Fabs, scFv or single domain antibodies into the N- or C-terminal of a convention IgG molecule or by heterodimerization strategies such as the “knobs-into-holes” technology developed by Genentech; see for details Neves et al. (2016) Trends Biotech. 34, 36-48. Thus, the anti- STING antibody of the present disclosure can be a bispecific antibody binding to STING and to a BBB RMT receptor.NAI-5002871589v188

[0240] As mentioned above, one of the main RMT receptors that have been studied is the transferrin receptor (TfR). Many platforms targeting TfR have been described including conventional high-affinity bivalent antibodies, bispecific antibodies, antibody fragments, peptides, antibody-fusion architectures, and most recently, a transport vehicle (TV) consisting of an Fc domain engineered to directly bind TfR; see Arguello et al., J Exp Med. 219 (2022), e20211057 as well as references cited therein. In Kariolis et al., Science translational medicine 12 (2020), 545, the development of a BBB transport vehicle (TV) comprising an engineered Fc fragment that exploits receptor-mediated transcytosis, i.e., that was engineered to bind the apical domain of hTfR, for CNS delivery of biotherapeutics by binding a highly expressed brain endothelial cell target is described. Thus, in certain embodiments, the antibody of the present disclosure comprises an engineered Fc domain which comprises at least the apical domain of hTfR.

[0241] In another approach, lipid nanoparticles / nanoexosomes can be used to deliver the antibodies or antigen-binding fragments of the present disclosure across the BBB. For example, dually decorated nanoliposomes with an anti-STING monoclonal antibody and an anti-RMT antibody, e.g., anti-TfR monoclonal antibody using biotin streptavidin conjugation can be used for improved delivery across the BBB. This principle is outlined in Markoutsa etal. (2012) Eur. J. Pharm. Biopharm. 81, 49-56) with an anti-A antibody instead of an anti-STING antibody.

[0242] Furthermore, as summarized in Tosi et al. (2013) (Curr. Med. Chem. 20, 2212-25), biodegradable nanoparticles formulated from poly(D,L-lactide-co-glycolide) (PLGA) have been extensively investigated for sustained and targeted delivery of different agents, including antibodies across the BBB. Thus, the antibodies and binding fragments of the present disclosure are conjugated to nanoparticles and nanoexosomes, respectively.

[0243] Accordingly, in certain embodiments, the anti-STING antibody of the present disclosure is capable of penetrating the BBB.

[0244] An antibody polypeptide of the invention may comprise, consist essentially of, or consist of a fusion protein. Fusion proteins are chimeric molecules which comprise, for example, an immunoglobulin STING-binding domain with at least one target binding site, and at least one heterologous portion, i.e., a portion with which it is not naturally linked in nature. The amino acid sequences may normally exist in separate proteins that are brought together in the fusion polypeptide or they may normally exist in the same protein but are placed in a new arrangement in the fusion polypeptide. Fusion proteins may be created, forNAI-5002871589v189example, by chemical synthesis, or by creating and translating a polynucleotide in which the peptide regions are encoded in the desired relationship.

[0245] The term “heterologous” as applied to a polynucleotide or a polypeptide, means that the polynucleotide or polypeptide is derived from a distinct entity from that of the rest of the entity to which it is being compared. For instance, as used herein, a “heterologous polypeptide” to be fused to an antibody, or an antigen-binding fragment, variant, or analog thereof is derived from a non-immunoglobulin polypeptide of the same species, or an immunoglobulin or non-immunoglobulin polypeptide of a different species.

[0246] The anti-STING antibody, optionally as fusion protein and / or labeled as described hereinbefore is then provided for various applications in accordance with standard techniques known in the art; see, e.g., Antibodies A Laboratory Manual 2nd edition, 2014 by Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA. Current advancements in therapeutic antibody design, manufacture, and formulation are described in Sifniotis et al., Antibodies 2019, 8(2), 36; https: / / doi.org / 10.3390 / antib8020036, wherein also developments in computational approaches for the strategic design of antibodies with modulated functions are discussed. 5.4 Pharmaceutical Compositions

[0247] The present disclosure also provides compositions comprising an anti-STING antibody or an antigen-binding fragment thereof disclosed herein (e.g., an anti-STING antibody or an antigen-binding fragment thereof of Section 5.2), or polynucleotides, vectors, or cells disclosed herein (e.g., polynucleotides, vectors, or cells of Section 5.3). In certain embodiments, the composition of the present disclosure is a pharmaceutical composition and further comprises a pharmaceutically acceptable carrier.

[0248] In certain embodiments, the pharmaceutical composition comprises an anti- STING antibody or an antigen-binding fragment thereof disclosed herein. In certain embodiments, the pharmaceutical composition comprises Ab1. In certain embodiments, the pharmaceutical composition comprises Ab2. In certain embodiments, the pharmaceutical composition comprises Ab3. In certain embodiments, the pharmaceutical composition comprises a therapeutically effective amount of the anti-STING antibody or antigen-binding fragment thereof.

[0249] The present disclosure also provides the pharmaceutical and diagnostic composition, respectively, in form of a pack or kit comprising one or more containers filled with one or more of the above-described ingredients, e.g., anti-STING antibody, polynucleotide(s), vector(s) or cell of the present disclosure. Associated with suchNAI-5002871589v190container(s) can be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, which notice reflects approval by the agency of manufacture, use or sale for human administration. In addition, or alternatively the kit comprises reagents and / or instructions for use in appropriate immuno- based diagnostic assays. The composition, e.g., kit of the present disclosure is of course particularly suitable for the treatment of autoimmune and (auto)inflammatory diseases discussed hereinbefore.

[0250] The pharmaceutical compositions of the present disclosure can be formulated according to methods well known in the art; see for example, Remington: The Science and Practice of Pharmacy (2000) by the University of Sciences in Philadelphia, ISBN 0-683- 306472. Examples of suitable pharmaceutical carriers are well known in the art and include standard buffer systems, like phosphate buffered saline solutions, water, emulsions, such as oil / water emulsions, various types of wetting agents, sterile solutions etc. Compositions comprising such carriers can be formulated by well-known conventional methods. These pharmaceutical compositions can be administered to the subject at a suitable dose. Administration of the suitable compositions may be effected by different ways, e.g., by intravenous, intraperitoneal, subcutaneous, intramuscular, intranasal, topical or intradermal administration or spinal or brain delivery. Aerosol formulations such as nasal spray formulations include purified aqueous or other solutions of the active agent with preservative agents and isotonic agents. Such formulations are preferably adjusted to a pH and isotonic state compatible with the nasal mucous membranes.

[0251] The present disclosure further provides a formulation suitable for in vivo administration (e.g., intravenous, intraperitoneal, subcutaneous, or intramuscular injection) comprising an anti-STING antibody or an antigen-binding fragment thereof disclosed herein (e.g., an anti-STING antibody or an antigen-binding fragment thereof of Section 5.2).

[0252] In certain embodiments, the concentration of the anti-STING antibody or antigen-binding fragment thereof in the presently disclosed formulation or pharmaceutical composition is at least 1 mg / ml. In certain embodiments, the presently disclosed formulation or pharmaceutical composition has a high purity of the anti-STING antibody or antigen- binding fragment thereof, for example, the monomeric purity of the anti-STING antibody is 80% or higher, 90% or higher, or 95% or higher in the formulation or pharmaceutical composition.NAI-5002871589v1915.5 Methods of Making and Using

[0253] As explained in detail above, the present disclosure also relates to use of the presently disclosed novel antagonistic anti-STING antibodies (e.g., anti-STING antibodies and antigen-binding fragments of Section 5.2) for treating a disease or a disorder. In certain embodiments, the disease or disorder is an autoimmune disease or an inflammatory disease. In certain embodiments, the inflammatory disease is an autoinflammatory disease. In certain embodiments, the inflammatory diseases is an acute inflammatory disease. In certain embodiments, the inflammatory diseases is a chronic inflammatory disease. Such antibodies preferably interfere with STING signaling resulting in reduction of inflammatory cytokine production. Thus, such antagonistic antibodies effect decreased STING signalling, i.e., reduce or prevent phosphorylation of TANK-binding kinase 1 (TBK1) and / or reduce the level ofhuman IFN- in differentiated human THP-1 derived macrophages, in particular in PMA-differentiated human THP-1-derived macrophages, which have been stimulated with the STING agonist cGAMP as well as in in vivo mouse models as illustrated in the appended Examples. The antagonistic anti-STING antibodies preferably bind to the C-terminal domain of STING.

[0254] The present disclosure also provides a method of treating a disease or a disorder (e.g., autoimmune and (auto)inflammatory diseases, for example acute and chronic inflammations), wherein the method comprises administering to a subject in need thereof a therapeutically effective amount of an antagonistic anti-STING antibody which preferably binds to the C-terminal domain of STING, and which is preferably the anti-STING antibody of the present disclosure. The present disclosure provides a method of treating an autoimmune and (auto)inflammatory disease in a subject by administering a therapeutically effective amount of said anti-STING antibody, wherein the administration of the antibody ameliorates, treats, or reduces the progression of at least one symptom of the disease in the subject.

[0255] The present disclosure further provides methods of inhibiting the phosphorylation of TBK1 in a cell with activated STING pathway, comprising contacting the cell with an anti-STING antibody or an antigen-binding fragment thereof disclosed herein (e.g., an anti-STING antibody or an antigen-binding fragment thereof of Section 5.2). In certain embodiments, the inhibition of the phosphorylation of TBK1 in the cells with activated STING pathway is indicated by a reduced pTBK1 level in the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof as compared to cells with activated STING pathway but without being contacted with theNAI-5002871589v192antibody or antigen-binding fragment thereof. In certain embodiments, the level of pTBK1 is measured by an immunocytochemistry assay or by Western blot. In certain embodiments, the cells are immune cells. In certain embodiments, the cells are lymphoid cells. In certain embodiments, the cells are myeloid cells. In certain embodiments, the cells are differentiated THP-1 cells. In certain embodiments, the cells are splenocytes. In certain embodiments, the differentiated THP-1 cells are macrophages. In certain embodiments, the THP-1 cells are human THP-1 cells. In certain embodiments, the differentiated THP-1 cells are PMA- differentiated human THP-1-derived macrophages. In certain embodiments, the cells are PBMCs. In certain embodiments, the splenocytes are mouse splenocytes. In certain embodiments, the STING pathway is activated by CMA or cGAMP. In certain embodiments, the method is performed in vitro, ex vivo or in vivo. In certain embodiments, the level of pTBK1 is measured as the ratio between pTBK1 and total TBK1 levels, or the ratio between pTBK1 and total GAPDH levels.

[0256] In certain embodiments, the pTBK1 level in the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less of the pTBK1 level in the cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof, as measured by Western Blot or ICC, and the cells are PMA-differentiated human THP-1- derived macrophages or mouse splenocytes with STING pathway activated by cGAMP or CMA. In certain embodiments, the pTBK1 level in the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof is about 40% or less of the pTBK1 level in the cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof as measured by ICC, wherein the cells are mouse splenocytes with STING pathway activated by CMA. In certain embodiments, the pTBK1 level in the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof is about 50% or less of the pTBK1 level in the cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof as measured by ICC, wherein the cells are mouse splenocytes with STING pathway activated by cGAMP. In certain embodiments, the pTBK1 level in the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof is about 60% or less of the pTBK1 level in the cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof as measured by Western Blot, wherein the cells are PMA-NAI-5002871589v193differentiated human THP-1-derived macrophages with STING pathway activated by cGAMP. In certain embodiments, the pTBK1 level in the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof is about 40% or less or 60% or less of the pTBK1 level in the cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof as measured by Western Blot, wherein the cells are PBMCs with STING pathway activated by cGAMP.

[0257] The present disclosure also provides methods inhibiting the release of an inflammatory cytokine in cells with activated STING pathway, comprising contacting the cells with an anti-STING antibody or an antigen-binding fragment thereof disclosed herein (e.g., an anti-STING antibody or an antigen-binding fragment thereof of Section 5.2). The present disclosure also provides methods inhibiting the release of an inflammatory cytokine in cells of a Trex1 null mouse, comprising administering to the mouse an anti-STING antibody or an antigen-binding fragment thereof disclosed herein (e.g., an anti-STING antibody or an antigen-binding fragment thereof of Section 5.2). In certain embodiments, the inhibition of the release of the inflammatory cytokine is indicated by a reduced level of the inflammatory cytokine released by the cells with activated STING pathway or the cells of the Trex1 null mouse and being contacted or administered with the antibody or antigen-binding fragment thereof as compared to cells with activated STING pathway or cells of a Trex1 null mouse but without being contacted or administered with the antibody or antigen-binding fragment thereof. In certain embodiments, the level of the inflammatory cytokine ismeasured by RT-qPCR. In certain embodiments, the inflammatory cytokine is IFN- . Incertain embodiments, the inflammatory cytokine is IL-6. In certain embodiments, the inflammatory cytokine is ISG-15. In certain embodiments, the cells are immune cells. In certain embodiments, the cells are lymphoid cells. In certain embodiments, the cells are myeloid cells. In certain embodiments, the cells are differentiated THP-1 cells. In certain embodiments, the cells are splenocytes. In certain embodiments, the differentiated THP-1 cells are macrophages. In certain embodiments, the THP-1 cells are human THP-1 cells. In certain embodiments, the differentiated THP-1 cells are PMA-differentiated human THP-1- derived macrophages. In certain embodiments, the splenocytes are mouse splenocytes. In certain embodiments, the cells of the Trex1 null mouse are tissue cells (e.g., heart tissue cells). In certain embodiments, the STING pathway is activated by CMA or cGAMP. In certain embodiments, the method is performed in vitro, ex vivo or in vivo.

[0258] In certain embodiments, the level of the inflammatory cytokine released by the cells with activated STING pathway and being contacted with the antibody or antigen-NAI-5002871589v194binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less of cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof, as measured by ELISA or HTRF, and the cells are PMA-differentiated human THP-1-derived macrophages or mouse splenocytes with STING pathway activated bycGAMP, and the inflammatory cytokine is IFN- . In certain embodiments, the IFN- levelreleased by the cells with activated STING pathway and being contacted with the antibody orantigen-binding fragment thereof is about 20% or less of the IFN- level released by the cellswith activated STING pathway but without being contacted with the antibody or antigen- binding fragment thereof as measured by ELISA, wherein the cells are mouse splenocyteswith STING pathway activated by cGAMP. In certain embodiments, the IFN- levelreleased by the cells with activated STING pathway and being contacted with the antibody orantigen-binding fragment thereof is about 50% or less or about 80% or less of the IFN- levelreleased by the cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof as measured by HTRF, wherein the cells are PMA-differentiated human THP-1-derived macrophages with STING pathway activated by cGAMP.

[0259] In certain embodiments, the IFN- mRNA level in a tissue sample (e.g., hearttissue) of a Trex1 null mouse and being administered with the antibody or antigen-bindingfragment thereof is about 20% or less or about 80% or less of the IFN- mRNA level in atissue sample (e.g., heart tissue) of a Trex1 null mouse but without being administered with the antibody or antigen-binding fragment thereof as measured by RT-qPCR. In certain embodiments, the IL-6 mRNA level in a tissue sample (e.g., heart tissue) of a Trex1 null mouse and being administered with the antibody or antigen-binding fragment thereof is about 60% or less or about 80% or less of the IL-6 mRNA level in a tissue sample (e.g., heart tissue) of a Trex1 null mouse but without being administered with the antibody or antigen- binding fragment thereof as measured by RT-qPCR. In certain embodiments, the ISG-15 mRNA level in a tissue sample (e.g., heart tissue) of a Trex1 null mouse being administered with the antibody or antigen-binding fragment thereof is about 80% or less of the ISG-15 mRNA level in a tissue sample (e.g., heart tissue) of a Trex1 null mouse without being administered with the antibody or antigen-binding fragment thereof as measured by RT- qPCR.

[0260] The present disclosure also provides methods of inhibiting STING pathway in a subject in need thereof, comprising administering to the subject a therapeutically effectiveNAI-5002871589v195amount of an anti-STING antibody or an antigen-binding fragment thereof disclosed herein (e.g., an anti-STING antibody or an antigen-binding fragment thereof of Section 5.2). The present disclosure also provides methods of inhibiting the phosphorylation of TBK1 in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an anti-STING antibody or an antigen-binding fragment thereof disclosed herein (e.g., an anti-STING antibody or an antigen-binding fragment thereof of Section 5.2). The present disclosure also provides methods of inhibiting the release of an inflammatory cytokine in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an anti-STING antibody or an antigen-binding fragment thereof disclosed herein (e.g., an anti-STING antibody or an antigen-binding fragment thereof of Section 5.2).

[0261] In certain embodiments, inhibiting STING pathway is indicated by a reduced level of pTBK1 and / or a reduced level of an inflammatory cytokine in a sample of the subject. In certain embodiments, inhibiting the phosphorylation of TBK1 is indicated by a reduced level of pTBK1 in a sample of the subject. In certain embodiments, inhibiting the release of an inflammatory cytokine is indicated by a reduced level of the inflammatory cytokine in a sample of the subject. In certain embodiments, the sample is a blood sample, a serum sample, a tissue sample, or a cell sample. In certain embodiments, the inflammatorycytokine is IL-6, IP-10, IFN- , and / or ISG-15. In certain embodiments, the reduced level ofpTBK1 and / or reduced level of the inflammatory cytokine is reduced as compared to the level of pTBK1 and / or the level of the inflammatory cytokine measured before the administration of the anti-STING antibody or antigen-binding fragment thereof or to a reference level. In certain embodiments, the level of pTBK1 is measured by an immunoassay. In certain embodiments, the immunoassay is immunocytochemistry assay or Western blot. In certain embodiments, the level of inflammatory cytokine is measured by an immunoassay or RT-PCR. In certain embodiments, the immunoassay is ELISA, HTRF, or MSD multiple assay. In certain embodiments, the level of pTBK1 is measured as the ratio between pTBK1 and total TBK1 levels, or the ratio between pTBK1 and total GAPDH levels. In certain embodiments, the subject has a disease or a disorder disclosed herein, for example, in Section 5.5.

[0262] The present disclosure also provides methods of treating a disease or a disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an anti-STING antibody or an antigen-binding fragment thereof disclosed herein (e.g., an anti-STING antibody or an antigen-binding fragment thereof of Section 5.2).NAI-5002871589v196In certain embodiments, the disease or disorder is an inflammatory disease or an autoimmune disease. In certain embodiments, the inflammatory disease is an autoinflammatory disease. In certain embodiments, the inflammatory diseases is an acute inflammatory disease. In certain embodiments, the inflammatory diseases is a chronic inflammatory disease.

[0263] The present disclosure also provides anti-STING antibodies or antigen-binding fragments thereof disclosed herein (e.g., anti-STING antibodies or antigen-binding fragments thereof of Section 5.2) for use in treating a disease or a disorder in a subject in need thereof. In certain embodiments, the disease or disorder is an inflammatory disease or an autoimmune disease. In certain embodiments, the inflammatory disease is an autoinflammatory disease. In certain embodiments, the inflammatory diseases is an acute inflammatory disease. In certain embodiments, the inflammatory diseases is a chronic inflammatory disease.

[0264] The present disclosure also provides use of anti-STING antibodies or antigen- binding fragments thereof disclosed herein (e.g., anti-STING antibodies or antigen-binding fragments thereof of Section 5.2) in the manufacture of a medicament for treating a disease or a disorder in a subject in need thereof. In certain embodiments, the disease or disorder is an inflammatory disease or an autoimmune disease. In certain embodiments, the inflammatory disease is an autoinflammatory disease. In certain embodiments, the inflammatory diseases is an acute inflammatory disease. In certain embodiments, the inflammatory diseases is a chronic inflammatory disease.

[0265] In certain embodiments, the inflammatory disease is Aicardi-Goutières syndrome (AGS), STING-associated vasculopathy of infancy (SAVI), systemic lupus erythematosus (SLE), COPA syndrome. In certain embodiments, the autoimmune diseases is amyotrophic lateral sclerosis (ALS), Niemann-Pick disease type C (NPC), multiple sclerosis, familial chilblain lupus (FCL), rheumatoid arthritis, or Sjögren’s syndrome. In certain embodiments, the disease or disorder is lupus. In certain embodiments, the disease or disorder is SLE. In certain embodiments, the disease or disorder is Sjögren’s syndrome. In certain embodiments, the disease or disorder is secondary Sjögren’s syndrome.

[0266] In certain embodiments, the inflammatory disease is Alzheimer disease, ischemic brain injury, Parkinson disease, neurodegeneration, Huntington disease, frontotemporal dementia, age-dependent macular degeneration, traumatic brain injury, nonalcoholic steatohepatitis, alcoholic liver disease, acute pancreatitis, silica- induced fibrosis, sepsis, myocardial infarction, chronic heart failure, or colorectal cancer.

[0267] Dosage regimen of the treatment will be determined by the attending physician and clinical factors. As is well known in the medical arts, dosages for any oneNAI-5002871589v197patient depends upon many factors, including the patient’s size, body surface area, age, the particular compound to be administered, sex, time and route of administration, general health, and other drugs being administered concurrently.

[0268] The present disclosure further provides methods of making an anti-STING antibody or an antigen-binding fragment thereof, comprising culturing a cell disclosed herein (e.g., a cell of Section 5.3) and isolating the antibody or antigen-binding fragment thereof.

[0269] The present disclosure further provides methods of detecting STING in a biological sample, comprising contacting the biological sample with an anti-STING antibody or an antigen-binding fragment thereof disclosed herein (e.g., an anti-STING antibody or an antigen-binding fragment thereof of Section 5.2); and detecting the binding between the antibody or antigen-binding fragment thereof and STING in the biological sample. In certain embodiments, the detecting comprises using flow cytometry, immunohistochemistry (IHC), Western Blot analysis, ELISA, or mass spectrometry. In certain embodiments, the antibody or antigen-binding fragment thereof is attached to a detectable label. In certain embodiments, the detectable label is an enzyme, a radioisotope, a fluorescent compound, a chemiluminescent compound, a bioluminescent compound, a tag, a flag or a heavy metal.

[0270] The present disclosure further provides methods for screening for an antibody or an antigen-binding fragment thereof that can specifically bind human STING, comprising measuring the binding of the antibody or antigen-binding fragment thereof to an epitope located within the C-terminal domain of STING, and selecting the antibody if the binding affinity is higher than an isotype control antibody. In certain embodiment, the binding is measured by ELISA. In certain embodiment, the C-terminal domain of STING comprises the amino acid sequence of SEQ ID NO:31. In certain embodiment, the epitope comprises amino acid residues S358, G367, M368, E369, and P371 of human STING, wherein the amino acid positions are numbered according to the positions in the amino acid sequence of SEQ ID NO:34. In certain embodiments, the epitope is a conformational epitope. In certain embodiments, the epitope is a linear epitope.

[0271] Several documents are cited throughout the text of this specification. The contents of all cited references (including literature references, issued patents, published patent applications as cited throughout this application including the background section and manufacturer’s specifications, instructions, etc.) are hereby expressly incorporated by reference; however, there is no admission that any document cited is indeed prior art as to the present disclosure.NAI-5002871589v198

[0272] A more complete understanding can be obtained by reference to the following specific examples which are provided herein for purposes of illustration only and are not intended to limit the scope of the invention. 6. EXAMPLES

[0273] The following is a description of various methods and materials used in the studies. They are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present disclosure, and are not intended to limit the scope of what the inventors regard as their invention, nor are they intended to represent that the experiments below were performed and are all of the experiments that may be performed. It is to be understood that exemplary descriptions written in the present tense were not necessarily performed, but rather that the descriptions can be performed to generate the data and the like associated with the teachings of the present disclosure. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, percentages, etc.), but some experimental errors and deviations should be accounted for. 6.1 Example 1: Isolation, identification, cloning and recombinant expression of anti-STING antibodies

[0274] The cDNA sequences encoding antibodies that bind to recombinant human STING protein (rhSTING) or recombinant mouse STING protein (rmSTING) were derived from a de-identified blood lymphocyte library collected from healthy elderly subjects applying the proprietary Reverse Translational Medicine™ (RTM) technology platform, Neurimmune AG, 8952 Schlieren / Zurich, Switzerland. Human blood samples were collected from healthy elderly subjects with written informed consent and study approval by the Ethics Committee of the Canton of Zurich. The study was conducted in compliance with the Helsinki Declaration. The isolated B cells were characterized as CD22+ CD27+ and IgD- IgM- CD3- CD56- and CD8a-. They were screened for the expression of antibodies binding to the initial target preparation using conventional ELISA technology. Positive hits were counter-screened to exclude clones cross-reacting with unrelated proteins. The antibodies generated by the library were in IgA1 allotype. STING-reactive B-cell clones were selected and subjected to cDNA cloning of IgG heavy and light-chain variable region sequences, which were sub-cloned into expression vectors encoding human IgG1 constant domainNAI-5002871589v199sequences. The resulting expression constructs were transiently expressed in the ExpiCHO cell line and purified by protein A affinity chromatography; see FIGS. 1A-1C details. 6.2 Example 2: Anti-STING antibody Ab1 selectively bound to the C-terminal domain of human and mouse recombinant STING

[0275] Antibody Ab1 was tested for its binding affinity to recombinant human and mouse STING protein and to C-terminus human STING peptide (SEQ ID NO:31), C- terminus cyno STING peptide (SEQ ID NO:32), and C-terminus rat STING peptide (SEQ ID NO:33) via a direct enzyme-linked immunosorbent assay (ELISA). The ELISA was conducted on recombinant human STING (AlexoTech) or recombinant mouse STING (LSBio) using 96 well half-area microplates (Corning), in order to assess binding efficacy and specificity of an anti-STING antibody. The microplates were coated with a solution containing 5 g / mL of the recombinant protein to immobilize it on the surface of the wells. This allows for the binding of specific antibodies to the protein of interest. The microplates were then blocked with 2% BSA in phosphate-buffered saline with Tween-20 (PBS-T) to prevent non-specific binding of antibodies to the plate wells. After blocking, serial dilutions of STING antibodies were added to each well and the plates were incubated for 2 hours at room temperature (RT). This incubation step allows for the binding of specific antibodies to the immobilized protein. The plates were then washed with PBS-T to remove any unbound antibodies. The bound antibodies were detected by incubating with a horseradish peroxidase (HRP)-conjugated anti-human Fc secondary antibody (Jackson ImmunoResearch) for 60 minutes at RT. This secondary antibody is specific for the constant region of human IgG and is conjugated to HRP which will generate a signal upon addition of the chromogenic substrate. The chromogenic substrate, 3,3’,5,5’-tetramethylbenzidine (TMB, Clinical Science) was added to each well and the plates were incubated for a short period of time. The HRP enzyme catalyses the conversion of TMB to a blue product, which can be quantified by measuring the absorbance at 450 nm using a plate reader (ThermoFisher). To estimate the half-maximal concentration (EC50) of the recombinant protein, log-transformed absorbance values were fitted to a four-parameter log(agonist) vs. response - variable slope curve using the formula Y=Bottom + (Top-Bottom) / (1+10^((LogEC50-X)*HillSlope)) in GraphPad Prism version 9.3. This curve fitting allows for the determination of the concentration at which the protein elicits half of the maximal response. A robust and specific binding affinity was detected on both recombinant human and mouse STING protein as well as on the C- terminal portion of the human, cyno and rat STING protein, with EC50 values of 844 pM, 475 pM, 257 pM, 233 pM and 648 pM respectively (FIG. 2).NAI-5002871589v1100

[0276] This data demonstrated the Ab1’s high specificity and affinity for the STING protein. On the other hand, no detectable binding was observed on the BSA and E. coli lysates, indicating the antibody’s selectivity for STING. Additionally, to ensure the specificity of the antibody, a human IgG control (isotype control antibody) was included as a negative control in the ELISA. The negative control demonstrated no binding, further confirming the specificity of the Ab1 antibody towards the STING protein. 6.3 Example 3: Anti-STING antibodies bound to human and mouse recombinant STING in a concentration-dependent manner

[0277] To further elucidate the binding selectivity and specificity of anti-STING antibodies, Western Blot analyses were performed on recombinant STING protein. Western Blot is the procedure used to detect binding of the anti-STING antibody to human recombinant STING, mouse recombinant STING, as well as BSA and E. coli lysates. For this analysis, 0.01, 0.05, 0.1 and 0.5 g of recombinant human STING from AlexoTech; 0.1 and 0.5 g of mouse STING (LSBio); as well as 0.1 and 0.5 g of BSA (ThermoScientific) and E. coli lysates were denatured using LDS sample buffer and 4% -Mercapto-Ethanol for 10 minutes at 95°C. The samples were loaded onto a 4-12% Bis-Tris Protein Gel and run for 45 minutes at 200V in 1 x MOPS SDS Running Buffer. The proteins were then transferred onto a nitrocellulose membrane using iBlot for 7 minutes and 20 V. The membrane was blocked with 5% skim milk in PBS-T for 60 minutes at room temperature, to avoid non-specific binding of the antibody to the membrane. Subsequently, the anti-STING antibody was added in blocking buffer at 10 g / mL and incubated overnight at 4°C. After 3 washes with PBS-T, the membrane was incubated with HRP-conjugated anti-human-Fc secondary antibody for 60 minutes at room temperature and washed again 3 times with PBS-T. Finally, the membrane was incubated with ECL (Amersham) for 2 minutes and the resulting chemiluminescence signal was detected using Image Quant LAS 4000. As depicted in FIGS. 3A-3C, the obtained results were in agreement with those obtained by ELISA in Example 2, as a strong binding affinity of anti-STING antibody Ab1 to both human and mouse recombinant STING protein. Furthermore, the analysis of the binding specificities of Ab1 anti-STING antibody to human STING protein, revealed a concentration dependent binding. Concentration dependent binding to human STING protein was also observed for Ab2 and Ab3 anti-STING antibodies. 6.4 Example 4: Anti-STING antibodies specifically bound to human cells

[0278] Immunocytochemistry (ICC) was performed to analyze the binding of the anti- STING antibodies in human peripheral blood mononuclear cells (PBMCs) and human derived splenocytes. Cryopreserved PBMCs obtained from StemCells were thawed in RPMINAI-5002871589v1101medium supplemented with 10% Fetal Bovine Serum (FBS), and human splenocytes from ZenBio were thawed in complete Hematogrow medium (Innoprot). The cells were counted and resuspended in PBS at a concentration of 1 million cells / ml, and 700 l of cell suspension was added to each glass coverslip placed inside the wells of a 24-well plate. The cells were allowed to settle for 30 minutes at room temperature (RT) before fixation in 4% paraformaldehyde (PFA) in PBS for 10 minutes at RT. After fixation, the cells were washed three times with PBS and blocked with 1% BSA in PBS for 60 minutes at RT. The primary anti-STING antibody was incubated at a concentration of 1 g / ml in PBS overnight at 4°C. The cells were then washed three times with PBS to remove unbound antibodies before incubation with Cy3-conjugated secondary anti-human antibody (Jackson ImmunoResearch) diluted 1:250 in PBS for 60 minutes at RT. The cells were washed three times with PBS to remove excess fluorescent antibody, and the nuclei were stained with DAPI. Finally, the cells were washed three times with PBS and the coverslip was mounted onto a glass slide for fluorescent signal detection by imaging (SlideView_VS200, Olympus). FIG. 4A shows specific binding of antibody Ab1 in human PBMCs, while no positive Cy3 signal was observed with a human IgG control (isotype control antibody) and with an anti-human control (secondary antibody control). Similar results were obtained in human derived splenocytes (FIG. 4B), demonstrating specific target binding of Ab1 to human cells. Same results were obtained with antibodies Ab2 and Ab3, i.e., both antibodies show specific binding to human cell lines. 6.5 Example 5: Anti-STING antibodies detected STING in human cancer tissue

[0279] To analyze the binding of the anti-STING antibodies in human lung adenocarcinoma, immunofluorescence was employed. Firstly, paraffin-fixed sections derived from lung adenocarcinoma and other tissues expressing STING, were subjected to a xylene / alcohol series to remove the wax, followed by washing in water. The sections were then treated with citrate buffer (0.1 M citric acid in water) for 15 minutes and microwaved. Blocking of the sections was carried out by incubating them in a blocking buffer (5% Goat serum, 5% Horse serum, 4% BSA in PBS) for 60 minutes at RT. The anti-STING antibody was incubated in PBS at a concentration of 50 g / ml, overnight at 4°C in the dark. Subsequently, the sections were washed in water to remove unbound antibody and then incubated for 60 minutes in the dark with a Cy3-conjugated anti-human IgG secondary antibody, which was diluted 1:400 in PBS. After washing in water, the nuclei were stained with DAPI for 7 minutes. Finally, coverslips were mounted on the slides, and the resulting stained sections were imaged using the SlideView_VS200 (Olympus) imaging system andNAI-5002871589v1102analyzed with Olyvia (Olympus). The specific binding of the anti-STING antibody Ab1 to human lung adenocarcinoma tissue is depicted in FIG. 5. The Ab1 staining showed a clear and specific perinuclear binding pattern as expected. The results were similar to those obtained with a commercially available anti-STING antibody, and no signal was detected with the secondary antibody controls, including anti-human and anti-rabbit controls. These findings confirmed the specificity of Ab1 for detecting STING in human lung adenocarcinoma tissue. Moreover, the specificity of the Ab1 was further confirmed by the lack of any significant signal detected in the anti-human and anti-rabbit control samples, demonstrating the reliability of the experimental results. Similarly, antibodies Ab2 and Ab3 were also able to detect STING in human lung adenocarcinoma tissue as well as in colon cancer tissue. 6.6 Example 6: Anti-STING antibody Ab1 bound STING on non-demented human brain tissue

[0280] To further elucidate the binding specificity on human tissue, Ab1 was tested on non-demented human brain tissue. The binding specificity of anti-STING antibodies in healthy brain tissues was analyzed using immunohistochemistry. To perform this analysis, paraffin-fixed sections derived from human brain tissue were initially subjected to a dewaxing procedure using a series of xylene / alcohol solutions, followed by washing with water. The sections were then pre-treated with a citrate buffer (0.1 M citric acid in water) for 15 minutes, and subsequently with EDTA for 10 minutes. To prevent non-specific binding, the sections were blocked in a blocking buffer (5% Goat serum, 5% Horse serum, 4% BSA in PBS) for 60 minutes at room temperature. To further block endogenous peroxidases, sections were then incubated with 3% hydrogen peroxide (H2O2) diluted in methanol (HRP-blocking) for 10 minutes. Ab1 anti-STING antibody at a concentration of 10 g / ml in PBS was then incubated overnight, followed by washing with water to remove unbound antibody. Next, the sections were incubated for 60 minutes with Biotin-SP goat anti-human IgG secondary antibody (Jackson ImmunoResearch) diluted 1:350 in PBS for detection. After washing off the excess antibody, the sections underwent an ABC step for 30 minutes using the Vectastain Elite ABC Kit (Vectorstain Laboratories), following the manufacturer’s instructions. Subsequently, the sections were washed in water and then stained with Hematoxylin (Roth) for 3 minutes to visualize the nuclei. Finally, coverslips were mounted on the slides and the resulting stained sections were imaged using the SlideView_VS200 (Olympus) imaging system and analyzed with Olyvia (Olympus). FIGS. 6A-6D show positive signal in tissues stained with Ab1 and with the positive control commercial anti-STING antibody, while noNAI-5002871589v1103signal was observed with human IgG control (isotype control antibody) and with anti-human IgG and anti-mouse IgG2a controls (secondary antibody controls). This data demonstrated that Ab1 binds STING on non-demented human brain sections. 6.7 Example 7: Anti-STING antibody Ab1 inhibited the release of IFN- inmouse splenocytes

[0281] To better understand the potential impact of the anti-STING antibody Ab1 onthe STING pathway, it was evaluated whether the levels of IFN- were altered in thepresence of the antibody. For this purpose, an enzyme-linked immunosorbent assay (ELISA)has been performed to quantify the levels of IFN- in the cell medium after activation of theSTING pathway with cGAMP. This assay enabled to investigate whether the anti-STING antibody has any effect on the downstream signaling of the STING pathway and subsequentproduction of IFN- . These findings provide insight into the potential immunomodulatoryproperties of the antibody and its potential for therapeutic use in modulating the immuneresponse. The VeriKine mouse IFN- ELISA immunoassay (Pbl Assay Science) wasemployed in this study to measure the levels of IFN- in the culture medium of mousesplenocytes cells that were treated with cGAMP (Invivogen) and anti-STING antibody. The aim was to investigate whether treatment with the STING antibody inhibits the STINGpathway triggered by cGAMP, as indicated by a decrease in IFN- released by cells.Cryopreserved mouse splenocytes obtained from ZenBio were thawed in Hematogrow medium, counted, and seeded at a concentration of 1 million cells / ml of medium in a 96-well plate. The cells were treated with 5 g / ml of anti-STING antibody and incubated at 37°C for 2 hours. Following incubation, the supernatant was replaced with a cGAMP solution (23.6 g / ml cGAMP in Hematogrow medium) to activate the STING pathway. The cells were then incubated for an additional 24 hours at 37°C, and the cell medium was collected and assessedfor IFN- levels using the VeriKine mouse IFN- ELISA immunoassay, according to themanufacturer’s instructions. The impact of the anti-STING antibody Ab1 on IFN- releasewas investigated in vitro using mouse splenocytes. To activate the STING pathway andinduce IFN- release, the small molecule cGAMP was used, and IFN- levels in the cellmedium were quantified by ELISA. FIG. 7 shows that stimulation with cGAMP resulted in asignificant increase in IFN- levels in the medium of mouse splenocytes. In contrast,treatment with Ab1 resulted in a notable decrease in the concentration of IFN- released bythe cells, while the human IgG control had no effect on IFN- levels. These results suggestedthat Ab1 was able to inhibit the release of IFN- in mouse splenocytes in vitro and indicateda potential therapeutic application of the antibody in STING-mediated autoimmune diseases.NAI-5002871589v11046.8 Example 8: Anti-STING antibody Ab1 inhibited IFN- release indifferentiated human THP-1 cells

[0282] In order to gain further insight into the impact of the anti-STING antibody Ab1 on cellular responses, Homogeneous Time-Resolved Fluorescence (HTRF) analysis hasbeen conducted to evaluate the effect of the antibody on IFN- levels in THP-1 cell medium.Activation of the STING pathway was induced using cGAMP. The HTRF assay provided ahighly sensitive method for detecting IFN- levels in the cell medium, allowing for a detailedassessment of the impact of Ab1 on STING pathway activation. The human IFN- HTRFassay from Cisbio Bioassays (Perkin Elmer) was utilized to measure the levels of IFN- inthe culture medium of differentiated THP-1 cells, which have been treated with cGAMP (Invivogen) and anti-STING antibody, to investigate if the treatment with the antibodyinhibits the STING pathway triggered by cGAMP (as indicated by a reduction in IFN-released by cells). For this purpose, THP-1 cells were plated at a concentration of 1 million cells / ml in a 96 well plate, using RPMI medium supplemented with 10% FBS. Next, cells were treated with 500 ng / ml PMA and incubated overnight at 37°C for differentiation. After differentiation, the supernatant was replaced with cGAMP solution (consisting of 23.6 g / ml cGAMP in RPMI) for 3 hours at 37°C. Then, the supernatant was removed, and the cells were provided with fresh RPMI supplemented with different concentrations of anti-STING antibody. The antibody was then incubated overnight at 37°C. Finally, the cell medium wascollected and assessed for IFN- content via HTRF assay following the instructions of themanufacturer. To gain a deeper understanding of the effects of the anti-STING antibody Ab1in vitro, IFN- levels have been assessed in differentiated THP-1 cells using HomogeneousTime-Resolved Fluorescence (HTRF) following activation of the STING pathway withcGAMP. As shown in FIG. 8, STING activation resulted in increased IFN- levels in the cellmedium. However, treatment with Ab1 led to a significant decrease in IFN- levels in a dose-dependent manner, indicating that the antibody effectively inhibits IFN- release indifferentiated THP-1 cells. Conversely, treatment with the human IgG control did not lead toany reduction in IFN- levels, confirming the specificity of the observed effect. These resultssuggested that Ab1 had the potential to modulate STING-mediated immune responses in vitro. 6.9 Example 9: Anti-STING antibodies antagonize STING activation by cGAMP and reduce pTBK1 levels in differentiated THP-1 cells

[0283] The STING pathway can be activated by incubation with cGAMP, resulting in the phosphorylation of TBK1. To investigate the impact of the anti-STING antibodies Ab1,NAI-5002871589v1105Ab2, and Ab3 on the phosphorylation status of TBK1 (pTBK1), THP-1 cell lysates were analyzed by Western Blot. TBK1 is a key protein involved in STING signaling pathway activation, and its phosphorylation is a critical event in this process. The inhibition of the STING pathway by different concentrations of STING antibody in differentiated THP-1 cells was investigated using Western Blot analysis, with a reduction in TBK1 phosphorylation serving as the readout. The level of pTBK1 was analyzed in cells that were either untreated or treated with cGAMP alone or in combination with an anti-STING antibody, i.e., Ab1, Ab2, or Ab3. THP-1 cells were initially plated at a concentration of 1 million cells / ml in T25 flasks containing RPMI medium supplemented with 10% FBS. The cells were then provided with 500 ng / ml PMA and incubated overnight at 37°C to allow differentiation. After this period, the cells were treated with cGAMP (BioLog) solution consisting of 750 ng / ml cGAMP in RPMI for 3 hours at 37°C. Following removal of the supernatant, the cells were incubated with fresh RPMI supplemented with different concentrations of anti-STING antibody overnight at 37°C. The subsequent preparation of THP-1 cell lysates involved washing and scraping the cells in PBS, collecting the cell suspension, and pelleting it for 5 minutes at 11’000 rpm, 4°C. The supernatant was then discarded, and the pellet was resuspended in about 20 l of RIPA buffer containing protease inhibitors. After incubation for 10 minutes with occasional vortexing, the lysates were pelleted at 13’000 rpm for 10 minutes and transferred to a new tube. The total protein content of the samples was determined using the BCA method (ThermoScientific), and 30 g of cell lysates were denatured using LDS sample buffer and 4% -Mercapto-Ethanol for 10 minutes at 95°C. The samples were loaded onto a 4-12% Bis-Tris Protein Gel and run for 45 minutes at 200V in 1X MOPS SDS Running Buffer. The proteins were subsequently transferred onto a PVDF membrane using iBlot for 7 minutes and 20 V. The membrane was blocked with 5% BSA in PBS-T for 60 minutes at room temperature to prevent non-specific binding of the antibody to the membrane. The anti- human pTBK1 antibody (Cell Signaling) was then added in blocking buffer at a 1:1000 dilution and incubated overnight at 4°C. After 3 washes with PBS-T, the membrane was incubated with HRP-conjugated anti-rabbit secondary antibody for 60 minutes at room temperature and washed again 3 times with PBS-T. Finally, the membrane was incubated with ECL (Amersham) for 2 minutes, and the resulting chemiluminescence signal was detected using Image Quant LAS 4000. The resulting ratio between pTBK1 and totTBK1 signals was quantified using ImageJ and reported on GraphPad after staining the same membrane with an anti-human total TBK1 antibody. The STING pathway can be activated by incubation with cGAMP, resulting in the phosphorylation of TBK1. To assess the effect ofNAI-5002871589v1106the anti-STING antibody Ab1 on TBK1 phosphorylation, THP-1 cells were incubated with different concentrations of the antibody after STING activation with cGAMP, and the levels of phospho-TBK1 (pTBK1) were analyzed by Western Blot. As shown in FIG. 9A and FIG. 9B, a positive control was included, consisting of cells treated with a small molecule STING antagonist (H-151). No significant changes in pTBK1 levels were observed in cells treated with a human IgG control (isotype control antibody), while a concentration-dependent decrease in pTBK1 levels was detected with increasing concentrations of Ab1 (FIG. 9B). Total TBK1 levels were used for normalization and quantification of pTBK1 levels. The results indicated that the incubation of differentiated THP-1 cells with Ab1 led to a concentration-dependent decrease in pTBK1 levels after cGAMP application.

[0284] Similarly, to assess the effect of the anti-STING antibodies Ab2 and Ab3 on TBK1 phosphorylation, THP-1 cells were incubated with different concentrations of the antibody after STING activation with cGAMP, and Western Blot analysis showed a decrease in pTBK1 levels during incubation with the respective antibody. 6.10 Example 10: Exploring Ab1 antibody-mediated inhibition of the STING- TBK1 pathway in PBMCs through quantitative analysis of phospho-TBK1 (pTBK1) levels via Western Blotting

[0285] The STING pathway can be activated by incubation with cGAMP, resulting in the phosphorylation of TBK1. To investigate the impact of the anti-STING antibody Ab1 on the phosphorylation status of TBK1 (pTBK1), PBMCs lysates were analyzed by Western Blot. TBK1 is a key protein involved in STING signaling pathway activation, and its phosphorylation is a critical event in this process. The inhibition of the STING pathway by different concentrations of STING antibody in PBMCs was investigated using Western Blot analysis, with a reduction in TBK1 phosphorylation serving as the readout.

[0286] The level of pTBK1 was analyzed in cells that were either untreated or treated with cGAMP alone or in combination with Ab1. PBMCs were plated at a concentration of 1.5 million cells / well in 6-well plates containing RPMI medium supplemented with 10% FBS and 1 mM sodium pyruvate. Cells were incubated with different concentrations of anti- STING antibody for 4 hours at 37°C followed by treatment with a cGAMP (BioLog) solution consisting of 2.5 μg / ml cGAMP in RPMI for 4 hours at 37°C. The subsequent preparation of PBMCs lysates involved scraping the cells in media, collecting the cell suspension, and pelleting it for 5 minutes at 11’000 rpm, 4°C. The pellet was resuspended in PBS and an additional centrifugation step was performed for 5 minutes at 11’000 rpm, 4°C. The resulting pellet was then resuspended in 15 l PhosphoSafe extraction reagent (Merck). AfterNAI-5002871589v1107incubation for 10 minutes with occasional vortexing, the lysates were pelleted at 13’000 rpm for 10 minutes and transferred to a new tube. The total protein content of the samples was determined using the BCA method (ThermoScientific), and 10 g of cell lysates were denatured using LDS sample buffer and Reducing Agent (ThermoScientific) for 5 minutes at 95°C. The samples were loaded onto a Bolt 4-12% Bis-Tris Plus Gel and run for 45 minutes at 150 V in 1X MOPS SDS Running Buffer. The proteins were subsequently transferred onto a PVDF membrane using iBlot for 7 minutes and 20 V. The membrane was blocked with 5% BSA in PBS-T for 60 minutes at room temperature to prevent non-specific binding of the antibody to the membrane. The anti-human pTBK1 antibody (Cell Signaling) was then added in blocking buffer at a 1:1000 dilution and incubated overnight at 4°C. After 3 washes with PBS-T, the membrane was incubated with HRP-conjugated anti-rabbit secondary antibody for 60 minutes at room temperature and washed again 3 times with PBS-T. Finally, the membrane was incubated with Femto Maximum Sensitivity Substrate (ThermoScientific) for 2 minutes, and the resulting chemiluminescence signal was detected using Image Quant LAS 4000. The resulting ratio between pTBK1 and totTBK1 or GAPDH signals was quantified using ImageJ and reported on GraphPad after staining the same membrane with an anti- human total TBK1 antibody or an anti-human GAPDH antibody.

[0287] The STING pathway can be activated by incubation with cGAMP, resulting in the phosphorylation of TBK1. To assess the effect of the anti-STING antibody Ab1 on TBK1 phosphorylation, PBMCs were incubated with different concentrations of the antibody after STING activation with cGAMP, and the levels of phospho-TBK1 (pTBK1) were analyzed by Western Blot.

[0288] As shown in FIGS. 10A-10C, a positive control was included, consisting of cells treated with a small molecule STING inhibitor (H151). No significant changes in pTBK1 levels were observed in cells treated with a human IgG control (isotype control antibody), while a decrease in pTBK1 levels was detected in cells incubated with Ab1 prior cGAMP treatment (FIGS. 10B & 10C). Total TBK1 and GAPDH levels were used for normalization and quantification of pTBK1 levels. The results indicated that the incubation of PBMCs with Ab1 led to a concentration-dependent decrease in pTBK1 levels after cGAMP application. 6.11 Example 11: Anti-STING antibody Ab1 antibody mediated decrease in pTBK1 levels in mouse splenocytes treated with CMA / cGAMP

[0289] The phosphorylation levels of TBK1 were evaluated in mouse splenocytes through immunocytochemistry to investigate the impact of antibody Ab1 on pTBK1 levelsNAI-5002871589v1108following ex vivo CMA-induced activation of the STING pathway. Immunocytochemistry (ICC) was employed to investigate the potential of different concentrations of STING antibody to attenuate CMA / cGAMP-mediated activation of the STING pathway in mouse splenocytes, using the reduction of TBK1 phosphorylation as a readout. To achieve this, splenocytes were freshly isolated from C57BL / 6J mouse spleens: upon harvesting, up to two spleens were placed in a 5 cm dish containing collagenase IV working solution (2 mg / ml of collagenase IV, diluted in RPMI supplemented with 2% FBS and 20 mM HEPES). To facilitate collagenase IV-mediated digestion, spleens were cut into pieces, and incubated for 30 min at 37°C. At the end of incubation time, spleens were smashed through a 40 m cell strainer to obtain a cell suspension, which was pelleted (300 g for 5 min) and subjected to red blood cell lysis, by incubation with ACK buffer (Gibco) for 5 minutes at room temperature. The reaction was stopped by addition of Hematogrow medium. After a filtration step, cells were pelleted, resuspended in Hematogrow medium and kept on ice until further use.

[0290] For the purpose of this experiment, the obtained mouse splenocytes were seeded in Hematogrow medium at a density of 1 million cells / ml in each well of a 6-well plate. Subsequently, CMA or cGAMP solutions (comprising Hematogrow supplemented with 250 g / ml CMA or 750 ng / ml cGAMP) were added to the appropriate wells and incubated at 37°C for 3 hours to activate the STING pathway. After the incubation period, the supernatant was replaced with 10 g / ml of anti-STING antibody in Hematogrow medium, and incubated overnight at 37°C. The cells were then subjected to ICC staining, beginning with adhesion to a coverslip, as previously described (see Deciphering Binding Specificities of Anti-STING Antibodies on human cells through Immunocytochemistry (ICC) analysis). Following fixation and 3 washes in PBS, the cells were permeabilized with 0.5% Triton X-100 in PBS for 5 minutes. Subsequent washing and blocking steps were carried out as previously described. The primary antibody incubation was conducted overnight at 4°C, utilizing an anti-human pTBK1 antibody (Cell Signaling) diluted 1:100 in PBS. The primary antibody was detected by incubation with a Cy5-conjugated anti-rabbit secondary antibody for 60 minutes at room temperature. DAPI was utilized as a nuclei stain. Finally, the coverslip was mounted onto a glass slide to allow imaging of the fluorescent signal using the SlideView_VS200 (Olympus) imaging system. Quantification of Cy5 (pTBK1) and DAPI positive signals was performed on ImageJ, using 5 individual areas per condition. The ratio between pTBK1 and DAPI signals was subsequently calculated and reported on GraphPad.

[0291] Upon ex vivo CMA-mediated STING pathway activation, incubation of mouse splenocytes with Ab1 resulted in a decrease in pTBK1 levels. The activation of the STINGNAI-5002871589v1109pathway by CMA led to phosphorylation of TBK1, as seen in FIG. 11B. Treatment with Ab1 and the small molecule STING inhibitor (H-151) both showed a strong reduction in pTBK1 signal; see FIG. 11E and 11C. In contrast, the human IgG control (isotype control antibody) did not affect pTBK1 levels; see FIG. 11D. The absence of signal in the anti-human control (secondary antibody control) confirmed the specificity of the pTBK1 signal. Quantification of pTBK1 signal was performed by normalizing the positive Cy5 signal with DAPI, as shown in FIG. 11F. These results demonstrated that Ab1 could inhibit the phosphorylation of TBK1 in mouse splenocytes upon ex vivo CMA-mediated STING pathway activation. The findings from Example 10 were further validated by assessing the effect of antibody Ab1 on pTBK1 levels in mouse splenocytes following cGAMP-mediated STING pathway activation. As depicted in FIG. 12B, cGAMP application led to an increase in pTBK1 signal, which was reduced upon treatment with Ab1 and the small molecule STING inhibitor (H-151); see FIGS. 12E and 12C. Similar to the results obtained in the previous experiment, the human IgG control (isotype control antibody) did not affect pTBK1 levels; see FIG. 12D. The quantification of pTBK1 signal was performed by normalizing the positive Cy5 signal (pTBK1 signal) with DAPI, as shown in FIG. 12F). Taken together, these data indicated that incubation of mouse splenocytes with Ab1 led to a decrease in pTBK1 levels upon ex vivo STING pathway activation, consistent with the findings obtained in the previous experiment.

[0292] Examples 10 and 11 demonstrated that the anti-STING antibodies of thepresent disclosure, besides inhibiting the release of IFN- , were capable of preventingphosphorylation of TBK1 similar to the well-established STING antagonist H-151 and thus acting as cGAS-STING pathway inhibitor. Accordingly, antagonistic anti-STING antibodies were provided which can be reasonably expected to be suitable for the treatment of disorders related to activated STING mediated cytokine production like in autoimmune and (auto)inflammatory diseases and particular for disease which have been shown to be amenable to the treatment with STING antagonists like H-151. 6.12 Example 12: Ab1 antibody in vivo efficacy in Trex1- / - mouse model

[0293] TREX1 (DNase III) is the principal 3 -5 DNA exonuclease in mammaliancells, playing a critical role in cell death and genomic DNA degradation. Dysfunction of TREX1 is suggested to activate immune responses against self-DNA, as evidenced by TREX1 null mice, which develop inflammatory myocarditis similar to autoimmune cardiomyopathy and produce type 1 IFN. Additionally, the TREX1 D18N mutation causes familial chilblain lupus, a monogenic cutaneous form of lupus, with the mutant enzyme exhibiting impaired dsDNA-degrading activity, thereby linking dsDNA degradation toNAI-5002871589v1110nucleic acid-mediated autoimmune disease. Trex1- / - mice (also known as Trex1 null mice), (GemPharmatech) underwent comprehensive phenotyping to assess the loss of function of TREX1 and its implications for systemic lupus erythematosus (SLE) research. This model demonstrated a lupus-like inflammatory autoimmune response, characterized by body weight loss and inflammation in multiple organs, thereby providing an established model for studying TREX1-mediated autoimmunity and evaluating potential treatments for lupus.

[0294] The in vivo experiment using a Trex1- / - mouse model of Aicardi-Goutieres syndrome, which serves as a model for Lupus-like autoimmune disease characterized by STING pathway activation leading to multi-organ inflammation and autoimmune disease, demonstrated significant improvements in body weight gain percentage in a dose-dependent manner with Ab1 treatment, compared to the untreated group. Specifically, Trex1- / - mice, aged 4-5 weeks, were weighed daily following the administration of Ab1 at dosages of 0.5 mg / kg, 5 mg / kg, and 50 mg / kg, with an established commercially available antibody serving as a positive control and an IgG antibody as a negative control. A total of 72 female mice were randomized into six groups (n=12 per group) to ensure balanced body weights at the study's start. Group 1 served as a naïve control with no treatment, Group 2 received the IgG control antibody, Groups 3 to 5 received anti-STING antibody Ab1 at the specified doses, and Group 6 received the positive control antibody. For group allocation see the Table 4 below. The study design is shown in FIG. 13. Table 4. Group Allocation Grou n Genotype Sex Age Treatment Dose Body p (weeks) Treatment weight Organ frequency measure harvest ment 112 Trex1- / - F 4-5 untreated n.a. - daily Studyendpoint212 Trex1- / - F 4-5 IgG2a 50Study mg / kgweekly dailyendpoint 312 Trex1- / - F 4-5 Ab1 0.5Stud mg / kgweekly dailyy endpoint 412 Trex1- / - F 4-5 Ab1 5 mg / kg weekly daily Studyendpoint512 Trex1- / - F 4-5 Ab1 50mg / kgweekly dailyStudy endpoint Positive 250 6 12 Trex1- / - F 4-5 control ug / mousbiweekly daily Studyantibody eendpointNAI-5002871589v1111

[0295] Body weight measurements, taken daily, indicated a dose-dependent increase in body weight gain with Ab1, with the highest dose proving most effective (FIG. 14).

[0296] Moreover, detailed analysis revealed that body weight change rate and heartmRNA IFN- levels correlated in a concentration-dependent manner upon administration ofthe STING antibody Ab1. Notably, the 5 mg / kg dose of Ab1 and the positive control antibody treatment groups exhibited comparable levels of body weight change rate, whereas the 50 mg / kg dose of Ab1 showed the most pronounced beneficial increase (FIG. 14). In the Trex1- / - mouse model, the Ab1 antibody demonstrates a statistically significant, concentration-dependent effect on body weight change rate compared to the untreated group and to IgG2a antibody control.

[0297] At the study endpoint (day 42), all animals were sacrificed, and their hearts were harvested, snap-frozen, and stored at -80°C. Quantitative RT-PCR (RT-qPCR) analysis of heart extracts revealed that Ab1 treatment significantly reduced the mRNA expressionlevels of inflammatory mediators IFN- , IL-6, and ISG-15 in a dose-dependent manner(FIGS. 15A-15C). Compared to the IgG2a control group, antibody Ab1 significantly reducedSTING-pathway dependent cytokine mRNA levels of IFN- and IL-6 in the heart. TheIgG2a control antibody and 0.5 mg / kg of Ab1 treatment groups displayed unaltered levels of the three analysed cytokines compared to the untreated control group, while IL-6 mRNA levels were significantly downregulated in the 0.5 and 5 mg / kg Ab1 and positive control groups compared to the IgG2a control group. Furthermore, ISG-15 mRNA levels were significantly downregulated in the positive control compared to the untreated and IgG2a control groups.

[0298] This phenotypic assessment and analysis of STING pathway-dependent heart mRNA levels underscored the potential therapeutic efficacy of Ab1 in mitigating inflammation and autoimmune responses in the Tr...

Claims

1. WHAT IS CLAIMED IS:

1. An antibody or an antigen-binding fragment thereof that can specifically bind human stimulator of interferon genes (STING), wherein the antibody or antigen-binding fragment thereof comprises: (i) a heavy chain variable region (VH) comprising a heavy chain variable region complementarity determining region 1 (VH-CDR1), a heavy chain variable region complementarity determining region 2 (VH-CDR2), and a heavy chain variable region complementarity determining region 3 (VH-CDR3) as set forth in a VH comprising the amino acid sequence of SEQ ID NO:2; and a light chain variable region (VL) comprising a light chain variable region complementarity determining region 1 (VL-CDR1), a light chain variable region complementarity determining region 2 (VL-CDR2), and a light chain variable region complementarity determining region 3 (VL-CDR3) as set forth in a VL comprising the amino acid sequence of SEQ ID NO:7; (ii) a VH comprising a VH-CDR1, a VH-CDR2, and a VH-CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO:12; and a VL comprising a VL-CDR1, a VL-CDR2, and a VL-CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO:17; or (iii) a VH comprising a VH-CDR1, a VH-CDR2, and a VH-CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO:22; and a VL comprising a VL-CDR1, a VL-CDR2, and a VL-CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO:

27.

2. The antibody or antigen-binding fragment thereof of claim 1, wherein the VH-CDR1 comprises the amino acid sequence of SEQ ID NO:3, the VH-CDR2 comprises the amino acid sequence of SEQ ID NO:4, the VH-CDR3 comprises the amino acid sequence of SEQ ID NO:5, the VL-CDR1 comprises the amino acid sequence of SEQ ID NO:8, the VL-CDR2 comprises the amino acid sequence of SEQ ID NO:9, and the VL-CDR3 comprises the amino acid sequence of SEQ ID NO:10, and wherein the amino acid sequences of the VH-CDR1, VH-CDR2, VH-CDR3, VL- CDR1, VL-CDR2, and VL-CDR3 are determined by Kabat definition.

3. The antibody or antigen-binding fragment thereof of claim 2, whereinNAI-5002871589v1126 (i) the VH comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:2, or the VL comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:7; (ii) the VH comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:2, and the VL comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:7; or (iii) the VH comprises the amino acid sequence of SEQ ID NO:2 or the VL comprises the amino acid sequence of SEQ ID NO:

7.

4. The antibody or antigen-binding fragment thereof of claim 2 or 3, wherein the VH comprises the amino acid sequence of SEQ ID NO:2 and the VL comprises the amino acid sequence of SEQ ID NO:

7.

5. The antibody or antigen-binding fragment thereof of claim 1, wherein the VH-CDR1 comprises the amino acid sequence of SEQ ID NO:13, the VH-CDR2 comprises the amino acid sequence of SEQ ID NO:14, the VH-CDR3 comprises the amino acid sequence of SEQ ID NO:15, the VL-CDR1 comprises the amino acid sequence of SEQ ID NO:18, the VL- CDR2 comprises the amino acid sequence of SEQ ID NO:19, and the VL-CDR3 comprises the amino acid sequence of SEQ ID NO:20, and wherein the amino acid sequences of the VH-CDR1, VH-CDR2, VH-CDR3, VL- CDR1, VL-CDR2, and VL-CDR3 are determined by Kabat definition.

6. The antibody or antigen-binding fragment thereof of claim 5, wherein (i) the VH comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:12, or the VL comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:17; (ii) the VH comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:12, and the VL comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:17; or (iii) the VH comprises the amino acid sequence of SEQ ID NO:12 or the VL comprises the amino acid sequence of SEQ ID NO:17.NAI-5002871589v11277. The antibody or antigen-binding fragment thereof of claim 5 or 6, wherein the VH comprises the amino acid sequence of SEQ ID NO:12 and the VL comprises the amino acid sequence of SEQ ID NO:

17.

8. The antibody or antigen-binding fragment thereof of claim 1, wherein the VH-CDR1 comprises the amino acid sequence of SEQ ID NO:23, the VH-CDR2 comprises the amino acid sequence of SEQ ID NO:24, the VH-CDR3 comprises the amino acid sequence of SEQ ID NO:25, the VL-CDR1 comprises the amino acid sequence of SEQ ID NO:28, the VL- CDR2 comprises the amino acid sequence of SEQ ID NO:29, and the VL-CDR3 comprises the amino acid sequence of SEQ ID NO:30, and wherein the amino acid sequences of the VH-CDR1, VH-CDR2, VH-CDR3, VL- CDR1, VL-CDR2, and VL-CDR3 are determined by Kabat definition.

9. The antibody or antigen-binding fragment thereof of claim 8, wherein (i) the VH comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:22, or the VL comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:27; (ii) the VH comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:22, and the VL comprises an amino acid sequence at least 80% or at least 90% identical to the amino acid sequence of SEQ ID NO:27; or (iii) the VH comprises the amino acid sequence of SEQ ID NO:22 or the VL comprises the amino acid sequence of SEQ ID NO:

27.

10. The antibody or antigen-binding fragment thereof of claim 8 or 9, wherein the VH comprises the amino acid sequence of SEQ ID NO:22 and the VL comprises the amino acid sequence of SEQ ID NO:

27.

11. An antibody or an antigen-binding fragment thereof that competes with the antibody or antigen-binding fragment thereof of any one of claims 1-10 for specifically binding to an epitope located within the C-terminal domain of STING.

12. The antibody or antigen-binding fragment thereof of claim 11, wherein the C-terminal domain of STING comprises the amino acid sequence of SEQ ID NO:31.NAI-5002871589v112813. The antibody or antigen-binding fragment thereof of claim 11 or 12, wherein the epitope is a conformational epitope or a linear epitope.

14. The antibody or antigen-binding fragment thereof of any one of claim 11-13, wherein the epitope comprises amino acid residues S358, G367, M368, E369, and P371 of human STING, wherein the amino acid positions are numbered according to the positions in the amino acid sequence of SEQ ID NO:

34.

15. The antibody or antigen-binding fragment thereof of any one of claims 11-14, wherein the (i) the antibody or antigen-binding fragment thereof can inhibit the phosphorylation of TANK binding kinase 1 (TBK1) in cells with activated STING pathway in vitro, ex vivo or in vivo; and / or (ii) the antibody or antigen-binding fragment thereof can inhibit the release of an inflammatory cytokine in cells with activated STING pathway in vitro, ex vivo or in vivo; (iii) the antibody or antigen-binding fragment thereof can inhibit the release of an inflammatory cytokine in cells of a Trex1 null mouse or reduce the level of an inflammatory cytokine of a NZB / W mouse.

16. An antagonistic antibody or an antigen-binding fragment thereof that can specifically bind human STING , wherein: (i) the antibody or antigen-binding fragment thereof can specifically bind an epitope located within the C-terminal domain of STING; (ii) the antibody or antigen-binding fragment thereof can inhibit the phosphorylation of TBK1 in cells with activated STING pathway in vitro, ex vivo or in vivo; (iii) the antibody or antigen-binding fragment thereof can inhibit the release of an inflammatory cytokine in cells with activated STING pathway in vitro, ex vivo or in vivo; and / or (iv) the antibody or antigen-binding fragment thereof can inhibit the release of an inflammatory cytokine in cells of a Trex1 null mouse or reduce the level of an inflammatory cytokine of a NZB / W mouse.NAI-5002871589v112917. The antibody or antigen-binding fragment thereof of claim 16, wherein the C-terminal domain of STING comprises the amino acid sequence of SEQ ID NO:

31.

18. The antibody or antigen-binding fragment thereof of claim 16 or 17, wherein the epitope is a conformational epitope or a linear epitope.

19. The antibody or antigen-binding fragment thereof of any one of claims 15-18, wherein the epitope comprises amino acid residues S358, G367, M368, E369, and P371 of human STING, wherein the amino acid positions are numbered according to the positions in the amino acid sequence of SEQ ID NO:

34.

20. The antibody or antigen-binding fragment thereof of any one of claims 15-19, wherein the inhibition of the phosphorylation of TBK1 in the cells with activated STING pathway is indicated by a reduced pTBK1 level in the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof as compared to cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof, optionally wherein the level of pTBK1 is measured by an immunohistochemistry assay, further optionally wherein the immunohistochemistry assay is Western Blot or immunocytochemistry (ICC).

21. The antibody or antigen-binding fragment thereof of any one of claims 15-19, wherein the inhibition of the release of the inflammatory cytokine is indicated by a reduced level of the inflammatory cytokine released by the cells with activated STING pathway or the cells of the Trex1 null mouse being contacted or administered with the antibody or antigen- binding fragment thereof as compared to cells with activated STING pathway or cells of a Trex1 null mouse but without being contacted or administered with the antibody or antigen- binding fragment thereof, optionally wherein the level of the inflammatory cytokine is measured by an immunoassay or RT-qPCR, optionally wherein the immunoassay is ELISA, HTRF (homogeneous time resolved fluorescence), or MSD (Meso Scale Discovery) multiplex assay.

22. The antibody or antigen-binding fragment thereof of any one of claims 15-19, wherein reducing the level of the inflammatory cytokine is indicated by a reduced level of the inflammatory cytokine in serum of the NZB / W mouse being administered with the antibody or antigen-binding fragment thereof as compared to the level of the inflammatory cytokine in serum of a NZB / W mouse but without being administered with the antibody or antigen-NAI-5002871589v1130binding fragment thereof, optionally wherein the level of the inflammatory cytokine is measured by an immunoassay, further optionally wherein the immunoassay is MSD multiplex assay.

23. The antibody or antigen-binding fragment thereof of any one of claims 15-22,wherein the inflammatory cytokine is IL-6, IP-10, IFN- , and / or ISG-15.

24. The antibody or antigen-binding fragment thereof of any one of claims 15-23, wherein the cells are immune cells, heart tissue cells of the Trex1 null mouse, differentiated THP-1 cells, or mouse splenocytes.

25. The antibody or antigen-binding fragment thereof of any one of claims 15-24, wherein the STING pathway is activated by 10-carboxymethyl-9-acridanone (CMA), cyclic GMP-AMP (cGAMP), or HT DNA (deoxyribonucleic acid from herring testes).

26. The antibody or antigen-binding fragment thereof of any one of claims 20-25, wherein the pTBK1 level in the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less of the pTBK1 level in the cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof, as measured by Western Blot or ICC, and the cells are PMA-differentiated human THP-1-derived macrophages or mouse splenocytes with STING pathway activated by cGAMP or CMA.

27. The antibody or antigen-binding fragment thereof of any one of claims 21-25, wherein the level of the inflammatory cytokine released by the cells with activated STING pathway and being contacted with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less of cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof, as measured by ELISA or HTRF, and the cells are PMA-differentiated human THP-1-derived macrophages or mouse splenocytes with STING pathway activated by cGAMP, and the inflammatorycytokine is IFN- .

28. The antibody or antigen-binding fragment thereof of any one of claims 21-25, wherein the level of the inflammatory cytokine released by the cells with activated STINGNAI-5002871589v1131pathway and being contacted with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less of cells with activated STING pathway but without being contacted with the antibody or antigen-binding fragment thereof, as measured by MSD multiplex assay, and the cells are PMA-differentiated human THP-1-derived macrophageswith STING pathway activated by HT DNA, and the inflammatory cytokine is IFN- , IL-6,and / or IP-10.

29. The antibody or antigen-binding fragment thereof of any one of claims 21-25, wherein the level of the inflammatory cytokine released by the cells from the Trex1 null mouse administered with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, or about 80% or less of cells from a Trex1 null mouse without being administered with the antibody or antigen-binding fragment thereof, as measured by RT-qPCR, and thecells are heart tissue cells, and the inflammatory cytokine is IFN- , IL-6, and / or ISG-15.

30. The antibody or antigen-binding fragment thereof of any one of claims 22-25, wherein the level of the inflammatory cytokine in serum of the NZB / W mouse being administered with the antibody or antigen-binding fragment thereof is about 20% or less, about 30% or less, about 40% or less, about 50% or less, about 60% or less, about 70% or less, about 80% or less, or about 90% or less of serum from a NZB / W mouse without being administered with the antibody or antigen-binding fragment thereof, as measured by MSD multiplex assay, and the inflammatory cytokine is IL-6.

31. The antibody or antigen-binding fragment thereof of any one of claims 1-30, which is an antagonistic antibody or antigen-binding fragment thereof.

32. The antibody or antigen-binding fragment thereof of any one of claims 1-31, wherein the antibody is a recombinant antibody and / or a monoclonal antibody.

33. The antibody or antigen-binding fragment thereof of any one of claims 1-32, wherein the antibody comprises a heavy chain constant region and / or a light chain constant region.

34. The antibody or antigen-binding fragment thereof of claim 33, wherein the heavy chain constant region is of human IgG type, optionally wherein the heavy chain constant region is of IgG1 type or human IgG4 type.NAI-5002871589v113235. The antibody or antigen-binding fragment thereof of claim 33 or 34, wherein the light chain constant region comprises a human kappa light chain constant region or a human lambda light chain constant region.

36. The antibody or antigen-binding fragment thereof of any one of claims 33-35, wherein the heavy chain constant region comprises the amino acid sequence of SEQ ID NO:129 or SEQ ID NO:132, and / or the light chain constant region comprises the amino acid sequence of SEQ ID NO:130 or SEQ ID NO:

131.

37. The antibody or antigen-binding fragment thereof of any one of claims 33-36, wherein the antibody comprises an Fc region comprising one or more mutations, optionally wherein the one or more mutations decrease immune effector function, increase immune effector function, or increase half-life of the antibody.

38. The antibody or antigen-binding fragment thereof of any one of claims 1-37, wherein the antigen-binding fragment a single chain Fv fragment (scFv), an F(ab’), an F(ab), an F(ab’)2, an Fd, an Fv, a single-chain antibody, a disulfide-linked Fv (sdFv), a VHH, or a nanobody.

39. The antibody or antigen-binding fragment thereof of any one of claims 1-37, wherein the antibody or antigen-binding fragment thereof is attached to a detectable label, wherein the detectable label is an enzyme, a radioisotope, a fluorescent compound, a chemiluminescent compound, a bioluminescent compound, a tag, a flag or a heavy metal.

40. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof of any one of claims 1-39 and 46 and a pharmaceutically acceptable carrier.

41. A formulation suitable for in vivo administration comprising the antibody or antigen- binding fragment thereof of any one of claims 1-39 and 46, optionally wherein the formulation is suitable for in vivo intravenous, intraperitoneal, subcutaneous, or intramuscular injection.

42. An isolated polynucleotide or a set of polynucleotides encoding the antibody or antigen-binding fragment thereof of any one of claims 1-39 and 46.

43. A vector or a set of vectors comprising the polynucleotide or set of polynucleotides of claim 42.NAI-5002871589v113344. An isolated cell comprising the polynucleotide or set of polynucleotides of claim 42 or the vector or set of vectors of claim 43.

45. A method of making an antibody or an antigen-binding fragment thereof, comprising: (i) culturing the cell of claim 44, and (ii) isolating the antibody or antigen-binding fragment thereof.

46. An antibody or an antigen-binding fragment thereof produced by the method of claim 45.

47. A method of detecting STING in a biological sample, comprising: (i) contacting the biological sample with the antibody or antigen-binding fragment thereof of any one of claims 1-39 and 46; and (ii) detecting the binding between the antibody or antigen-binding fragment thereof and STING in the biological sample, optionally wherein the detecting comprises using flow cytometry, immunohistochemistry (IHC), Western Blot analysis, ELISA, or mass spectrometry.

48. A method for screening for an antibody or an antigen-binding fragment thereof that can specifically bind human STING, comprising measuring the binding of the antibody or antigen-binding fragment thereof to an epitope located within the C-terminal domain of STING, and selecting the antibody if the binding affinity is higher than an isotype control antibody, optionally wherein the binding is measured by ELISA.

49. The method of claim 48, wherein the C-terminal domain of STING comprises the amino acid sequence of SEQ ID NO:

31.

50. The method of claim 48 or 49, wherein the epitope is a conformational epitope or a linear epitope.

51. The method of any one of claim 48 -50, wherein the epitope comprises amino acid residues S358, G367, M368, E369, and P371 of human STING, wherein the amino acid positions are numbered according to the positions in the amino acid sequence of SEQ ID NO:

34.

52. A method of treating a disease or a disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the antibody or antigen-NAI-5002871589v1134binding fragment thereof of any one of claims 1-39 and 46, the pharmaceutical composition of claim 40, or the formulation of claim 41.

53. The antibody or antigen-binding fragment thereof of any one of claims 1-39 and 46, the pharmaceutical composition of claim 40, or the formulation of claim 41 for use in treating a disease or a disorder in a subject in need thereof 54. Use of the antibody or antigen-binding fragment thereof of any one of claims 1-39 and 46, the pharmaceutical composition of claim 40, or the formulation of claim 41 in the manufacture of a medicament for treating a disease or a disorder in a subject in need thereof.

55. The method of claim 52, the antibody or antigen-binding fragment thereof for use in accordance with claim 53, or the use of claim 54, wherein the disease or disorder is an autoimmune disease or an inflammatory disease.

56. The method of claim 52 or 55, the antibody or antigen-binding fragment thereof for use in accordance with claim 53 or 55, or the use of claim 54 or 55, wherein the disease or disorder is Aicardi-Goutières syndrome (AGS), STING-associated vasculopathy of infancy (SAVI), lupus, systemic lupus erythematosus (SLE), COPA syndrome, amyotrophic lateral sclerosis (ALS), Niemann-Pick disease type C (NPC), multiple sclerosis, familial chilblain lupus (FCL), rheumatoid arthritis, Sjögren’s syndrome, secondary Sjögren’s syndrome, Alzheimer disease, ischemic brain injury, Parkinson disease, neurodegeneration, Huntington disease, frontotemporal dementia, age-dependent macular degeneration, traumatic brain injury, nonalcoholic steatohepatitis, alcoholic liver disease, acute pancreatitis, silica- induced fibrosis, sepsis, myocardial infarction, chronic heart failure, or colorectal cancer.

57. A method of inhibiting STING pathway, inhibiting the phosphorylation of TBK1, and / or inhibiting the release of an inflammatory cytokine in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the antibody or antigen-binding fragment thereof of any one of claims 1-39 and 46, the pharmaceutical composition of claim 40, or the formulation of claim 41.

58. The method of claim 57, wherein (i) inhibiting STING pathway is indicated by a reduced level of pTBK1 and / or a reduced level of an inflammatory cytokine in a sample of the subject; (ii) inhibiting the phosphorylation of TBK1 is indicated by a reduced level of pTBK1 in a sample of the subject; and / or (iii) inhibiting the release of an inflammatoryNAI-5002871589v1135cytokine is indicated by a reduced level of the inflammatory cytokine in a sample of the subject, optionally wherein the reduced level of pTBK1 and / or reduced level of the inflammatory cytokine is reduced as compared to the level of pTBK1 and / or the level of the inflammatory cytokine measured before the administration of the anti-STING antibody or antigen-binding fragment thereof.

59. The method of claim 57 or 58, wherein the inflammatory cytokine is IL-6, IP-10,IFN- , and / or ISG-15.

60. The method of claim 58 or 59, wherein the sample is a blood sample, a serum sample, a tissue sample, or a cell sample.NAI-5002871589v1136

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