Protein formulations and uses thereof

Abstract

The present disclosure relates to protein formulations and uses thereof. In particular, the present disclosure relates to formulations comprising a protein comprising an antigen binding domain that binds to or specifically binds to granulocyte colony- stimulating factor receptor (G-CSFR).

Description

[0001] PROTEIN FORMULATIONS AND USES THEREOF

[0002] RELATED APPLICATION DATA

[0003] The present application claims priority from Australian provisional application No. 2024901051 entitled “Protein formulations and uses thereof’ filed 15 April 2024, the entire contents of which are hereby incorporated by reference.

[0004] SEQUENCE LISTING

[0005] The present application is filed together with a Sequence Listing in electronic form. The entire contents of the Sequence Listing are hereby incorporated by reference.

[0006] FIELD

[0007] The present disclosure relates to protein formulations and uses thereof. In particular, the present disclosure relates to formulations comprising a protein comprising an antigen binding domain that binds to or specifically binds to granulocyte colonystimulating factor receptor (G-CSFR).

[0008] BACKGROUND

[0009] Granulocyte colony- stimulating factor (G-CSF) is a major regulator of granulocyte production. G-CSF is produced by bone marrow stromal cells, endothelial cells, macrophages, and fibroblasts, and production is induced by inflammatory stimuli. G-CSF acts through the G-CSF receptor (G-CSFR), which is expressed predominantly on neutrophils, but also on myeloid progenitors, endothelial cells, monocytes / macrophages, and T and B lymphocytes. Both G-CSF and G-CSFR are therapeutic targets for a range of diseases.

[0010] Although antibodies and inhibitors against G-CSF and G-CSFR exist, there are an increasing number of challenges in formulation development for drug manufacturers. For example, there are numerous challenges associated with formulating high concentration antibody formulations (e.g., >25 mg / mL protein) suitable for subcutaneous administration. Formulations for subcutaneous administration typically require higher concentrations of product so as to achieve smaller injection volumes, yet increasing protein concentration often negatively impacts protein aggregation and degradation, solubility, stability, and viscosity. Reducing protein concentrations to try to mitigate these issues can also lead to reduced stability and / or reduced long term potency.

[0011] In addition to changes in intrinsic protein properties, manufacturing and supply chain challenges also exist including difficulties with processing and storage to ensure that the formulated protein remains stable for long periods of time (e.g., greater than six months) and at higher temperatures (e.g., room temperature). Other challenges include optimising the rheological and syringeability properties of the final formulation. For example, viscous solutions typically require a higher injection force to administer, therefore a prolonged injection time may also be required contributing to patient pain and discomfort.

[0012] Various solutions to manufacturing high concentration antibody formulations include lyophilised formulations for reconstitution, bufferless formulations and the addition of high concentrations of salt or other additives to reduce aggregation and / or the viscosity of the formulation. However, the use of excessive amounts of such excipients, may lead to hypertonic preparations or changes in ionic strength of the formulation and related protein aggregation issues.

[0013] Thus, there is a need for formulations comprising protein therapeutics that bind to G-CSFR which are stable and suitable for administration to a subject for treating neutrophil-mediated conditions.

[0014] SUMMARY

[0015] The present disclosure is based on the identification of a stable pharmaceutical formulation for a protein comprising an antigen binding domain that specifically binds to G-CSFR.

[0016] The inventors found that they can produce liquid formulations comprising 45-55 mg / mL of a protein comprising an antigen binding domain that binds to G-CSFR, which remained stable, soluble, and had a viscosity suitable for subcutaneous injection. In particular, formulations of the disclosure comprising histidine, proline, arginine, and polysorbate 80 were found to remain stable, potent, and with minimal indicators of protein aggregation for at least 24 months at high temperatures (i.e. room temperature) and without the need for stabilizing agents, such as sodium chloride (NaCl). Advantageously, the formulations of the disclosure allow the administration of practical volumes, which are beneficial for fixed dosing regimens and / or self-administration.

[0017] The present disclosure thus provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of between 45 mg / mL and 55 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises: (i) a heavy chain variable region (VH) comprising three complementarity determining regions (CDRs) of a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a light chain variable region (VL) comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 5; or

[0018] (ii) a VH comprising three CDRs of a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 3.

[0019] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of between 45 mg / mL and 55 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 m proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises a heavy chain variable region (VH) comprising three complementarity determining regions (CDRs) of a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a light chain variable region (VL) comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 5.

[0020] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of between 45 mg / mL and 55 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises a VH comprising three CDRs of a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 3.

[0021] In one example, the protein is at a concentration of about 50 mg / mL. In one example, the protein is at a concentration of 50 mg / mL.

[0022] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of about 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises:

[0023] (i) a heavy chain variable region (VH) comprising three complementarity determining regions (CDRs) of a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a light chain variable region (VL) comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 5; or

[0024] (ii) a VH comprising three CDRs of a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 3.

[0025] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of about 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises: a heavy chain variable region (VH) comprising three complementarity determining regions (CDRs) of a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a light chain variable region (VL) comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 5.

[0026] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of about 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises: a VH comprising three CDRs of a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 3.

[0027] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises: (i) a heavy chain variable region (VH) comprising three complementarity determining regions (CDRs) of a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a light chain variable region (VL) comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 5; or a VH comprising three CDRs of a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 3.

[0028] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises: a heavy chain variable region (VH) comprising three complementarity determining regions (CDRs) of a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a light chain variable region (VL) comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 5.

[0029] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises: a VH comprising three CDRs of a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 3.

[0030] The present disclosure thus provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of between 45 mg / mL and 55 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and consists of 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises:

[0031] (i) a heavy chain variable region (VH) comprising three complementarity determining regions (CDRs) of a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a light chain variable region (VL) comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 5; or

[0032] (ii) a VH comprising three CDRs of a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 3.

[0033] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of between 45 mg / mL and 55 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and consists of 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises a heavy chain variable region (VH) comprising three complementarity determining regions (CDRs) of a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a light chain variable region (VL) comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 5.

[0034] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of between 45 mg / mL and 55 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and consists of 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises a VH comprising three CDRs of a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 3.

[0035] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of about 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and consists of 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises:

[0036] (i) a heavy chain variable region (VH) comprising three complementarity determining regions (CDRs) of a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a light chain variable region (VL) comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 5; or

[0037] (ii) a VH comprising three CDRs of a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 3.

[0038] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of about 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and consists of 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises: a heavy chain variable region (VH) comprising three complementarity determining regions (CDRs) of a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a light chain variable region (VL) comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 5.

[0039] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of about 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and consists of 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises: a VH comprising three CDRs of a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 3.

[0040] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and consists of 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises:

[0041] (i) a heavy chain variable region (VH) comprising three complementarity determining regions (CDRs) of a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a light chain variable region (VL) comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 5; or (ii) a VH comprising three CDRs of a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 3.

[0042] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and consists of 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises: a heavy chain variable region (VH) comprising three complementarity determining regions (CDRs) of a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a light chain variable region (VL) comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 5.

[0043] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and consists of 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises: a VH comprising three CDRs of a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 3.

[0044] In one example, the formulation is an aqueous formulation.

[0045] In one example, the formulation has not previously been lyophilised. In one example, the formulation is not a reconstituted formulation.

[0046] In one example, the formulation comprises:

[0047] (i) 20 mM histidine buffer;

[0048] (ii) 0.03% (w / v) polysorbate 80;

[0049] (iii) 100 mM arginine; and

[0050] (iv) 100 mM proline.

[0051] In one example, the formulation consists of:

[0052] (i) 20 mM histidine buffer;

[0053] (ii) 0.03% (w / v) polysorbate 80;

[0054] (iii) 100 mM arginine; and

[0055] (iv) 100 mM proline. In one example, the formulation comprises 15 mM to 25 mM histidine buffer. For example, the formulation comprises 17 mM to 23 mM histidine buffer. For example, the formulation comprises 18 mM to 22 mM histidine buffer. In one example, the formulation comprises 15 mM histidine buffer. In one example, the formulation comprises 16 mM histidine buffer. In one example, the formulation comprises 17 mM histidine buffer. In one example, the formulation comprises 18 mM histidine buffer. In one example, the formulation comprises 19 mM histidine buffer. In one example, the formulation comprises 20 mM histidine buffer. In one example, the formulation comprises 21 mM histidine buffer. In one example, the formulation comprises 22 mM histidine buffer. In one example, the formulation comprises 23 mM histidine buffer. In one example, the formulation comprises 24 mM histidine buffer. In one example, the formulation comprises 25 mM histidine buffer.

[0056] In one example, the formulation comprises 0.02% (w / v) polysorbate 80. In one example, the formulation comprises 0.03% (w / v) polysorbate 80. In one example, the formulation comprises 0.04% (w / v) polysorbate 80.

[0057] In one example, the formulation comprises 95 mM to 105 mM arginine. In one example, the formulation comprises 95 mM arginine. In one example, the formulation comprises 96 mM arginine. In one example, the formulation comprises 97 mM arginine. In one example, the formulation comprises 98 mM arginine. In one example, the formulation comprises 99 mM arginine. In one example, the formulation comprises 100 mM arginine. In one example, the formulation comprises 101 mM arginine. In one example, the formulation comprises 102 mM arginine. In one example, the formulation comprises 103 mM arginine. In one example, the formulation comprises 104 mM arginine. In one example, the formulation comprises 105 mM arginine.

[0058] In one example, the formulation comprises 100 mM to 105 mM proline. In one example, the formulation comprises 96 mM proline. In one example, the formulation comprises 97 mM proline. In one example, the formulation comprises 98 mM proline. In one example, the formulation comprises 99 mM proline. In one example, the formulation comprises 100 mM proline. In one example, the formulation comprises 101 mM proline. In one example, the formulation comprises 102 mM proline. In one example, the formulation comprises 103 mM proline. In one example, the formulation comprises 104 mM proline. In one example, the formulation comprises 105 mM proline.

[0059] In one example, the formulation has a pH of 5.6 to 5.8. In one example, the formulation has a pH of 5.6. In one example, the formulation has a pH of 5.7. In one example, the formulation has a pH of 5.8. In one example, the formulation is a stable liquid formulation. The stability of the formulation may be assessed by any means known in the art. For example, the stability of the formulation may be assessed by measuring total high molecular weight species (HMWS) and / or monomer content. Methods for assessing accumulation of HMWS and monomer content of the formulation will be apparent to the skilled person and / or described herein. In one example, the percent HMWS of the protein in the formulation is determined by size-exclusion chromatography or size exclusion high performance liquid chromatography (e.g., SEC or SE-HPLC).

[0060] In another example, the formation of HMWS of the protein is assessed using dynamic light scattering (DLS). For example, the fluctuation of light intensity using a digital correlator (e.g., Malvern Zetasizer software) is measured and the Z-average hydrodynamic diameter and polydispersity index (using e.g., a cumulants analysis) are determined. In one example, one or more or all of the following apply:

[0061] (i) the formulation comprises no more than 5% HMWS, or no more than 3.5% HMWS, as determined by SE-HPLC;

[0062] (ii) at least 95% of the protein in the formulation is a monomer, as determined by SE-HPLC;

[0063] (iii)the formulation comprises no more than 50% acidic species, as determined by cation exchange high performance liquid chromatography (CEX-HPLC);

[0064] (iv)the formulation comprises no more than 20% basic species, as determined by CEX-HPLC; and

[0065] (v) the formulation comprises no more than 10% low molecular weight species (LMWS), as determined by capillary electrophoresis with sodium dodecylsulfate (CE-SDS) under non-reducing conditions.

[0066] In one example, all of the following apply:

[0067] (i) the formulation comprises no more than 5% HMWS, or no more than 3.5% HMWS, as determined by SE-HPLC;

[0068] (ii) at least 95% of the protein in the formulation is a monomer, as determined by SE-HPLC;

[0069] (iii)the formulation comprises no more than 50% acidic species, as determined by CEX-HPLC;

[0070] (iv)the formulation comprises no more than 20% basic species, as determined by CEX-HPLC; and

[0071] (v) the formulation comprises no more than 10% LMWS, as determined by CE- SDS under non-reducing conditions. In one example, the formulation comprises no more than 5% HMWS, as determined by SE-HPLC. For example, the formulation comprises no more than 5%, or 4.5%, or 4%, or 2.5% HMWS as determined by SE-HPLC. In one example, the formulation comprises no more than 3.5% HMWS, as determined by SE-HPLC. For example, the formulation comprises no more than 3.5%, or 3%, or 2.5%, or 2%, or 1.5%, or 1% HMWS as determined by SE-HPLC.

[0072] In one example, at least 95% of the protein in the formulation is a monomer, as determined by SE-HPLC. For example, at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99% of the protein in the formulation is a monomer, as determined by SE-HPLC.

[0073] In one example, the formulation of the present disclosure comprises at least 95% monomer protein and less than (i.e., no more than) 5% HMWS. In another example, the formulation comprises at least 95% monomer protein and less than (i.e., no more than) 3.5% HMWS.

[0074] Another method for assessing the stability of the formulation includes measuring the accumulation of acidic and / or basic species of the protein. The amount of acidic and / or basic species of a protein can be measured using cation exchange chromatography (e.g., CEX-HPLC), for example.

[0075] In one example, the formulation comprises no more than 50% acidic species, as determined by CEX-HPLC. For example, the formulation comprises no more than 45%, or 40%, or 35%, or 30%, or 25%, or 20%, or 10%, or 5% acidic species, as determined by CEX-HPLC.

[0076] In one example, the formulation comprises no more than 20% basic species, as determined by CEX-HPLC. For example, the formulation comprises no more than 18%, or 15%, or 12%, or 10%, or 8%, or 5%, or 2% basic species, as determined by CEX- HPLC

[0077] In one example, the formulation comprises no more than 10% LMWS, as determined by CE-SDS under non-reducing conditions. For example, the formulation comprises no more than 10%, or 8%, or 6%, or 5%, or 3%, or 2%, or 1% LMWS, as determined by CE-SDS under non-reducing conditions.

[0078] In one example, the formulation comprises less than 6000 particles of > 10 pm per container as determined by light obscuration. For example, the formulation comprises less than 3000, or less than 1000, or less than 300, or less than 200, or less than 100, or less than 50, or less than 20, or less than 10, or less than 5 particles of > 10 pm per container as determined by light obscuration. In one example, the formulation comprises less than 600 particles of > 25 pm per container as determined by light obscuration. For example, the formulation comprises less than 300, or less than 100, or less than 50, or less than 25, or less than 10, or less than 5 particles of > 25 pm per container as determined by light obscuration.

[0079] In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 1 month, at least 3 months, at least 6 months, at least 9 months, at least 12 months, at least 18 months, or at least 24 months at a temperature in the range of 2 °C to 30 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 1 month, at least 3 months, at least 6 months, at least 9 months, at least 12 months, at least 18 months, at least 24 months, at least 30 months, at least 36 months, or at least 42 months at a temperature in the range of 2 °C to 30 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 1 month at a temperature in the range of 2 °C to 30 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 3 months at a temperature in the range of 2 °C to 30 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 6 months at a temperature in the range of 2 °C to 30 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 9 months at a temperature in the range of 2 °C to 30 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 12 months at a temperature in the range of 2 °C to 30 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 18 months at a temperature in the range of 2 °C to 30 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 24 months at a temperature in the range of 2 °C to 30 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 30 months at a temperature in the range of 2 °C to 30 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 36 months at a temperature in the range of 2 °C to 30 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 42 months at a temperature in the range of 2 °C to 30°C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 1 month, at least 3 months, at least 6 months, at least 9 months, at least 12 months, at least 18 months, or at least 24 months at a temperature in the range of 2 °C to 28 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 1 month, at least 3 months, at least 6 months, at least 9 months, at least 12 months, at least 18 months, at least 24 months, at least 30 months, at least 36 months, or at least 42 months at a temperature in the range of 2 °C to 28 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 1 month at a temperature in the range of 2 °C to 28 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 3 months at a temperature in the range of 2 °C to 28 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 6 months at a temperature in the range of 2 °C to 28 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 9 months at a temperature in the range of 2 °C to 28 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 12 months at a temperature in the range of 2 °C to 28 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 18 months at a temperature in the range of 2 °C to 28 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 24 months at a temperature in the range of 2 °C to 28 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 30 months at a temperature in the range of 2 °C to 28 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 36 months at a temperature in the range of 2 °C to 28 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of at least 42 months at a temperature in the range of 2 °C to 28 °C.

[0080] In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of 3 months at a temperature in the range of 5 °C ± 3 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of 6 months at a temperature in the range of about 5 °C ± 3 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of 9 months at a temperature in the range of about 5 °C ± 3 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of 12 months at a temperature in the range of about 5 °C ± 3 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of 18 months at a temperature in the range of about 5 °C ± 3 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of 24 months at a temperature in the range of about 5 °C ± 3 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of 30 months at a temperature in the range of about 5 °C ± 3 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of 36 months at a temperature in the range of about 5 °C ± 3 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of 42 months at a temperature in the range of about 5 °C ± 3 °C.

[0081] In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of 3 months at a temperature in the range of about 25 °C ± 2 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of 6 months at a temperature in the range of about 25 °C ± 2 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of 9 months at a temperature in the range of about 25 °C ± 2 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of 12 months at a temperature in the range of about 25 °C ± 2 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of 18 months at a temperature in the range of about 25 °C ± 2 °C. In one example, the amount of HMWS, monomer, acidic species, basic species, and / or LMWS is determined after storage for a period of 24 months at a temperature in the range of about 25 °C ± 2 °C.

[0082] In one example, after storage of the formulation for a period of at least 18 months at a temperature of 5 °C ± 3 °C, one or more or all of the following apply:

[0083] (i) the formulation comprises no more than 1.5% HMWS as determined by SE- HPLC; (ii) the formulation comprises no more than 1% LMWS as determined by CE-SDS under non-reducing conditions;

[0084] (iii)at least 97% of the protein in the formulation is a monomer, as determined by SE- HPLC;

[0085] (iv)the formulation comprises no more than 30% acidic species, as determined by CEX-HPLC;

[0086] (v) the formulation comprises no more than 20% basic species, as determined by CEX-HPLC;

[0087] (vi)the formulation comprises less than 20 particles of > 10 pm per container as determined by light obscuration; and

[0088] (vii) the formulation comprises less than 5 particles of > 25 pm per container as determined by light obscuration.

[0089] In one example, after storage of the formulation for a period of at least 18 months at a temperature of 5 °C ± 3 °C, the formulation comprises no more than 1.5% HMWS as determined by SE-HPLC.

[0090] In one example, after storage of the formulation for a period of at least 18 months at a temperature of 5 °C ± 3 °C, the formulation comprises no more than 1% LMWS as determined by CE-SDS under non-reducing conditions.

[0091] In one example, after storage of the formulation for a period of at least 18 months at a temperature of 5 °C ± 3 °C, at least 97% of the protein in the formulation is a monomer, as determined by SE-HPLC.

[0092] In one example, after storage of the formulation for a period of at least 18 months at a temperature of 5 °C ± 3 °C, the formulation comprises no more than 30% acidic species, as determined by CEX-HPLC.

[0093] In one example, after storage of the formulation for a period of at least 18 months at a temperature of 5 °C ± 3 °C, the formulation comprises no more than 20% basic species, as determined by CEX-HPLC.

[0094] In one example, after storage of the formulation for a period of at least 18 months at a temperature of 5 °C ± 3 °C, the formulation comprises less than 20 particles of > 10 pm per container as determined by light obscuration.

[0095] In one example, after storage of the formulation for a period of at least 18 months at a temperature of 5 °C ± 3 °C, the formulation comprises less than 5 particles of > 25 pm per container as determined by light obscuration.

[0096] In one example, after storage of the formulation for a period of at least 24 months at a temperature of 5 °C ± 3 °C, one or more or all of the following apply: (i) the formulation comprises no more than 1.6% HMWS as determined by SE- HPLC;

[0097] (ii)the formulation comprises no more than 1% LMWS as determined by CE-SDS under non-reducing conditions;

[0098] (iii)at least 97% of the protein in the formulation is a monomer, as determined by SE- HPLC;

[0099] (iv)the formulation comprises no more than 31% acidic species, as determined by CEX-HPLC;

[0100] (v) the formulation comprises no more than 20% basic species, as determined by CEX-HPLC;

[0101] (vi)the formulation comprises less than 20 particles of > 10 pm per container as determined by light obscuration; and

[0102] (vii) the formulation comprises less than 5 particles of > 25 pm per container as determined by light obscuration.

[0103] In one example, after storage of the formulation for a period of at least 24 months at a temperature of 5 °C ± 3 °C, the formulation comprises no more than 1.6% HMWS as determined by SE-HPLC.

[0104] In one example, after storage of the formulation for a period of at least 24 months at a temperature of 5 °C ± 3 °C, the formulation comprises no more than 1% LMWS as determined by CE-SDS under non-reducing conditions.

[0105] In one example, after storage of the formulation for a period of at least 24 months at a temperature of 5 °C ± 3 °C, at least 97% of the protein in the formulation is a monomer, as determined by SE-HPLC.

[0106] In one example, after storage of the formulation for a period of at least 24 months at a temperature of 5 °C ± 3 °C, the formulation comprises no more than 31% acidic species, as determined by CEX-HPLC.

[0107] In one example, after storage of the formulation for a period of at least 24 months at a temperature of 5 °C ± 3 °C, the formulation comprises no more than 20% basic species, as determined by CEX-HPLC.

[0108] In one example, after storage of the formulation for a period of at least 24 months at a temperature of 5 °C ± 3 °C, the formulation comprises less than 20 particles of > 10 pm per container as determined by light obscuration,

[0109] In one example, after storage of the formulation for a period of at least 24 months at a temperature of 5 °C ± 3 °C, the formulation comprises less than 5 particles of > 25 pm per container as determined by light obscuration. In one example, after storage of the formulation for a period of at least 30 months at a temperature of 5 °C ± 3 °C, one or more or all of the following apply:

[0110] (i) the formulation comprises no more than 1.7% HMWS as determined by SE- HPLC;

[0111] (ii)the formulation comprises no more than 1% LMWS as determined by CE-SDS under non-reducing conditions;

[0112] (iii)at least 97% of the protein in the formulation is a monomer, as determined by SE- HPLC;

[0113] (iv)the formulation comprises no more than 31% acidic species, as determined by CEX-HPLC;

[0114] (v) the formulation comprises no more than 20% basic species, as determined by CEX-HPLC;

[0115] (vi)the formulation comprises less than 20 particles of > 10 pm per container as determined by light obscuration; and

[0116] (vii) the formulation comprises less than 5 particles of > 25 pm per container as determined by light obscuration.

[0117] In one example, after storage of the formulation for a period of at least 30 months at a temperature of 5 °C ± 3 °C, the formulation comprises no more than 1.7% HMWS as determined by SE-HPLC.

[0118] In one example, after storage of the formulation for a period of at least 30 months at a temperature of 5 °C ± 3 °C, the formulation comprises no more than 1% LMWS as determined by CE-SDS under non-reducing conditions.

[0119] In one example, after storage of the formulation for a period of at least 30 months at a temperature of 5 °C ± 3 °C, at least 97% of the protein in the formulation is a monomer, as determined by SE-HPLC.

[0120] In one example, after storage of the formulation for a period of at least 30 months at a temperature of 5 °C ± 3 °C, the formulation comprises no more than 31% acidic species, as determined by CEX-HPLC.

[0121] In one example, after storage of the formulation for a period of at least 30 months at a temperature of 5 °C ± 3 °C, the formulation comprises no more than 20% basic species, as determined by CEX-HPLC.

[0122] In one example, after storage of the formulation for a period of at least 30 months at a temperature of 5 °C ± 3 °C, the formulation comprises less than 20 particles of > 10 pm per container as determined by light obscuration, In one example, after storage of the formulation for a period of at least 30 months at a temperature of 5 °C ± 3 °C, the formulation comprises less than 5 particles of > 25 pm per container as determined by light obscuration.

[0123] In one example, after storage of the formulation for a period of at least 36 months at a temperature of 5 °C ± 3 °C, one or more or all of the following apply:

[0124] (i) the formulation comprises no more than 1.8% HMWS as determined by SE- HPLC;

[0125] (ii)the formulation comprises no more than 2% LMWS as determined by CE-SDS under non-reducing conditions;

[0126] (iii)at least 97% of the protein in the formulation is a monomer, as determined by SE- HPLC;

[0127] (iv)the formulation comprises no more than 32% acidic species, as determined by CEX-HPLC;

[0128] (v) the formulation comprises no more than 20% basic species, as determined by CEX-HPLC;

[0129] (vi)the formulation comprises less than 20 particles of > 10 pm per container as determined by light obscuration; and

[0130] (vii) the formulation comprises less than 5 particles of > 25 pm per container as determined by light obscuration.

[0131] In one example, after storage of the formulation for a period of at least 36 months at a temperature of 5 °C ± 3 °C, the formulation comprises no more than 1.8% HMWS as determined by SE-HPLC.

[0132] In one example, after storage of the formulation for a period of at least 36 months at a temperature of 5 °C ± 3 °C, the formulation comprises no more than 2% LMWS as determined by CE-SDS under non-reducing conditions.

[0133] In one example, after storage of the formulation for a period of at least 36 months at a temperature of 5 °C ± 3 °C, at least 97% of the protein in the formulation is a monomer, as determined by SE-HPLC.

[0134] In one example, after storage of the formulation for a period of at least 36 months at a temperature of 5 °C ± 3 °C, the formulation comprises no more than 32% acidic species, as determined by CEX-HPLC.

[0135] In one example, after storage of the formulation for a period of at least 36 months at a temperature of 5 °C ± 3 °C, the formulation comprises no more than 20% basic species, as determined by CEX-HPLC. In one example, after storage of the formulation for a period of at least 36 months at a temperature of 5 °C ± 3 °C, the formulation comprises less than 20 particles of > 10 pm per container as determined by light obscuration,

[0136] In one example, after storage of the formulation for a period of at least 36 months at a temperature of 5 °C ± 3 °C, the formulation comprises less than 5 particles of > 25 pm per container as determined by light obscuration.

[0137] In one example, after storage of the formulation for a period of at least 42 months at a temperature of 5 °C ± 3 °C, one or more or all of the following apply:

[0138] (i) the formulation comprises no more than 1.8% HMWS as determined by SE- HPLC;

[0139] (ii)the formulation comprises no more than 2% LMWS as determined by CE-SDS under non-reducing conditions;

[0140] (iii)at least 97% of the protein in the formulation is a monomer, as determined by SE- HPLC;

[0141] (iv)the formulation comprises no more than 32% acidic species, as determined by CEX-HPLC;

[0142] (v) the formulation comprises no more than 20% basic species, as determined by CEX-HPLC;

[0143] (vi)the formulation comprises less than 20 particles of > 10 pm per container as determined by light obscuration; and

[0144] (vii) the formulation comprises less than 5 particles of > 25 pm per container as determined by light obscuration.

[0145] In one example, after storage of the formulation for a period of at least 42 months at a temperature of 5 °C ± 3 °C, the formulation comprises no more than 1.8% HMWS as determined by SE-HPLC.

[0146] In one example, after storage of the formulation for a period of at least 42 months at a temperature of 5 °C ± 3 °C, the formulation comprises no more than 2% LMWS as determined by CE-SDS under non-reducing conditions.

[0147] In one example, after storage of the formulation for a period of at least 42 months at a temperature of 5 °C ± 3 °C, at least 97% of the protein in the formulation is a monomer, as determined by SE-HPLC.

[0148] In one example, after storage of the formulation for a period of at least 42 months at a temperature of 5 °C ± 3 °C, the formulation comprises no more than 32% acidic species, as determined by CEX-HPLC. In one example, after storage of the formulation for a period of at least 42 months at a temperature of 5 °C ± 3 °C, the formulation comprises no more than 20% basic species, as determined by CEX-HPLC.

[0149] In one example, after storage of the formulation for a period of at least 42 months at a temperature of 5 °C ± 3 °C, the formulation comprises less than 20 particles of > 10 pm per container as determined by light obscuration,

[0150] In one example, after storage of the formulation for a period of at least 42 months at a temperature of 5 °C ± 3 °C, the formulation comprises less than 5 particles of > 25 pm per container as determined by light obscuration.

[0151] In one example, after storage of the formulation for a period of at least 18 months at a temperature of 25 °C ± 2 °C, one or more or all of the following apply:

[0152] (i) the formulation comprises no more than 3.5% HMWS as determined by SE- HPLC;

[0153] (ii)the formulation comprises no more than 6% LMWS as determined by CE-SDS under non-reducing conditions;

[0154] (iii)at least 95% of the protein in the formulation is a monomer, as determined by SE- HPLC;

[0155] (iv)the formulation comprises no more than 53% acidic species, as determined by CEX-HPLC;

[0156] (v) , the formulation comprises no more than 20% basic species, as determined by CEX-HPLC;

[0157] (vi)the formulation comprises less than 105 particles of > 10 pm per container as determined by light obscuration; and

[0158] (vii) the formulation comprises less than 5 particles of > 25 pm per container as determined by light obscuration.

[0159] In one example, after storage of the formulation for a period of at least 18 months at a temperature of 25 °C ± 2 °C, the formulation comprises no more than 3.5% HMWS as determined by SE-HPLC.

[0160] In one example, after storage of the formulation for a period of at least 18 months at a temperature of 25 °C ± 2 °C, the formulation comprises no more than 6% LMWS as determined by CE-SDS under non-reducing conditions.

[0161] In one example, after storage of the formulation for a period of at least 18 months at a temperature of 25 °C ± 2 °C, at least 95% of the protein in the formulation is a monomer, as determined by SE-HPLC. In one example, after storage of the formulation for a period of at least 18 months at a temperature of 25 °C ± 2 °C, the formulation comprises no more than 53% acidic species, as determined by CEX-HPLC.

[0162] In one example, after storage of the formulation for a period of at least 18 months at a temperature of 25 °C ± 2 °C, the formulation comprises no more than 20% basic species, as determined by CEX-HPLC.

[0163] In one example, after storage of the formulation for a period of at least 18 months at a temperature of 25 °C ± 2 °C, the formulation comprises less than 105 particles of > 10 pm per container as determined by light obscuration.

[0164] In one example, after storage of the formulation for a period of at least 18 months at a temperature of 25 °C ± 2 °C, the formulation comprises less than 5 particles of > 25 pm per container as determined by light obscuration.

[0165] In one example, after storage of the formulation for a period of at least 24 months at a temperature of 25 °C ± 2 °C, one or more or all of the following apply:

[0166] (i) the formulation comprises no more than 4% HMWS as determined by SE-HPLC;

[0167] (ii)the formulation comprises no more than 10% LMWS as determined by CE-SDS under non-reducing conditions;

[0168] (iii)at least 95% of the protein in the formulation is a monomer, as determined by SE- HPLC;

[0169] (iv)the formulation comprises no more than 51% acidic species, as determined by CEX-HPLC;

[0170] (v) the formulation comprises no more than 20% basic species, as determined by CEX-HPLC;

[0171] (vi)the formulation comprises less than 20 particles of > 10 pm per container as determined by light obscuration; and

[0172] (vii) the formulation comprises less than 5 particles of > 25 pm per container as determined by light obscuration.

[0173] In one example, after storage of the formulation for a period of at least 24 months at a temperature of 25 °C ± 2 °C, the formulation comprises no more than 4% HMWS as determined by SE-HPLC.

[0174] In one example, after storage of the formulation for a period of at least 24 months at a temperature of 25 °C ± 2 °C, the formulation comprises no more than 10% LMWS as determined by CE-SDS under non-reducing conditions.

[0175] In one example, after storage of the formulation for a period of at least 24 months at a temperature of 25 °C ± 2 °C, at least 95% of the protein in the formulation is a monomer, as determined by SE-HPLC. In one example, after storage of the formulation for a period of at least 24 months at a temperature of 25 °C ± 2 °C, the formulation comprises no more than 51% acidic species, as determined by CEX-HPLC.

[0176] In one example, after storage of the formulation for a period of at least 24 months at a temperature of 25 °C ± 2 °C, the formulation comprises no more than 20% basic species, as determined by CEX-HPLC.

[0177] In one example, after storage of the formulation for a period of at least 24 months at a temperature of 25 °C ± 2 °C, the formulation comprises less than 20 particles of > 10 pm per container as determined by light obscuration.

[0178] In one example, after storage of the formulation for a period of at least 24 months at a temperature of 25 °C ± 2 °C, the formulation comprises less than 5 particles of > 25 pm per container as determined by light obscuration.

[0179] In one example, after storage of the formulation for a period of between 1 month and 6 months at a temperature of 35 °C ± 2 °C, one or more or all of the following apply:

[0180] (i) the formulation comprises no more than 5% HMWS as determined by SE-HPLC;

[0181] (ii) the formulation comprises no more than 10% LMWS as determined by CE-SDS under non-reducing conditions;

[0182] (iii)at least 93% of the protein in the formulation is a monomer, as determined by SE- HPLC;

[0183] (iv)the formulation comprises no more than 58% acidic species, as determined by CEX-HPLC;

[0184] (v) the formulation comprises no more than 22% basic species, as determined by CEX-HPLC;

[0185] (vi)the formulation comprises less than 6 particles of > 10 pm per container as determined by light obscuration; and

[0186] (vii) the formulation comprises less than 5 particles of > 25 pm per container as determined by light obscuration.

[0187] In one example, after storage of the formulation for a period of at least 12 months at a temperature of 35 °C ± 2 °C, one or more or all of the following apply:

[0188] (i) the formulation comprises no more than 8% HMWS as determined by SE-HPLC;

[0189] (ii) the formulation comprises no more than 18% LMWS as determined by CE-SDS under non-reducing conditions;

[0190] (iii)at least 88% of the protein in the formulation is a monomer, as determined by SE- HPLC;

[0191] (iv)the formulation comprises no more than 61% acidic species, as determined by CEX-HPLC; (v) the formulation comprises no more than 21% basic species, as determined by CEX-HPLC;

[0192] (vi)the formulation comprises less than 6 particles of > 10 pm per container as determined by light obscuration; and

[0193] (vii) the formulation comprises less than 5 particles of > 25 pm per container as determined by light obscuration.

[0194] In one example, the formulation has a colour value of B5 or B6 and 0 to 1.2% haze as determined using Vista Transmission colorimeter Fa. Hunterlab.

[0195] In one example, the formulation has a volume in the range of 0.4 mL to 2 mL. In one example, the formulation has a volume in the range of 0.5 mL to 2 mL. In one example, the formulation has a volume in the range of 0.5 mL to 1.6 mL. In one example, the formulation has a volume of 2 mL. In one example, the formulation has a volume of 1.8 mL. In one example, the formulation has a volume of 1.6 mL. In a one example, the formulation has a volume of 1.5 mL. In one example, the formulation has a volume of 1.4 mL. In one example, the formulation has a volume of 1.2 mL. In another example, the formulation has a volume of 1 mL. In one example, the formulation has a volume of 0.8 mL. In one example, the formulation has a volume of 0.6 mL. In another example, the formulation has a volume of 0.5 mL. In one example, the formulation has a volume of 0.4 mL.

[0196] In one example, the formulation has a volume of 1.6 mL and 80 mg protein. In a one example, the formulation has a volume of 1.5 mL and 75 mg protein. In one example, the formulation has a volume of 1.4 mL and 70 mg protein. In one example, the formulation has a volume of 1.2 mL and 60 mg protein. In another example, the formulation has a volume of 1 mL and 50 mg protein. In one example, the formulation has a volume of 0.8 mL and 40 mg protein. In one example, the formulation has a volume of 0.6 mL and 30 mg protein. In another example, the formulation has a volume of 0.5 mL and 25 mg protein. In one example, the formulation has a volume of 0.4 mL and 20 mg protein.

[0197] In one example, the protein inhibits granulocyte colony stimulating factor (G- CSF) signaling.

[0198] In one example, the protein comprises an antigen binding domain of an antibody.

[0199] In one example, the protein is selected from the group consisting of:

[0200] (i) a single chain Fv fragment (scFv);

[0201] (ii) a dimeric scFv (di-scFv);

[0202] (iii) a diabody;

[0203] (iv) a triabody; (v) a tetrabody;

[0204] (vi) a Fab;

[0205] (vii) a F(ab’)2;

[0206] (viii) a Fv;

[0207] (ix) one of (i) to (viii) is linked to a constant region of an antibody, Fc or a heavy chain constant domain (CH) CH2 and / or CH3; and

[0208] (x) an antibody.

[0209] In one example, the protein is selected from the group consisting of:

[0210] (i) a single chain Fv fragment (scFv);

[0211] (ii) a dimeric scFv (di-scFv);

[0212] (iii) a diabody;

[0213] (iv) a triabody;

[0214] (v) a tetrabody;

[0215] (vi) a Fab;

[0216] (vii) a F(ab’)2;

[0217] (viii) a Fv;

[0218] (ix) one of (i) to (viii) is linked to a constant region of an antibody, Fc or a heavy chain constant domain (CH) CH2 and / or CH3;

[0219] (x) one of (i) to (viii) linked to albumin, functional fragments or variants thereof or a protein (e.g., antibody or antigen binding fragment thereof) that binds to albumin; and

[0220] (xi) an antibody.

[0221] In one example, the protein is an antibody.

[0222] In one example, the protein comprises:

[0223] (i) a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 5; or

[0224] (ii) a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 3; or

[0225] (iii) a VH comprising:

[0226] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 6;

[0227] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 7; and

[0228] (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 8; and a VL comprising:

[0229] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 9;

[0230] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 10; and

[0231] (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 11. In one example, the protein comprises a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 5.

[0232] In one example, the protein comprises a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 3.

[0233] In one example, the protein comprises: a VH comprising:

[0234] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 6;

[0235] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 7; and

[0236] (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 8; and a VL comprising:

[0237] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 9;

[0238] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 10; and

[0239] (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 11.

[0240] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of between 45 mg / mL and 55 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises:

[0241] (i) a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 5; or

[0242] (ii) a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 3; or

[0243] (iii) a VH comprising:

[0244] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 6;

[0245] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 7; and

[0246] (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 8; and a VL comprising:

[0247] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 9;

[0248] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 10; and

[0249] (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 11.

[0250] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of between 45 mg / mL and 55 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises: a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 5.

[0251] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of between 45 mg / mL and 55 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises: a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 3.

[0252] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of between 45 mg / mL and 55 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises: a VH comprising:

[0253] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 6;

[0254] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 7; and

[0255] (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 8; and a VL comprising:

[0256] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 9;

[0257] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 10; and

[0258] (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 11.

[0259] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of about 50 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises:

[0260] (i) a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 5; or (ii) a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 3; or

[0261] (iii) a VH comprising:

[0262] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 6;

[0263] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 7; and

[0264] (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 8; and a VL comprising:

[0265] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 9;

[0266] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 10; and

[0267] (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 11.

[0268] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of about 50 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 5.

[0269] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of about 50 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 3.

[0270] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of about 50 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises: a VH comprising:

[0271] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 6;

[0272] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 7; and (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 8; and a VL comprising:

[0273] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 9;

[0274] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 10; and

[0275] (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 11.

[0276] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises:

[0277] (i) a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 5; or

[0278] (ii) a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 3; or

[0279] (iii) a VH comprising:

[0280] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 6;

[0281] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 7; and

[0282] (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 8; and a VL comprising:

[0283] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 9;

[0284] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 10; and

[0285] (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 11.

[0286] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 5.

[0287] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 3.

[0288] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises: a VH comprising:

[0289] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 6;

[0290] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 7; and

[0291] (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 8; and a VL comprising:

[0292] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 9;

[0293] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 10; and

[0294] (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 11.

[0295] The present disclosure also provides a pharmaceutical formulation consisting of a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises:

[0296] (iv)a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 5; or

[0297] (v) a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 3; or

[0298] (vi) a VH comprising:

[0299] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 6;

[0300] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 7; and

[0301] (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 8; and a VL comprising:

[0302] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 9;

[0303] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 10; and

[0304] (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 11. The present disclosure also provides a pharmaceutical formulation consisting of a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 5.

[0305] The present disclosure also provides a pharmaceutical formulation consisting of a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 3.

[0306] The present disclosure also provides a pharmaceutical formulation consisting of a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises: a VH comprising:

[0307] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 6;

[0308] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 7; and

[0309] (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 8; and a VL comprising:

[0310] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 9;

[0311] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 10; and

[0312] (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 11.

[0313] In one example, the CDRs are positioned according to the numbering system of Kabat.

[0314] In one example, the protein comprises a VH and a VL, wherein:

[0315] (i) the VH comprises:

[0316] (a) a CDR1 comprising a sequence set forth between amino acids 31-35 of SEQ ID NO: 2; (b) a CDR2 comprising a sequence set forth between amino acids 50-65 of SEQ ID NO: 2; and

[0317] (c) a CDR3 comprising a sequence set forth between amino acids 99-107 of SEQ ID NO: 2; and

[0318] (ii)the VL comprises:

[0319] (a) a CDR1 comprising a sequence set forth between amino acids 24-34 of SEQ ID NO: 3;

[0320] (b) a CDR2 comprising a sequence set forth between amino acids 51-56 of SEQ ID NO: 3; and

[0321] (c) a CDR3 comprising a sequence set forth between amino acids 89-97 of SEQ ID NO: 3.

[0322] In one example, the protein comprises a VH and a VL, wherein:

[0323] (i) the VH comprises:

[0324] (a) a CDR1 comprising a sequence set forth between amino acids 31-35 of SEQ ID NO: 4;

[0325] (b) a CDR2 comprising a sequence set forth between amino acids 50-65 of SEQ ID NO: 4; and

[0326] (c) a CDR3 comprising a sequence set forth between amino acids 99-107 of SEQ ID NO: 4; and

[0327] (ii)the VL comprises:

[0328] (a) a CDR1 comprising a sequence set forth between amino acids 24-34 of SEQ ID NO: 5;

[0329] (b) a CDR2 comprising a sequence set forth between amino acids 51-56 of SEQ ID NO: 5; and

[0330] (c) a CDR3 comprising a sequence set forth between amino acids 89-97 of SEQ ID NO: 5.

[0331] In one example, the protein comprises a VH and a VL, wherein:

[0332] (i) the VH comprises:

[0333] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 6,

[0334] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 7, and

[0335] (c) a CDR3 comprising a sequence set forth in LGELGX1X2X3X4 (SEQ ID NO: 12), wherein:

[0336] Xi is selected from the group consisting of tryptophan, glutamine, methionine, serine, phenylalanine, glutamic acid and histidine;

[0337] X2 is an amino acid selected from the group consisting of phenylalanine, tyrosine, methionine, serine, glycine and isoleucine; Xs is an amino acid selected from the group consisting of aspartic acid, methionine, glutamine, serine, leucine, valine, arginine and histidine; and X4 is any amino acid or an amino acid selected from the group consisting of proline, glutamic acid, alanine, leucine, phenylalanine, tyrosine, threonine, asparagine, aspartic acid, serine, glycine, arginine, and lysine; and

[0338] (ii) a light chain variable region (VL) comprising:

[0339] (a) a CDR1 comprising a sequence set forth in SEQID NO: 9,

[0340] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 10, and

[0341] (c) a CDR3 comprising a sequence set forth in X1X2X3X4X5X6X7X8X9, (SEQ ID NO: 13) wherein:

[0342] Xi is an amino acid selected from the group consisting of glutamine, glutamic acid, histidine, alanine and serine;

[0343] X2 is an amino acid selected from the group consisting of glutamine, valine, phenylalanine, asparagine and glutamic acid;

[0344] X3 is an amino acid selected from the group consisting of serine and glycine; X4 is an amino acid selected from the group consisting of tryptophan, methionine, phenylalanine, tyrosine, isoleucine and leucine;

[0345] X5 is an amino acid selected from the group consisting of glutamic acid, methionine, glutamine, tryptophan, serine, valine, asparagine, glycine, alanine, arginine, histidine, tyrosine, lysine and threonine;

[0346] Xe is an amino acid selected from the group consisting of tyrosine, methionine, isoleucine and threonine;

[0347] X7 is an amino acid selected from the group consisting of proline, alanine, histidine, glycine and lysine;

[0348] Xs is an amino acid selected from the group consisting of leucine, glutamine, methionine, alanine, phenylalanine, isoleucine, lysine, histidine and glycine; and

[0349] X9 is an amino acid selected from the group consisting of threonine, phenylalanine, tyrosine, methionine, lysine, serine, histidine, proline, tryptophan, isoleucine, glutamine, glycine and valine.

[0350] In one example, the protein comprises a VH and a VL, wherein:

[0351] (i) the VH comprises:

[0352] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 6;

[0353] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 7; and

[0354] (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 8; and (ii)the VL comprises:

[0355] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 9;

[0356] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 10; and

[0357] (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 11.

[0358] In one example, the protein comprises a VH and a VL, wherein:

[0359] (A) (i) the VH comprises a CDR1 comprising a sequence set forth between amino acids 31-35 of SEQ ID NO: 2; a CDR2 comprising a sequence set forth between amino acids 50-65 of SEQ ID NO: 2; and a CDR3 comprising a sequence set forth between amino acids 99-107 of SEQ ID NO: 2; and

[0360] (ii) the VL comprises a CDR1 comprising a sequence set forth between amino acids 24-34 of SEQ ID NO: 3; a CDR2 comprising a sequence set forth between amino acids 51-56 of SEQ ID NO: 3; and a CDR3 comprising a sequence set forth between amino acids 89-97 of SEQ ID NO: 3; or

[0361] (B) (i) the VH comprises a CDR1 comprising a sequence set forth between amino acids 31-35 of SEQ ID NO: 4; a CDR2 comprising a sequence set forth between amino acids 50-65 of SEQ ID NO: 4; and a CDR3 comprising a sequence set forth between amino acids 99-107 of SEQ ID NO: 4; and

[0362] (ii) the VL comprises a CDR1 comprising a sequence set forth between amino acids 24-34 of SEQ ID NO: 5; a CDR2 comprising a sequence set forth between amino acids 51-56 of SEQ ID NO: 5; and a CDR3 comprising a sequence set forth between amino acids 89-97 of SEQ ID NO: 5; or

[0363] (C) (i) the VH comprises a CDR1 comprising a sequence set forth in SEQ ID NO: 6; a CDR2 comprising a sequence set forth in SEQ ID NO: 7; and a CDR3 comprising a sequence set forth in SEQ ID NO: 8; and

[0364] (ii) the VL comprises a CDR1 comprising a sequence set forth in SEQ ID NO: 9; a CDR2 comprising a sequence set forth in SEQ ID NO: 10; and CDR3 comprising a sequence set forth in SEQ ID NO: 11.

[0365] In one example, the protein comprises:

[0366] (i) a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22;

[0367] (ii) a VH comprising three CDRs of a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising three CDRs of a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22;

[0368] (iii) a VH comprising: (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 23;

[0369] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 24; and

[0370] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 25; and a VL comprising:

[0371] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 26;

[0372] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 27; and

[0373] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ

[0374] ID NO: 28.

[0375] In one example, the protein is any form of the protein encoded by a nucleic acid encoding any of the foregoing proteins.

[0376] In one example, the protein comprises a VH expressed by a nucleic acid encoding an amino acid of SEQ ID NO: 4 and a VL expressed by a nucleic acid encoding an amino acid of SEQ ID NO: 5.

[0377] In one example, the protein comprises a VH expressed by a nucleic acid encoding an amino acid of SEQ ID NO: 2 and a VL expressed by a nucleic acid encoding an amino acid of SEQ ID NO: 3.

[0378] In one example, the protein comprises:

[0379] (i) a VH comprising:

[0380] (a) a CDR1 expressed by a nucleic acid encoding an amino acid of SEQ ID NO: 6;

[0381] (b) a CDR2 expressed by a nucleic acid encoding an amino acid of SEQ ID NO: 7; and

[0382] (c) a CDR3 expressed by a nucleic acid encoding an amino acid of SEQ ID NO: 8; and

[0383] (ii) a VL comprising:

[0384] (a) a CDR1 expressed by a nucleic acid encoding an amino acid of SEQ ID NO: 9;

[0385] (b) a CDR2 expressed by a nucleic acid encoding an amino acid of SEQ ID NO: 10; and

[0386] (c) a CDR3 expressed by a nucleic acid encoding an amino acid of SEQ ID NO: 11. In one example, the protein comprises:

[0387] (i) a VH comprising:

[0388] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 23;

[0389] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 24; and

[0390] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 25; and

[0391] (ii) a VL comprising:

[0392] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 26;

[0393] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 27; and

[0394] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 28.

[0395] In one example, the protein comprises:

[0396] (i) a VH comprising:

[0397] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 23;

[0398] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 24; and

[0399] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 25; and

[0400] (ii) a VL comprising:

[0401] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 26;

[0402] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 27; and

[0403] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 28.

[0404] In one example, the protein comprises a VH comprising three CDRs of a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising three CDRs of a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22. In one example, the protein comprises a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21; and a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22.

[0405] In one example, the protein comprises a heavy chain comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 19; and a light chain comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 20.

[0406] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of between 45 mg / mL and 55 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises:

[0407] (i) a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO:

[0408] 21 and a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22;

[0409] (ii) a VH comprising three CDRs of a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising three CDRs of a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22; or

[0410] (iii) a VH comprising:

[0411] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 23;

[0412] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 24; and

[0413] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 25; and a VL comprising:

[0414] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 26;

[0415] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 27; and

[0416] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 28.

[0417] The present disclosure thus provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of between 45 mg / mL and 55 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22.

[0418] The present disclosure thus provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of between 45 mg / mL and 55 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises a VH comprising three CDRs of a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising three CDRs of a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22.

[0419] The present disclosure thus provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of between 45 mg / mL and 55 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises: a VH comprising:

[0420] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 23;

[0421] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 24; and

[0422] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 25; and a VL comprising:

[0423] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 26;

[0424] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 27; and (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 28.

[0425] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of about 50 mg / mE, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises:

[0426] (i) a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VE comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22;

[0427] (ii) a VH comprising three CDRs of a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising three CDRs of a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22; or

[0428] (iii) a VH comprising:

[0429] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 23;

[0430] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 24; and

[0431] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 25; and a VL comprising:

[0432] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 26;

[0433] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 27; and

[0434] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 28.

[0435] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of about 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22.

[0436] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of about 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises a VH comprising three CDRs of a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising three CDRs of a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22.

[0437] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of about 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises: a VH comprising:

[0438] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 23;

[0439] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 24; and

[0440] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 25; and a VL comprising:

[0441] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 26;

[0442] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 27; and

[0443] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 28.

[0444] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises:

[0445] (i) a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22;

[0446] (ii) a VH comprising three CDRs of a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising three CDRs of a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22; or

[0447] (iii) a VH comprising:

[0448] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 23;

[0449] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 24; and

[0450] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 25; and a VL comprising:

[0451] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 26;

[0452] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 27; and

[0453] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 28.

[0454] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22.

[0455] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises a VH comprising three CDRs of a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising three CDRs of a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22.

[0456] The present disclosure also provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises: a VH comprising:

[0457] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 23;

[0458] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 24; and

[0459] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 25; and a VL comprising:

[0460] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 26;

[0461] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 27; and

[0462] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 28.

[0463] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of between 45 mg / mL and 55 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises: (i) a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22;

[0464] (ii) a VH comprising three CDRs of a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising three CDRs of a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22; or

[0465] (iii) a VH comprising:

[0466] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 23;

[0467] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 24; and

[0468] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 25; and a VL comprising:

[0469] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 26;

[0470] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 27; and

[0471] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 28.

[0472] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of between 45 mg / mL and 55 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22.

[0473] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of between 45 mg / mL and 55 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises a VH comprising three CDRs of a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising three CDRs of a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22.

[0474] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of between 45 mg / mL and 55 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises: a VH comprising:

[0475] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 23;

[0476] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 24; and

[0477] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 25; and a VL comprising:

[0478] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 26;

[0479] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 27; and

[0480] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 28.

[0481] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of about 50 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises:

[0482] (i) a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO:

[0483] 21 and a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22;

[0484] (ii) a VH comprising three CDRs of a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising three CDRs of a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22; or

[0485] (iii) a VH comprising: (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 23;

[0486] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 24; and

[0487] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 25; and a VL comprising:

[0488] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 26;

[0489] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 27; and

[0490] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 28.

[0491] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of about 50 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to

[0492] 5.9, and wherein the antigen binding domain comprises a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22.

[0493] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of about 50 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to

[0494] 5.9, and wherein the antigen binding domain comprises a VH comprising three CDRs of a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising three CDRs of a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22.

[0495] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of about 50 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to

[0496] 5.9, and wherein the antigen binding domain comprises: a VH comprising:

[0497] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 23;

[0498] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 24; and

[0499] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 25; and a VL comprising:

[0500] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 26;

[0501] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 27; and

[0502] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 28.

[0503] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises:

[0504] (i) a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO:

[0505] 21 and a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22;

[0506] (ii) a VH comprising three CDRs of a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising three CDRs of a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22; or

[0507] (iii) a VH comprising:

[0508] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 23;

[0509] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 24; and

[0510] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 25; and a VL comprising: (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 26;

[0511] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 27; and

[0512] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 28.

[0513] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22.

[0514] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises a VH comprising three CDRs of a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising three CDRs of a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22.

[0515] The present disclosure also provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises: a VH comprising:

[0516] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 23;

[0517] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 24; and

[0518] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 25; and a VL comprising:

[0519] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 26;

[0520] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 27; and

[0521] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 28.

[0522] In one example, the protein as described herein comprises a human constant region, e.g., an IgG constant region, such as an IgGi, IgGi, IgGs or IgG4 constant region or mixtures thereof. In the case of a protein comprising a VH and a VL, the VH can be linked to a heavy chain constant region and the VL can be linked to a light chain constant region.

[0523] In one example, the protein as described herein comprises a constant region of an IgG4antibody or a stabilized constant region of an IgG4 antibody. In one example, the protein comprises an IgG4 constant region with a proline at position 241 (according to the numbering system of Kabat (Kabat et al., Sequences of Proteins of Immunological Interest Washington DC United States Department of Health and Human Services, 1987 and / or 1991)).

[0524] The C-terminal lysine of the heavy chain constant region of a whole antibody (or a protein or antibody comprising a constant region or a CH3) of the disclosure may be removed, for example, during production or purification of the protein, or by recombinantly engineering the nucleic acid encoding a heavy chain of the antibody. Accordingly, whole antibodies may comprise populations with all C-terminal lysine residues removed, populations with no C-terminal lysine residues removed, and / or populations having a mixture of protein with and without the C-terminal lysine residue. In some examples, the populations may additionally comprise protein in which the C- terminal lysine residue is removed in one of the heavy chain constant regions. Similarly, a composition of whole antibodies may comprise the same or a similar mix of antibody populations with or without the C-terminal lysine residue.

[0525] In one example the protein as described herein or a composition of the protein as described herein, comprises a heavy chain constant region, including a stabilized heavy chain constant region, comprising a mixture of sequences fully or partially with or without the C-terminal lysine residue.

[0526] In one example, the protein comprises a VH disclosed herein linked or fused to an IgG4 constant region or stabilized IgG4 constant region (e.g., as discussed above) and the VL is linked to or fused to a kappa light chain constant region. In one example, the protein comprises:

[0527] (i) a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 14 or 18 and a light chain comprising an amino acid sequence set forth in SEQ ID NO: 15; or

[0528] (ii) one heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 14 and one heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 18 and two light chains comprising an amino acid sequence set forth in SEQ ID NO: 15.

[0529] In one example, the protein comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 14 or 18 and a light chain comprising an amino acid sequence set forth in SEQ ID NO: 15. For example, the protein comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 14 and a light chain comprising an amino acid sequence set forth in SEQ ID NO: 15. In another example, the protein is comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 18 and a light chain comprising an amino acid sequence set forth in SEQ ID NO: 15.

[0530] In one example, the protein comprises one heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 14 and one heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 18 and two light chains comprising an amino acid sequence set forth in SEQ ID NO: 15.

[0531] In one example, the pharmaceutical formulation of the present disclosure is:

[0532] (i) formulated for subcutaneous administration; and / or

[0533] (ii) formulated for self-administration; and / or

[0534] (iii) formulated for fixed-dose administration.

[0535] In one example, the pharmaceutical formulation of the present disclosure is formulated for subcutaneous administration. In one example, the pharmaceutical formulation of the present disclosure is formulated for self-administration. In one example, the pharmaceutical formulation of the present disclosure is formulated for fixed-dose administration.

[0536] In one example, the pharmaceutical formulation of the present disclosure is for use as a medicament.

[0537] The present disclosure also provides a method of reducing circulating neutrophils in a human subject suffering from a neutrophil-mediated condition, the method comprising administering the pharmaceutical formulation of the disclosure to the subject. The present disclosure also provides a method of treating or preventing a neutrophil-mediated condition in a subject, the method comprising administering the pharmaceutical formulation of the disclosure to the subject.

[0538] The present disclosure also provides a composition for use in reducing circulating neutrophils in a human subject suffering from a neutrophil-mediated condition, the composition comprising the pharmaceutical formulation of the disclosure.

[0539] The present disclosure also provides a composition for use in treating or preventing a neutrophil-mediated condition in a subject, the composition comprising the pharmaceutical formulation of the disclosure.

[0540] The present disclosure also provides the use of a pharmaceutical formulation of the disclosure in the manufacture of a medicament for reducing circulating neutrophils in a human subject suffering from a neutrophil-mediated condition.

[0541] The present disclosure also provides the use of a pharmaceutical formulation of the disclosure in the manufacture of a medicament for treating or preventing a neutrophil- mediated condition.

[0542] In one example, the neutrophil-mediated condition is

[0543] (i) a neutrophilic skin condition; an autoimmune disease; an inflammatory disease; ischemia-reperfusion injury; a pulmonary disease; a neurological condition; or a neurodegenerative condition; or

[0544] (ii) a neutrophilic dermatosis; amicrobial pustulosis of the folds (APF); plaque psoriasis; CARD 14 -mediated pustular psoriasis (CAMPS); cryopyrin associated periodic syndromes (CAPS); deficiency of interleukin- 1 receptor (DIRA); deficiency of interleukin- 36 receptor antagonist(DIRTA); hidradenitis suppurativa (HS); palmoplantar pustulosis; pyogenic arthritis; pyoderma gangrenosum and acne (PAPA); pyoderma gangrenosum; acne; and hidradenitis suppurativa (PASH); pyoderma gangrenosum(PG); skin lesions of Behcet’s disease; Still’s disease; Sweet syndrome; subcorneal pustulosis (Sneddon- Wilkinson); pustular psoriasis; palmoplantar pustulosis; acute generalized exanthematic pustulosis; infantile acropustulosis; synovitis; acne; pustulosis; hyperostosis and osteitis (SAPHO) syndrome; bowel-associated dermatosisarthritis syndrome (BAD AS); neutrophilic dermatosis of the dorsal hands; neutrophilic eccrine hidradenitis; erythema elevatum diutinum; Pyoderma gangrenosum; systemic inflammatory response syndrome (SIRS); chronic obstructive pulmonary disease (COPD); emphysema; bronchitis; pneumonia; acute respiratory distress syndrome; acute lung injury; asthma; multiple Sclerosis(MS); Devic's disease; a viral infection in the brain; uveitis; or arthritis. In one example, the neutrophil-mediated condition is a neutrophilic skin condition; an autoimmune disease; an inflammatory disease; ischemia-reperfusion injury; a pulmonary disease; a neurological condition; or a neurodegenerative condition. In one example, the neutrophil-mediated condition is a neutrophilic skin condition. In one example, the neutrophil-mediated condition is an autoimmune disease. In one example, the neutrophil-mediated condition is an inflammatory disease. In one example, the neutrophil-mediated condition is ischemia-reperfusion injury. In one example, the neutrophil-mediated condition is a pulmonary disease. In one example, the neutrophil- mediated condition is a neurological condition. In one example, the neutrophil-mediated condition is a neurodegenerative condition.

[0545] In one example, the neutrophil mediated condition is a neutrophilic dermatosis; amicrobial pustulosis of the folds (APF); plaque psoriasis; CARD14-mediated pustular psoriasis (CAMPS); cryopyrin associated periodic syndromes (CAPS); deficiency of interleukin- 1 receptor (DIRA); deficiency of interleukin-36 receptor antagonist(DIRTA); hidradenitis suppurativa (HS); palmoplantar pustulosis; pyogenic arthritis; pyoderma gangrenosum and acne (PAPA); pyoderma gangrenosum; acne; and hidradenitis suppurativa (PASH); pyoderma gangrenosum(PG); skin lesions of Behcet’s disease; Still’s disease; Sweet syndrome; subcorneal pustulosis (Sneddon-Wilkinson); pustular psoriasis; palmoplantar pustulosis; acute generalized exanthematic pustulosis; infantile acropustulosis; synovitis; acne; pustulosis; hyperostosis and osteitis (SAPHO) syndrome; bowel-associated dermatosis-arthritis syndrome (BAD AS); neutrophilic dermatosis of the dorsal hands; neutrophilic eccrine hidradenitis; erythema elevatum diutinum; Pyoderma gangrenosum; systemic inflammatory response syndrome (SIRS); chronic obstructive pulmonary disease (COPD); emphysema; bronchitis; pneumonia; acute respiratory distress syndrome; acute lung injury; asthma; multiple Sclerosis(MS); Devic's disease; a viral infection in the brain; uveitis; or arthritis

[0546] In one example, neutrophil-mediated condition is neutrophilic dermatosis or systemic inflammatory response syndrome (SIRS). In one example, neutrophil-mediated condition is neutrophilic dermatosis. In one example, neutrophil-mediated condition is systemic inflammatory response syndrome (SIRS).

[0547] In one example, the neutrophilic dermatosis is hidradenitis suppurativa (HS).

[0548] In one example, the systemic inflammatory response syndrome (SIRS) is due to or associated with surgery.

[0549] In one example, (i) the formulation is administered subcutaneously to the subject; (ii) the formulation is self-administered to the subject; and / or (iii) the formulation is administered to the subject using a fixed-dose administration regimen. In one example, (i) the formulation is administered subcutaneously to the subject; (ii) the formulation is self-administered to the subject; or (iii) the formulation is administered to the subject using a fixed-dose administration regimen. In one example, (i) the formulation is administered subcutaneously to the subject; (ii) the formulation is self-administered to the subject; and (iii) the formulation is administered to the subject using a fixed-dose administration regimen. In one example, the formulation is administered subcutaneously to the subject. In one example, the formulation is self-administered to the subject. In one example, the formulation is administered to the subject using a fixed-dose administration regimen.

[0550] The present disclosure also provides a kit for use in treating or preventing a neutrophil-mediated condition in a subject, the kit comprising:

[0551] (a) at least one formulation or pharmaceutical formulation of the disclosure;

[0552] (b) instructions for using the kit in treating or preventing the neutrophil-mediated condition in the subject; and

[0553] (c) optionally, at least one further therapeutically active compound or drug.

[0554] In one example, the formulation or pharmaceutical formulation is present in a vial, a prefilled syringe or an autoinjector device.

[0555] The present disclosure also provides a prefilled syringe comprising the formulation or pharmaceutical formulation of the disclosure.

[0556] The present disclosure also provides an autoinjector device comprising the formulation or pharmaceutical formulation of the disclosure.

[0557] BRIEF DESCRIPTION OF THE FIGURES

[0558] Figure 1 is a table showing excipient concentrations of 24 test formulations.

[0559] Figure 2 is a graph showing turbidity, thermal unfolding, % acidic species, and low molecular weight species in formulations. Figure 2A is a graph showing the % of turbidity of formulations with increasing protein, histidine, arginine, and proline concentrations and pH. Data is shown for protein concentration and pH only and is representative of evaluation at the initial timepoint. Figure 2B is a graph showing thermal unfolding of formulations with increasing protein, histidine, arginine, and proline concentrations and pH, as measured by nanoDSF. Data is shown for protein concentration and pH only and is representative of evaluation at the initial timepoint. Figure 2C-2D is a graph showing % of acidic species of formulations with increasing protein, histidine, arginine, and proline concentrations and pH, as measured by CEX. (2C) Data is shown for protein concentration and pH only and is representative of evaluation after 3 months at +35 °C. (2D) Data is shown for protein concentration and pH only and is representative of evaluation after 3 months at +5 °C. Figure 2E is a graph showing low molecular weight species of formulations with increasing protein, histidine, arginine, and proline concentrations and pH, as measured by CE-SDS under nonreducing conditions. Data is shown for protein concentration and pH only and is representative of evaluation after 3 months at +35 °C.

[0560] Figure 3 is a graph showing % of high molecular weight species of formulations with increasing protein, histidine, arginine, and proline concentrations and pH, as measured by SE-HPLC. (A) Data is shown for protein concentration and pH only and representative of evaluation after 3 months at +35°C. (B) Data is representative of evaluation after 3 months at +5 °C. (C) Data is shown for protein concentration and pH only and is representative of evaluation after 3 months at +5 °C. (D) Data is shown for protein concentration and pH only and is representative of evaluation after 3 months at +25°C.

[0561] Figure 4 is a graph showing %PD, Rh and subvisible particle characterisation. Figure 4A is a graph showing %PD and Rh of formulations with increasing protein, histidine, arginine, and proline concentrations and pH, as measured by dynamic light scattering (DLS). Data is shown for protein concentration and pH only and is representative of evaluation at the initial time point. Figure 4B is a graph showing subvisible particle characterisation of formulations with increasing protein, histidine, arginine, and proline concentrations and pH, as measured by BMI using Horizon. Data is shown for protein concentration and pH only and is representative of evaluation after 3 months at +35°C.

[0562] KEY TO SEQUENCE LISTING

[0563] SEQ ID NO: 1 amino acids 25-335 of Homo sapiens G-CSFR (hG-CSFR) with a C-terminal polyhistidine tag

[0564] SEQ ID NO: 2 Vn of Cl.2

[0565] SEQ ID NO: 3 VL 0f C1.2

[0566] SEQ ID NO: 4 VH of C1.2G

[0567] SEQ ID NO: 5 VLof C1.2G

[0568] SEQ ID NO: 6 HCDR1 of C 1.2

[0569] SEQ ID NO: 7 HCDR2 of C 1.2

[0570] SEQ ID NO: 8 HCDR3 of C 1.2

[0571] SEQ ID NO: 9 LCDR1 of Cl.2

[0572] SEQ ID NO: 10 LCDR2 of C1.2

[0573] SEQ ID NO: 11 LCDR3 of Cl.2 SEQ ID NO: 12 consensus sequence of HCDR3 of Cl.2

[0574] SEQ ID NO: 13 consensus sequence of LCDR3 of Cl.2

[0575] SEQ ID NO: 14 Heavy chain of C1.2G with stabilized IgG4 constant region

[0576] SEQ ID NO: 15 Light chain of C1.2G with kappa constant region

[0577] SEQ ID NO: 16 sequence of exemplary h-G-CSFR

[0578] SEQ ID NO: 17 polypeptide comprising Ig and CRH domains of Macaca fascicularis G-CSFR (cynoG-CSFR) with a C-terminal polyhistidine tag

[0579] SEQ ID NO: 18 Heavy chain of C1.2G with stabilized IgG4 constant region and lacking C-terminal lysine

[0580] SEQ ID NO: 19 nucleotide sequence of heavy chain of C1.2G

[0581] SEQ ID NO: 20 nucleotide sequence of light chain of C1.2G

[0582] SEQ ID NO: 21 nucleotide sequence of VH of C1.2G

[0583] SEQ ID NO: 22 nucleotide sequence of VL of C1.2G

[0584] SEQ ID NO: 23 nucleotide sequence of HCDR1 of C1.2G

[0585] SEQ ID NO: 24 nucleotide sequence of HCDR2 of C1.2G

[0586] SEQ ID NO: 25 nucleotide sequence of HCDR3 of C1.2G

[0587] SEQ ID NO: 26 nucleotide sequence of LCDR1 of C1.2G

[0588] SEQ ID NO: 27 nucleotide sequence of LCDR2 of C1.2G

[0589] SEQ ID NO: 28 nucleotide sequence of LCDR3 of C1.2G

[0590] DETAILED DESCRIPTION

[0591] General

[0592] Throughout this specification, unless specifically stated otherwise or the context requires otherwise, reference to a single step, composition of matter, group of steps or group of compositions of matter shall be taken to encompass one and a plurality (i.e. one or more) of those steps, compositions of matter, groups of steps or groups of compositions of matter.

[0593] Those skilled in the art will appreciate that the present disclosure is susceptible to variations and modifications other than those specifically described. It is to be understood that the disclosure includes all such variations and modifications. The disclosure also includes all of the steps, features, compositions and compounds referred to or indicated in this specification, individually or collectively, and any and all combinations or any two or more of said steps or features. The present disclosure is not to be limited in scope by the specific examples described herein, which are intended for the purpose of exemplification only. Functionally-equivalent products, compositions and methods are clearly within the scope of the present disclosure.

[0594] Any example of the present disclosure herein shall be taken to apply mutatis mutandis to any other example of the disclosure unless specifically stated otherwise. Stated another way, any specific example of the present disclosure may be combined with any other specific example of the disclosure (except where mutually exclusive).

[0595] Any example of the present disclosure disclosing a specific feature or group of features or method or method steps will be taken to provide explicit support for disclaiming the specific feature or group of features or method or method steps.

[0596] Unless specifically defined otherwise, all technical and scientific terms used herein shall be taken to have the same meaning as commonly understood by one of ordinary skill in the art (for example, in cell culture, molecular genetics, immunology, immunohistochemistry, protein chemistry, and biochemistry).

[0597] Unless otherwise indicated, the recombinant protein, cell culture, and immunological techniques utilized in the present disclosure are standard procedures, well known to those skilled in the art. Such techniques are described and explained throughout the literature in sources such as, J. Perbal, A Practical Guide to Molecular Cloning, John Wiley and Sons (1984), J. Sambrook et al. Molecular Cloning: A Laboratory Manual, Cold Spring Harbour Laboratory Press (1989), T.A. Brown (editor), Essential Molecular Biology: A Practical Approach, Volumes 1 and 2, IRL Press (1991), D.M. Glover and B.D. Hames (editors), DNA Cloning: A Practical Approach, Volumes 1-4, IRL Press (1995 and 1996), and F.M. Ausubel et al. (editors), Current Protocols in Molecular Biology, Greene Pub. Associates and Wiley-Interscience (1988, including all updates until present), Ed Harlow and David Lane (editors) Antibodies: A Laboratory Manual, Cold Spring Harbour Laboratory, (1988), and J.E. Coligan et al. (editors) Current Protocols in Immunology, John Wiley & Sons (including all updates until present).

[0598] The description and definitions of variable regions and parts thereof, antibodies and fragments thereof herein may be further clarified by the discussion in Kabat Sequences of Proteins of Immunological Interest, National Institutes of Health, Bethesda, Md., 1987 and 1991.

[0599] The term “EU numbering system of Kabat” will be understood to mean the numbering of an antibody heavy chain is according to the EU index as taught in Kabat et al., 1991, Sequences of Proteins of Immunological Interest, 5th Ed., United States Public Health Service, National Institutes of Health, Bethesda. The EU index is based on the residue numbering of the human IgGl EU antibody. Any discussion of a protein or antibody herein will be understood to include any variants of the protein or antibody produced during manufacturing and / or storage. For example, during manufacturing or storage an antibody can be deamidated (e.g., at an asparagine or a glutamine residue) and / or have misincorporated amino acid residues (e.g., a serine misincorporated in place of an asparagine residue) and / or have altered glycosylation and / or have a glutamine residue converted to pyroglutamine and / or have a N-terminal or C-terminal residue removed or “clipped” and / or have part or all of a signal sequence incompletely processed and, as a consequence, remain at the terminus of the antibody. It is understood that a composition comprising a particular amino acid sequence may be a heterogeneous mixture of the stated or encoded sequence and / or variants of that stated or encoded sequence.

[0600] The term “and / or”, e.g., “X and / or Y” shall be understood to mean either “X and Y” or “X or Y” and shall be taken to provide explicit support for both meanings or for either meaning.

[0601] As used herein, the term “about”, unless stated to the contrary, refers to + / - 10% of the designated value. In some examples, the term “about” refers to + / - 5% of the designated value.

[0602] Throughout this specification the word “comprise”, or variations such as “comprises” or “comprising”, will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps.

[0603] As used herein the term “derived from” shall be taken to indicate that a specified integer may be obtained from a particular source albeit not necessarily directly from that source.

[0604] All publications cited herein are hereby incorporated by reference in their entirety. Where reference is made to a URL or other such identifier or address, it is understood that such identifiers can change and particular information on the internet can come and go, but equivalent information can be found by searching the internet. Reference thereto evidences the availability and public dissemination of such information.

[0605] Any discussion of documents, acts, materials, devices, articles or the like that has been included in the present specification is solely for the purpose of providing a context for the present invention. It is not to be taken as an admission that any or all of these matters form part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed before the priority date of each claim of this application. Selected Definitions

[0606] Reference herein to “granulocyte colony-stimulating factor” (G-CSF) includes native forms of G-CSF, mutant forms thereof, e.g., filgrastim and pegylated forms of G- CSF or filgrastim. This term also encompasses mutant forms of G-CSF retaining activity to bind to G-CSFR (e.g., human G-CSFR) and induce signaling.

[0607] G-CSF is a major regulator of granulocyte production. G-CSF is produced by bone marrow stromal cells, endothelial cells, macrophages, and fibroblasts, and production is induced by inflammatory stimuli. G-CSF acts through the G-CSF receptor (G-CSFR), which is expressed on early myeloid progenitors, mature neutrophils, monocytes / macrophages, T and B lymphocytes and endothelial cells.

[0608] For the purposes of nomenclature only and not limitation, an exemplary sequence of a human G-CSFR is set out in NCBI Reference Sequence: NP_000751.1 (and set out in SEQ ID NO: 16). The sequence of G-CSFR from other species can be determined using sequences provided herein and / or in publically available databases and / or determined using standard techniques (e.g., as described in Ausubel et al., (editors), Current Protocols in Molecular Biology, Greene Pub. Associates and Wiley-Interscience (1988, including all updates until present) or Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press (1989)) Reference to human G-CSFR may be abbreviated to hG-CSFR and reference to cynomolgus monkey G- CSFR may be abbreviated to cynoG-CSFR. Reference to soluble G-CSFR refers to polypeptides comprising the ligand binding region of G-CSFR. The Ig and CRH domains of the G-CSFR are involved in ligand binding and receptor dimerization (Layton et al., J. Biol Chem., 272: 29735-29741, 1997 and Fukunaga et al, EMBO J. 10: 2855- 2865, 1991). Soluble forms of G-CSFR comprising these portions of the receptor have been used in various studies of the receptor and mutation of the free cysteines at positions 78, 163, and 228 of the receptor assists in expression and isolation of the soluble receptor polypeptide (Mine et al., Biochem., 43: 2458-2464 2004) without affecting ligand binding.

[0609] As used herein, the term “G-CSF signaling” refers to biological activities mediated via the G-CSF receptor (G-CSFR). It will be understood that reference to inhibiting G-CSF signaling encompasses inhibition of G-CSF activity, including downstream pathways, mediated via the receptor.

[0610] A “stable” formulation is one in which the protein in the formulation essentially retains its physical stability and / or chemical stability and / or biological activity upon storage. In the context of the present disclosure, the term “monomer” or “monomeric” refers to the correctly folded protein (e.g., antibody or antigen binding fragment thereof). For example, a monomer of an antibody according to the present disclosure relates to the standard tetrameric antibody comprising two identical, glycosylated heavy and light chains respectively. An “aggregate” is a non-specific association of two or more protein molecules (e.g., high molecular weight species).

[0611] As used herein, the term “specifically binds” or “binds specifically” shall be taken to mean that a protein described herein reacts or associates more frequently, more rapidly, with greater duration and / or with greater affinity with a particular molecule (e.g., antigen) than it does with alternative molecules. For example, a protein may bind to G- CSFR (e.g., hG-CSFR) with materially greater affinity (e.g., 20 fold or 40 fold or 60 fold or 80 fold to 100 fold or 150 fold or 200 fold) than it does to other cytokine receptor or to antigens commonly recognized by polyreactive natural antibodies (i.e., by naturally occurring antibodies known to bind a variety of antigens naturally found in humans). Generally, but not necessarily, reference to binding means specific binding, and each term shall be understood to provide explicit support for the other term.

[0612] For the purposes of clarification and as will be apparent to the skilled artisan based on the exemplified subject matter herein, reference to “affinity” in this specification is a reference to KD of a protein or antibody. For the purposes of clarification and as will be apparent to the skilled artisan based on the description herein, reference to an “affinity of at least about” will be understood to mean that the affinity (or KD) is equal to the recited value or higher (i.e., the value recited as the affinity is lower), i.e., an affinity of 2 nM is greater than an affinity of 3 nM. Stated another way, this term could be “an affinity of X or less”, wherein X is a value recited herein.

[0613] The term “recombinant” shall be understood to mean the product of artificial genetic recombination. Accordingly, in the context of a protein comprising an antigen binding domain described herein, this term does not encompass an antibody naturally occurring within a subject’s body that is the product of natural recombination that occurs during B cell maturation. However, if such an antibody is isolated, it is to be considered an isolated protein comprising an antigen binding domain. Similarly, if nucleic acid encoding the protein is isolated and expressed using recombinant means, the resulting protein is a recombinant protein comprising an antibody antigen binding domain. A recombinant protein also encompasses a protein expressed by artificial recombinant means when it is within a cell, tissue or subject, e.g., in which it is expressed. The term “protein” shall be taken to include a single polypeptide chain, i.e., a series of contiguous amino acids linked by peptide bonds or a series of polypeptide chains covalently or non-covalently linked to one another (i.e., a polypeptide complex). For example, the series of polypeptide chains can be covalently linked using a suitable chemical or a disulfide bond. Examples of non-covalent bonds include hydrogen bonds, ionic bonds, Van der Waals forces, and hydrophobic interactions.

[0614] The term “polypeptide” or “polypeptide chain” will be understood from the foregoing paragraph to mean a series of contiguous amino acids linked by peptide bonds.

[0615] As used herein, the term “antigen binding domain” or “antigen binding site” shall be taken to mean a structure formed by a protein that is capable of binding or specifically binding to an antigen. The antigen binding domain need not be a series of contiguous amino acids, or even amino acids in a single polypeptide chain. For example, in a Fv produced from two different polypeptide chains the antigen binding domain is made up of a series of amino acids of a VL and a VH that interact with the antigen and that are generally, however not always in the one or more of the CDRs in each variable region. In some examples, an antigen binding domain is or comprises a VH or a VL or a Fv. In some examples, the antigen binding domain comprises one or more CDRs of an antibody.

[0616] The skilled artisan will be aware that an “antibody” is generally considered to be a protein that comprises a variable region made up of a plurality of polypeptide chains, e.g., a polypeptide comprising a VL and a polypeptide comprising a VH. An antibody also generally comprises constant domains, some of which can be arranged into a constant region, which includes a constant fragment or fragment crystallizable (Fc), in the case of a heavy chain. A VH and a VL interact to form a Fv comprising an antigen binding region that is capable of specifically binding to one or a few closely related antigens. Generally, a light chain from mammals is either a K light chain or a X light chain and a heavy chain from mammals is a, 5, a, y, or p. Antibodies can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgGi, IgGi. IgGs, IgG4, IgAi and IgAi) or subclass. The term “antibody” also encompasses humanized antibodies, primatized antibodies, human antibodies and chimeric antibodies.

[0617] The terms "full-length antibody," "intact antibody" or "whole antibody" are used interchangeably to refer to an antibody in its substantially intact form, as opposed to an antigen binding fragment of an antibody. Specifically, whole antibodies include those with heavy and light chains including an Fc region. The constant domains may be wildtype sequence constant domains (e.g., human wild-type sequence constant domains) or amino acid sequence variants thereof. As used herein, “variable region" refers to the portions of the light and / or heavy chains of an antibody as defined herein that is capable of specifically binding to an antigen and includes amino acid sequences of complementarity determining regions (CDRs); i.e., CDR1, CDR2, and CDR3, and framework regions (FRs). Exemplary variable regions comprise three or four FRs (e.g., FR1, FR2, FR3 and optionally FR4) together with three CDRs. In the case of a protein derived from an IgNAR, the protein may lack a CDR2. VH refers to the variable region of the heavy chain. VL refers to the variable region of the light chain.

[0618] As used herein, the term "complementarity determining regions” (syn. CDRs; i.e., CDR1, CDR2, and CDR3) refers to the amino acid residues of an antibody variable region the presence of which are necessary for antigen binding. Each variable region typically has three CDR regions identified as CDR1, CDR2 and CDR3. The amino acid positions assigned to CDRs and FRs can be defined according to Kabat Sequences of Proteins of Immunological Interest, National Institutes of Health, Bethesda, Md., 1987 and 1991 or other numbering systems in the performance of this disclosure, e.g., the canonical numbering system of Chothia and Eesk J. Mol Biol. 196: 901-917, 1987; Chothia et al. Nature 342, 877-883, 1989; and / or Al-Eazikani et al., J Mol Biol 273: 927-948, 1997; the IMGT numbering system of Eefranc et al., Devel. And Compar. Immunol., 27: 55- 77 , 2003; or the AHO numbering system of Honnegher and Pliikthun J. Mol. Biol., 309: 657-670, 2001. For example, according to the numbering system of Kabat, VH framework regions (FRs) and CDRs are positioned as follows: residues 1-30 (FR1 ), 31- 35 (CDR1), 36-49 (FR2), 50-65 (CDR2), 66-94 (FR3), 95-102 (CDR3) and 103- 113 (FR4). According to the numbering system of Kabat, VL FRS and CDRs are positioned as follows: residues 1-23 (FR1), 24-34 (CDR1), 35-49 (FR2), 50-56 (CDR2), 57-88 (FR3), 89-97 (CDR3) and 98-107 (FR4). The present disclosure is not limited to FRs and CDRs as defined by the Kabat numbering system, but includes all numbering systems, including those discussed above. In one example, reference herein to a CDR (or a FR) is in respect of those regions according to the Kabat numbering system.

[0619] "Framework regions" (FRs) are those variable region residues other than the CDR residues.

[0620] As used herein, the term “Fv” shall be taken to mean any protein, whether comprised of multiple polypeptides or a single polypeptide, in which a VL and a VH associate and form a complex having an antigen binding site, i.e., capable of specifically binding to an antigen. The VH and the VL which form the antigen binding site can be in a single polypeptide chain or in different polypeptide chains. Furthermore, an Fv of the disclosure (as well as any protein of the disclosure) may have multiple antigen binding sites which may or may not bind the same antigen. This term shall be understood to encompass fragments directly derived from an antibody as well as proteins corresponding to such a fragment produced using recombinant means. In some examples, the VH is not linked to a heavy chain constant domain (CH) 1 and / or the VL is not linked to a light chain constant domain (CL). Exemplary Fv containing polypeptides or proteins include a Fab fragment, a Fab’ fragment, aF(ab’) fragment, a scFv, a diabody, atriabody, a tetrabody or higher order complex, or any of the foregoing linked to a constant region or domain thereof, e.g., CH2 or CH3 domain, e.g., a minibody. A "Fab fragment" consists of a monovalent antigen-binding fragment of an immunoglobulin, and can be produced by digestion of a whole antibody with the enzyme papain, to yield a fragment consisting of an intact light chain and a portion of a heavy chain or can be produced using recombinant means. A "Fab' fragment" of an antibody can be obtained by treating a whole antibody with pepsin, followed by reduction, to yield a molecule consisting of an intact light chain and a portion of a heavy chain comprising a VH and a single constant domain. Two Fab' fragments are obtained per antibody treated in this manner. A Fab’ fragment can also be produced by recombinant means. A "F(ab')2 fragment” of an antibody consists of a dimer of two Fab' fragments held together by two disulfide bonds, and is obtained by treating a whole antibody molecule with the enzyme pepsin, without subsequent reduction. A “Fabi” fragment is a recombinant fragment comprising two Fab fragments linked using, for example a leucine zipper or a CH3 domain. A “single chain Fv” or “scFv” is a recombinant molecule containing the variable region fragment (Fv) of an antibody in which the variable region of the light chain and the variable region of the heavy chain are covalently linked by a suitable, flexible polypeptide linker.

[0621] The term “fragment crystallisable” or “Fc” or “Fc region” or “Fc portion” (which can be used interchangeably herein) refers to a region of an antibody comprising at least one constant domain and which is generally (though not necessarily) glycosylated and which is capable of binding to one or more Fc receptors and / or components of the complement cascade. The heavy chain constant region can be selected from any of the five isotypes: a, 5, a, y, or p. Furthermore, heavy chains of various subclasses (such as the IgG subclasses of heavy chains) are responsible for different effector functions and thus, by choosing the desired heavy chain constant region, proteins with desired effector function can be produced. Exemplary heavy chain constant regions are gamma 1 (IgGi), gamma 2 (IgGi) , gamma 3 (IgGa) and gamma 4 (IgGa). or hybrids thereof.

[0622] The term “constant region” as used herein, refers to a portion of heavy chain or light chain of an antibody other than the variable region. In a heavy chain, the constant region generally comprises a plurality of constant domains and a hinge region, e.g., a IgG constant region comprises the following linked components, a constant heavy CH CH 1, a linker, a CH2 and a CH3. In a light chain, a constant region generally comprises one constant domain (a CLI).

[0623] The term “stabilised IgG4 constant region” will be understood to mean an IgG4 constant region that has been modified to reduce Fab arm exchange or the propensity to undergo Fab arm exchange or formation of a half-antibody or a propensity to form a half antibody. “Fab arm exchange” refers to a type of protein modification for human IgG4, in which an IgG4 heavy chain and attached light chain (half-molecule) is swapped for a heavy-light chain pair from another IgG4 molecule. Thus, IgG4 molecules may acquire two distinct Fab arms recognising two distinct antigens (resulting in bispecific molecules). Fab arm exchange occurs naturally in vivo and can be induced in vitro by purified blood cells or reducing agents such as reduced glutathione.

[0624] The phrase “conservative amino acid substitution” refers to replacement or substitution of an amino acid residue with an amino acid residue having a similar side chain and / or hydropathicity and / or hydrophilicity. Families of amino acid residues having similar side chains have been defined in the art, including basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), / -branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Hydropathic indices are described, for example in Kyte and Doolittle J. Mol. Biol., 157: 105-132, 1982 and hydrophilic indices are described in, e.g., US4554101.

[0625] As used herein, the terms “disease”, “disorder” or “condition” refers to a disruption of or interference with normal function, and is not to be limited to any specific condition, and will include diseases or disorders.

[0626] As used herein, the terms “treating”, “treat” or “treatment” include administering a protein described herein to thereby reduce or eliminate at least one symptom of a specified disease or condition or to slow progression of the disease or condition.

[0627] As used herein, the terms “preventing”, “prevent” or “prevention” includes providing prophylaxis with respect to occurrence or recurrence of a specified disease or condition in an individual. An individual may be predisposed to or at risk of developing the disease or disease relapse but has not yet been diagnosed with the disease or the relapse.

[0628] As used herein, a subject “at risk” of developing a disease or condition or relapse thereof or relapsing may or may not have detectable disease or symptoms of disease, and may or may not have displayed detectable disease or symptoms of disease prior to the treatment according to the present disclosure. “At risk” denotes that a subject has one or more risk factors, which are measurable parameters that correlate with development of the disease or condition, as known in the art and / or described herein.

[0629] As used herein, the term “subject” shall be taken to mean any animal including humans, for example a mammal. Exemplary subjects include but are not limited to humans and non-human primates. For example, the subject is a human.

[0630] Proteins of the Pharmaceutical Formulation

[0631] As discussed herein, the present disclosure provides a liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain that binds to or specifically binds to G-CSFR.

[0632] Proteins comprising antigen binding domains

[0633] The present disclosure provides a pharmaceutical formulation comprising a protein comprising an antigen binding domain that specifically binds to G-CSFR, wherein the protein is an antibody or antigen binding fragment. For example, the protein is an antibody or antigen binding fragment that binds to G-CSFR.

[0634] Methods for generating antibodies are known in the art and / or described in Harlow and Fane (editors) Antibodies: A Eaboratory Manual, Cold Spring Harbor Eaboratory, (1988). Generally, in such methods G-CSFR or a region thereof (e.g., an extracellular domain) or immunogenic fragment or epitope thereof or a cell expressing and displaying same (i.e., an immunogen), optionally formulated with any suitable or desired carrier, adjuvant, or pharmaceutically acceptable excipient, is administered to a non-human animal, for example, a mouse, chicken, rat, rabbit, guinea pig, dog, horse, cow, goat or pig. The immunogen may be administered intranasally, intramuscularly, subcutaneously, intravenously, intradermally, intraperitoneally, or by other known route.

[0635] Monoclonal antibodies are one exemplary form of an antibody contemplated by the present disclosure. The term “monoclonal antibody" or “mAb” refers to a homogeneous antibody population capable of binding to the same antigen(s), for example, to the same epitope within the antigen. This term is not intended to be limited as regards to the source of the antibody or the manner in which it is made.

[0636] For the production of mAbs any one of a number of known techniques may be used, such as, for example, the procedure exemplified in US4196265 or Harlow and Lane (1988), supra. Alternatively, ABL-MYC technology (NeoClone, Madison WI 53713, USA) is used to produce cell lines secreting MAbs (e.g., as described in Largaespada et al, J. Immunol. Methods. 197: 85-95, 1996).

[0637] Antibodies can also be produced or isolated by screening a display library, e.g., a phage display library, e.g., as described in US6300064 and / or US5885793. For example, the present inventors have isolated fully human antibodies from a phage display library.

[0638] The antibody of the present disclosure may be a synthetic antibody. For example, the antibody is a chimeric antibody, a humanised antibody, a human antibody or a deimmunised antibody.

[0639] In one example an antibody or antigen binding fragment thereof of the disclosure is a chimeric antibody or fragment. The term “chimeric antibody” or “chimeric antigen binding fragment” refers to an antibody or fragment in which one or more of the variable domains is from a particular species (e.g., murine, such as mouse or rat) or belonging to a particular antibody class or subclass, while the remainder of the antibody or fragment is from another species (such as, for example, human or non-human primate) or belonging to another antibody class or subclass. In one example, a chimeric antibody comprising a VH and / or a VL from a non-human antibody (e.g., a murine antibody) and the remaining regions of the antibody are from a human antibody. The production of such chimeric antibodies and antigen binding fragments thereof is known in the art, and may be achieved by standard means (as described, e.g., in US6331415; US5807715; US4816567 and US4816397).

[0640] The antibodies or antigen binding fragments of the present disclosure may be humanised. The term "humanized antibody” shall be understood to refer to a subclass of chimeric antibodies having an antigen binding site or variable region derived from an antibody from a non-human species (e.g., mouse or rat or non-human primate) and the remaining antibody structure based upon the structure and / or sequence of a human antibody. In a humanized antibody, the antigen-binding site generally comprises the complementarity determining regions (CDRs) from the non-human antibody grafted onto appropriate FRs in the variable regions of a human antibody and the remaining regions from a human antibody (this type of antibody is also referred to a “CDR-grafted antibody”). Antigen binding sites may be wild-type (i.e., identical to those of the non- human antibody) or modified by one or more amino acid substitutions. In some instances, FR residues of the human antibody are replaced by corresponding non-human residues.

[0641] Humanised antibodies also include antibodies in which one or more residues of the human protein are modified by one or more amino acid substitutions and / or one or more FR residues of the human antibody are replaced by corresponding non-human residues. Humanised antibodies may also comprise residues which are found in neither the human antibody or in the non-human antibody. Any additional regions of the antibody (e.g., Fc region) are generally human. Humanisation can be performed using a method known in the art, e.g., US5225539, US6054297, US7566771 or US5585089. The term “humanised antibody” also encompasses a super-humanised antibody, e.g., as described in US7732578. A similar meaning will be taken to apply to the term “humanised antigen binding fragment”.

[0642] The antibodies or antigen binding fragments thereof of the present disclosure may be human antibodies or antigen binding fragments thereof. The term “human antibody” as used herein refers to antibodies having variable and, optionally, constant antibody regions found in humans, e.g. in the human germline or somatic cells or from libraries produced using such regions. The “human” antibodies can include amino acid residues not encoded by human sequences, e.g. mutations introduced by random or site directed mutations in vitro (in particular mutations which involve conservative substitutions or mutations in a small number of residues of the protein, e.g. in 1, 2, 3, 4 or 5 of the residues of the protein). These “human antibodies” do not necessarily need to be generated as a result of an immune response of a human, rather, they can be generated using recombinant means (e.g., screening a phage display library) and / or by a transgenic animal (e.g., a mouse) comprising nucleic acid encoding human antibody constant and / or variable regions and / or using guided selection (e.g., as described in or US5565332). This term also encompasses affinity matured forms of such antibodies. For the purposes of the present disclosure, a human antibody will also be considered to include a protein comprising FRs from a human antibody or FRs comprising sequences from a consensus sequence of human FRs and in which one or more of the CDRs are random or semirandom, e.g., as described in US6300064 and / or US6248516. A similar meaning will be taken to apply to the term “human antigen binding fragment”.

[0643] The antibodies or antigen binding fragments thereof of the present disclosure may be synhumanised antibodies or antigen binding fragments thereof. The term “synhumanised antibody” refers to an antibody prepared by a method described in W02007019620. A synhumanised antibody includes a variable region of an antibody, wherein the variable region comprises FRs from a New World primate antibody variable region and CDRs from a non-New World primate antibody variable region.

[0644] The antibody or antigen binding fragment thereof of the present disclosure may be primatised. A “primatised antibody” comprises variable region(s) from an antibody generated following immunisation of a non-human primate (e.g., a cynomolgus macaque). Optionally, the variable regions of the non-human primate antibody are linked to human constant regions to produce a primatised antibody. Exemplary methods for producing primatised antibodies are described in US6113898.

[0645] The present disclosure also contemplates a deimmunised antibody or antigen binding fragment thereof, e.g., as described in W02000034317 and W02004108158. De-immunised antibodies and fragments have one or more epitopes, e.g., B cell epitopes or T cell epitopes removed (i.e., mutated) to thereby reduce the likelihood that a subject will raise an immune response against the antibody or protein. For example, an antibody of the disclosure is analysed to identify one or more B or T cell epitopes and one or more amino acid residues within the epitope is mutated to thereby reduce the immunogenicity of the antibody.

[0646] Exemplary human antibodies are described herein and include C1.2 and C1.2G and / or variable regions thereof. These human antibodies provide an advantage of reduced immunogenicity in a human compared to non-human antibodies. Exemplary antibodies are described in W02012 / 171057, which is incorporated herein by reference.

[0647] Antibody Fragments

[0648] As described herein, a protein of the disclosure comprises an antigen binding fragment of an antibody. Exemplary antigen binding fragments for use in the present disclosure are described below.

[0649] Diabodies, Triabodies, Tetrabodies

[0650] In some examples, an antigen binding fragment of the disclosure is or comprises a diabody, triabody, tetrabody or higher order protein complex such as those described in W098 / 044001 and / or W094 / 007921.

[0651] For example, a diabody is a protein comprising two associated polypeptide chains, each polypeptide chain comprising the structure VL-X-VH or VH-X-VL, wherein X is a linker comprising insufficient residues to permit the VH and VL in a single polypeptide chain to associate (or form an Fv) or is absent, and wherein the VH of one polypeptide chain binds to a VL of the other polypeptide chain to form an antigen binding site, i.e., to form a Fv molecule capable of specifically binding to one or more antigens. The VL and VH can be the same in each polypeptide chain or the VL and VH can be different in each polypeptide chain so as to form a bispecific diabody (i.e., comprising two Fvs having different specificity). Single Chain Fv (scFv) Fragments

[0652] The skilled artisan will be aware that scFvs comprise VH and VL regions in a single polypeptide chain and a polypeptide linker between the VH and VL which enables the scFv to form the desired structure for antigen binding (i.e., for the VH and VL of the single polypeptide chain to associate with one another to form a Fv). For example, the linker comprises in excess of 12 amino acid residues with (Gly4Ser)3 being one of the more favoured linkers for a scFv.

[0653] In one example, the linker comprises the sequence SGGGGSGGGGSGGGGS.

[0654] The present disclosure also contemplates a disulfide stabilized Fv (or diFv or dsFv), in which a single cysteine residue is introduced into a FR of VH and a FR of VL and the cysteine residues linked by a disulfide bond to yield a stable Fv.

[0655] Alternatively, or in addition, the present disclosure encompasses a dimeric scFv, i.e., a protein comprising two scFv molecules linked by a non-covalent or covalent linkage, e.g., by a leucine zipper domain (e.g., derived from Fos or Jun). Alternatively, two scFvs are linked by a peptide linker of sufficient length to permit both scFvs to form and to bind to an antigen, e.g., as described in US20060263367.

[0656] Half-antibodies

[0657] In some examples, the antigen binding fragment of the present disclosure is a halfantibody or a half-molecule. The skilled artisan will be aware that a half antibody refers to a protein comprising a single heavy chain and a single light chain. The term “half antibody” also encompasses a protein comprising an antibody light chain and an antibody heavy chain, wherein the antibody heavy chain has been mutated to prevent association with another antibody heavy chain. In one example, a half antibody forms when an antibody dissociates to form two molecules each containing a single heavy chain and a single light chain.

[0658] Methods for generating half antibodies are known in the art and exemplary methods are described herein.

[0659] In one example, the half antibody can be secreted by introducing into cells genes of the single heavy chain and single light chain that constitute the IgG of interest for expression. In one example, a constant region (e.g., an IgG4 constant region) comprises a “key or hole” (or “knob or hole”) mutation to prevent heterodimer formation. In one example, a constant region (e.g., an IgG4 constant region) comprises a T366W mutation (or knob). In another example, a constant region (e.g., an IgG4 constant region) comprises a T366S, L368A and Y407V mutation (or hole). In another example, the constant region comprises T350V, T366L, K392L and T394W mutations (knob). In another example, the constant region comprises T350V, L351Y, F405A and Y407V mutations (hole). Exemplary constant region amino acid substitutions are numbered according to the EU numbering system.

[0660] Other Antibodies and Antibody Fragments

[0661] The present disclosure also contemplates other antibodies and antibody fragments, such as:

[0662] (i) “key and hole” bispecific proteins as described in US5, 731,168;

[0663] (ii) minibodies, e.g., as described in US5837821;

[0664] (iii) heteroconjugate proteins, e.g., as described in US4676980;

[0665] (iv) heteroconjugate proteins produced using a chemical cross-linker, e.g., as described in US4676980; and

[0666] (v) Fabs (e.g., as described in EP19930302894).

[0667] Constant Regions

[0668] The present disclosure encompasses proteins and / or antibodies described herein comprising a constant region of an antibody. This includes antigen binding fragments of an antibody fused to a Fc.

[0669] Sequences of constant regions useful for producing the proteins of the present disclosure may be obtained from a number of different sources. In some examples, the constant region or portion thereof of the protein is derived from a human antibody. The constant region or portion thereof may be derived from any antibody class, including IgM, IgG, IgD, IgA and IgE, and any antibody isotype, including IgGl, IgG2, IgG3 and IgG4. In one example, the constant region is human isotype IgG4 or a stabilized IgG4 constant region.

[0670] In one example, the Fc region of the constant region has a reduced ability to induce effector function, e.g., compared to a native or wild-type human IgGl or IgG3 Fc region. In one example, the effector function is antibody-dependent cell-mediated cytotoxicity (ADCC) and / or antibody -dependent cell-mediated phagocytosis (ADCP) and / or complement-dependent cytotoxicity (CDC). Methods for assessing the level of effector function of an Fc region containing protein are known in the art and / or described herein.

[0671] In one example, the Fc region is an IgG4 Fc region (i.e., from an IgG4 constant region), e.g., a human IgG4 Fc region. Sequences of suitable IgG4 Fc regions will be apparent to the skilled person and / or available in publically available databases (e.g., available from National Center for Biotechnology Information). In one example, the constant region is a stabilized IgG4 constant region. The term “stabilized IgG4 constant region” will be understood to mean an IgG4 constant region that has been modified to reduce Fab arm exchange or the propensity to undergo Fab arm exchange or formation of a half-antibody or a propensity to form a half antibody. “Fab arm exchange" refers to a type of protein modification for human IgG4, in which an IgG4 heavy chain and attached light chain (half-molecule) is swapped for a heavy-light chain pair from another IgG4 molecule. Thus, IgG4 molecules may acquire two distinct Fab arms recognizing two distinct antigens (resulting in bispecific molecules). Fab arm exchange occurs naturally in vivo and can be induced in vitro by purified blood cells or reducing agents such as reduced glutathione. A “half antibody” forms when an IgG4 antibody dissociates to form two molecules each containing a single heavy chain and a single light chain.

[0672] In one example, a stabilized IgG4 constant region comprises a proline at position 241 of the hinge region according to the system of Kabat (Kabat et al., Sequences of Proteins of Immunological Interest Washington DC United States Department of Health and Human Services, 1987 and / or 1991). This position corresponds to position 228 of the hinge region according to the EU numbering system (Kabat et al., Sequences of Proteins of Immunological Interest Washington DC United States Department of Health and Human Services, 2001 and Edelman et al., Proc. Natl. Acad. USA, 63, 78-85, 1969). In human IgG4, this residue is generally a serine. Following substitution of the serine for proline, the IgG4 hinge region comprises a sequence CPPC. In this regard, the skilled person will be aware that the “hinge region” is a proline-rich portion of an antibody heavy chain constant region that links the Fc and Fab regions that confers mobility on the two Fab arms of an antibody. The hinge region includes cysteine residues which are involved in inter-heavy chain disulfide bonds. It is generally defined as stretching from Glu226 to Pro243 of human IgGl according to the numbering system of Kabat. Hinge regions of other IgG isotypes may be aligned with the IgGl sequence by placing the first and last cysteine residues forming inter-heavy chain disulphide (S-S) bonds in the same positions (see for example W02010 / 080538).

[0673] Additional examples of stabilized IgG4 antibodies are antibodies in which arginine at position 409 in a heavy chain constant region of human IgG4 (according to the EU numbering system) is substituted with lysine, threonine, methionine, or leucine (e.g., as described in W02006 / 033386). The Fc region of the constant region may additionally or alternatively comprise a residue selected from the group consisting of: alanine, valine, glycine, isoleucine and leucine at the position corresponding to 405 (according to the EU numbering system). Optionally, the hinge region comprises a proline at position 241 (i.e., a CPPC sequence) (as described above).

[0674] In another example, the Fc region is a region modified to have reduced effector function, i.e., a “non-immunostimulatory Fc region”. For example, the Fc region is an IgGl Fc region comprising a substitution at one or more positions selected from the group consisting of 268, 309, 330 and 331. In another example, the Fc region is an IgGl Fc region comprising one or more of the following changes E233P, L234V, L235A and deletion of G236 and / or one or more of the following changes A327G, A33OS and P33 IS (Armour et al., Eur J Immunol. 29:2613-2624, 1999; Shields et al., J Biol Chem. 276(9):6591-604, 2001). Additional examples of non-immunostimulatory Fc regions are described, for example, in Dall'Acqua et al., J Immunol. 177 : 1129-1138 2006; and / or Hezareh J Virol ;75 12161-12168, 2001).

[0675] In another example, the Fc region is a chimeric Fc region, e.g., comprising at least one CH2 domain from an IgG4 antibody and at least one CH3 domain from an IgGl antibody, wherein the Fc region comprises a substitution at one or more amino acid positions selected from the group consisting of 240, 262, 264, 266, 297, 299, 307, 309, 323, 399, 409 and 427 (EU numbering) (e.g., as described in W02010 / 085682). Exemplary substitutions include 240F, 262L, 264T, 266F, 297Q, 299A, 299K, 307P, 309K, 309M, 309P, 323F, 399S, and 427F.

[0676] Mutations to Proteins

[0677] The present disclosure also contemplates mutant forms of a protein of the disclosure. In this regard, data presented herein indicate sites within a CDR of a protein of the disclosure that can be changed in addition to exemplary changes that can be made. The skilled person will understand that changes can additionally or alternatively be made within a framework region of a variable region containing protein without inhibiting or significantly reducing its function in the context of the present disclosure.

[0678] For example, such a mutant protein comprises one or more conservative amino acid substitutions compared to a sequence set forth herein. In some examples, the protein comprises 30 or fewer or 20 or fewer or 10 or fewer, e.g., 9 or 8 or 7 or 6 or 5 or 4 or 3 or 2 conservative amino acid substitutions. A "conservative amino acid substitution" is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain and / or hydropathicity and / or hydrophilicity.

[0679] In one example, a mutant protein has only, or not more than, one or two or three or four or five or six conservative amino acid changes when compared to a naturally occurring protein. Details of conservative amino acid changes are provided below. As the skilled person would be aware, e.g., from the disclosure herein, such minor changes can reasonably be predicted not to alter the activity of the protein.

[0680] Families of amino acid residues having similar side chains have been defined in the art, including basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), P-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine).

[0681] The present disclosure also contemplates non-conservative amino acid changes (e.g., substitutions) in a protein of the present disclosure, e.g., in a CDR, such as CDR3. For example, the present inventors have identified several non-conservative amino acid substitutions that can be made while retaining an activity of a protein of the disclosure. In one example, the protein comprises fewer than 6 or 5 or 4 or 3 or 2 or 1 non- conservative amino acid substitutions, e.g., in a CDR3, such as in a CDR3.

[0682] The present disclosure also contemplates one or more insertions or deletions compared to a sequence set forth herein. In some examples, the protein comprises 10 or fewer, e.g., 9 or 8 or 7 or 6 or 5 or 4 or 3 or 2 insertions and / or deletions.

[0683] Additional Modifications / Variants

[0684] The present disclosure also contemplates additional modifications or variants to an antibody or protein of the disclosure.

[0685] For example, the antibody comprises one or more amino acid substitutions that increase the half-life of the protein. For example, the antibody comprises a Fc region comprising one or more amino acid substitutions that increase the affinity of the Fc region for the neonatal Fc region (FcRn). For example, the Fc region has increased affinity for FcRn at lower pH, e.g., about pH 6.0, to facilitate Fc / FcRn binding in an endosome. In one example, the Fc region has increased affinity for FcRn at about pH 6 compared to its affinity at about pH 7.4, which facilitates the re-release of Fc into blood following cellular recycling. These amino acid substitutions are useful for extending the half life of a protein, by reducing clearance from the blood.

[0686] Exemplary amino acid substitutions include T250Q and / or M428L or T252A, T254S and T266F or M252Y, S254T and T256E or H433K and N434F according to the EU numbering system. Additional or alternative amino acid substitutions are described, for example, in US20070135620 or US7083784. In one example, the protein of the disclosure additionally comprises albumin, a functional fragment or variant thereof. In one example, the albumin, functional fragment or variant thereof is serum albumin, such as human serum albumin. In one example, the albumin, functional fragment or variant thereof, comprises one or more amino acid substitutions, deletions or insertions, e.g., no more than 5 or 4 or 3 or 2 or 1 substitutions. Amino acid substitutions suitable for use in the present disclosure will be apparent to the skilled person and include naturally-occurring substitutions and engineered substitutions such as those described, for example, in WO2011051489, WO2014072481, WO201 1103076, WO2012112188, W02013075066, W02015063611 and

[0687] WO2014179657.

[0688] In one example, the protein or antibody of the disclosure comprises one or more variants. For example, the variant is a post-translationally modified variant.

[0689] In one example, the protein or antibody comprises a variant missing an encoded C-terminal lysine residue, a deamidated variant, a miscorporated amino acid residue, a glycosylated variant, a variant comprising a pyroglutamate, a variant lacking a N- terminal residue, and / or a variant comprising all or part of a secretion signal.

[0690] In one example, the protein or antibody comprises a variant missing an encoded C-terminal lysine residue.

[0691] In one example, the protein or antibody comprises a deamidated variant. Deamidated variants of encoded asparagine residues may result in isoaspartic acid and / or aspartic acid being generated or even a succinamide involving an adjacent amino acid residue. Deamidated variants of encoded glutamine residues may result in glutamic acid being formed. Compositions comprising a heterogeneous mixture of such sequences and variants are intended to be included when reference is made to a particular amino acid sequence.

[0692] In one example, the protein or antibody comprises a miscorporated amino acid residue. For example, methionine residues are substituted with nor-leucine, asparagine residues are substituted with serine and / or phenylalanine residues are substituted with tyrosine.

[0693] In one example, the protein or antibody comprises a variant comprising a pyroglutamate. For example, at the N-terminus of a protein.

[0694] In one example, the protein or antibody comprises a glycosylated variant.

[0695] In one example, the protein or antibody comprises a variant lacking a N-terminal residue. For example, a N-terminal glutamine in an antibody or V region.

[0696] In one example, the protein or antibody comprises a variant comprising all or part of a secretion signal. Protein Production

[0697] Methods of producing and obtaining proteins for use in the formulation described herein will be known to those skilled in the art. For example, in the case of a recombinant protein, nucleic acid encoding same can be cloned into expression constructs or vectors, which are then transfected into host cells, such as E. coli cells, yeast cells, insect cells, or mammalian cells, such as simian COS cells, Chinese Hamster Ovary (CHO) cells, human embryonic kidney (HEK) cells, or myeloma cells that do not otherwise produce the protein. Exemplary cells used for expressing an protein are CHO cells, myeloma cells or HEK cells. Molecular cloning techniques to achieve these ends are known in the art and described, for example in Ausubel et al., (editors), Current Protocols in Molecular Biology, Greene Pub. Associates and Wiley-Interscience (1988, including all updates until present) or Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press (1989). A wide variety of cloning and in vitro amplification methods are suitable for the construction of recombinant nucleic acids. Methods of producing recombinant proteins are also known in the art, see, e.g., US4816567 or US5530101.

[0698] Following isolation, the nucleic acid is inserted operably linked to a promoter in an expression construct or expression vector for further cloning (amplification of the DNA) or for expression in a cell-free system or in cells.

[0699] As used herein, the term “promoter” is to be taken in its broadest context and includes the transcriptional regulatory sequences of a genomic gene, including the TATA box or initiator element, which is required for accurate transcription initiation, with or without additional regulatory elements (e.g., upstream activating sequences, transcription factor binding sites, enhancers and silencers) that alter expression of a nucleic acid, e.g., in response to a developmental and / or external stimulus, or in a tissue specific manner. In the present context, the term “promoter” is also used to describe a recombinant, synthetic or fusion nucleic acid, or derivative which confers, activates or enhances the expression of a nucleic acid to which it is operably linked. Exemplary promoters can contain additional copies of one or more specific regulatory elements to further enhance expression and / or alter the spatial expression and / or temporal expression of said nucleic acid.

[0700] As used herein, the term “operably linked to" means positioning a promoter relative to a nucleic acid such that expression of the nucleic acid is controlled by the promoter. Many vectors for expression in cells are available. The vector components generally include, but are not limited to, one or more of the following: a signal sequence, a sequence encoding an protein (e.g., derived from the information provided herein), an enhancer element, a promoter, and a transcription termination sequence. The skilled artisan will be aware of suitable sequences for expression of an protein. Exemplary signal sequences include prokaryotic secretion signals (e.g., pelB, alkaline phosphatase, penicillinase, Ipp, or heat-stable enterotoxin II), yeast secretion signals (e.g., invertase leader, a factor leader, or acid phosphatase leader) or mammalian secretion signals (e.g., herpes simplex gD signal).

[0701] Exemplary promoters active in mammalian cells include cytomegalovirus immediate early promoter (CMV-IE), human elongation factor 1-a promoter (EFl), small nuclear RNA promoters (Ula and Ulb), a-myosin heavy chain promoter, Simian virus 40 promoter (SV40), Rous sarcoma virus promoter (RSV), Adenovirus major late promoter, P-actin promoter; hybrid regulatory element comprising a CMV enhancer / P- actin promoter or an immunoglobulin promoter or active fragment thereof. Examples of useful mammalian host cell lines are monkey kidney CV1 line transformed by SV40 (COS-7, ATCC CRL 1651); human embryonic kidney line (293 or 293 cells subcloned for growth in suspension culture; baby hamster kidney cells (BHK, ATCC CCL 10); or Chinese hamster ovary cells (CHO).

[0702] Typical promoters suitable for expression in yeast cells such as for example a yeast cell selected from the group comprising Pichia pastoris, Saccharomyces cerevisiae and S. pombe, include, but are not limited to, the ADH1 promoter, the GALI promoter, the GAIA promoter, the CUP1 promoter, the PHO 5 promoter, the nmt promoter, the RPR1 promoter, or the TEF1 promoter.

[0703] Means for introducing the isolated nucleic acid or expression construct comprising same into a cell for expression are known to those skilled in the art. The technique used for a given cell depends on the known successful techniques. Means for introducing recombinant DNA into cells include microinjection, transfection mediated by DEAE-dextran, transfection mediated by liposomes such as by using lipofectamine (Gibco, MD, USA) and / or cellfectin (Gibco, MD, USA), PEG-mediated DNA uptake, electroporation and microparticle bombardment such as by using DNA-coated tungsten or gold particles (Agracetus Inc., WI, USA) amongst others.

[0704] The host cells used to produce the protein may be cultured in a variety of media, depending on the cell type used. Commercially available media such as Ham's F10 (Sigma), Minimal Essential Medium ((MEM), (Sigma), RPM1-1640 (Sigma), and Dulbecco's Modified Eagle's Medium ((DMEM), Sigma) are suitable for culturing mammalian cells. Media for culturing other cell types discussed herein are known in the art.

[0705] Isolation of Proteins

[0706] Where a protein (e.g., antibody) is secreted into culture medium, supernatants from such expression systems can be first concentrated using a commercially available protein concentration filter, for example, an Amicon or Millipore Pellicon ultrafiltration unit. A protease inhibitor such as PMSF may be included in any of the foregoing steps to inhibit proteolysis and antibiotics may be included to prevent the growth of adventitious contaminants. Alternatively, or additionally, supernatants can be filtered and / or separated from cells expressing the protein, e.g., using continuous centrifugation.

[0707] The protein prepared from the cells can be purified using, for example, ion exchange, hydroxyapatite chromatography, hydrophobic interaction chromatography, gel electrophoresis, dialysis, affinity chromatography (e.g., protein A affinity chromatography or protein G chromatography), or any combination of the foregoing. These methods are known in the art and described, for example in WO99 / 57134 or Ed Harlow and David Lane (editors) Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, (1988).

[0708] The skilled artisan will also be aware that a protein can be modified to include a tag to facilitate purification or detection, e.g., a poly-histidine tag, e.g., a hexa-histidine tag, or a influenza virus hemagglutinin (HA) tag, or a Simian Virus 5 (V5) tag, or a FLAG tag, or a glutathione S-transferase (GST) tag. The resulting protein is then purified using methods known in the art, such as, affinity purification. For example, a protein comprising a hexa-his tag is purified by contacting a sample comprising the protein with nickel-nitrilotriacetic acid (Ni-NTA) that specifically binds a hexa-his tag immobilized on a solid or semi-solid support, washing the sample to remove unbound protein, and subsequently eluting the bound protein. Alternatively, or in addition a ligand or antibody that binds to a tag is used in an affinity purification method.

[0709] Preparation of the Pharmaceutical Formulation

[0710] As described herein, the formulations of the present disclosure comprise a histidine buffer, polysorbate 80, arginine and proline, and has a pH of 5.7 to 5.9. Preparation of the pharmaceutical formulation is performed according to standard methods known in the art and / or according to methods described herein. Histidine Buffer

[0711] It will be apparent to the skilled person that histidine buffers suitable for use in the present disclosure will be stable and effective at the desired pH and will provide sufficient buffer capacity to maintain the desired pH over the range of conditions to which it will be exposed during formulation and storage of the product.

[0712] Suitable histidine buffers for use in the present disclosure will be apparent to the skilled person and include, for example L-histidine, histidine chloride, histidine acetate, histidine phosphate, histidine sulfate, etc. In one example, the histidine buffer is L- histidine.

[0713] In one example of the present disclose, the formulation comprises 12 mM to 30 mM histidine buffer. For example, the formulation comprises 15 mM to 25 mM histidine buffer. For example, the formulation comprises 17 mM to 23 mM histidine buffer. For example, the formulation comprises 18 mM to 22 mM histidine buffer. For example, the formulation comprises 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, or 25 mM histidine buffer. For example, the formulation comprises 20 mM histidine buffer.

[0714] Methods of assessing the suitability of buffers will be apparent to the skilled person and / or described herein and include, for example, differential scanning fluorimetry and dynamic light scattering.

[0715] Polysorbate 80

[0716] Polysorbate 80 (polyoxyethylene sorbitan monooleate) is a non-ionic surfactant that can be added to pharmaceutical formulations in an amount such that it suppresses aggregation (e.g., by preventing surface denaturation), increases stabilisation (e.g., during thermal and / or physical stress), minimises the formation of particulates in the formulation (e.g., sub-visible and / or visible particle formation), reduces surface adsorption and / or assists in protein refolding.

[0717] In one example of the present disclosure, the formulation comprises 0.01 % to 0.05 % (w / v) polysorbate 80. For example, the formulation comprises of 0.02% (w / v) to 0.04 %(w / v) polysorbate 80. For example, the formulation comprises 0.01% (w / v), 0.02% (w / v), 0.03% (w / v), 0.04% (w / v), or 0.05% (w / v) polysorbate 80. For example, the formulation comprises 0.03% (w / v) polysorbate 80.

[0718] Amino Acid Stabilisers

[0719] Formulations of the disclosure comprise two amino acid stablisers: proline and arginine. The amino acid stabilisers are added to the pharmaceutical formulation in an amount that such that it reduces thermal and / or physical stress (e.g., freeze / thaw or agitation), and / or confers or enhances stability of the protein.

[0720] In one example, the amino acid stabilisers are proline and arginine or a salt form thereof.

[0721] In one example, the formulation comprises 95 mM to 110 mM proline. For example, the formulation comprises 100 mM to 110 mM proline. For example, the formulation comprises 100 mM, 101 mM, 102 mM, 103 mM, 104 mM, 105 mM, 106 mM, 107 mM, 108 mM, 109 mM, or 110 mM proline. For example, the formulation comprises 100 mM proline.

[0722] In one example, the formulation comprises 60 mM to 125 mM arginine. For example, the formulation 90 mM to 110 mM arginine. For example, the formulation comprises 95 mM to 105 mM arginine. For example, the formulation comprises 95 mM, 96 mM, 97 mM, 98 mM, 99 mM, 100 mM, 101 mM, 102 mM, 103 mM, 104 mM, or 105 mM arginine. For example, the formulation comprises 100 mM arginine.

[0723] In one example, the formulation comprises 100 mM to 125 mM proline and 60 mM to 125 mM arginine. For example, the formulation comprises 100 mM to 110 mM proline and 90 mM to 110 mM arginine. For example, the formulation comprises 100 mM, 101 mM, 102 mM, 103 mM, 104 mM, 105 mM, 106 mM, 107 mM, 108 mM, 109 mM, or 110 mM proline; and 95 mM, 96 mM, 97 mM, 98 mM, 99 mM, 100 mM, 101 mM, 102 mM, 103 mM, 104 mM, or 105 mM arginine. For example, the formulation comprises 100 mM proline and 100 mM arginine.

[0724] Protein

[0725] Pharmaceutical formulations of the disclosure comprise a protein concentration of 45 mg / mL to 55 mg / mL. For example, the formulation comprises a protein concentration of 45 mg / mL, 46 mg / mL, 47 mg / mL, 48 mg / mL, 49 mg / mL, 50 mg / mL, 51 mg / mL, 52 mg / mL, 53 mg / mL, 54 mg / mL, or 55 mg / mL. In one example, the formulation comprises a protein concentration of 50 mg / mL.

[0726] Assaying the Pharmaceutical Formulation and Proteins of the Disclosure

[0727] Pharmaceutical formulations of the present disclosure are readily screened for physical and biological activity and / or stability using methods known in the art and / or as described below. Visual Appearance

[0728] Pharmaceutical formulations encompassed by the present disclosure can be assessed for visual appearance to determine, for example, the colour and clarity or for the presence of visible particles.

[0729] Dynamic Light Scattering

[0730] In one example, the particle size distribution is assessed using dynamic light scattering (DLS). DLS measures light scattered from particles based on Brownian motion and relies on differences in the index of refraction between the particle and the formulation. For example, the fluctuation of light intensity using a digital correlator is measured. The correlation functions are fitted into an analytical program (e.g., Malvern Zetasizer software) to calculate the particle size distribution. For the determination of Z- average hydrodynamic diameter, a cumulants analysis and the Stokes Einstein equation is performed using e.g., the viscosity of water (0.8872 mPa*s) at 25°C. The polydispersity index can also be obtained from the same cumulants analysis. Modality of fit is evaluated based on plots of size distribution versus intensity: modality can be described as monomodal (i.e., one peak) or multimodal (i.e., two or more peaks).

[0731] Micro-Flow Imaging

[0732] In one example, sub-visible particles are assessed using micro-flow imaging (MFI). For example, digital images of particles suspended in a fluid are captured and automatically analysed for particle parameters, such as aspect ratio (AR) and intensity. The size (e.g., in pm) and count (i.e., number of particles per ml) can also be obtained. According to this method the data are morphologically categorised as proteinaceous (i.e., circular) and non-proteinaceous (i.e., non -proteinaceous particles such as air bubbles or silicone oil droplets) and a ratio of the non-proteinaceous particles to proteinaceous particles (i.e., the circular fraction) can be determined. A low circular fraction value indicates that the test article is comprised of mostly non-circular, likely proteinaceous particles.

[0733] Size Exclusion Chromatography

[0734] In one example, aggregates / HMWS are assessed using size exclusion chromatography (SEC or SE-HPLC) which separates lower and higher molecular mass variants of the protein, as well as any impurities. According to this method, the results are described as the summation of aggregation peaks (APs) and summation of degradation peaks (DPs). For example, the identity of a pharmaceutical formulation of the present disclosure can be determined by comparing the chromatographic retention time of the major peaks with the retention time of the major peak of a reference standard.

[0735] Differential Scanning Fluorimetry ( DSF)

[0736] In one example, thermal stability of the pharmaceutical formulation of the present disclosure is assessed using differential scanning fluorimetry (DSF). DSF is a fluorescence-based assay using real-time PCR to monitor thermally induced protein denaturation by measuring changes fluorescence of a dye that binds preferentially to unfolded protein. For example, thermal unfolding and aggregation are monitored by changes in intrinsic protein fluorescence and static light scattering, respectively, as a function of temperature. According to this method, the midpoint of thermal transition (Tm) and onset of melting temperature (Tonset) are determined by monitoring intrinsic fluorescence. The onset of aggregation temperature (Tagg) are determined by monitoring static light scattering, e.g., at 266 nm and 473 nm. Samples of the pharmaceutical formulation can be assessed across a range of temperatures, (e.g., 20°C - 95°C) with a temperature increase at the rate of e.g., 0.5°C / min.

[0737] Capillary Gel Electrophoresis

[0738] In one example, the pharmaceutical formulation of the present disclosure is assessed for stability and / or total accumulation of impurities using capillary gel electrophoresis (CGE). For example, both reduced-CGE (R-CGE) and non-reduced- CGE (NR-CGE) may be performed. In one example, R-GCE and NR-CGE are carried out using a capillary electrophoresis system (e.g., Beckman P / ACE MDQ or PA800) with a capillary length of e.g., 20.2 cm and 10 cm respectively from inlet to detection window, temperature control from e.g., 20 to 40°C (±2°C) and detector at e.g., 488 nm excitation.

[0739] Cation Exchange Chromatography

[0740] In one example, the pharmaceutical formulation of the present disclosure is assessed for total charged variants (i.e. acid and basic species) using cation exchange (CEX) chromatography. CEX chromatography separates proteins according to their overall charge under native conditions. The CEX analysis is used to determine the purity of the product by separating the acidic and basic variants. The protein of interest must have a charge opposite to that of the functional group attached to the resin of the column in order to bind. Elution of the protein is achieved by increasing the ionic strength breaking the ionic interaction between the protein and the resin. The chromatographic technique separates the acidic, neutral and basic variants of a sample based on ionic strength. The peaks of interest are observed by UV detection at 280nm where the acidic variants eluting first followed by neutral and basic variants. In one example CEX chromatography is carried out using a high performance liquid chromatography (HPLC) system (e.g. Dionex UltiMate 3000 BioRS (U) HPLC).

[0741] Gibbs Free Energy (AGtrend; HUNK)

[0742] In one example, the chemical stability and aggregation behaviour of a pharmaceutical formulation of the present disclosure is evaluated by the change in the Gibbs free energy or AGtrend (HUNK) analysis. The AGtrend analysis measures the relationship between AG of protein unfolding and protein aggregation as a function of protein concentration. In the absence of aggregation, the AG of protein unfolding is a unimolecular process independent of protein concentration. If a change in AG is observed as a function of protein concentration, it signifies presence of aggregation. According to this method, there are two possible relationships between AG of protein unfolding and protein concentration if aggregation occurs:

[0743] 1. AGtrend increases with protein concentration: This relationship indicates the presence of native state aggregation - the AG of protein unfolding increases (becomes more positive) as a function of protein concentration (i.e., concentration of native protein aggregates increases as a function of protein concentration); or

[0744] 2. AGtrend decreases with protein concentration: This relationship indicates the presence of denatured state aggregation - the AG of protein unfolding decreases (become less positive) as a function of protein concentration (i.e., concentration of denatured protein aggregates increases as a function of protein concentration). In a HUNK experiment the AG of protein unfolding is determined isothermally by measuring changes in a protein’s intrinsic fluorescence spectrum (i.e., emission from tryptophan residues) as it unfolds in the presence of increasing amounts of denaturant.

[0745] In one example, AGtrend is determined by measuring AG of the protein unfolding at varying concentrations (e.g., 0.25, 0.6, 2.5, 6.0, 25.0 mg / ml) diluted to target concentration in a buffer of the pharmaceutical formulation of the disclosure. Each concentration level is titrated with increasing denaturant concentration (e.g., 32-point curve spanning urea concentration 2.00-8.74 M) while fluorescence spectra is measured from 300-500 nm (excitation 280 nm) with a slit width of 10 nm. The emission spectrum wavelength ratio of 350nm / 330nm is plotted against urea concentration for each sample concentration level, and AG of protein unfolding determined using a 2 state (i.e., one transition) model fit. Determined AG values are plotted against sample concentration to determine AGtrend.

[0746] Capillary electrophoresis

[0747] In some examples, the formulation is assessed by capillary electrophoresis (CE). For example, the formulation may be assessed by capillary electrophoresis with sodium dodecylsulfate (CE-SDS) under non-reducing conditions to determine the proportion of LMWS present. Capillary electrophoresis is a separation method performed in submillimeter diameter capillaries and in micro- and nanofluidic channels. Proteins migrate through electrolyte solutions under the influence of an electric field. In the presence of SDS, proteins are denatured and are separated on the basis of their molecular weight. This enables the detection of LMWS present in the formulation, for example LMWS produced upon degradation (e.g., proteolytic degradation) of the protein.

[0748] Turbidity Assessed by Absorbance at 550 nm

[0749] In one example, the turbidity of the pharmaceutical formulation of the present disclosure is assessed. For example, the turbidity is assessed using a spectrophotometer and measuring the absorbance at 550 nm.

[0750] Syringeability

[0751] In one example, the syringeability of the pharmaceutical formulation of the present disclosure is assessed. For example, the formulation is expelled with a 2 ml syringe, 10 ml syringe, or left untreated as a pre-expulsion control. According to this method, the syringe plunger is pushed through the 2 ml syringes at a linear speed of 0.2 in / min and through the 10 ml syringes at 0.6 in / min until the plunger reaches the bottom and reaches the force of 30 N. Break-loose (BF) and glide (GF) forces are measured during expulsion and used to assess application suitability. Break-loose force describes the force required to initiate movement of the plunger (the initial 0.3 mm for 2 ml syringe and 0.5 mm for 10 ml syringe). Glide force Max refers to the maximum friction force required to sustain plunger movement. The maximum force value is measured from the end of the break loose region to the end of the glide force region (26 mm for 2 ml syringe and 24 mm for 10 ml syringe) prior to the point where the force reaches 30 N).

[0752] Uses of the Pharmaceutical Formulation

[0753] As discussed herein, the present disclosure provides a method of treating or preventing a neutrophil-mediated disease or condition in a subject, comprising administering a pharmaceutical formulation of the present disclosure to the subject. In one example, the present disclosure provides a method of treating or preventing a neutrophil-mediated disease or condition in a subject in need thereof.

[0754] The present disclosure also provides for use of a pharmaceutical formulation of the present disclosure for treating or preventing a neutrophil-mediated disease or condition in a subject comprising administering the pharmaceutical formulation of the present disclosure to the subject. In one example, the present disclosure provides for use of a pharmaceutical formulation of the present disclosure for treating or preventing a neutrophil-mediated disease or condition in a subject in need thereof.

[0755] In some examples, the neutrophil-mediated condition is a neutrophilic skin condition; an autoimmune disease; an inflammatory disease; ischemia-reperfusion injury; a pulmonary disease; a neurological condition; or a neurodegenerative condition.

[0756] Exemplary autoimmune conditions include, arthritis (such as rheumatoid arthritis, psoriatic arthritis and or idiopathic arthritis, e.g., juvenile idiopathic arthritis) or psoriasis.

[0757] Exemplary inflammatory conditions include inflammatory neurological conditions (e.g., Devic's disease, a viral infection in the brain, multiple sclerosis and neuromyelitis optica), an inflammatory lung disease (e.g., chronic obstructive pulmonary disease (COPD), acute respiratory distress syndrome (ARDS) or asthma), systemic inflammatory response syndrome (SIRS), or an inflammatory eye condition (e.g., uveitis).

[0758] Exemplary pulmonary conditions include chronic obstructive pulmonary disease (COPD); emphysema; bronchitis; pneumonia; acute respiratory distress syndrome; acute lung injury; or asthma.

[0759] Exemplary neurological conditions include multiple sclerosis (MS); Devic's disease; or a viral infection in the brain.

[0760] Exemplary neurodegenerative conditions include multiple sclerosis (MS).

[0761] In one example, the neutrophil-mediated condition is asthma.

[0762] In one example, the neutrophil-mediated condition is ARDS.

[0763] In one example, the neutrophil-mediated condition is ischemia-reperfusion injury. For example, the ischemia-reperfusion injury is due to or associated with tissue or organ transplantation (e.g., kidney transplantation). For example, the antibody is administered to a tissue or organ transplantation recipient, e.g., prior to organ collection and / or to a tissue or organ prior to transplantation or is administered to a harvested tissue or organ ex vivo. In some examples, the neutrophil-mediated condition is psoriasis. In one example, the neutrophil-mediated condition is plaque psoriasis (also known in the art as “psoriasis vulgaris” or “common psoriasis”).

[0764] In one example, the neutrophil-mediated condition is neutrophilic skin condition. In one example, the neutrophil-mediated condition is neutrophilic dermatosis. For example, the neutrophilic dermatosis is a pustular psoriasis. In another example, the neutrophilic dermatosis is hidradenitis suppurativa.

[0765] In one example, the neutrophilic dermatosis is selected from the group consisting of amicrobial pustulosis of the folds (APF); plaque psoriasis; CARD14- mediated pustular psoriasis (CAMPS); cryopyrin associated periodic syndromes (CAPS); deficiency of interleukin- 1 receptor (DIRA); deficiency of interleukin- 36 receptor antagonist(DIRTA); hidradenitis suppurativa (HS); palmoplantar pustulosis; pyogenic arthritis; pyoderma gangrenosum and acne (PAPA); pyoderma gangrenosum, acne, and hidradenitis suppurativa (PASH); pyoderma gangrenosum(PG); skin lesions of Behcet’s disease; Still’s disease; Sweet syndrome; subcorneal pustulosis (Sneddon- Wilkinson); pustular psoriasis; palmoplantar pustulosis; acute generalized exanthematic pustulosis; infantile acropustulosis; synovitis, acne, pustulosis; hyperostosis and osteitis (SAPHO) syndrome; bowel-associated dermatosis-arthritis syndrome (BADAS); neutrophilic dermatosis of the dorsal hands; neutrophilic eccrine hidradenitis; erythema elevatum diutinum; and Pyoderma gangrenosum. In one example, the neutrophilic dermatosis is hidradenitis suppurativa (HS) or palmoplantar pustulosis (PPP).

[0766] In one example, the neutrophil-mediated condition is systemic inflammation. In one example, the systemic inflammation is due to or associated with surgery. In one example, the systemic inflammation is systemic inflammatory response syndrome.

[0767] The present disclosure also provides a method of reducing circulating neutrophils in a subject, the method comprising administering the formulation of the present disclosure. Such methods are useful in circumstances in which the subject is suffering from a disease or condition that is associated with neutrophils (e.g., neutrophil-mediated conditions).

[0768] In some examples, the subject is administered an effective amount of the protein in the formulation of the present disclosure. An “effective amount” refers to at least an amount effective, at dosages and for periods of time necessary, to achieve the desired result. For example, the desired result may be a therapeutic or prophylactic result. An effective amount can be provided in one or more administrations. In some examples of the present disclosure, the term “effective amount” is meant an amount necessary to effect treatment of a disease or condition as hereinbefore described. In some examples of the present disclosure, the term “effective amount” is meant an amount necessary to effect a change in a factor associated with a disease or condition as hereinbefore described. The effective amount may vary according to the disease or condition to be treated or factor to be altered and also according to the weight, age, racial background, sex, health and / or physical condition and other factors relevant to the mammal being treated. Typically, the effective amount will fall within a relatively broad range (e.g. a “dosage” range) that can be determined through routine trial and experimentation by a medical practitioner. Accordingly, this term is not to be construed to limit the disclosure to a specific quantity, e.g., weight or number. The effective amount can be administered in a single dose or in a dose repeated once or several times over a treatment period.

[0769] In some examples, the subject is administered a therapeutically effective amount of the protein in the formulation of the present disclosure. A “therapeutically effective amount” is at least the minimum concentration required to effect a measurable improvement of a particular disease or condition. A therapeutically effective amount herein may vary according to factors such as the disease state, age, sex, and weight of the patient, and the ability of the antibody or antigen binding fragment thereof to elicit a desired response in the individual. A therapeutically effective amount is also one in which any toxic or detrimental effects of the protein are outweighed by the therapeutically beneficial effects.

[0770] In one example, the pharmaceutical formulation of the present disclosure is administered to the subject in an amount to reduce the severity of the disease or condition in the subject.

[0771] In one example, the subject is at risk of developing a neutrophil-mediated condition. A subject is at risk if he or she has a higher risk of developing a neutrophil- mediated condition than a control population. The control population may include one or more subjects selected at random from the general population (e.g., matched by age, gender, race and / or ethnicity) who have not suffered from or have a family history of a neutrophil-mediated condition. A subject can be considered at risk for a disease or condition if a "risk factor" associated with a neutrophil-mediated condition is found to be associated with that subject. A risk factor can include any activity, trait, event or property associated with a given disorder, for example, through statistical or epidemiological studies on a population of subjects. A subject can thus be classified as being at risk for a neutrophil-mediated condition even if studies identifying the underlying risk factors did not include the subject specifically.

[0772] In one example, the subject is at risk of developing a neutrophil-mediated condition and the pharmaceutical formulation of the present disclosure is administered before or after the onset of symptoms of a neutrophil-mediated condition. In one example, the pharmaceutical formulation is administered before the onset of symptoms of a neutrophil-mediated condition. In one example, the pharmaceutical formulation is administered after the onset of symptoms of a neutrophil-mediated condition. In one example, the pharmaceutical formulation of the present disclosure is administered at a dose that alleviates or reduces one or more of the symptoms of a neutrophil-mediated condition in a subject at risk.

[0773] The methods of the present disclosure can be readily applied to any form of a neutrophil-mediated condition in a subject. In one example, a method of the disclosure reduces any symptom of a neutrophil-mediated condition known in the art and / or described herein. As will be apparent to the skilled person a “reduction” in a symptom of a disorder in a subject will be comparative to another subject who also suffers from a disorder but who has not received treatment with a method described herein. This does not necessarily require a side-by-side comparison of two subjects. Rather population data can be relied upon. For example, a population of subjects suffering from a neutrophil-mediated condition who have not received treatment with a method described herein (optionally, a population of similar subjects to the treated subject, e.g., age, weight, race) are assessed and the mean values are compared to results of a subject or population of subjects treated with a method described herein.

[0774] In some examples, a pharmaceutical formulation as described herein can be administered orally, parenterally, by inhalation spray, adsorption, absorption, topically, rectally, nasally, buccally, vaginally, intraventricularly, via an implanted reservoir in dosage formulations containing conventional non-toxic pharmaceutically-acceptable carriers, or by any other convenient dosage form. The term “parenteral” as used herein includes subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrastemal, and intracranial injection or infusion techniques. In one example, the formulation as described herein is administered subcutaneously. In one example, the formulation as described herein is self-administered.

[0775] Methods for preparing a formulation into a suitable form for administration (e.g. a pharmaceutical composition) are known in the art and include, for example, methods as described in Remington's Pharmaceutical Sciences (18th ed., Mack Publishing Co., Easton, Pa., 1990) and U.S. Pharmacopeia: National Formulary (Mack Publishing Company, Easton, Pa., 1984).

[0776] The pharmaceutical formulations of the present disclosure are particularly useful for parenteral administration, such as intravenous administration or subcutaneous administration or administration into a body cavity or lumen of an organ or joint. Upon formulation, compounds of the present disclosure will be administered in a manner compatible with the dosage formulation and in such amount as is prophylactically effective.

[0777] One skilled in the art would be able, by routine experimentation, to determine what an effective dose of the compound of the disclosure would be for the purpose of delaying progression or reducing or inhibiting or hindering development of a neutrophil- mediated condition described herein. For example, an effective dose may vary according to factors such as the disease stage, age, sex and weight of the subject, and the ability of the compound to elicit a desired response in the subject. The dosage regimen may be adjusted to provide the optimum response.

[0778] In other examples, formulations of the disclosure can be administered as a fixed- dose regimen. As used herein, the term “fixed dose” refers to a dosage that is administered in a particular amount, irrespective of the subject’s body weight.

[0779] It will be appreciated by persons skilled in the art that numerous variations and / or modifications may be made to the above-described embodiments, without departing from the broad general scope of the present disclosure. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive.

[0780] A method of the present disclosure may also include co-administration of the pharmaceutical formulation according to the disclosure together with the administration of another therapeutically effective agent for the prevention or treatment of a neutrophil- mediated condition.

[0781] In one example, the pharmaceutical formulation of the disclosure is used in combination with at least one additional known compound or therapy which is currently being used or is in development for preventing or treating neutrophil-mediated condition, or reducing circulating neutrophils. For example, the other compound is an antiinflammatory compound, e.g, methotrexate or a non-steroidal anti-inflammatory compound. Alternatively, or additionally, the other compound is an immunosuppressant. Alternatively, or additionally, the other compound is a corticosteroid, such as prednisone and / or prednisolone. In on example, the other compound is methotrexate. Alternatively, or additionally, the other compound is cyclophosphamide.

[0782] In some examples, the formulation is administered in combination with a cell. In some examples, the cell is a stem cell, such as a mesenchymal stem cell.

[0783] In some examples, the formulation is administered in combination with a gene therapy. In some examples, the formulation is administered in combination with a nonpharmaceutical intervention, for example, apharesis, such as plasmapheresis, cytapheresis, leukapheresis, granulocyte and / or monocyte apheresis. In this context, the formulation can be administered during the period of time in which the nonpharmaceutical intervention is being performed and will be considered “in combination with” the non-pharmaceutical intervention. For example, the non-pharmaceutical intervention may be granulocyte and / or monocyte apheresis, which is performed once per week for five weeks and the formulation can be administered over this time period. In one example, the formulation is administered before the non-pharmaceutical intervention. In one example, the formulation is administered after the non- pharmaceutical intervention.

[0784] Another non-pharmaceutical intervention is light therapy. Light therapy is used to treat some neutrophilic dermatoses.

[0785] As will be apparent from the foregoing, the present disclosure provides methods of concomitant therapeutic treatment of a subject, comprising administering to a subject in need thereof an effective amount of a first agent and a second agent or therapy, wherein the first agent is a pharmaceutical formulation of the present disclosure, and the second agent or therapy is also for the prevention or treatment of a neutrophil-mediated condition.

[0786] As used herein, the term "concomitant" as in the phrase "concomitant therapeutic treatment" includes administering a first agent in the presence of a second agent or therapy. A concomitant therapeutic treatment method includes methods in which the first, second, third or additional agents / therapies are co-administered. A concomitant therapeutic treatment method also includes methods in which the first or additional agents are administered in the presence of a second or additional agent or therapy, wherein the second or additional agent or therapy, for example, may have been previously administered. A concomitant therapeutic treatment may be executed step- wise by different actors. For example, one actor may administer to a subject a first agent and as a second actor may administer to the subject a second agent or therapy and the administering steps may be executed at the same time, or nearly the same time, or at distant times, so long as the first agent (and / or additional agents) are after administration in the presence of the second agent or therapy (and / or additional agents or therapies). The actor and the subject may be the same entity (e.g. a human). Kits and Other Compositions of Matter

[0787] Another example of the disclosure provides kits containing a pharmaceutical formulation of the present disclosure useful for the treatment or prevention of a disease or condition as described above.

[0788] In one example, the kit comprises (a) a container comprising a pharmaceutical formulation of the present disclosure; and (b) a package insert with instructions for treating or preventing a neutrophil-mediated condition in a subject.

[0789] In one example, the kit comprises (a) at least one pharmaceutical formulation of the present disclosure; (b) instructions for using the kit in treating or preventing the disease or condition in the subject; and (c) optionally, at least one further therapeutically active compound or drug.

[0790] In accordance with this example of the disclosure, the package insert is on or associated with the container. Suitable containers include, for example, bottles, vials, syringes, etc. The containers may be formed from a variety of materials such as glass or plastic. The container holds or contains a composition that is effective for treating a neutrophil-mediated condition and may have a sterile access port (for example, the container may be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). The label or package insert indicates that the composition is used for treating a subject eligible for treatment, e.g., one having or predisposed to developing a neutrophil-mediated condition, with specific guidance regarding dosing amounts and intervals of the pharmaceutical formulation and any other medicament being provided. The kit may further include other materials desirable from a commercial and user standpoint, including filters, needles, and syringes. In some examples of the present disclosure, the formulation can be present in an injectable device (e.g., an injectable syringe, e.g., a prefilled injectable syringe). The syringe may be adapted for individual administration, e.g., as a single vial system including an autoinjector (e.g., a pen-injector device). In one example, the injectable device is a prefilled pen or other suitable autoinjectable device, optionally with instruction for use and administration.

[0791] The kit optionally further comprises a container comprising a second medicament, wherein the pharmaceutical formulation is a first medicament, and which article further comprises instructions on the package insert for treating the subject with the second medicament, in an effective amount. The second medicament may be a therapeutic protein set forth above.

[0792] In one example, the disclosure provides a prefilled syringe or autoinjector comprising a formulation of the present disclosure. In one example, the prefilled syringe is a glass luer syringe with plunger. In one example, the disclosure provides a vial comprising a formulation of the disclosure.

[0793] The present disclosure includes the following non-limiting Examples.

[0794] EXAMPLES

[0795] Example 1: Methods

[0796] Visual Appearance

[0797] Visual appearance was conducted in an inspection station equipped with a white and black background and fluorescent light. Formulations in vials were gently swirled without producing bubbles then inspected for colour, clarity and the presence of visible particles. Inspections were conducted by two independent inspectors. In some experiments, colour / haze was determined using Vista Transmission colorimeter Fa. Hunterlab. pH measurements

[0798] The pH of the formulations was measured using a Mettler Toledo SevenExcellence pH meter equipped with a InLab®Ultra Micro ISM electrode.

[0799] UV Spectroscopy

[0800] Protein concentration was measured by using A280 / UV determination on the formulations via two methods:

[0801] • neat on an IMPLEN P360 Nanophotometer Measurements were conducted in triplicate and the mean value of the measurement calculated

[0802] • via gravimetric dilution to on the Shimadzu UV-1700 Spectrophotometer and performed in duplicate.

[0803] Size exclusion chromatography (SEC)-high performance liquid chromatography (HPLC)

[0804] SEC-HPLC was used to determine the protein aggregation profile of the formulations. Intact protein was detected at 280 nm with monomer species, high molecular weight species (HMWS, aggregates) and low molecular weight species (LMWS, fragments) reported as a relative area %. Internal and external references were used to validate the run. This was performed with a Dionex system (Ultimate 3000) via two methods:

[0805] (i) first method was performed with an Acquity BEH200 column (Waters, 1.7 pm, 4.6x150mm) to analyse the samples. Samples were diluted to 5 g / L in appropriate buffer, 3 pL was injected or 10 g / L in appropriate buffer and 1.5 pL was injected. Separation was performed under isocratic conditions at a flow rate of 0.3 mL / min. Mobile phase consisted Bis-Tris Propane buffer (pH 7.0) with a run time of 12 min.

[0806] (ii) second method was equipped with a TSkgel G3000SWxL column (TOSOH, 5 pm, 7.8 x 300 mm 250A) to analyse the samples. Samples were diluted to 5 g / L in appropriate buffer, 10.0 pL was injected and the separation was performed under isocratic conditions at a flow rate of 1.0 mL / min. Mobile phase consisted of sodium phosphate buffer (pH 7.0) with a run time of 15 min.

[0807] Cation exchange chromatography (CEX)

[0808] CEX-HPLC was used to determine the proportions of proteinaceous acidic, main and basics species. A Dionex system (Ultimate 3000) equipped with a Waters Acquity ProteinPak™ HiRes CM 7 pm 4.6x100 mm column was used to analyse the samples. Samples were diluted to 10 g / L in appropriate buffer, 2.5 pL injection volume & / or 5 g / L in appropriate buffer, 5 pL injection volume was used and separation was conducted with a gradient method at 0.7 ml / min. Briefly, two aqueous MES buffers at pH 6.2 with an increasing salt gradient over a run period of 24 minutes. Species were detected at 280 nm, identified against a reference standard and reported as relative Area percentage over the integrated area.

[0809] Capillary Gel Electrophoresis (CGE),

[0810] The protein “banding pattern” was obtained by Capillary Gel Electrophoresis. Analysis was performed using a microfluidic LabChip GXII system (Perkin Elmer Australia Pty Ltd) or PA800 (Beckman Coulter). The protein electrophoresis on the microfluidic chip was achieved by integration of the main features of one-dimensional SDSPAGE: these include the separation, staining, de-staining, and detection. Denatured proteins were loaded onto the chip directly from a microtiter plate through a capillary sipper. The samples were then electrokinetically loaded and injected into the 14 mm long separation channel containing a low viscosity matrix of entangled polymer solution. The entire sample preparation procedures were performed according to the manufacturers protocol. For non-reducing samples, protein solution were diluted to 2 g / L with nonreducing buffer and Milli-Q water. Reducing samples were diluted with kit buffer containing DTT. Denaturation occurred at 40°C for 20 min for non-reduced samples and at 80°C for 15 min for reduced samples. The PA800 method separates protein species based on their molecular weight, and detection occurs using a UV detector at 214 nm. Under non-reducing conditions, prior to analysis the sample is denatured by addition of Sodium Dodecyl Sulphate (SDS) and heat, followed by alkylation of free cysteines using N-ethylmaleimide (NEM). The relative main peak (purity) and low molecular weight species (LMWS; impurity) are measured. Under reducing conditions, prior to analysis the sample is denatured by addition of SDS and heat, followed by reduction of disulphide bonds with -mercaptoethanol (BME). Results were reported in relative area percentage for LMWS Intact and HMWS for non-reduced samples. For reduced samples, heavy and short chain fractions were considered.

[0811] Sub-visible particle count testing

[0812] Sub-visible particle counting was performed by Light obscuration using HIAC 9703+ utilising a low volume method of 4 x 1 mL, with the average of the final 3 runs being calculated and reported as particles > 2 pm, > 5 pm, > 10 pm and > 25 pm. Analysis of sub-visible particles morphology, size distribution and counts was also performed using a FlowCam Biologies instrument (a Dynamic / flow Imaging Particle Analysis - DIPA - technique) on selected formulations of interest. A minimum sample volume of 0.5 mL was used. Measurements were conducted in triplicate per formulation and the mean values of the measurements calculated and particles counted as 2 to 5 pm, 5 to 10 pm, 10 to 25 pm and > 25 pm.

[0813] In some experiments, the HORIZON® system from Halo Labs was used which employs Backgrounded Membrane Imaging (BMI) technology to measure and characterize subvisible particles in a high-throughput, low volume format. The system utilizes disposable membrane plates and automated image processing. BMI is also insensitive to solution refractive index, enabling reliable measurement of translucent protein aggregates.

[0814] Reverse Phase HPLC

[0815] A RP-HPLC method was used to determine the total amount of oxidized species as a percentage of the total area, and the relative amount of oxidation of the HC FC / 2, Light Chain region and HC Fd’domain. The sample is initially diluted with PBS to 10 mg / ml. The sample is digested with IdeS enzyme (Genovis FabRICATOR), which performs a site-specific cleavage below the hinge region of the IgG followed by an incubation step of an hour at 37°C. This is followed by denaturation and reduction of the sample with the addition of 20 mM DTT, 1 mM EDTA, 100 mM MES, pH 5.5, 3 M Guanidine-HCl and an incubation of 30 minutes at 56°C. The sample is then diluted with 25:75 v / v sample: MPA (0.1% TFA) to adjust the pH of the sample to enhance the sample stability. A Thermo Ultimate 3000 (or equivalent) equipped with an Acquity UPLC BEH300 C4 1.7 pm, 50mm x 2.1mm column was used to analyse the samples. A target loading of 5 pg was used with the final sample concentration and separation was conducted with a gradient method at 0.30 ml / min. Column temperature was set to 70°C. Briefly, two buffers (0.1% trifluoroacetic acid in water and 0.08% TFA in acetonitrile) were alternated over a period of 30 minutes. Species were reported at 280 nm, identified against a reference standard and reported as Relative Area percentage over the integrated area. The chromatograms of the samples contain three main peaks of light chain (LC), Fd’ and monomeric Fc (Fc / 2), the respective oxidation products associated with each domain elutes slightly earlier than each of the main peaks listed. The oxidised species for each domain is reported separately, as the percentage area relative to the area of the total peaks in that domain.

[0816] Closed Container Integrity

[0817] The closed container integrity of the formulations in the vials is performed via vacuum decay method using the glass vial VeriPac 455.

[0818] Endotoxin

[0819] A limulus amebocyte lysate method is used to measure endotoxin by the kinetic chromogenic method. Samples were required to be tested at 4 different dilutions, increasing by 10 fold and results reported from the valid result which has achieved an end-point result and has a PPC recovery of at closest to 100%.

[0820] Example 2: Stability: design of experiments

[0821] The aim of the experiments described in the following examples was to produce a formulation of CSL324 (anumigilimab), an antibody that binds to GCSF-R, which had long term stability and was suitable for subcutaneous administration.

[0822] To test the impact of several excipients on robustness and stability, 24 formulations were prepared across the ranges shown in Table 1. Details of each formulation are shown in Figure 1. Table 1: 5- factorial, customized design

[0823] Stability of each test formulation was measured at varying temperatures and intervals as shown in Table 2.

[0824] Table 2: Stability conditions Example 3: Stability results

[0825] Visual description

[0826] All vials at all time points were free of visible particles. All formulations showed colour B5 or B6 and were clear (0 to 1.2 % Haze) at all time points. Colour was determined using the Vista Transmission colorimeter Fa. Hunterlab; EP colour scale. Haze was determined according to ASTM D1003. Slightly increasing turbidity values were associated with an increasing protein concentration up to 80 mg / ml. Above 80 mg / ml, turbidity did not significantly increase further. (Figure 2A). No differences in turbidity under all stability conditions were observed due to increases in concentration of any of L-histidine, Arginine-HCl or L-proline (data not shown), or in formulations ranging in pH from 5.4-6.0 (Figure 2A).

[0827] Nano Differential Scanning Fluorimetry (nanoDSF) thermal unfolding

[0828] Mostly three prominent unfolding events were detected in line with capDSC (capillary differential scanning colorimetry) measurements. For most antibodies, the first transition resembles to the unfolding of the Fc fragment (~+60°C), the second one to the Fab domain (~+68°C) and the third one to the CH3 domain (~+78°C). Onset of scattering signal is in line with first unfolding event. Strong increases in scattering signal with Tm2 were observed, suggesting that unfolding of Fab domain mainly leads to aggregation.

[0829] Increasing pH was found to increase the thermal stability of CSE324 (Figure 2B). Increasing protein concentration increased Tml (Time to melting temperature) and decreased time to onset of scattering (Figure 2B). There was otherwise only minor effects observed for other excipients (data not shown).

[0830] Acidic / Basic species by CEX

[0831] Acidic species were found to increase with an increasing pH (Figure 2C, 2D). This increase in acidic species was associated with a corresponding decrease in basic and main species with an increasing pH. Acidic species were also found to increase after storage with increasing temperature (and pH). Other excipients showed only minor or no effects.

[0832] CE-SDS (non-reducing and reducing conditions)

[0833] Under non-reducing conditions, increasing pH was found to decrease low molecular weight species (EMWS) and AFMWS at +35°C (Figure 2E). No effect associated with changing pH was observed at +5 or +25°C (data not shown). A slight increase in LMWS with an increasing protein concentration was observed (Figure 2E). Other excipients showed only minor or no effects.

[0834] Under reducing conditions, no formulations were observed to impact LMWS. No significant changes after stability compared to initial time point and no main effects were detected (data not shown).

[0835] High molecular weight species (HMWS) by SE-HPLC

[0836] HMWS and change (A) HMWS increased mainly with an increasing protein concentration, particularly at >80 mg / mL (Figure 3). HMWS and A HMWS increased slightly with an increasing pH at real time and accelerated conditions (Figure 3A), but HMWS and A HMWS decreased with an increasing pH at +35°C (Figure 3B) and +5 (Figure 3D). Other excipients showed only minor effects. Figure 3A is representative of evaluation after 3 months at +35°C, Figures 3B and 3C are representative of evaluation after 3 months at +5°C, and Figure 3D is representative of evaluation after 3 months at +25°C. Quantitation is shown in the table below.

[0837] HMWS: Nominal protein concentration 40 to 120 mg / ml

[0838] Dynamic light scattering

[0839] All formulations showed a monomodal size distribution and no changes to the initial time point. Rh (hydrodynamic radius) and %PD (polydispersity index) increased with an increasing protein concentration and a decreasing Arginine-HCl concentration (Figure 4A). These increases might be caused by an increasing solution viscosity. A slightly decreased Rh & %PD was observed with increasing pH which was more pronounced at higher protein concentrations. Subvisible particles (SVP) by BMI using Horizon

[0840] Overall, all particle counts were far below US Pharmacopeia (USP)<787> limits. Slightly higher particle counts were observed at higher protein concentrations (2-25 pm) (Figure 4B). Minor effects of other excipients and pH were detected (Figure 4B).

[0841] Conclusions

[0842] Increasing protein concentrations were found to increase indicators of protein aggregation, as observed by HMWC, Rh, %PD, onset temperature of scattering and particle counts. Increasing pH-value was found to increase acidic species. pH-values showed different effects on aggregation depending on temperature; increasing pH increased HMWS at real time conditions.

[0843] Example 4: Exemplary formulation and long term stability

[0844] An exemplary antibody formulation, based on the results described above, contains the following components: 50 mg / mL CSL324, 20 mM L-histidine, 100 mM L-Arginine-HCl, 100 mM L-Proline, and 0.03% w / vPS80, at pH 5.7.

[0845] The long-term stability of the exemplary formulation was assessed by holding the formulation at 5°C (±3°C) (i.e. 2°C to 8°C) for 42 months, 25°C (±2°C) for 24 months, and 35°C (±3°C) for 12 months. The results are shown in Tables 3, 4 and 5, respectively.

[0846] Table 3: Long-term stability of exemplary CSL324 formulation after 42 months at 5°C ± 3°C

[0847] NS: Not scheduled

[0848] Table 4: Long-term stability of exemplary CSL324 formulation after 24 months at 25° C ± 2°C

[0849] NS: Not scheduled

[0850] Table 5: Long-term stability of exemplary CSL324 formulation after 12 months at 35°C ± 2°C

[0851] NS: Not scheduled

Claims

CLAIMS1. A liquid pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 m proline, and 90 mM to 110 mM arginine, and wherein the antigen binding domain comprises:(i) a heavy chain variable region (VH) comprising three complementarity determining regions (CDRs) of a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a light chain variable region (VL) comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 5; or(ii) a VH comprising three CDRs of a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 3.

2. The pharmaceutical formulation of claim 1, wherein the formulation comprises:(i) 20 mM histidine buffer;(ii) 0.03% (w / v) polysorbate 80;(iii) 100 mM arginine; and / or(iv) 100 mM proline.

3. The pharmaceutical formulation of claim 1 or 2, wherein the formulation has a pH of 5.7.

4. The pharmaceutical formulation of any one of claims 1 to 3, wherein one or more or all of the following apply:(i) the formulation comprises no more than 5% high molecular weight species (HMWS), or no more than 3.5% HMWS, as determined by size exclusion high performance liquid chromatography (SE-HPLC);(ii) at least 95% of the protein in the formulation is a monomer, as determined by SE-HPLC;(iii)the formulation comprises no more than 50% acidic species, as determined by cation exchange high performance liquid chromatography (CEX-HPLC);(iv)the formulation comprises no more than 20% basic species, as determined by CEX-HPLC; and(v) the formulation comprises no more than 10% low molecular weight species (LMWS), as determined by capillary electrophoresis with sodium dodecylsulfate (CE-SDS) under non-reducing conditions.

5. The pharmaceutical formulation of claim 4, wherein the amount of HMWS, monomer, acidic species, basic species, or LMWS is determined after storage for a period of at least 1 month, at least 3 months, at least 6 months, at least 9 months, at least 12 months, at least 18 months, at least 24 months, at least 30 months, at least 36 months, or at least 42 months at a temperature in the range of 2 °C to 30 °C.

6. The pharmaceutical formulation of any one of claims 1 to 5, wherein the formulation has a volume in the range of 0.5 mL to 5 mL.

7. The pharmaceutical formulation of any one of claims 1 to 6, wherein the formulation has a volume of 1 mL.

8. The pharmaceutical formulation of any one of claims 1 to 7, wherein the protein comprises:(i) a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 5; or(ii) a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 3; or(iii) a VH comprising:(a)a CDR1 comprising a sequence set forth in SEQ ID NO: 6;(b)a CDR2 comprising a sequence set forth in SEQ ID NO: 7; and(c)a CDR3 comprising a sequence set forth in SEQ ID NO: 8; and a VL comprising:(a)a CDR1 comprising a sequence set forth in SEQ ID NO: 9;(b)a CDR2 comprising a sequence set forth in SEQ ID NO: 10; and(c)a CDR3 comprising a sequence set forth in SEQ ID NO: 11.

9. A pharmaceutical formulation comprising a protein comprising an antigen binding domain of an antibody that specifically binds to granulocyte colony stimulating factor receptor (G-CSFR) and inhibits G-CSF signaling at a concentration of 50 mg / mL, 20 mM histidine buffer, 0.3 % (w / v) polysorbate 80, 100 mM proline, and 100 mMarginine, wherein the formulation has a pH of 5.5 to 5.9, and wherein the antigen binding domain comprises:(i) a heavy chain variable region (VH) comprising an amino acid sequence set forth in SEQ ID NO: 4 and a light chain variable region (VL) comprising an amino acid sequence set forth in SEQ ID NO: 5; or(ii) a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 3; or(iii) a VH comprising:(a) a complementarity determining region (CDR)l comprising a sequence set forth in SEQ ID NO: 6;(b) a CDR2 comprising a sequence set forth in SEQ ID NO: 7; and(c) a CDR3 comprising a sequence set forth in SEQ ID NO: 8; and a VL comprising:(a) a CDR1 comprising a sequence set forth in SEQ ID NO: 9;(b) a CDR2 comprising a sequence set forth in SEQ ID NO: 10; and(c) a CDR3 comprising a sequence set forth in SEQ ID NO: 11.

10. The pharmaceutical formulation of any one of claims 1 to 9, wherein the protein comprises:(i) a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22;(ii) a VH comprising three CDRs of a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising three CDRs of a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22; or(iii) a VH comprising:(a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 23;(b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 24; and(c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 25; and a VL comprising:(a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 26;(b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 27; and(c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 28.

11. The pharmaceutical formulation of any one of claims 1 to 10, wherein the protein comprises:(i) a heavy chain comprising a sequence set forth in SEQ ID NO: 14 and a light chain comprising a sequence set forth in SEQ ID NO: 15; or(ii) a heavy chain comprising a sequence set forth in SEQ ID NO: 16 and a light chain comprising a sequence set forth in SEQ ID NO: 15.

12. The pharmaceutical formulation of any one of claims 1 to 11, wherein(i) the formulation is formulated for subcutaneous administration;(ii) the formulation is formulated for self-administration; and / or(iii) the formulation is formulated for fixed-dose administration.

13. The pharmaceutical formulation of any one of claims 1 to 12, for use as a medicament.

14. A method of reducing circulating neutrophils in a human subject suffering from a neutrophil-mediated condition, the method comprising administering the pharmaceutical formulation of any one of claims 1 to 12 to the subject.

15. A method of treating or preventing a neutrophil-mediated condition in a subject, the method comprising administering the pharmaceutical formulation of any one of claims 1 to 12. to the subject.

16. A composition for use in reducing circulating neutrophils in a human subject suffering from a neutrophil-mediated condition, the composition comprising the pharmaceutical formulation of any one of claims 1 to 12.

17. A composition for use in treating or preventing a neutrophil-mediated condition in a subject, the composition comprising the pharmaceutical formulation of any one of claims 1 to 12.

18. The method of claim 14 or 15, or the composition for use of claim 16 or 17, wherein the neutrophil-mediated condition is(i) a neutrophilic skin condition; an autoimmune disease; an inflammatory disease; ischemia-reperfusion injury; a pulmonary disease; a neurological condition; or a neurodegenerative condition; or(ii) a neutrophilic dermatosis; amicrobial pustulosis of the folds (APF); plaque psoriasis; CARD 14 -mediated pustular psoriasis (CAMPS); cryopyrin associated periodic syndromes (CAPS); deficiency of interleukin- 1 receptor (DIRA); deficiency of interleukin- 36 receptor antagonist(DIRTA); hidradenitis suppurativa (HS); palmoplantar pustulosis; pyogenic arthritis; pyoderma gangrenosum and acne (PAPA); pyoderma gangrenosum; acne; and hidradenitis suppurativa (PASH); pyoderma gangrenosum(PG); skin lesions of Behcet’s disease; Still’s disease; Sweet syndrome; subcorneal pustulosis (Sneddon- Wilkinson); pustular psoriasis; palmoplantar pustulosis; acute generalized exanthematic pustulosis; infantile acropustulosis; synovitis; acne; pustulosis; hyperostosis and osteitis (SAPHO) syndrome; bowel-associated dermatosisarthritis syndrome (BAD AS); neutrophilic dermatosis of the dorsal hands; neutrophilic eccrine hidradenitis; erythema elevatum diutinum; Pyoderma gangrenosum; systemic inflammatory response syndrome (SIRS); chronic obstructive pulmonary disease (COPD); emphysema; bronchitis; pneumonia; acute respiratory distress syndrome; acute lung injury; asthma; multiple Sclerosis(MS); Devic's disease; a viral infection in the brain; uveitis; or arthritis.

19. The method of any one of claims 14, 15, 17 or 18, or the composition for use of any one of claims 16 to 18, wherein(i) the formulation is administered subcutaneously to the subject;(ii) the formulation is self-administered to the subject; and / or(iii) the formulation is administered to the subject using a fixed-dose administration regimen.

20. A kit for use in treating or preventing a neutrophil-mediated condition in a subject, the kit comprising:(a) at least one pharmaceutical formulation of any one of claims 1 to 12;(b) instructions for using the kit in treating or preventing the neutrophil-mediated condition in the subject; and(c) optionally, at least one further therapeutically active compound or drug.

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