Stable aqueous antibody formulations

Abstract

The present invention relates to stable aqueous antibody formulations. In some embodiments, the stable aqueous formulation comprises an anti-IL5R antibody in the range of from about 2 mg / mL to about 100 mg / mL, and a polysorbate-20 in the range of from about 0.002% to about 0.01%. The invention also provides methods of making and methods of using such antibody formulations.

Description

This application is a divisional application of Chinese patent application 201480055263.0, filed on October 23, 2014, “Stable Aqueous Antibody Formulation”. Technical Field

[0001] The present invention relates to a stable aqueous antibody formulation. In some embodiments, the stable aqueous formulation comprises: about 2 mg / mL to about 100 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises a CDR1, CDR2, and CDR3 sequence defined by Kabat of SEQ ID NO: 5-7, and wherein the light chain variable region comprises a CDR1, CDR2, and CDR3 sequence defined by Kabat of SEQ ID NO: 8-10; and about 0.002% to about 0.01% of polysorbate-20. The present invention also provides methods for preparing and using such antibody formulations. Background Art

[0002] Antibodies are used to treat different diseases and conditions due to the specificity of their target recognition, thereby producing highly selective results after systemic administration. In order to keep the antibody effective, the antibody must maintain its biological activity during production, purification, transportation and storage. New production and purification technologies have been developed to allow the production of a large amount of highly purified monoclonal antibodies. However, there are still problems that make these antibodies stable for transportation and storage, and there is even a problem of providing antibodies in a dosage form suitable for administration.

[0003] Denaturation, aggregation, contamination and particle formation can constitute a major obstacle in antibody formulation and storage. Due to the diversity of antibodies, there is no universal formula or conditions suitable for the storage of all antibodies. The best formula and conditions suitable for the storage of an antibody are usually specific to that antibody. Therefore, antibody storage formulas and methods are usually an important part of the commercial antibody research and development process.

[0004] Different methods have been proposed to overcome the difficult problems associated with antibody stability. For example, in some cases, antibodies are often lyophilized and then rehydrated shortly before administration. However, rehydration is usually undesirable because it adds extra steps in the administration process and may introduce contaminants into the formulation. In addition, even rehydrated antibodies may also suffer from aggregation and particle formation. Therefore, there is a need to provide a stable aqueous antibody formulation that can overcome the difficult problems associated with transportation and storage. Summary of the invention

[0005] The present invention relates to a stable aqueous antibody formulation, comprising: (a) about 2 mg / mL to about 100 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; and (b) about 0.002% to about 0.01% polysorbate-20.

[0006] In some embodiments, the aqueous formulation further comprises an uncharged excipient. In some embodiments, the uncharged excipient is trehalose. In some embodiments, the concentration of the uncharged excipient is about 20mM to about 80mM. In some embodiments, the concentration of the uncharged excipient is about 200mM to about 400mM.

[0007] The antibody can be present in different concentrations. In some embodiments, the formulation comprises about 2 to about 20 mg / ml of the antibody. In some embodiments, the formulation comprises about 20 to about 100 mg / ml of the antibody. In one embodiment, the formulation comprises 30 mg / ml of the antibody.

[0008] The formulation may further comprise arginine. In some embodiments, the arginine is L-arginine. In some embodiments, the formulation comprises about 100mM to about 200mM L-arginine. In some embodiments, the formulation comprises about 120mM to about 140mM L-arginine, and about 40mM to about 60mM uncharged excipient. In one embodiment, the formulation comprises about 125mM L-arginine. In one embodiment, the formulation comprises about 130mM L-arginine.

[0009] In some embodiments, the formulation further comprises histidine. In some embodiments, the concentration of the histidine is about 15 mM to about 30 mM. In one embodiment, the concentration of the histidine is about 20 mM.

[0010] In some embodiments, the antibody is not subjected to lyophilization.

[0011] In some embodiments, the present invention relates to a stable aqueous antibody formulation comprising about 2 mg / mL to about 100 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10, wherein the formulation is stable after storage at about 40°C for at least 1 month. In some embodiments, the formulation is stable after storage at about 25°C for at least 3 months. In some embodiments, the formulation is stable after storage at about 5°C for at least 18 months. In some embodiments, the antibody stored at about 40°C for at least 1 month retains at least 80% of the binding ability to the IL-5R polypeptide compared to a reference antibody that has not been stored. In some embodiments, the antibody stored at about 5°C for at least 6 months retains at least 80% of the binding ability to the IL-5R polypeptide compared to a reference antibody that has not been stored. In some embodiments, an antibody stored at about 40°C for at least 1 month retains at least 95% of the binding ability to the IL-5R polypeptide compared to a reference antibody that has not been stored. In some embodiments, an antibody stored at about 5°C for at least 6 months retains at least 95% of the binding ability to the IL-5R polypeptide compared to a reference antibody that has not been stored. In some embodiments, less than 2% of the antibodies form aggregates after storage at about 40°C for at least 1 month as determined by HPSEC. In some embodiments, less than 2% of the antibodies form aggregates after storage at about 5°C for at least 12 months as determined by HPSEC.

[0012] In some embodiments, the formulation is substantially free of particles as determined by visual inspection after storage at about 40° C. for at least 1 month. In some embodiments, the formulation is substantially free of particles as determined by visual inspection after storage at about 5° C. for at least 12 months.

[0013] In some embodiments, the formulation is an injectable formulation. In some embodiments, the formulation is suitable for intravenous, subcutaneous or intramuscular administration.

[0014] In some embodiments, the present invention relates to a sealed container containing an antibody formulation as described herein. In some embodiments, the present invention relates to a pharmaceutical unit dosage form suitable for parenteral administration to humans, the pharmaceutical unit dosage form being included in an antibody formulation as described herein in a suitable container. In some embodiments, the antibody formulation is administered intravenously, subcutaneously or intramuscularly. In some embodiments, the suitable container is a prefilled syringe. In some embodiments, the prefilled syringe includes an injection needle. In some embodiments, the injection needle is a 29G thin tube injection needle. In some embodiments, the prefilled syringe is a plastic syringe or a glass syringe. In some embodiments, the prefilled syringe is made of a material that is substantially free of tungsten.

[0015] In some embodiments, the pre-filled syringe is coated with silicone. In some embodiments, the pre-filled syringe includes a plunger having a fluoropolymer resin disk. In some embodiments, the pre-filled syringe comprises: (a) about 2 mg / mL to about 20 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises CDR1, CDR2, and CDR3 sequences defined by Kabat of SEQ ID NO: 5-7, and wherein the light chain variable region comprises CDR1, CDR2, and CDR3 sequences defined by Kabat of SEQ ID NO: 8-10; and (b) about 0.002% to about 0.01% polysorbate-20. In some embodiments, the pre-filled syringe further comprises: (c) about 40 mM to about 60 mM trehalose, and (d) about 110 mM to about 150 mM L-arginine. In some embodiments, the present invention relates to a prefilled syringe comprising: (a) about 20 mg / mL to about 100 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; and (b) about 0.002% to about 0.01% polysorbate-20. In some embodiments, the formulation further comprises: (c) about 200 mM to about 300 mM trehalose.

[0016] In some embodiments, the invention relates to a kit comprising a formulation described herein, a container described herein, a unit dosage form described herein, or a pre-filled syringe described herein.

[0017] In some embodiments, the present invention relates to a stable aqueous antibody formulation comprising: (a) about 2 mg / mL to about 20 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; (b) about 0.002% to about 0.01% polysorbate-20; (c) about 40 mM to about 60 mM trehalose; (d) about 110 mM to about 150 mM L-arginine; and about 15 mM to about 30 mM histidine. In one embodiment, the present invention relates to a stable aqueous antibody formulation comprising: (a) about 2 mg / mL to about 20 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; (b) about 0.006% polysorbate-20; (c) about 50 mM trehalose; (d) about 130 mM L-arginine; and (e) about 20 mM histidine.

[0018] In some embodiments, the present invention relates to a stable aqueous antibody formulation comprising: (a) about 20 mg / mL to about 100 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; (b) about 0.002% to about 0.01% polysorbate-20; (c) about 200 mM to about 300 mM trehalose; and (d) about 15 mM to about 30 mM histidine. In one embodiment, the present invention relates to a stable aqueous antibody formulation comprising: (a) about 20 mg / mL to about 100 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; (b) about 0.006% polysorbate-20; (c) about 250 mM trehalose; and (d) about 20 mM histidine. In another embodiment, the present invention relates to a stable aqueous antibody formulation comprising: (a) about 30 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; (b) about 0.006% polysorbate-20; (c) about 250 mM trehalose; and (d) about 20 mM histidine.

[0019] In some embodiments, the present invention relates to a method for preparing a stable aqueous antibody formulation, the method comprising: (a) purifying an antibody to about 1 mg / mL to about 400 mg / mL, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; (b) placing the isolated antibody in a stabilizing formulation to form the stable aqueous antibody formulation, wherein the resulting stable aqueous antibody formulation comprises: (i) about 2 mg / mL to about 100 mg / mL of the antibody and (ii) about 0.002% to about 0.01% polysorbate-20.

[0020] In some embodiments, the present invention relates to a method for preparing a stable aqueous antibody formulation, the method comprising: (a) purifying an antibody to about 1 mg / mL to about 400 mg / mL, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; (b) diluting the antibody to about 2 mg / mL to about 20 mg / mL of the antibody in a solution comprising: (i) about 0.002% to about 0.01% polysorbate-20; (ii) about 40 mM to about 60 mM trehalose; and (iii) about 110 mM to about 150 mM L-arginine.

[0021] In some embodiments, the present invention relates to a method for preparing a stable aqueous antibody formulation, the method comprising: (a) purifying an antibody to about 1 mg / mL to about 400 mg / mL, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; (b) diluting the antibody to about 20 mg / mL to about 100 mg / mL of the antibody in a solution comprising: (i) about 0.002% to about 0.01% polysorbate-20; and (ii) about 200 mM to about 300 mM trehalose.

[0022] In some embodiments, the present invention relates to a method of preparing a rehydrated antibody formulation comprising an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10, the method comprising: (a) purifying the antibody from cell culture; (b) lyophilizing the isolated antibody; (c) adding the lyophilized antibody to an aqueous solution to form a rehydrated antibody formulation, wherein the rehydrated antibody formulation comprises: (i) about 2 mg / mL to about 100 mg / mL of the antibody and (ii) about 0.002% to about 0.01% polysorbate-20.

[0023] In some embodiments, the present invention relates to an antibody formulation comprising an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the CDR1, CDR2 and CDR3 sequences defined by Kabat of SEQ ID NO: 5-7, and wherein the light chain variable region comprises the CDR1, CDR2 and CDR3 sequences defined by Kabat of SEQ ID NO: 8-10, wherein the antibody formulation is substantially free of particles. In some embodiments, the antibody formulation comprises an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the CDR1, CDR2 and CDR3 sequences defined by Kabat of SEQ ID NO: 5-7, and wherein the light chain variable region comprises the CDR1, CDR2 and CDR3 sequences defined by Kabat of SEQ ID NO: 8-10, wherein the antibody formulation is substantially free of active glutathione S-transferase (GST). In some embodiments, the antibody formulation is substantially free of GST. In some embodiments, the antibody formulation is substantially free of particles when stored at 38°C-42°C for at least 1 month. In some embodiments, the antibody formulation is substantially free of particles when stored at 2° C.-6° C. for at least 6 months. In some embodiments, the antibody formulation is substantially free of particles when stored at 2° C.-6° C. for at least 18 months.

[0024] In some embodiments, the present invention relates to a method for purifying an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises a CDR1, CDR2 and CDR3 sequence defined by Kabat of SEQ ID NO: 5-7, and wherein the light chain variable region comprises a CDR1, CDR2 and CDR3 sequence defined by Kabat of SEQ ID NO: 8-10, the method comprising: (a) obtaining a cell culture comprising the antibody; (b) performing affinity chromatography on the antibody; (c) performing cation exchange on the antibody; (d) performing mixed mode chromatography on the antibody. In some embodiments, the method further comprises a virus inactivation process and / or a diafiltration process.

[0025] In some embodiments, the present invention relates to a method for treating a lung disease or condition in a subject, the method comprising administering a therapeutically effective amount of an antibody formulation described herein, a container described herein, a unit dosage form described herein, or a prefilled syringe described herein. In some embodiments, the lung disease or condition is an eosinophilic disease or condition. In some embodiments, the lung disease or condition is asthma, COPD, eosinophilic asthma, eosinophilic and neutrophilic combined asthma, aspirin-sensitive asthma, allergic bronchopulmonary aspergillosis, acute and chronic eosinophilic bronchitis, acute and chronic eosinophilic pneumonia, Churg-Strauss syndrome, hypereosinophilic syndrome, drugs, irritants, and radiation-induced pulmonary eosinophilia, infection-induced pulmonary eosinophilia (fungal, tuberculosis, parasites), autoimmune-related pulmonary eosinophilia, eosinophilic esophagitis, Crohn's disease, or a combination thereof. In some embodiments, the lung disease or condition is asthma. In some embodiments, the pulmonary disease or disorder is chronic obstructive pulmonary disease (COPD). BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 .Amino acid sequence of anti-IL5R antibody.

[0027] Figure 2 The effect of polysorbate-20 in solution on the monomer fraction is shown. Polysorbate concentrations above 0.005% are required to fully maintain monomer levels.

[0028] Figure 3 The effect of polysorbate-20 on sub-visible particle counts in 2 g / L solution is shown. Data for particles >2 μm are not shown, but exhibit a similar pattern to larger particles. The data indicate that sub-visible particle levels in 2 g / L solution are not controlled by any polysorbate level.

[0029] Figure 4The effect of polysorbate-20 on sub-visible particle counts in a 100 g / L solution is shown. Data for particles >2 μm are not shown, but exhibit a similar pattern to larger particles. The data indicate that polysorbate levels above 0.003% in a 100 g / L solution control sub-visible particle levels.

[0030] Figure 5 HPLC monomer (%) and others (%) as a function of solution pH and protein concentration. Monomer loss is minimized in the pH range of 5.5-6.5.

[0031] Figure 6 Particle formation as a function of solution pH and protein concentration, including sub-visible particles >10 μm measured by MFI and visible particles assessed by comparison with standards. Sub-visible particle counts depended on protein concentration but did not show a trend with pH. In the pH range of 5.5-6.5, more visible particles were observed in lower protein concentration solutions.

[0032] Figure 7 .Sub-visible particle counts >10 μm filtered with aspect ratio to remove silicone oil droplets. This data compares SVP counts immediately after shipping to those after 1 month storage at 25°C. Independent of PS-20, high counts were observed for low protein concentrations.

[0033] Figure 8 . Particle counts >10 μm after simulated transport as determined by MFI for different protein concentrations and formulations. Formulations with higher ionic strength were more stable, like formulations ≥10 g / L trehalose.

[0034] Fig. 9 . Particle counts >10 μm after simulated transport determined by MFI for 2 g / L protein and different formulations. Arginine concentration should be >50 mM and NaCl concentration should be >75 mM.

[0035] Fig.10 . Particle counts >10 μm after simulated transport as determined by MFI for 2 g / L protein and different formulations. Any excipient concentration within this range is acceptable.

[0036] Fig.11 . Particle counts >10 μm after simulated transport as determined by MFI for 2 g / L protein and different formulations.

[0037] Fig.12 . Shown are the monomer losses of the 2 mg / mL, 20 mg / mL, and 100 mg / mL formulations in the vials and prefilled syringes. All PFSs showed similar losses to the vials and to each other.

[0038] Fig.13 . A graphical representation of the binning strategy is shown. Blue indicates formulations containing arginine or NaCl. Green indicates trehalose formulations. The data points are prepared samples and the lines indicate binning options that can be used to obtain intermediate doses.

[0039] Fig.14 .Test plan for stability study #2. All tests marked are performed on the antibody formulation. Yellow shading indicates tests that will be performed on placebo. The symbol "ABC" indicates submissions for the following tests: potency (bioassay), RP-HPLC, cIEF, non-reduced bioanalyzer, and reduced bioanalyzer.

[0040] Fig.15 is a graph showing samples prepared to define a design space as a function of protein concentration and polysorbate-20 ("PS-20") concentration for two formulation intervals.

[0041] Fig.16 The graph is of sub-visible particles at time 0 measured by MFI after delivery. The results show that particles are formed in the absence of PS-20, but 0.002% PS-20 is sufficient to inhibit particle formation after delivery.

[0042] Fig.17 (AD). Visible particle observations were scored against appearance criteria and are shown here at the 9 month time point. Observations were made in close proximity to light. The samples shown included a pre-filled syringe ("PFS") containing a trehalose formulation ( Fig.17 A), vial containing trehalose formulation ( Fig.17 B) PFS containing arginine preparations ( Fig.17 C), and vials containing arginine formulation ( Fig.17 D). The data supports the target of 0.006% PS-20 and an acceptable range of 0.002% - 0.01% PS-20 in PFS. The vials are shown as a worst case comparison.

[0043] Fig.18 Different SVP methods were compared for capturing the increase in sample time in association with visible particles. Flow cytometry and small particle counts (>1 μm and >2 μm) determined by MFI could also capture this trend.

[0044] Fig.19 Is PFS( Fig.19 A) and vials ( Fig.19Comparison of appearance standard scores and MFI results (particles > 1 μm) for the trehalose formulations in B). Good agreement of both methods was observed, both indicating an acceptable PS-20 range of 0.002% - 0.01%.

[0045] Fig. 20 Represents the interval design as outlined in Example 3 and the advance batch stability study performed in ABC. The orange shaded area indicates the arginine formulation and the blue shaded area indicates the trehalose formulation, all formulations having 0.006% PS-20.

[0046] Fig.21 A schematic example of the antibody purification process is shown.

[0047] Fig. 22 2D gel analysis showing the flow-through of the Protein A column used in the purification of anti-IL5R antibodies. DETAILED DESCRIPTION

[0048] It should be appreciated that the specific implementations shown and described herein are examples, and are not intended to otherwise limit the scope of the present application in any way.It should also be appreciated that the various embodiments and features of the invention described herein may be combined in any and all ways.

[0049] These disclosed patents, patent applications, websites, company names, and scientific literature mentioned herein are hereby incorporated by reference in their entirety to the same extent as if each were specifically and individually indicated to be incorporated by reference. Any conflict between any reference cited herein and the specific teaching of this specification sheet should be resolved in a manner that is favorable to the latter. Likewise, any conflict between the definition of a word or phrase as understood in the field and the definition of the word or phrase as specifically taught in this specification sheet should be resolved in a manner that is favorable to the latter.

[0050] As used in the present specification, the singular forms "a", "an" and "the" specifically also include the plural forms of the terms they refer to, unless the content clearly indicates otherwise.

[0051] Throughout this disclosure, unless otherwise indicated, all percentages, ratios, and the like are expressed "by weight." As used herein, "by weight" is synonymous with the term "by mass" and indicates that the ratios or percentages defined herein are expressed in terms of weight rather than volume, thickness, or some other measurement.

[0052] The term "about" is used herein to mean approximately, in the region of, roughly, or around. When the term "about" is used in conjunction with a numerical range, it modifies the range by extending the upper and lower limits of the recited numerical values. Generally, the term "about" is used herein to modify a numerical value by a variation of 10% above and below the stated value.

[0053] Technical and scientific terms used herein have the meanings commonly understood by one of ordinary skill in the art to which this application relates unless defined otherwise. Reference is made herein to various methods and materials known to one of ordinary skill in the art. Standard reference works setting forth the general principles of recombinant DNA technology include Sambrook et al., "Molecular Cloning: A Laboratory Manual," 2nd Ed., Cold Spring Harbor Laboratory Press, New York (1989); Kaufman et al., Eds., "Handbook of Molecular and Cellular Methods in Biology in Medicine," CRC Press, Boca Raton (1995); and McPherson, Ed., "Directed Mutagenesis: A Practical Approach," IRL Press, Oxford (1991), the disclosures of each of which are incorporated herein by reference in their entirety.

[0054] The present invention relates to stable aqueous antibody formulations. As described herein, the term "antibody formulation" refers to a composition comprising one or antibody molecules. In the present invention, the term "antibody" is not specifically limited. For clarity, "antibody" takes its broadest meaning and includes any immunoglobulin (Ig), its active or desired variant and its active or desired fragment (e.g., Fab fragment, camelid antibody (single-chain antibody) and nanobody). The term "antibody" can also refer to a dimer or polymer. Antibodies can be polyclonal or monoclonal, and can be naturally occurring or recombinantly produced. Therefore, human antibodies, non-human antibodies, humanized antibodies and chimeric antibodies are all included in the term "antibody". Typically, an antibody is a monoclonal antibody of one of the following categories: IgG, IgE, IgM, IgD and IgA; and more typically IgG or IgA.

[0055] The antibodies of the present invention can be from any animal source, including birds and mammals. In certain embodiments, the antibodies of the methods of the present invention are human, murine (e.g., mouse and rat), donkey, sheep, rabbit, goat, guinea pig, camel, horse or chicken. As used herein, "human" antibodies include antibodies with the amino acid sequence of human immunoglobulins, and include antibodies separated from human immunoglobulin libraries or from transgenic animals for one or more human immunoglobulins and that do not express endogenous immunoglobulins. See, for example, U.S. Pat. No. 5,939,598 to Kucherlapati et al.

[0056] Antibodies of the invention may include, for example, natural antibodies, intact monoclonal antibodies, polyclonal antibodies, multispecific antibodies formed from at least two intact antibodies (e.g., bispecific antibodies), antibody fragments (e.g., antibody fragments that bind and / or recognize one or more antigens), humanized antibodies, human antibodies (Jakobovits et al., Proc. Natl. Acad. Sci. USA 90:2551 (1993); Jakobovits et al., Nature 362:255-258 (1993); Bruggermann et al., Year in Immunology 7:33 (1993). Immunol. 7:33 (1993); U.S. Pat. Nos. 5,591,669 and 5,545,807), antibodies and antibody fragments isolated from antibody phage libraries (McCafferty et al., Nature 348:552-554 (1990); Clarkson et al., Nature 352:624-628 (1991); Marks et al., J. Mol. Biol. 222:581-597 (1991); Marks et al., Biotechnol. 10:779-783 (1992); et al., Bio / Technology 10:779-783 (1992); Waterhouse et al., Nucl. Acids Res. 21:2265-2266 (1993). The antibodies purified by the methods of the present invention can be recombinantly fused to the N- or C-terminus of a heterologous polypeptide or chemically conjugated (including covalently and non-covalently conjugated) to a polypeptide or other composition. For example, the antibodies purified by the methods of the present invention can be recombinantly fused or conjugated to molecules useful as labels and effector molecules (such as heterologous polypeptides, drugs or toxins) in detection assays. See, e.g., PCT Publications WO 92 / 08495; WO 91 / 14438; WO 89 / 12624; U.S. Pat. No. 5,314,995; and EP 396,387.

[0057] In some embodiments, the antibody formulations of the present invention comprise anti-IL5 receptor (anti-IL5R) antibodies. The antibodies of the present invention specifically bind to an antibody of interest or a fragment thereof, and do not specifically bind to other antigens or fragments thereof. For example, an anti-IL5R antibody will immunospecifically bind to an interleukin-5 receptor polypeptide and do not specifically bind to other polypeptides. Preferably, the antibody or antibody fragment that immunospecifically binds to the IL-5 receptor has a higher affinity for the IL-5 receptor or a fragment of the IL-5 receptor polypeptide than for other polypeptides or other polypeptide fragments. The affinity of an antibody is a measure of its binding to a specific antigen at a single antigen-antibody site, and is essentially the sum of all attractive and repulsive forces present in the interaction between an antibody and an antigen binding site of a specific epitope. The affinity of an antibody for a particular antigen (e.g., an IL-5 polypeptide or a fragment of an IL-5 polypeptide) can be represented by the equilibrium constant K, which is defined by the equation K = [Ag Ab] / [Ag] [Ab], which is the affinity of the antibody combining site, where [Ag] is the concentration of free antigen, [Ab] is the concentration of free antibody and [Ag Ab] is the concentration of antigen-antibody complexes. When the antigen and antibody react strongly together there will be very little free antigen or free antibody, and therefore the equilibrium constant or affinity of the antibody will be high. High affinity antibodies occur when there is a good match between antigen and antibody (for a discussion of antibody affinity, see Sigal and Ron ed., 1994, Immunology and Inflammation-Basic Mechanisms and Clinical Consequences, McGraw-Hill, Inc. New York at pages 56-57; and Seymour et al., 1995, Immunology-An Introduction for the Health Sciences, McGraw-Hill Book Company, Australia at pages 31-32). Preferably, the antibody or antibody fragment that immunospecifically binds to an IL-5 polypeptide or fragment thereof does not cross-react with other antigens.That is, the antibody or antibody fragment immunospecifically binds to the IL-5 polypeptide or fragment thereof with a higher energy than to other polypeptides or fragments of other polypeptides (for a discussion of antibody specificity, see, e.g., Paul ed., 1989, Fundamental Immunology, 2nd ed., Raven Press, New York, pp. 332-336). nd ed., Raven Press, New York at pages 332-336)). Antibodies or antibody fragments that immunospecifically bind to IL-5 polypeptides can be identified, for example, by immunoassays such as radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), and BIAcore assays or other techniques known to those skilled in the art (for a discussion of different assays used to determine antibody-antigen interactions in vivo, see, for example, Seymour et al, 1995, Immunology-An Introduction for the Health Sciences, McGraw-Hill Book Company, Australia at pages 33-41). Antibodies or antibody fragments that immunospecifically bind to IL-5 polypeptides or fragments thereof only act against the IL-5 polypeptide and do not significantly act against other activities.

[0058] In one embodiment, the IL-5R polypeptide is human IL-5R, an analog, derivative or fragment thereof. The nucleotide sequence of human IL-5R can be found in the GenBank database (see, e.g., accession number M96652.1). The amino acid sequence of human IL-5R can be found in the GenBank database (see, e.g., accession number Q01344). Each of these accession numbers is expressly incorporated herein by reference.

[0059] In some embodiments, the antibody formulation comprises an anti-IL5R antibody, such as a human anti-IL5R antibody. In some embodiments, the anti-IL5R antibody comprises: a light chain comprising SEQ ID NO: 2 and a heavy chain comprising SEQ ID NO: 4. In other embodiments, the anti-IL5R antibody comprises: a light chain variable region comprising SEQ ID NO: 1 and a heavy chain variable region comprising SEQ ID NO: 3. In another embodiment, the anti-IL5R antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10. One of ordinary skill in the art will readily be able to identify Chothia-defined, Abm-defined, or other CDRs.

[0060] In one embodiment, the anti-IL5R antibody is benralizumab. Information about benralizumab (or fragments thereof) for use in the methods provided herein can be found in U.S. Patent Application Publication No. US2010 / 0291073 A1, the disclosure of which is incorporated herein by reference in its entirety.

[0061] As used herein, the term "analog" or "antibody analog" in the context of antibodies refers to a second antibody that has a function similar to or identical to that of an antibody, but does not necessarily contain an amino acid sequence similar to or identical to that of an antibody or possess a structure similar to or identical to that of an antibody, i.e., an antibody analog. An antibody having an analogous amino acid sequence refers to an antibody analog that satisfies at least one of the following: (a) an antibody analog having an amino acid sequence that is at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identical to the amino acid sequence of the antibody; (b) an antibody analog encoded by a nucleotide sequence that hybridizes under stringent conditions to a nucleotide sequence encoding an antibody having at least 5 consecutive amino acid residues, at least 10 consecutive amino acid residues, at least 15 consecutive amino acid residues, at least 20 consecutive amino acid residues, at least 25 consecutive amino acid residues, at least 30 consecutive amino acid residues, at least 35 consecutive amino acid residues, at least 40 consecutive amino acid residues, at least 45 consecutive amino acid residues, at least 50 consecutive amino acid residues, at least 55 consecutive amino acid residues, at least 60 consecutive amino acid residues, at least 65 consecutive amino acid residues, at least 70 consecutive amino acid residues, at least 75 consecutive amino acid residues, at least 80 consecutive amino acid residues, at least 85 consecutive amino acid residues, at least 90 consecutive amino acid residues, at least 95 consecutive amino acid residues, at least 99 consecutive amino acid residues, at least 10 consecutive amino acid residues, at least 15 consecutive amino acid residues, at least 10 consecutive amino acid residues, at least 15 consecutive amino acid residue amino acid residues, at least 40 consecutive amino acid residues, at least 50 consecutive amino acid residues, at least 60 consecutive amino acid residues, at least 70 consecutive amino acid residues, at least 80 consecutive amino acid residues, at least 90 consecutive amino acid residues, at least 100 consecutive amino acid residues, at least 125 consecutive amino acid residues, or at least 150 consecutive amino acid residues; and (c) antibody analogs encoded by nucleotide sequences that are at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identical to the nucleotide sequence encoding the antibody. Antibody analogs having a structure similar to an antibody refer to protein agents having a secondary structure, tertiary structure or quaternary structure similar to an antibody. The structure of an antibody analog or antibody can be determined by methods known to those skilled in the art, including but not limited to peptide sequencing, X-ray crystallography, nuclear magnetic resonance, circular dichroism, and crystallographic electron microscopy.

[0062] In order to determine the percent identity of two amino acid sequences or two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in the first amino acid sequence or nucleic acid sequence for optimal alignment with the second amino acid or nucleic acid sequence). The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, the molecule is considered to be consistent at that position. The percent identity between the two sequences is a function of the number of consistent positions shared by the sequences (i.e., % identity = number of consistent overlapping positions / total number of positions × 100%). In one embodiment, the two sequences are the same length.

[0063] The determination of the percentage identity between two sequences can also be completed using a mathematical algorithm. A non-limiting example of a mathematical algorithm for comparing two sequences is Karlin and Altschul, 1990, Proc. Natl. Acad. Sci. USA 87: 2264-2268, such as in Karlin and Altschul, 1993, Proc. Natl. Acad. Sci. USA 90: 5873-5877 (Karlin and Altschul, 1993, Proc. Natl. Acad. Sci. USA 90: 5873-5877) modified algorithm. This algorithm is incorporated into the NBLAST and XBLAST programs of Altschul et al., 1990, J. Mol. Biol. 215: 403 (Altschul et ah, 1990, J. Mol. Biol. 215: 403). The BLAST nucleotide search is performed using NBLAST nucleotide program parameter settings such as score = 100 and word length = 12 to obtain nucleotide sequences homologous to the nucleic acid molecules of the present invention. The BLAST protein search is performed using XBLAST program parameter settings such as score = 50 and word length = 3 to obtain amino acid sequences homologous to the protein molecules of the present invention. In order to obtain gap comparisons for comparison purposes, the gap BLAST program described in Alchur et al., 1997, Nucleic Acids Res. 25: 3389-3402 (Altschul et al, 1997, Nucleic Acids Res. 25: 3389-3402) can be used. Alternatively, PSI-BLAST can be used to perform an iterative search to detect distant relationships between molecules (the same). When using BLAST programs, gap BLAST programs, and PSI-Blast programs, the default parameters of each program (e.g., XBLAST and NBLAST) can be used (see, e.g., NCBI website). Another preferred, non-limiting example of a mathematical algorithm for the comparison of sequences is the algorithm of Myers and Miller, 1988, CABIOS 4: 11-17. This algorithm is incorporated into the ALIGN program (version 2.0), which is part of the GCG sequence alignment software package. When utilizing the ALIGN program to compare amino acid sequences, a PAM 120 weight residue table, a gap length penalty of 12, and a gap penalty of 4 may be used.

[0064] In some embodiments, the antibody in the antibody formulation is purified before being added to the antibody formulation. The terms "isolate" and "purify" refer to separating an antibody from impurities or other contaminants in a composition in which the antibody is present (e.g., a composition comprising host cell proteins). In some embodiments, at least 50%, 70%, 80%, 90%, 95%, 98%, 99%, 99.5%, or 99.9% (w / w) of the impurities are purified away from the antibody. For example, in some embodiments, purification of an antibody, such as an anti-IL5R antibody, includes separating the antibody from 99% (w / w) of the host cell proteins initially present in the composition.

[0065] In some embodiments, the terms "isolate" and "purify" refer to separating an antibody (e.g., an anti-IL5R antibody) from impurities or other contaminants in a composition to a degree consistent with the guidelines of a governmental organization (e.g., the World Health Organization or the U.S. Food and Drug Administration).

[0066] Methods of purifying antibodies are known to those skilled in the art. Suitable techniques for performing purification include different types of chromatography, such as affinity chromatography, hydrophobic interaction, ion exchange (such as cation exchange chromatography or mixed mode chromatography), and diafiltration.

[0067] Affinity chromatography refers to the separation method of the affinity ligand of the antibody by the specific binding property of the antibody. Functional affinity ligands can be fixed on a solid or semi-solid support, so that when the composition comprising the antibody passes through a ligand and a solid support, the antibody with the specific binding affinity for the ligand is adsorbed on the ligand, and one or more other impurities are not adsorbed (or combined with lower affinity), and can be separated from the antibody. Examples of impurities that are not typically combined (or not well combined) include process-related impurities (e.g., host cell proteins, DNA, culture medium components) and some product-related impurities (e.g., antibody fragments). In certain embodiments, the solid support comprising the ligand is washed with a buffer solution once or multiple times to remove additional impurities, and then the adsorbed antibody is removed from the ligand and support. After one or more impurities have been removed, the adsorbed antibody can be removed (eluted) from the ligand and support, resulting in the separation of the antibody and the original composition. The method of removing the antibody from the part and support depends on the part, and is known to those skilled in the art, and can include, for example, changes in environmental aspects (e.g., pH), the addition of a chaotropic agent or a denaturing agent, or the addition of a commercially available elution buffer. In certain embodiments, more than one affinity purification process can be used to the composition. Different affinity ligands are known in the art, including protein A and protein G (and combinations thereof). Fixed ligands are commercially available. For example, protein A affinity systems include MabSelect, MabSelect SuRe, MabSelect Xtra, MabSelect SuRe LX, Sepaharose CL-4B, ProSep vA, ProSep vA Ultra, and Ceramic HyperD.

[0068] Ion exchange chromatography includes cation exchange chromatography and mixed chromatography. Cation exchange chromatography refers to any method in which antibodies and some impurities or multiple impurities can be separated based on charge difference using a cation exchange matrix. The cation exchange matrix generally includes covalently bound negatively charged groups. Weak or strong cation exchange resins can be used. Typically, strong cation exchange resins include loaded organic groups containing sulfonic acid or sulfonate groups (depending on pH). Weak cation exchange resins typically include loaded organic groups containing carboxylic acid or carboxylate groups (depending on pH). In certain embodiments, multimodal cation exchange resins can be used, which combine additional binding mechanisms and ionic interactions, such as one or more of hydrogen bond interactions and hydrophobic interactions. Examples of suitable cation exchange resins are well known in the art and may include, but are not limited to, Fractogel, carboxymethyl (CM), sulfoethyl (SE), sulfopropyl (SP), phosphate (P) and sulfonate (S), PROPAC WCX-10TM (Dionex), Capto S, S-Sepharose FF, FractogelEMD SO3M, Toyopearl Megacap II SP 550C, Poros 50HS, and SP-Sepharose Matrix. In some embodiments, more than one cation exchange chromatography process may be used on the composition.

[0069] Mixed mode chromatography refers to a method that utilizes more than one form of interaction between the stationary phase and the analyte to achieve their separation from impurities (e.g., process-related impurities such as host cell proteins, DNA and / or endogenous or adventitious viruses). Examples of suitable anion exchange matrices are known in the art and may include, but are not limited to, Capto Adhere, Sartobind Q, Natrix Q, Chromasorb Q, and Mustang Q.

[0070] In certain embodiments, additional filtering steps can be used to remove impurities. For example, in certain embodiments, nanofiltration or ultrafiltration is used. Nanofiltration includes making compositions pass through a matrix with a pore size of, for example, less than 75nm, less than 50nm and even less than 15nm to separate impurities such as viruses from antibodies. Available commercially available nanofilters and ultrafilters are produced by different suppliers, such as Millipore (Billerica, Massachusetts, such as ViresolvePro and Viresolve Pro+), Pall Corporation (East Hills, New York) (Pall Corporation (East Hills, NY)), GE Healthcare Sciences (Piscataway, New Jersey) (GE Healthcare Sciences (Piscataway, NJ)) and Sartorius Corporation (Goettingen, Germany) (Sartorius Corporation (Goettingen, Germany)).

[0071] In some embodiments, the antibody of the invention (e.g., anti-IL5R, comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10) is 1 mg / ml to 200 mg / ml, 2 mg / ml to 100 mg / ml, 2 mg / ml to 30 mg / ml, 2 mg / ml to 25 mg / ml, 2 mg / ml to 20 mg / ml, 3 mg / ml, 4 mg / ml, 5 mg / ml, 6 mg / ml, 7 mg / ml, 8 mg / ml, 9 mg / ml, 10 mg / ml, 11 mg / ml, 12 mg / ml, 13 mg / ml, 14 mg / ml, 15 mg / ml, 16 mg / ml, 17 mg / ml, 18 mg / ml, 19 mg / ml, or 20 mg / ml. In some embodiments, an antibody of the invention, e.g., anti-IL5R, is at a concentration of about 20 mg / ml, 25 mg / ml, 30 mg / ml, 40 mg / ml, 45 mg / ml, 50 mg / ml, 55 mg / ml, 60 mg / ml, 65 mg / ml, 70 mg / ml, 75 mg / ml, 80 mg / ml, 85 mg / ml, 90 mg / ml, 95 mg / ml, or 100 mg / ml.

[0072] The antibody formulations of the present invention may include uncharged excipients. The term excipient refers to a pharmacologically inactive substance formulated with the antibodies described herein. In some embodiments, the excipient may help prevent denaturation or otherwise help stabilize the antibody. Suitable excipients that can be used for pharmaceutical compositions are well known in the art. Examples can be taken from, for example, handbooks: Gennaro, Alfonso R.: "Remington's Pharmaceutical Sciences", Mack Publishing Company, Easton, Pa., 1990. In some embodiments, the excipient is an "uncharged" excipient, i.e., the excipient does not carry a positive "+" charge or a negative "-" charge. In some embodiments, the excipient is selected from the group consisting of fructose, glucose, mannose, sorbitol, xylose, lactose, maltose, sucrose, dextran, amylase, dextrin, cyclodextrin, soluble starch, trehalose, sorbitol, erythritol, isomalt, lactitol, maltitol, xylitol, glycerol, lactitol, hydroxyethyl starch, and water-soluble dextran.

[0073] In some embodiments, in antibody formulations, the uncharged excipient is about 1mM to about 1M, about 2mM to about 500mM, about 5mM to about 400mM, about 10mM to about 300mM, or about 20mM to about 250mM. In some embodiments, in antibody formulations (e.g., antibody formulations comprising 2 to 20mg / mL antibody), the uncharged excipient is about 5mM to about 150mM, about 10mM to about 100mM, about 20mM to about 80mM, about 30mM, about 40mM, about 50mM, about 60mM, or about 70mM. In one embodiment, in antibody formulations, the uncharged excipient is about 50mM. In some embodiments, in an antibody formulation (e.g., an antibody formulation comprising 20 to 100 mg / mL of antibody), the uncharged excipient is about 50 mM to about 800 mM, about 100 mM to about 500 mM, about 150 mM to about 400 mM, about 200 mM, about 400 mM, about 200 mM, about 300 mM, or about 250 mM. In one embodiment, in an antibody formulation, the uncharged excipient is about 250 mM.

[0074] In some embodiments, the uncharged excipient is trehalose, as represented by the following formula:

[0075] In some embodiments, in an antibody formulation, the trehalose is about 1mM to about 1M, about 2mM to about 500mM, about 5mM to about 400mM, about 10mM to about 300mM, or about 20mM to about 250mM. In some embodiments, in an antibody formulation (e.g., an antibody formulation comprising 2 to 20mg / mL of antibody), the trehalose is about 5mM to about 150mM, about 10mM to about 100mM, about 20mM to about 80mM, about 30mM, about 40mM, about 50mM, about 60mM, or about 70mM. In one embodiment, in an antibody formulation, the trehalose is about 50mM. In some embodiments, in an antibody formulation (e.g., an antibody formulation comprising 20 to 100 mg / mL of antibody), the trehalose is about 50 mM to about 800 mM, about 100 mM to about 500 mM, about 150 mM to about 400 mM, about 200 mM, about 400 mM, about 200 mM, about 300 mM, or about 250 mM. In one embodiment, in an antibody formulation, the trehalose is about 250 mM.

[0076] The antibody formulation of the present invention comprises arginine. Arginine is a conditionally non-essential amino acid, which can be represented by the following chemical formula: As used herein, arginine can include arginine in the form of free alkali, and all its salts. In certain embodiments, arginine includes its pharmaceutically acceptable salts. For example, arginine will include arginine hydrochloride. As used herein, arginine also includes all enantiomers (L-arginine and S-arginine), and any combination of enantiomers (e.g., 50% L-arginine and 50% S-arginine; 90%-100% L-arginine and 10%-0% S-arginine, etc.). In certain embodiments, the term "arginine" includes greater than 99% L-arginine and less than 1% S-arginine. In certain embodiments, the term "arginine" includes enantiomerically pure L-arginine. In certain embodiments, arginine is pharmaceutical grade arginine.

[0077] Arginine can be present in different concentrations in the antibody formulation. In some embodiments, the antibody formulation comprises greater than 50 mM arginine, greater than 75 mM arginine, greater than 100 mM arginine, greater than 125 mM arginine, greater than 130 mM arginine, greater than 150 mM arginine, greater than 175 mM arginine, or greater than 200 mM arginine.

[0078] In some embodiments, the antibody formulation comprises up to 800mM arginine, up to 600mM arginine, up to 400mM arginine, up to 200mM arginine, up to 150mM arginine, up to 130mM arginine, or up to 125mM arginine. In some embodiments, the antibody formulation comprises 50mM to 300mM, 75mM to 250mM, 100mM to 200mM, 110mM to 160mM, 120mM to 150mM, or about 125mM arginine. In some embodiments, the antibody formulation comprises 125mM arginine. In some embodiments, the antibody formulation comprises 130mM arginine. In some embodiments, arginine is added in an amount sufficient to maintain the weight molar osmotic pressure concentration of the formulation. In some embodiments, arginine is added in an amount sufficient to obtain a hypertonic solution. Applicants have discovered that, in some embodiments, increased ionic strength of the antibody formulation provides increased stability and reduced particle formation.

[0079] The antibody formulations described herein can have different viscosities. The method for measuring the viscosity of antibody formulations is known to those skilled in the art, and may include, for example, a rheometer (e.g., Anton Paar MCR301 rheometer with a 50mm, 40mm, or 20mm plate attachment). In some embodiments of the invention, the viscosity is reported under the high shear limit of 1000 shear rates per second. In certain embodiments, the antibody formulation has a viscosity less than 20 centipoise (cP), less than 18cP, less than 15cP, less than 13cP, or less than 11cP. In certain embodiments, the antibody formulation has a viscosity less than 13cP. Those skilled in the art will appreciate that viscosity depends on temperature, so unless otherwise stated, the viscosity provided herein is measured at 25°C, unless otherwise stated.

[0080] The term "injection force" is the amount of pressure (Newtons) required to pass the antibody formulation through the injection needle. The injection force is related to the amount of resistance provided by the antibody formulation when the antibody formulation is administered to a subject. The injection force will depend on the specifications of the administration injection needle, as well as the temperature. In some embodiments, the antibody formulation has an injection force of less than 15N, 12N, 10N or 8N when passing through a 27Ga thin tube PFS injection needle. In some embodiments, the antibody formulation has an injection force of less than 15N, 12N, 10N or 8N when passing through a 29Ga thin tube PFS injection needle.

[0081] The antibody preparation can have different molar osmotic pressure concentrations. The method for measuring the molar osmotic pressure concentration of the antibody preparation is known to those skilled in the art, and may include, for example, an osmometer (e.g., Advanced Instrument Inc 2020 freezing point depression osmometer). In certain embodiments, the preparation has a molar osmotic pressure concentration between 200 and 600mosm / kg, between 260 and 500mosm / kg, or between 300 and 450mosm / kg.

[0082] The antibody formulations of the present invention can have different pH levels. In some embodiments, the pH of the antibody formulation is between 4 and 7, between 4.5 and 6.5, or between 5 and 6. In some embodiments, the pH of the antibody formulation is 5.0. In some embodiments, the pH of the antibody formulation is 6.0. In some embodiments, the pH of the antibody formulation is ≥7.0. The desired pH level can be achieved using different means, including but not limited to adding appropriate buffers.

[0083] Other different components may be included in the antibody formulation. In certain embodiments, the antibody formulation may include a buffer (e.g., histidine, acetate, phosphate or citrate buffer), a surfactant (e.g., polysorbate) and / or a stabilizer (e.g., human albumin) etc. In certain embodiments, the antibody formulation may include a pharmaceutically acceptable carrier, including, for example, an ion exchanger, aluminum oxide, aluminum stearate, lecithin, serum protein (such as human serum albumin), a buffer substance (such as phosphate), sucrose, glycine, sorbic acid, potassium sorbate, a partial glyceride mixture of saturated vegetable fatty acids, water, salt or electrolyte (such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salt), silica sol, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based materials, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylate, polyethylene-polyoxypropylene-block polymers and polyethylene glycol. In certain embodiments, the antibody formulation further includes a surfactant. In some embodiments, the surfactant is selected from the group consisting of polysorbates, sodium lauryl sulfate, and nonionic surfactants.

[0084] In some embodiments, the surfactant is polysorbate 20, i.e., polyoxyethylene (20) sorbitan monolaurate, as represented by the following chemical formula: Polysorbate 20 (PS-20) is commercially available from several commercial suppliers, such as TW 20 (Oxiteno, Brazil) and 20 (Pierce, Rockford IL). Applicants have discovered that by carefully controlling the concentration of PS-20 in the antibody formulation, the antibody has increased stability and reduced particle formation when stored for extended periods of time.

[0085] In some embodiments, PS-20 is about 0.001% to about 0.02%, about 0.002% to about 0.015%, about 0.002% to about 0.01%, about 0.004% to about 0.009%, about 0.005% to about 0.008%, about 0.007%, or about 0.006% of the antibody formulation.

[0086] In some embodiments, the antibody formulation further comprises histidine. In some embodiments, the antibody formulation comprises about 1 mM to about 100 mM, about 5 mM to about 80 mM histidine, about 10 mM to about 60 mM histidine, about 15 mM to about 50 mM histidine, about 15 mM to about 30 mM histidine, or about 20 mM histidine.

[0087] In some embodiments, various components may be omitted from the antibody formulation, or may be "substantially free" of that component. As used herein, the term "substantially free" refers to an antibody formulation that contains less than 0.01%, less than 0.001%, less than 0.0005%, less than 0.0003%, or less than 0.0001% of a specified component.

[0088] In some embodiments, the antibody formulation is substantially free of sugars, i.e., the antibody formulation contains less than 0.01%, less than 0.001%, less than 0.0005%, less than 0.0003%, or less than 0.0001% sugars. As used herein, the term "sugar" refers to a class of molecules that are derivatives of polyols. Sugars are often referred to as carbohydrates and can contain varying amounts of sugar (sugar / saccharide) units, such as monosaccharides, disaccharides, and polysaccharides. In some embodiments, the formulation is substantially free of disaccharides. In some embodiments, the formulation is substantially free of reducing sugars, non-reducing sugars, or sugar alcohols. In some embodiments, the antibody formulation is substantially free of proline, glutamate, sorbitol, divalent metal ions, and / or succinate.

[0089] In some embodiments, the present invention relates to a stable aqueous antibody formulation comprising: (a) about 2 mg / mL to about 20 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; (b) about 0.002% to about 0.01% polysorbate-20; (c) about 40 mM to about 60 mM trehalose; and (d) about 110 mM to about 150 mM L-arginine. In some embodiments, the formulation further comprises about 20 mM histidine. In one embodiment, the present invention relates to a stable aqueous antibody formulation comprising: (a) about 2 mg / mL to about 20 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; (b) about 0.006% polysorbate-20; (c) about 50 mM trehalose; (d) about 130 mM L-arginine; and about 20 mM histidine.

[0090] In some embodiments, the present invention relates to a stable aqueous antibody formulation comprising: (a) about 20 mg / mL to about 100 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; (b) about 0.002% to about 0.01% polysorbate-20; (c) about 200 mM to about 300 mM trehalose; and (d) about 20 mM histidine. In one embodiment, the present invention relates to a stable aqueous antibody formulation comprising: (a) about 20 mg / mL to about 100 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; (b) about 0.006% polysorbate-20; (c) about 250 mM trehalose; and (d) about 20 mM histidine. In another embodiment, the present invention relates to a stable aqueous antibody formulation comprising: (a) about 30 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; (b) about 0.006% polysorbate-20; (c) about 250 mM trehalose; and (d) about 20 mM histidine.

[0091] The antibody formulations of the present invention are aqueous solutions. In some embodiments, the antibody formulations have not been subjected to freezing temperatures, and / or have not been frozen, i.e., they remain liquid. In some embodiments, the antibodies in the antibody formulations have not been subjected to lyophilization.

[0092] As used herein, the term "stability" generally relates to maintaining the integrity of a biologically active agent (such as a protein, peptide or another biologically active macromolecule) or minimizing degradation, denaturation, aggregation or unfolding of the biologically active agent. As used herein, "improved stability" generally means that under conditions known to cause degradation, denaturation, aggregation or unfolding, the protein of interest (e.g., such as an antibody against IL5R), peptide or another biologically active macromolecule maintains greater stability than a control protein, peptide or another biologically active macromolecule.

[0093] In some embodiments, stability refers to an antibody formulation having a low to undetectable level of particle formation. As used herein, the phrase "low to undetectable level of particle formation" refers to a sample containing less than 30 particles / mL, less than 20 particles / ml, less than 20 particles / ml, less than 15 particles / ml, less than 10 particles / ml, less than 5 particles / ml, less than 2 particles / ml, or less than 1 particle / ml, as determined by HIAC analysis or visual analysis. In some embodiments, no particles are detected in the antibody formulation, whether by HIAC analysis or by visual analysis.

[0094] In certain embodiments, stability refers to reduced antibody fragmentation. As used herein, the term "low to undetectable fragmentation level" refers to a sample measured by HPSEC equal to or greater than 80%, 85%, 90%, 95%, 98% or 99% of total protein in (for example) a single peak, or measured by reduced capillary gel electrophoresis (rCGE) in two peaks (for example, heavy chain and light chain) (or the same number of peaks as the number of subunits), representing non-degraded antibodies or non-degraded fragments, and not containing each having more than 5%, more than 4%, more than 3%, more than 2%, more than 1% or more than 0.5% of total protein Other single peaks. As used herein, the term "reduced capillary gel electrophoresis" refers to capillary gel electrophoresis under reducing conditions sufficient to reduce the disulfide bonds in the antibody.

[0095] Those skilled in the art will appreciate that, in addition to the composition of the formulation, the stability of the protein depends on other properties. For example, stability can be affected by temperature, pressure, humidity, pH and radiation external forms. Therefore, unless otherwise stated, the stability mentioned herein is considered to be measured at 5°C, one atmosphere, 50% relative humidity, pH of 6.0 and normal radiation background levels. The stability of the antibody in the antibody formulation can be determined by different means. In some embodiments, antibody stability is determined by size exclusion chromatography (SEC). SEC separates analytes (e.g., macromolecules such as proteins and antibodies) based on the hydrodynamic size, diffusion coefficient and surface properties of the analyte. Therefore, for example, SEC can separate antibodies in their native three-dimensional conformation from antibodies in different denatured states and / or degraded antibodies. In SEC, the stationary phase is usually composed of inert particles, which are packed into a dense three-dimensional matrix in a glass column or steel column. The mobile phase can be pure water, an aqueous buffer, an organic solvent, a mixture of these substances, or other solvents. The stationary phase particles have small holes and / or channels that will only allow substances below a certain size to enter. Therefore, large particles are excluded from these holes and channels, and smaller particles are removed from the flowing mobile phase. The time that particles spend immobilized in the stationary phase holes depends in part on the speed at which the particles can penetrate the holes. The removal of particles from the mobile phase flow causes them to spend longer time eluting from the column and results in separation between particles based on differences in particle size.

[0096] In some embodiments, SEC is combined with identification techniques to identify or characterize proteins or fragments thereof. Protein identification and characterization can be accomplished by different techniques, including but not limited to chromatographic techniques (e.g., high performance liquid chromatography (HPLC)), immunoassays, electrophoresis, UV / visible / infrared spectroscopy, Raman spectroscopy, surface enhanced Raman spectroscopy, mass spectrometry, gas chromatography, static light scattering (SLS), Fourier transform infrared spectroscopy (FTIR), circular dichroism (CD), urea-induced protein unfolding techniques, intrinsic tryptophan fluorescence, differential scanning calorimetry, and / or ANS protein binding.

[0097] In some embodiments, protein identification is achieved by high pressure liquid chromatography. Different instruments and equipment for performing HPLC are known to those skilled in the art. Typically HPLC involves loading a liquid solvent containing the protein of interest onto a separation column, where separation occurs. The HPLC separation column is filled with solid particles (e.g., silica, polymers, or adsorbents), and the sample mixture is separated into a variety of compounds when interacting with the column particles. HPLC separation is subject to the conditions of the liquid solvent (e.g., pressure, temperature), chemical interactions between the sample mixture and the liquid solvent (e.g., hydrophobicity, protonation, etc.), and chemical interactions between the sample mixture and the solid particles filled in the separation column (e.g., ligand affinity, ion exchange, etc.).

[0098] In some embodiments, SEC and protein identification occur in the same instrument or occur simultaneously. For example, SEC and HPLC can be combined, often referred to as SE-HPLC.

[0099] In some embodiments, the aqueous formulation comprises about 2 mg / mL to about 100 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10, wherein the formulation is stable after storage at about 40°C for at least 1 month. In some embodiments, the formulation is stable after storage at about 25°C for at least 3 months. In some embodiments, the formulation is stable after storage at about 5°C for at least 6 months. In some embodiments, the formulation is stable after storage at about 5°C for at least 12 months. In some embodiments, the formulation is stable after storage at about 5°C for at least 18 months. In some embodiments, the formulation is stable after storage at about 5°C for at least 24 months or 36 months.

[0100] The term "stable" can be relative and not absolute. Therefore, in some embodiments, when the antibody is stored at 2 ° C to 8 ° C for 6 months, if less than 20%, less than 15%, less than 10%, less than 5% or less than 2% of the antibody degradation, denaturation, aggregation or unfolding is determined by SEC HPLC, the antibody is stable. In some embodiments, when the antibody is stored at 2 ° C to 8 ° C for 12 months, if less than 20%, less than 15%, less than 10%, less than 5% or less than 2% of the antibody degradation, denaturation, aggregation or unfolding is determined by SEC HPLC, the antibody is stable. In some embodiments, when the antibody in the antibody formulation is stored at 2 ° C to 8 ° C for 18 months, if less than 20%, less than 15%, less than 10%, less than 5% or less than 2% of the antibody degradation, denaturation, aggregation or unfolding is determined by SEC HPLC, the antibody is stable. In some embodiments, the antibody is stable if less than 20%, less than 15%, less than 10%, less than 5%, or less than 2% of the antibody is degraded, denatured, aggregated, or unfolded when the antibody in the antibody formulation is stored at 2°C to 8°C for 24 months as determined by SEC HPLC.

[0101] In some embodiments, the antibody is stable if less than 20%, less than 15%, less than 10%, less than 5%, or less than 2% of the antibody is degraded, denatured, aggregated, or unfolded as determined by SEC HPLC when the antibody is stored at 23°C to 27°C for 3 months. In some embodiments, the antibody is stable if less than 20%, less than 15%, less than 10%, less than 5%, or less than 2% of the antibody is degraded, denatured, aggregated, or unfolded as determined by SEC HPLC when the antibody is stored at 23°C to 27°C for 6 months. In some embodiments, the antibody is stable if less than 20%, less than 15%, less than 10%, less than 5%, or less than 2% of the antibody is degraded, denatured, aggregated, or unfolded as determined by SEC HPLC when the antibody is stored at 23°C to 27°C for 12 months. In some embodiments, the antibody is stable if less than 20%, less than 15%, less than 10%, less than 5%, or less than 2% of the antibody is degraded, denatured, aggregated, or unfolded when stored at 23°C to 27°C for 24 months as determined by SEC HPLC.

[0102] In some embodiments, the antibody is stable if less than 6%, less than 4%, less than 3%, less than 2%, or less than 1% of the antibody is degraded, denatured, aggregated, or unfolded as determined by SEC HPLC when the antibody is stored at 40° C. In some embodiments, the antibody is stable if less than 6%, less than 4%, less than 3%, less than 2%, or less than 1% of the antibody is degraded, denatured, aggregated, or unfolded as determined by SEC HPLC when the antibody is stored at 5° C.

[0103] In some embodiments, the antibody formulations of the invention are considered stable if the antibody exhibits little loss of binding activity of the antibody (including antibody fragments thereof) of the formulation compared to a reference antibody as measured by an antibody binding assay known to those skilled in the art, such as ELISA, over a period of 8 weeks, 4 months, 6 months, 9 months, 12 months, or 24 months. In some embodiments, an antibody stored at about 40°C for at least 1 month retains at least 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, or at least about 99% of the binding capacity to IL-5 receptor polypeptide compared to a reference antibody that has not been stored. In some embodiments, an antibody stored at about 5°C for at least 6 months retains at least 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, or at least about 99% of the binding capacity to IL-5 receptor polypeptide compared to a reference antibody that has not been stored. In some embodiments, an antibody stored at about 40°C for at least 1 month retains at least 95% of the binding capacity to IL-5 receptor polypeptide compared to a reference antibody that has not been stored. In some embodiments, the antibody stored at about 5°C for at least 6 months retains at least 95% of its binding ability to IL-5 receptor polypeptide compared to a reference antibody that has not been stored.

[0104] The antibody formulation can provide low to undetectable antibody aggregation levels. As used herein, the phrase "low to undetectable aggregation levels" refers to as measured by high performance size exclusion chromatography (HPSEC) or static light scattering (SLS) technology, by the weight of the protein antibody contains no more than about 5%, no more than about 4%, no more than about 3%, no more than about 2%, no more than about 1% and no more than about 0.5% aggregation. In some embodiments, as measured by HPSEC, after being stored at about 40 ° C for at least 4 weeks, less than 2% of the antibody forms aggregates. In some embodiments, as measured by HPSEC, after being stored at about 5 ° for at least 3 months, at least 6 months, at least 9 months, at least 12 months, at least 15 months, at least 18 months, at least 24 months or at least 36 months, less than 2% of the antibody forms aggregates.

[0105] Applicants have found that the antibody formulations provided herein result in a substantial reduction in particle formation, as determined by visual inspection, microfluidic imaging (MFI), or size exclusion chromatography (SEC). In some embodiments, the formulation is substantially free of particles after storage at about 40°C for at least 1 month, as determined by visual inspection. In some embodiments, the formulation is substantially free of particles after storage at about 5° for at least 6 months, at least 9 months, at least 12 months, at least 15 months, at least 18 months, at least 24 months, or at least 36 months, as determined by visual inspection.

[0106] In some embodiments, the antibody formulation of the present invention can be used for pharmaceutical purposes. Antibodies used in pharmaceutical applications must have high purity, especially for contaminants from cell cultures, which include cell protein contaminants, cell DNA contaminants, viruses and other infectious agents. See "WHO Requirements for the use of animal cells as in vitro substrates for the production of biologicals: Requirements for Biological Substances No.50 in the production of biologicals: No. 878, Annex 1, 1998. In response to concerns about contaminants, the World Health Organization (WHO) sets limits on the levels of multiple contaminants. For example, WHO recommends that the DNA limit for each dose be less than 10 ng for protein products. Similarly, the U.S. Food and Drug Administration (FDA) sets the DNA limit to be less than or equal to 0.5 pg / mg protein. Thus, in some embodiments, the invention relates to antibodies that meet or exceed contaminant limits as defined by one or more governmental agencies, such as the U.S. Food and Drug Administration and / or the World Health Organization.

[0107] In some embodiments, the antibody formulations described herein are pharmaceutically acceptable. "Pharmaceutically acceptable" refers to antibody formulations that are suitable for use in contact with human and animal tissues within the scope of sound medical judgment without excessive toxicity or other complications commensurate with a reasonable benefit / risk ratio.

[0108] The purity of the antibody formulation can vary. In some embodiments, the therapeutic antibody of interest (e.g., anti-IL5R antibody) is greater than 90% (w / w) of the total polypeptide present in the antibody formulation. In some embodiments, the therapeutic antibody of interest (e.g., anti-IL5R) is greater than 95% (w / w), 98% (w / w), 99% (w / w), 99.5% (w / w), 99.9% (w / w) of the total polypeptide present in the antibody formulation.

[0109] The formulations provided herein can be suitable for treating a subject. As used herein, "subject" can be used interchangeably with "patient" and refers to any animal classified as a mammal, including humans and non-humans, such as but not limited to domestic and farm animals, zoo animals, sports animals, and pets. In some embodiments, the subject refers to a human.

[0110] The terms "treat" and "treatment" refer to both therapeutic treatment and prophylactic, maintenance or preventative measures, wherein the goal is to prevent or slow down (lessen) an undesirable physiological condition, disorder or disease, or to obtain a beneficial or desired clinical result. The terms "treat," "treatment," and "treating" refer to reducing or ameliorating the progression, severity and / or duration of such a disease or disorder (e.g., a disease or disorder characterized by abnormal expression and / or activity of an IL-5 polypeptide, a disease or disorder characterized by abnormal expression and / or activity of an IL-5 polypeptide or one or more of its subunits, an autoimmune disease, an inflammatory disease, a proliferative disease, or an infection), or ameliorating one or more symptoms of such a disease or disorder by administering one or more therapies (including, but not limited to, administering one or more prophylactic or therapeutic agents). In certain embodiments, such terms refer to the reduction of inflammation associated with eosinophil-mediated inflammation. In other embodiments, such terms refer to the reduction of inflammatory factors released by mast cells, or the reduction of the biological effects of such inflammatory factors. In other embodiments, such terms refer to a reduction in the growth, formation and / or increase in number of hyperproliferative cells (e.g., cancerous cells). In yet other embodiments, such terms refer to a reduction in inflammation of the airways, skin, gastrointestinal tract, or a combination thereof. In yet other embodiments, such terms refer to a reduction in symptoms associated with asthma. In yet other embodiments, such terms refer to a reduction in symptoms associated with chronic obstructive pulmonary disease (COPD).

[0111] The antibody formulations of the present invention can be administered to a subject by different means. In certain embodiments, the antibody formulations are suitable for parenteral administration, such as via inhalation (e.g., powders or sprays), transmucosal, intravenous, subcutaneous or intramuscular administration. In certain embodiments, the formulations are injectable formulations. In certain embodiments, the present invention relates to a sealed container comprising any antibody formulation as described herein.

[0112] In some aspects, the present invention relates to different pharmaceutical dosage forms. Different dosage forms can be applied to the formulations provided herein. See, for example, Pharmaceutical Dosage Form: Parenteral Medications, Volume 1, 2nd Edition. ndIn one embodiment, the pharmaceutical unit dosage form of the present invention comprises an antibody formulation in a suitable container (e.g., a vial or syringe). In one embodiment, the pharmaceutical unit dosage form of the present invention comprises an antibody formulation for intravenous, subcutaneous, or intramuscular delivery. In another embodiment, the pharmaceutical unit dosage form of the present invention comprises an antibody formulation for aerosol delivery. In a specific embodiment, the pharmaceutical unit dosage form of the present invention comprises an antibody formulation for subcutaneous delivery. In another embodiment, the pharmaceutical unit dosage form of the present invention comprises an antibody formulation for aerosol delivery. In a further embodiment, the pharmaceutical unit dosage form of the present invention comprises an antibody formulation for intranasal administration.

[0113] The antibody formulations of the present invention can be prepared as unit dosage forms by preparing vials containing aliquots of aqueous antibody formulations for single use. For example, the unit dose of each vial can contain 1 ml, 2 ml, 3 ml, 4 ml, 5 ml, 6 ml, 7 ml, 8 ml, 9 ml, 10 ml, 15 ml or 20 ml of different concentrations of antibodies specifically binding to IL5 receptors, and the concentration range of the antibody is from about 0.1 mg / ml to about 300 mg / ml. If necessary, these preparations can be adjusted to the desired concentration by adding a sterile diluent to each vial. In specific embodiments, an aqueous antibody formulation of the invention is formulated into a single-dose vial as a sterile liquid containing: about 2 mg / mL to about 20 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; about 0.002% to about 0.01% polysorbate-20; about 40 mM to about 60 mM trehalose; and about 110 mM to about 150 mM L-arginine. In another specific embodiment, the aqueous antibody formulation of the present invention is formulated into a single-dose vial as a sterile liquid containing: about 20 mg / mL to about 100 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises CDR1, CDR2, and CDR3 sequences defined by Kabat of SEQ ID NO: 5-7, and wherein the light chain variable region comprises CDR1, CDR2, and CDR3 sequences defined by Kabat of SEQ ID NO: 8-10; about 0.002% to about 0.01% polysorbate-20; and about 200 mM to about 300 mM trehalose. In one embodiment, the antibody of the present invention is provided in a 3cc USPI type borosilicate amber vial (West Pharmaceutical Services—Part No. 6800-0675) at 2 to 20 mg / ml. In one embodiment, an antibody of the invention is provided at 20 to 100 mg / ml in a 3cc USPI type borosilicate amber vial. The target fill volume is 1.2 mL.

[0114] The antibody formulation of the present invention can be prepared as a unit dosage form by preparing a pre-filled syringe containing an aliquot of the aqueous antibody formulation for a single use. For example, each unit dose of the pre-filled syringe can contain 0.1 ml, 0.2 ml, 0.3 ml, 0.4 ml, 0.5 ml, 0.6 ml, 0.7 ml, 0.8 ml, 0.9 ml, 1 ml, 2 ml, 3 ml, 4 ml, 5 ml, 6 ml, 7 ml, 8 ml, 9 ml, 10 ml, 15 ml or 20 ml of an antibody that specifically binds to an IL-5 polypeptide at a concentration ranging from about 2 mg / ml to about 100 mg / ml. In a specific embodiment, an aqueous antibody formulation of the invention is formulated into a single-dose prefilled syringe as a sterile liquid containing: about 2 mg / mL to about 20 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; about 0.002% to about 0.01% polysorbate-20; about 40 mM to about 60 mM trehalose; and about 110 mM to about 150 mM L-arginine. In a specific embodiment, an aqueous antibody formulation of the invention is formulated into a single-dose prefilled syringe as a sterile liquid containing: about 20 mg / mL to about 100 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; about 0.002% to about 0.01% polysorbate-20; and about 200 mM to about 300 mM trehalose.

[0115] The amount of different dosages can be administered in a single use. For example, in some embodiments, 0.1 mg, 0.2 mg, 0.3 mg, 0.4 mg, 0.5 mg, 0.6 mg, 0.7 mg, 0.8 mg, 0.9 mg, 1.0 mg, 1.1 mg, 1.2 mg, 1.3 mg, 1.4 mg, 1.5 mg, 1.6 mg, 1.7 mg, 1.8 mg, 1.9 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 12 mg, 14 mg, 16 mg, 18 mg, 20 mg, 30 mg, 40 mg, 50 mg, 70 mg or 100 mg of the antibody can be administered in a single dose.

[0116] Different types of syringes can be used. Can be before giving the experimenter immediately, for example, before giving the experimenter less than 1 week, 1 day, 6 hours, 3 hours, 2 hours, 1 hour, 30 minutes, 20 minutes or 10 minutes, syringe is filled with antibody preparations. In certain embodiments, can be at retail outlets, or by the facility that the experimenter is treated, syringe is filled with antibody preparations. In certain embodiments, before giving the experimenter greater than 1 day, 2 days, 4 days, 1 week, 2 weeks, 1 month, 2 months, 3 months, 6 months, 12 months, 18 months, 24 months, 3 years or 4 years, syringe is pre-filled, for example, syringe is filled with antibody preparations. In certain embodiments, this pre-filled syringe comprises injection needle, for example 27G conventional thin tube injection needle, 27G thin tube injection needle, 29G conventional thin tube injection needle or 29G thin tube injection needle. In certain embodiments, this pre-filled syringe comprises 29G thin tube injection needle.

[0117] In some embodiments, any syringe suitable for administration to a desired subject may be used. In some embodiments, the syringe is a plastic syringe or a glass syringe. In some embodiments, the syringe is made of a material that is substantially free of tungsten. In some embodiments, the syringe is coated with silicone. In some embodiments, the prefilled syringe includes a plunger having a fluoropolymer resin disc. Examples of syringes may include, but are not limited to, a 1 ml long Hypak TM for Biotech (Becton Dickinson), with 1 mL long Becton Dickinson Hypak plunger stopper 4023 Flurotec Daikyo Si1000 (Cat. No. 47271919), C3Pin (Lot No. E912701); 0.8 mg silicone oil in Hypak TM for Biotech (Becton Dickinson); and CZ syringe (West Corporation, catalog number 19550807).

[0118] The aqueous antibody formulations of the present invention can be sterilized by various sterilization methods, including sterile filtration, irradiation, etc. In a specific embodiment, the diafiltered antibody formulation is sterile filtered with a pre-sterilized 0.2 micron filter. The sterilized aqueous antibody formulations of the present invention can be administered to a subject to prevent, treat and / or manage an immune response, such as an inflammatory response.

[0119] In some embodiments, the pre-filled syringe comprises: (a) about 2 mg / mL to about 20 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; and (b) about 0.002% to about 0.01% polysorbate-20. In some embodiments, the pre-filled syringe further comprises: (c) about 40 mM to about 60 mM trehalose, and (d) about 110 mM to about 150 mM L-arginine. In some embodiments, the pre-filled syringe comprises: (a) about 20 mg / mL to about 100 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; and (b) about 0.002% to about 0.01% polysorbate-20. In some embodiments, the pre-filled syringe further comprises: (c) about 200 mM to about 300 mM trehalose.

[0120] In some embodiments, the invention relates to a kit comprising any of the antibody formulations described herein, the containers described herein, the unit dosage forms described herein, or the pre-filled syringes described herein.

[0121] In some embodiments, the present invention may also be directed to a method for preparing a stable aqueous antibody formulation comprising an antibody, the method comprising: (a) purifying the antibody to about 1 mg / mL to about 400 mg / mL, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; and (b) placing the isolated antibody in a stabilizing formulation to form the stable aqueous antibody formulation, wherein the resulting stable aqueous antibody formulation comprises: (i) about 2 mg / mL to about 100 mg / mL of the antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10. NO: 8-10 of the Kabat-defined CDR1, CDR2 and CDR3 sequences; and (ii) about 0.002% to about 0.01% polysorbate-20. In some embodiments, the invention relates to a method for preparing a stable aqueous antibody formulation, the method comprising: (a) purifying an antibody to about 1 mg / mL to about 400 mg / mL, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; (b) diluting the antibody to about 2 mg / mL to about 20 mg / mL of the antibody in a solution comprising: (i) about 0.002% to about 0.01% polysorbate-20; (ii) about 40 mM to about 60 mM trehalose; and (iii) about 110 mM to about 150 mM L-arginine, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10. The light chain variable region comprises the Kabat-defined CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 8-10.In some embodiments, the invention relates to a method for preparing a stable aqueous antibody formulation, the method comprising: (a) purifying an antibody to about 1 mg / mL to about 400 mg / mL, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; (b) diluting the antibody to about 20 mg / mL to about 100 mg / mL of the antibody in a solution comprising: (i) about 0.002% to about 0.01% polysorbate-20; (ii) about 200 mM to about 300 mM trehalose; and (iii) about 20 mM histidine, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10. The light chain variable region comprises the Kabat-defined CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 8-10.

[0122] Although many aspects of the invention relate to aqueous formulations, it should be noted that for equivalent purposes, the antibodies or antibody formulations of the invention can be lyophilized, if desired. Thus, the invention encompasses lyophilized formulations of the invention, or lyophilized antibodies that are later rehydrated into an aqueous form. In some embodiments, the present invention relates to a method for preparing a rehydrated antibody formulation, the rehydrated antibody formulation comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10, the method comprising: (a) purifying the antibody from a cell culture; (b) lyophilizing the isolated antibody; (c) adding the lyophilized antibody to an aqueous solution to form a rehydrated antibody formulation, wherein the rehydrated antibody formulation comprises: (i) about 2 mg / mL to about 100 mg / mL of the antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; NO: 8-10 of the Kabat-defined CDR1, CDR2 and CDR3 sequences; and (ii) about 0.002% to about 0.01% polysorbate-20.

[0123] In some embodiments, the inventors have found that anti-IL5R antibody formulations with increased glutathione S-transferase (GST) result in reduced (e.g., undetectable) particle formation. Removal of particles is important for avoiding potential immunogenicity and limiting the impact on product quality. In some embodiments, the GST concentration is reduced by affinity chromatography. In some embodiments, the GST concentration is reduced by using a protein A column. In some embodiments, the protein A column is MabSelect Sure (GE Healthcare Life Sciences). In some embodiments, the GST concentration is reduced by using mixed mode chromatography. In some embodiments, the mixed mode column is Capto TM Adhere (GE Healthcare Life Sciences).

[0124] In some embodiments, the present invention relates to an antibody formulation comprising an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises CDR1, CDR2 and CDR3 sequences defined by Kabat of SEQ ID NO: 5-7, and wherein the light chain variable region comprises CDR1, CDR2 and CDR3 sequences defined by Kabat of SEQ ID NO: 8-10, wherein the antibody formulation is substantially free of particles. In some embodiments, the term "substantially free of particles" refers to the absence of visible particles when observed under a light box. In some embodiments, the term "substantially free of particles" is synonymous with the phrase "low to undetectable particle formation levels" as previously described. In some embodiments, substantially free of particles refers to a sample containing less than 30 particles / mL, less than 20 particles / ml, less than 20 particles / ml, less than 15 particles / ml, less than 10 particles / ml, less than 5 particles / ml, less than 2 particles / ml or less than 1 particle / ml, wherein the particles are greater than 25 μm and the particle count is measured by HIAC analysis or visual analysis. In some embodiments, substantially free of particles refers to a sample containing 1 to 50 particles / mL, 2 to 40 particles / ml, 3-30 particles / ml, 4 to 25 particles / ml, or 5 to 20 particles / ml, wherein the particles are larger than 25 μm and the particle count is measured by HIAC analysis or visual analysis. In some embodiments, the term "visible particles" refers to particles larger than 25 μm.

[0125] In some embodiments, substantially free of particles refers to a sample containing 1 to 200 particles / mL, 10 to 150 particles / ml, 30-100 particles / ml, or 40 to 80 particles / ml, wherein the particles are greater than 5 μm and the particle count is measured by HIAC analysis or visual analysis. In some embodiments, the term "visible particles" refers to particles greater than 5 μm. In some embodiments, no particles are detected in the antibody formulation, whether by HIAC analysis or by visual analysis.

[0126] In some embodiments, the present invention relates to an antibody formulation comprising an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10, wherein the antibody formulation is substantially free of glutathione S-transferase (GST). Unless otherwise specified, the term "substantially free of glutathione S-transferase" or "substantially free of GST" will encompass compositions lacking active GST (but may contain inactive GST) and compositions without GST protein (whether in active or inactive form). In some embodiments, the present invention relates to an antibody formulation comprising an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10, wherein the antibody formulation is substantially free of active GST. The term "active GST" refers to a GST that is capable of catalyzing the formation of a thiol group of glutathione (GSH) to an electrophilic compound such as 1-chloro-2,4-dinitrobenzene (CDNB) to form a GS-DNB conjugate. GST or glutathione S-transferase refers to a family of enzymes that are capable of catalyzing a variety of reactions, but primarily the conjugation of reduced glutathione to electrophilic centers (e.g., aromatic compounds, double bonds, C-Cl x The GST monomers are generally in the range of 22-29 kDa, and they can appear as dimers, trimers and also heterodimers (with other proteins). In some embodiments, the term GST refers to a protein that can catalyze the formation of the thiol group of glutathione (GSH) to 1-chloro-2,4-dinitrobenzene (CDNB) to form a GS-DNB conjugate.

[0127] In some embodiments, the invention relates to an antibody formulation comprising an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10, wherein the antibody formulation is substantially free of particles when stored at 38°C-42°C for at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 8 months, at least 10 months, at least 12 months, or at least 18 months. In some embodiments, the invention relates to an antibody formulation comprising an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10, wherein the antibody formulation is substantially free of particles when stored at 2°C-6°C for at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 8 months, at least 10 months, at least 12 months, at least 18 months, at least 24 months, at least 30 months, at least 36 months, or at least 48 months.

[0128] In some embodiments, the antibody formulation is substantially free of GST. In some embodiments, the term "substantially free of GST" refers to an antibody formulation having a GST activity of less than about 0.5 units / mg antibody, less than about 0.3 units / mg antibody, less than about 0.1 units / mg antibody, less than about 0.08 units / mg antibody, less than about 0.05 units / mg antibody, less than about 0.03 units / mg antibody, less than about 0.01 units / mg antibody, less than about 0.005 units / mg antibody, less than about 0.001 units / mg antibody, less than about 5×10 -3 units / mg antibody, less than about 1×10 -4 units / mg antibody, less than about 1×10 -5 units / mg antibody, or less than about 1 1×10 -6 In some embodiments, the term "substantially free" means that the level of GST is undetectable using common GST detection techniques.

[0129] Different methods for determining GST activity are known to those skilled in the art. In some embodiments, GST activity is determined using a glutathione (GSH / GSSG / total) fluorescence assay kit (BioVision, San Francisco CA).

[0130] In some embodiments, the present invention relates to a method for purifying an antibody comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 8-10, the method comprising: (i) obtaining a cell culture comprising the antibody; (ii) performing affinity chromatography on the antibody; (iv) performing cation exchange on the antibody; (v) performing mixed mode chromatography on the antibody. In some embodiments, the present invention relates to a method for purifying an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises CDR1, CDR2 and CDR3 sequences defined by Kabat of SEQ ID NO: 5-7, and wherein the light chain variable region comprises CDR1, CDR2 and CDR3 sequences defined by Kabat of SEQ ID NO: 8-10, the method comprising: (i) obtaining a cell culture comprising the antibody; (ii) binding the antibody to a protein A column; (iii) eluting the antibody from the protein A column; (iv) performing cation exchange on the antibody; (v) performing mixed mode chromatography on the antibody. In some embodiments, the method for purifying antibodies further comprises a virus inactivation process. In some embodiments, the virus inactivation step is performed by reducing the pH to less than 4.0. In some embodiments, the method further comprises a diafiltration process. In some embodiments, the method further comprises a filtration process. In some embodiments, the filtration process is sufficient to remove active viral particles.

[0131] In some embodiments, the invention relates to a method of treating a patient. In some embodiments, the invention comprises administering to a subject in need thereof an antibody formulation described herein, a container described herein, a unit dosage form described herein, or a prefilled syringe described herein.

[0132] In some embodiments, the invention is suitable for treating a pulmonary disease or condition by administering an antibody formulation described herein. In some embodiments, the invention relates to a method of treating a patient suffering from an eosinophilic disease or condition by administering an antibody formulation described herein. In some embodiments, the invention relates to a method of treating a pulmonary disease or condition in a subject comprising administering an antibody formulation described herein. In some embodiments, the invention relates to a method of treating an eosinophilic disease or condition in a subject comprising administering an antibody formulation described herein. In some embodiments, the present invention relates to the treatment of the following pulmonary diseases or conditions in a subject: for example, asthma, COPD, eosinophilic asthma, combined eosinophilic and neutrophilic asthma, aspirin-sensitive asthma, allergic bronchopulmonary aspergillosis, acute and chronic eosinophilic bronchitis, acute and chronic eosinophilic pneumonia, Churg-Strauss syndrome, hypereosinophilic syndrome, drug, irritant and radiation-induced pulmonary eosinophilia, infection-induced pulmonary eosinophilia (fungal, tuberculosis, parasites), autoimmune-related pulmonary eosinophilia, eosinophilic esophagitis, or Crohn's disease or a combination thereof, the method comprising administering an antibody formulation described herein. In some embodiments, the present invention relates to the treatment of asthma in a subject, the method comprising administering an antibody formulation described herein. In some embodiments, the present invention relates to the treatment of COPD in a subject, the method comprising administering an antibody formulation described herein.

[0133] In some embodiments, a therapeutically effective amount of an antibody formulation described herein is administered to treat a condition. As used herein, the term "therapeutically effective amount" refers to an amount of a therapy (e.g., an antibody that immunospecifically binds to an IL-5 receptor polypeptide) that is sufficient to reduce the severity of a disease or condition (e.g., a disease or condition characterized by abnormal expression and / or activity of an IL-5 polypeptide, a disease or condition characterized by abnormal expression and / or activity of an IL-5 polypeptide or one or more of its subunits, an autoimmune disease, an inflammatory disease, a proliferative disease, or an infection (preferably a respiratory infection) or one or more symptoms thereof), reduce the duration of a respiratory condition, improve one or more symptoms of such a disease or condition, prevent the progression of such a disease or condition, cause regression of such a disease or condition, or enhance or improve the therapeutic effect of another therapy. In some embodiments, a therapeutically effective amount cannot be specified in advance and can be determined by a caregiver (e.g., by a physician or other healthcare provider) using various methods (e.g., dosage adjustments). An appropriate therapeutically effective amount can also be determined by routine experimentation using, for example, an animal model.

[0134] The terms "therapies" and "therapy" may refer to any regimen, method and / or agent that can be used to prevent, treat, manage or ameliorate a disease or condition (e.g., a disease or condition characterized by aberrant expression and / or activity of an IL-5 polypeptide, a disease or condition characterized by aberrant expression and / or activity of an IL-5 polypeptide or one or more subunits thereof, an autoimmune disease, an inflammatory disease, a proliferative disease or an infection (preferably a respiratory tract infection) or one or more symptoms thereof). In certain embodiments, the terms "therapies" and "therapy" refer to biological therapies, supportive therapies and / or other therapies known to skilled medical personnel that can be used to treat, manage, prevent or ameliorate such a disease or condition or one or more symptoms.

[0135] As used herein, the term "therapeutic regimen" refers to a regimen for scheduling the dosage and timing of administration of one or more therapies (eg, therapeutic agents) to have a therapeutic effect.

[0136] The route of administration of the antibody formulations of the present invention can be, for example, via oral, parenteral, inhalation or topical modes of administration. The term parenteral as used herein includes, for example, intravenous, intraarterial, intraperitoneal, intramuscular, subcutaneous, rectal or vaginal administration. In some embodiments, the antibody is an anti-IL5R antibody and the route of administration is intramuscular injection. Although all of these forms of administration are clearly considered to be within the scope of the present invention, in some embodiments, the antibody formulation is suitable for administration via injection, particularly via intravenous or intraarterial injection or infusion.

[0137] In some embodiments, the compositions and methods of the invention enable manufacturers to produce antibody formulations suitable for administration to humans in a more efficient manner by reducing costs, reducing process steps, reducing the chance of errors, reducing the chance of introduction of unsafe or inappropriate additives, reducing waste, increasing storage time, etc. The present invention also relates to the following embodiments: 1. A stable aqueous antibody formulation comprising: a. about 2 mg / mL to about 100 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; and b. From about 0.002% to about 0.01% polysorbate-20. 2. The antibody formulation of embodiment 1, further comprising an uncharged excipient. 3. The antibody formulation of embodiment 2, wherein the uncharged excipient is trehalose. 4. The antibody formulation of any one of embodiments 1 to 3, comprising about 2 to about 20 mg / ml of the antibody. 5. The antibody formulation of any one of embodiments 1 to 3, comprising about 20 to about 100 mg / ml of the antibody. 6. The antibody formulation of embodiment 4, wherein the concentration of the uncharged excipient is about 20 mM to about 80 mM. 7. The antibody formulation of embodiment 5, wherein the concentration of the uncharged excipient is about 200 mM to about 400 mM. 8. The antibody formulation of embodiment 4, further comprising arginine. 9. The antibody formulation of embodiment 8, wherein the arginine is L-arginine. 10. The antibody formulation of embodiment 8, comprising about 100 mM to about 200 mM L-arginine. 11. The antibody formulation of embodiment 8, comprising about 120 mM to about 140 mM L-arginine, and about 40 mM to about 60 mM uncharged excipient. 12. The antibody formulation of any one of embodiments 1 to 11, further comprising histidine. 13. The antibody formulation of embodiment 12, wherein the concentration of the histidine is about 15 mM to about 30 mM. 14. The antibody formulation of any one of embodiments 1 to 13, wherein the antibody is not lyophilized. 15. A stable aqueous antibody formulation comprising about 2 mg / mL to about 100 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10, wherein the formulation is stable after storage at about 40°C for at least 1 month. 16. A stable aqueous antibody formulation comprising: a. about 2 mg / mL to about 20 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; b. about 0.002% to about 0.01% polysorbate 20; c. about 40 mM to about 60 mM trehalose; d. about 110 mM to about 150 mM L-arginine; and e. About 15 mM to about 30 mM histidine. 17. A stable aqueous antibody formulation comprising: a. about 20 mg / mL to about 100 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; b. about 0.002% to about 0.01% polysorbate 20; c. about 200 mM to about 300 mM trehalose; and d. About 15 mM to about 30 mM histidine. 18. A stable aqueous antibody formulation comprising: a. about 2 mg / mL to about 20 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; b. about 0.006% polysorbate 20; c. about 50 mM trehalose; d. about 130 mM L-arginine; and e. About 20 mM histidine. 19. A stable aqueous antibody formulation comprising: a. about 20 mg / mL to about 100 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; b. about 0.006% polysorbate 20; c. about 250 mM trehalose; and d. About 20 mM histidine. 20. A stable aqueous antibody formulation comprising: a. about 30 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; b. about 0.006% polysorbate 20; c. about 250 mM trehalose; and d. About 20 mM histidine. 21. The antibody formulation of any one of embodiments 1 to 20, wherein the formulation is stable after storage at about 25°C for at least 3 months. 22. The antibody formulation of any one of embodiments 1 to 21, wherein the formulation is stable after storage at about 5°C for at least 18 months. 23. The antibody formulation of any one of embodiments 1 to 22, wherein the antibody stored at about 40°C for at least 1 month retains at least 80% of its binding ability to the IL-5R polypeptide compared to a reference antibody that has not been stored. 24. The antibody formulation of any one of embodiments 1 to 23, wherein the antibody stored at about 5°C for at least 6 months retains at least 80% of its binding ability to the IL-5R polypeptide compared to a reference antibody that has not been stored. 25. The antibody formulation of any one of embodiments 1 to 24, wherein the antibody stored at about 40°C for at least 1 month retains at least 95% of its binding ability to the IL-5R polypeptide compared to a reference antibody that has not been stored. 26. The antibody formulation of any one of embodiments 1 to 25, wherein the antibody stored at about 5°C for at least 6 months retains at least 95% of its binding ability to the IL-5R polypeptide compared to a reference antibody that has not been stored. 27. The antibody formulation of any one of embodiments 1 to 26, wherein less than 2% of the antibody forms aggregates after storage at about 40°C for at least 1 month as determined by HPSEC. 28. The antibody formulation of any one of embodiments 1 to 27, wherein less than 2% of the antibody forms aggregates after storage at about 5°C for at least 12 months as determined by HPSEC. 29. The antibody formulation of any one of embodiments 1 to 28, wherein the formulation is substantially free of particles after storage at about 40°C for at least 1 month as determined by visual inspection. 30. The antibody formulation of any one of embodiments 1 to 29, wherein the formulation is substantially free of particles after storage at about 5°C for at least 12 months as determined by visual inspection. 31. The antibody formulation of any one of embodiments 1 to 30, wherein the formulation is an injectable formulation. 32. The antibody formulation of any one of embodiments 1 to 31, wherein the formulation is suitable for intravenous, subcutaneous or intramuscular administration. 33. A sealed container containing the antibody formulation of any one of embodiments 1 to 32. 34. A pharmaceutical unit dosage form suitable for parenteral administration to humans, comprising the antibody formulation of any one of embodiments 1 to 32 in a suitable container. 35. The pharmaceutical unit dosage form of embodiment 34, wherein the antibody formulation is administered intravenously, subcutaneously or intramuscularly. 36. The pharmaceutical unit dosage form of embodiment 34 or 35, wherein the suitable container is a prefilled syringe. 37. The pharmaceutical unit dosage form of embodiment 36, wherein the prefilled syringe comprises an injection needle. 38. The pharmaceutical unit dosage form of embodiment 37, wherein the injection needle is a 29G thin-barrel injection needle. 39. The pharmaceutical unit dosage form of any one of embodiments 36 to 38, wherein the prefilled syringe is a plastic syringe or a glass syringe. 40. The pharmaceutical unit dosage form of any one of embodiments 36 to 39, wherein the prefilled syringe is made of a material that is substantially free of tungsten. 41. The pharmaceutical unit dosage form of any one of embodiments 361 to 40, wherein the prefilled syringe is coated with silicone. 42. The pharmaceutical unit dosage form of any one of embodiments 36 to 41, wherein the prefilled syringe comprises a plunger having a fluoropolymer resin disk. 43. A prefilled syringe comprising: a. about 2 mg / mL to about 20 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; and b. From about 0.002% to about 0.01% polysorbate-20. 44. The prefilled syringe of embodiment 38, further comprising: c. about 40 mM to about 60 mM trehalose; and d. About 110 mM to about 150 mM L-arginine. 45. A prefilled syringe comprising: a. about 20 mg / mL to about 100 mg / mL of an antibody, wherein the antibody comprises the amino acid sequence of SEQ ID NO: 1, b. From about 0.002% to about 0.01% polysorbate-20. 46. ​​The prefilled syringe of embodiment 40, further comprising: c. about 200 mM to about 300 mM trehalose. 47. A prefilled syringe comprising: a. about 2 mg / mL to about 20 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; and b. About 0.006% polysorbate 20. 48. The prefilled syringe of embodiment 47, further comprising: c. about 50 mM trehalose; and d. About 130 mM L-arginine. 49. A prefilled syringe comprising: a. about 20 mg / mL to about 100 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; b. About 0.006% polysorbate 20. 50. The prefilled syringe of embodiment 49, further comprising: c. About 250 mM trehalose. 51. A prefilled syringe comprising: a. about 30 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; and b. About 0.006% polysorbate 20. 52. The prefilled syringe of embodiment 51, further comprising: c. About 250 mM trehalose. 53. A kit comprising the formulation of any one of embodiments 1 to 32, the container of embodiment 33, the unit dosage form of any one of embodiments 34 to 42, or the prefilled syringe of any one of embodiments 43 to 52. 54. A method for preparing a stable aqueous antibody formulation, the method comprising: a. purifying the antibody to about 1 mg / mL to about 400 mg / mL, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; b. placing the isolated antibody in a stabilizing formulation to form the stable aqueous antibody formulation, wherein the resulting stable aqueous antibody formulation comprises: i. about 2 mg / mL to about 100 mg / mL of the antibody; and ii. from about 0.002% to about 0.01% polysorbate-20. 55. A method for preparing a stable aqueous antibody formulation, the method comprising: a. purifying the antibody to about 1 mg / mL to about 400 mg / mL, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; b. diluting the antibody to about 2 mg / mL to about 20 mg / mL of the antibody in a solution comprising: i. about 0.002% to about 0.01% polysorbate 20; ii. about 40 mM to about 60 mM trehalose; and iii. about 110 mM to about 150 mM L-arginine. 56. A method for preparing a stable aqueous antibody formulation, the method comprising: a. purifying the antibody to about 1 mg / mL to about 400 mg / mL, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; b. diluting the antibody to about 20 mg / mL to about 100 mg / mL of the antibody in a solution comprising: i. from about 0.002% to about 0.01% polysorbate 20; and ii. about 200 mM to about 300 mM trehalose. 57. A method of preparing a rehydrated antibody formulation comprising an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10, the method comprising: a. purifying the antibody from cell culture; b. lyophilizing the separated antibody; c. adding the lyophilized antibody to an aqueous solution to form a reconstituted antibody formulation, wherein the reconstituted antibody formulation comprises: i. about 2 mg / mL to about 100 mg / mL of the antibody; and ii. from about 0.002% to about 0.01% polysorbate-20. 58. An antibody formulation comprising an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10, wherein the antibody formulation is substantially free of particles. 59. An antibody formulation comprising an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10, wherein the antibody formulation is substantially free of active glutathione S-transferase (GST). 60. The antibody formulation of embodiment 59, wherein the antibody sequence is substantially free of GST. 61. The antibody formulation of embodiment 58, wherein the antibody formulation is substantially free of particles when stored at 38°C-42°C for at least 1 month. 62. The antibody formulation of embodiment 58, wherein the antibody formulation is substantially free of particles when stored at 2°C-6°C for at least 6 months. 63. A method for purifying an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10, the method comprising: a. obtaining a cell culture containing the antibody; b. performing affinity chromatography on the antibody; c. performing a cation exchange on the antibody; d. Perform mixed mode chromatography on the antibody. 64. The method of embodiment 63 further comprises a virus inactivation process. 65. The method of embodiment 63 further comprises a diafiltration process. 66. A method of treating a pulmonary disease or condition in a subject, the method comprising administering a therapeutically effective amount of the antibody formulation of any one of embodiments 1 to 32, the container of embodiment 33, the unit dosage form of any one of embodiments 34 to 42, or the pre-filled syringe of any one of embodiments 43 to 52. 67. The method of embodiment 49, wherein the pulmonary disease or disorder is an eosinophilic disease or disorder. 68. The method of embodiment 49, wherein the pulmonary disease or disorder is asthma, COPD, eosinophilic asthma, combined eosinophilic and neutrophilic asthma, aspirin-sensitive asthma, allergic bronchopulmonary aspergillosis, acute and chronic eosinophilic bronchitis, acute and chronic eosinophilic pneumonia, Churg-Strauss syndrome, hypereosinophilic syndrome, drug-, irritant- and radiation-induced pulmonary eosinophilia, infection-induced pulmonary eosinophilia (fungal, tuberculosis, parasitic), autoimmune-associated pulmonary eosinophilia, eosinophilic esophagitis, Crohn's disease, or a combination thereof. 69. The method of embodiment 49, wherein the pulmonary disease or disorder is asthma. 70. The method of embodiment 49, wherein the pulmonary disease or disorder is chronic obstructive pulmonary disease. Examples Example 1

[0138] Formulation studies were performed to develop an anti-IL5R antibody formulation suitable for delivery via subcutaneous delivery from a prefilled syringe (or via a ready-to-use configuration) at a dose of 2-100 mg. Specifically, formulations were developed at two separate antibody concentrations, a 2-20 mg / mL formulation and a 20-100 mg / mL formulation. 1. Materials and Methods a. Anti-IL5R Sources and Formulation Preparation

[0139] Multiple batches of anti-IL5R were used in these studies. All of these batches were produced by MedImmune at different scales and delivered after diafiltration and concentration to a concentration of approximately 130 g / L in 20 mM histidine / histidine hydrochloride at pH 6. Some batches also had 250 mM trehalose in the diafiltration buffer.

[0140] Anti-IL5R was also formulated by spiking in excipient addition buffer (EAB) to achieve an anti-IL5R concentration of 100 g / L and appropriate concentrations of buffer and excipient substances. Lower concentration drug products were prepared from the 100 g / L formulation. 2. Accelerated stress method a.Storage at increased temperature

[0141] The vials and syringes were stored in a controlled stability chamber to maintain a constant temperature during storage. The chambers were maintained at 2°C-8°C, 23°C-27°C / 60% RH, or 38°C-42°C / 75% RH, but their midpoint temperatures of 5°C, 25°C, or 40°C will be referred to below. Unless otherwise noted, the vials were stored upright and the pre-filled syringes were stored with the tips down. b. Freeze-thaw cycle

[0142] Both controlled and uncontrolled freeze-thaw cycles were used in these studies. Uncontrolled freeze-thaw was accomplished by freezing the vials in a -40°C chamber and thawing at room temperature. c. Transport and shaking

[0143] A variety of methods were used to investigate the effects of shipping on anti-IL5R. The vials were shaken benchtop at 150 rpm for 24 hours. Actual shipping was simulated by shipping the product to an off-site location. The product underwent two round trips and was shipped by ground and air within 4 days. A combination of freezer bags and ice packs was used to maintain product temperature at 2°C-8°C and monitored by sensors that indicated temperatures below 0°C or above 9°C.

[0144] For the screening study, a vibration table (transport simulator) was used to simulate transport. The product was subjected to the same "air" and "ground" transport modes as during round trip transport. The process lasted 12 hours and the temperature was again controlled at 2°C-8°C with ice packs; no sensors were used. The horizontal orientation was chosen as the worst orientation during actual or simulated transport because of the possibility of air bubble travel and the possibility of drug contact with the entire barrel, needle and stopper. 3. Experimental Methods a. Methods that follow or are derived from SOP

[0145] Visual inspection was performed by comparison with particle and opalescent standards. Aggregation and fragmentation were monitored by SE-HPLC. For anti-IL5R concentrations below 10 g / L, larger volume injections were used to achieve similar total protein amounts per injection. Some samples were also used for cIEF measurements and RP-HPLC to monitor fragmentation. b. Protein concentration

[0146] Protein concentration was measured by serial gravimetric dilutions to approximately 0.5 g / L and measuring absorbance at 280 nm. Concentrations were calculated from the attenuation coefficient and the dilution factor, and the effect of density on gravimetric dilution was corrected for initial concentrations above 50 g / L. c. Sub-visible particle counting

[0147] Sub-visible particle counts were produced using MFI and HIAC. For MFI, 0.9 mL of solution was run neat after running daylight illumination with water. The first 0.2 mL was used to purge the system and was not included in the analysis. A filter with an aspect ratio of <0.85 was used to remove spherical bubbles or silicone oil droplets. For HIAC, solutions with concentrations >5 g / L were diluted to approximately 5 g / L and the diluted samples were run neat. Dilutions were performed in a laminar flow clean bench with 20 mM histidine / histidine hydrochloride pH 6 buffer and filtered immediately prior to use. The samples were degassed under vacuum for at least 30 minutes prior to testing. The average of the three runs was multiplied by the dilution factor to obtain the final result. Silicone oil droplets were not distinguished from protein particles by HIAC. 4. Data and Discussion a. Polysorbate 20 concentration screening

[0148] The first goal is to optimize the PS-20 concentration in the aqueous antibody formulation. Polysorbate is included in the solution to prevent protein denaturation and aggregation at the interface, and the concentration required in the expected liquid product and the lyophilized product is different. The main interfacial stress occurs during freeze-thaw and transportation, so the experimental plan focuses on mimicking these. Previous experiments have shown that 0.02% polysorbate 20 is sufficient to completely avoid these stresses (data not shown). As a verification, these stresses are continuously combined in the order expected for clinical production, and the incubation period is extended after the stress to enhance the growth of any protein particles. First choice, the drug undergoes three uncontrolled freeze-thaw cycles, and the material is filtered and filled into a vial. It is then shaken on a shaking table and incubated at 40°C for one week, and then tested by SE-HPLC and MFI. The test conditions are the limits of the concentration range 2 and 100g / L, in 240mM trehalose, 20mM histidine / histidine hydrochloride (pH 6), and the PS-20 with a level varying from 0 to 0.03% w / v. The results are shown in Figure 2 , 3 and 4 in.

[0149] The monomer fraction data show that the 2 g / L solution remains pure regardless of the polysorbate level, but small amounts of polysorbate-20 (less than about 0.005%) cause a small amount of aggregation at 100 g / L, but these results themselves do not indicate "edge of failure". The stress utilized is severe and the degradation level is minimal, so any PS-20 level tested will be sufficient from a SEC aggregation perspective. At 2 g / L, the sub-visible particle counts are quite high and are not controlled by polysorbate-20 within the tested range. At 100 g / L, the high sub-visible particle counts are controlled by the presence of 0.003% or more PS-20. Overall, the data indicate that the PS-20 level should be maintained at or above 0.003%.

[0150] For low concentration solutions, alternative methods of controlling subvisible particles are needed, as discussed below. b. pH screening

[0151] The effect of solution pH was studied in 2, 20 and 100 g / L solutions, with pH values ​​ranging from 5 to 7.5. The remainder of the formulation was constant and included 240 mM trehalose, 20 mM histidine / histidine hydrochloride and 0.02% PS-20. Solutions were prepared and stored at 40°C for at least one month before testing. For all samples, monomer loss as determined by SE-HPLC, sub-visible particles as determined by MFI, and visible particles found by visual inspection were evaluated. Additional testing was performed on the 100 g / L sample, including RP-HPLC and cIEF. The results are provided in Tables 5 and 6.

[0152] In the pH range of 5.5-6.5, aggregation and fragmentation are minimized. The results obtained from cIEF are consistent with the reference standards at pH 7.0 and above. The sub-visible particle counts are neither low nor show any pattern with the solution pH. At pH 5.5-6.5, the visible particle fraction is higher. Even though these scores are high, these samples are close to the light when higher particle counts are routinely seen upon inspection. It is possible that the source material also causes high particle levels because the HCP level of this material is reduced due to further purification on protein A. This study shows that from the perspective of particle formation, the optimal pH will be pH 5 or pH ≥ 7. However, since the visible particle fraction is understood to be overestimated here, the pH does not change from pH 6.0 due to this study. c. Shipping impact

[0153] The formulation needed to be robust to withstand shipping, so it was tested in pre-filled syringes at different protein and PS-20 concentrations. The formulations varied from 2–100 g / L anti-IL5R and 0–0.05% PS-20, with other conditions held constant, i.e., 240 mM trehalose, 20 mM histidine / histidine HCl, pH 6. Several other conditions were tested at 2 g / L, including glycine, calcium chloride, pH 5.5, pH 6.5, 0.02% polysorbate-80. None of these conditions showed improvement over the pH 6 trehalose formulation with polysorbate-20, and so are not discussed further below.

[0154] 1 mL of sample was filled into a flatbed PFS with 0.4 mg of silicone oil. The samples were shipped, stored at 5°C, 25°C, and 40°C, and tested within two months by visual inspection, MFI, and HIAC. The results are presented in Figure 7middle.

[0155] This figure shows the sub-visible particle counts from MFI after storage at 25°C for 1 month. Sub-visible particle counts from HIAC or after storage at other temperatures show similar trends to the data sets shown. Visual inspection did not indicate high visible particle counts for any sample, except for samples containing calcium chloride. High protein concentration solutions (≥20 g / L) are robust and able to withstand shipping as long as there is some PS-20. Therefore, for solutions of 20-100 g / L, trehalose formulations were used in long-term stability studies.

[0156] The shipping data confirmed that the low concentration formulation was not robust, as observed by high and highly variable sub-visible particle counts. The data also showed that the problem was not solved by polysorbate alone, and therefore reformulation of the low concentration solution was required. d. Reconstitution of low concentration DS

[0157] Low concentration reconstitution screened by simulated shipping pressurization and by MFI testing. Subvisible particle counts shown are for aspect ratio filtered particles >10 μm; trends were similar for other particle sizes.

[0158] During the manufacturing process, a high concentration (≥100 g / L) unformulated drug product (UDS) containing trehalose will be produced and frozen. Storage of this high concentration intermediate is necessary to enable dilution into different formulations spanning the 2-100 mg / mL formulation range. Dilution from UDS may result in some trehalose residue; for uniformity of composition, a single trehalose concentration will be used for the entire low dose range. Buffer and pH are unchanged.

[0159] A primary screen was used to determine the minimum protein concentration at which the trehalose formulation was stable, and the effect of increased ionic strength was determined by formulating in 150 mM trehalose and 75 mM arginine hydrochloride or calcium chloride. The results shown below indicate that protein concentrations ≥ 10 g / L were stable, but for robustness, the low concentration range was set to 2–20 g / L. Increasing ionic strength produced more stable solutions in both excipient cases. See, e.g. Figure 8 .

[0160] Subsequent studies focused on optimizing arginine or NaCl concentrations; for all studies, a protein concentration of 2 g / L was used as a worst-case scenario. For each excipient, broad and narrow excipient concentration screens were run at 0.02% PS-20. The initial arginine screen was run with varying amounts of trehalose, where solutions were prepared by mixing 270 mM arginine with 250 mM trehalose. The goal was to maintain osmolality, but there was an error in the calculation (arginine HCl is divalent), so the solutions containing arginine were hypertonic. The remainder of the excipient concentration screens were run at constant trehalose concentrations of 40 or 50 mM, based on residual trehalose at 20 g / L after dilution from the 100 g / L stock. The results are provided in Fig. 9 and 10 middle.

[0161] Broad arginine and NaCl screening results indicate that concentrations need to be greater than 50 mM arginine or 75 mM NaCl in order to produce a stable formulation. Narrow concentration screening of 75-150 mM arginine or 100-200 mM NaCl resulted in low particle counts across the entire range. This indicates that concentrations in the middle of these ranges should produce a robust formulation; 130 mM was selected to be isotonic in combination with 50 mM residual trehalose.

[0162] The same approach was used to examine PS-20 concentration optimization for the new formulation. These experiments were performed in parallel with the narrow excipient concentration screen, so midpoint concentrations were used. The test conditions were 0.01-0.1% PS-20, 115mM arginine HCl, 40mM trehalose; and 0.01-0.05% PS-20, 150mM NaCl, 50mM trehalose. Several samples were tested with 0.02% PS-80, but produced higher counts than the corresponding PS-20 results (data not shown). Particle counts were low for all polysorbate levels tested, indicating that it was not necessary to change the level from 0.02% in the new formulation. See, e.g. Fig.11 .

[0163] The results of reconstitution of low concentration anti-IL5R indicated that either arginine or NaCl were able to stabilize the formulation in the short term. For protein concentrations of 2–20 g / L, the formulations considered were 130 mM arginine HCl, or 130 mM NaCl, with 50 mM trehalose, 20 mM histidine / histidine HCl, 0.02% PS-20, pH 6. e. Vials and PFS Considerations

[0164] The three formulations formed above (20-100 mg / mL in trehalose, 2-20 mg / mL in trehalose / arginine, and 2-20 mg / mL in trehalose / NaCl) are all appropriate for both the vial and PFS configurations. The vial configuration is a 3cc Schott vial with a 4432 / 50 West stopper. The biggest risk with the vial configuration is the silicone oil level on the stopper, so the long term stability studies were performed with the silicone oil level (0.039 mg / cm 2 ) is higher than the commonly used silicone oil level (0.007–0.024 mg / cm 2 )'s plug.

[0165] The syringe tested for the anti-IL5R formulation was a flatbed syringe, BD 1 mL long PFS, with a chamfered flange, staked 29G thin-barrel injection needle, containing 0.4 mg silicone oil, and covered with a BD260 rigid needle shield (Cat. No. 47363119). f. Long-term stability study Two long-term stability studies were performed to validate the decisions made based on the screening studies. Stability study No. 1 investigated the long-term stability of trehalose and arginine / trehalose formulations in PFS and vials. In addition, a PFS comparison was performed, which is not discussed here. Stability study No. 2 was initiated for the configuration tested in study No. 1 to provide data from another batch of material, and also investigated the NaCl / trehalose formulation interval and the effect of fill volume from 1 / 2 mL to 1 mL in PFS and vials. i. Stability Study No. 1: Anti-IL5R PFS Stability Description

[0166] Stability studies were performed in syringes, using vials as controls. Each endpoint of the formulation interval was tested in each primary container. The syringes tested were platform Hypak TM for Biotech; this is a 1 mL long BD glass syringe, virtually tungsten free, with a 29G thin barrel injection needle and 0.4 mg silicone oil. The vials used are 3cc Schott vials with West 4423 / 50 stoppers and closures.

[0167] The filling preparations are as follows: 2 and 20 mg / mL anti-IL5R antibody, 125 mM arginine HCl, 50 mM trehalose, 20 mM histidine / histidine HCl, 0.02% PS-20, pH 6; and 20 and 100 mg / mL anti-IL5R antibody, 250 mM trehalose, 20 mM histidine / histidine hydrochloride, 0.02% PS-20, pH 6. A. Purity Statement for Anti-IL5R Prefilled Syringes

[0168] There was no significant effect of primary container on monomer loss for any formulation. Some effect of protein concentration was observed, but monomer loss rates were consistently low. Fig.12 A. B. Particle Analysis Description of Anti-IL5R Prefilled Syringes Particle formation is considered to be the major degradation pathway of anti-IL5R, so it is thought to play a major role in determining the appropriate PFS. Subvisible particle measurement by HIAC showed an increase in the number of particles in PFS, which may be attributed to silicone oil droplets (see, Fig.12 B and 12C). However, for all configurations, the total particle counts remained well below the USP limits of 6000 particles / mL >10 μm and 600 particles / mL >25 μm. MFI was used as an orthogonal method and showed similar results, although less variability was observed between vessels because silicone oil droplets can be filtered out of the results in the MFI software. ii. At 5°C in Hypak TM for Biotech Overview of Stability in Syringes

[0169] Table 1 shows the available materials for use in Hypak TM Summary of stability data (Stability Study #1) for anti-IL5R in Biotech syringes up to 16 months (arginine formulation) and 24 months (trehalose formulation). Visible particles were detected, which resulted in a reduction in PS-20 concentration from 0.02% to 0.006%. No other high risks were identified, but sub-visible particle counts were variable. Table 1 Stability overview of anti-IL5R in various formulations in Hypak for Biotech syringes at 5°C for 16 months. Appearance results included particles, which were mitigated by reducing PS-20 concentration (covered in a separate report). Subvisible particle results were variable, but no trend was observed. All other results were within expectations for a stable product.* iii. Stability study No. 2

[0170] Stability Study No. 2 was an identical stability study in pre-filled syringes and vials designed to determine the low dose formulation and fill volume, as well as to verify placebo stability.

[0171] Previous stability studies used to select PFS had only a 1 mL fill volume. However, there are multiple potential benefits of lower fill volumes, including reduced injection pain, reduced subcutaneous mass, more rapid administration, and less leakage from the administration site.

[0172] The interval strategy for the 1 mL fill option is two protein concentration intervals, 2-20 g / L for low doses, and 20-100 g / L for high doses. The same intervals for the 1 / 2 mL fill cover the dose range of 1-50 mg, and a 100 g / L fill volume interval from 1 / 2 mL to 1 mL is also required for the 50-100 mg dose. For clarity, this result is shown graphically in Fig.13 In both cases, the lowest dose interval was studied in the arginine and NaCl based formulations.

[0173] The drugs and placebos filled were as follows: All solutions: 20 mM histidine / histidine hydrochloride, 0.02% PS-20, pH 6.0 2Arg / 20Arg: 2 or 20 g / L anti-IL5R, 125 mM arginine hydrochloride, 50 mM trehalose 2NaCl / 20NaCl: 2 or 20 g / L anti-IL5R, 130 mM NaCl, 50 mM trehalose 20Tre / 100Tre: 20 or 100 g / L anti-IL5R, 250 mM trehalose Arg placebo: 125 mM arginine hydrochloride, 50 mM trehalose NaCl placebo: 130 mM NaCl, 50 mM trehalose Tre placebo: 250 mM trehalose

[0174] The 1 mL fill volume configuration was not placed on test for any vial configuration or placebo in the PFS in order to reduce the overall size of the study, since the 1 / 2 mL fill volume was most likely the worst case configuration due to the higher surface area-to-volume ratio.

[0175] All of these samples were shipped and then placed in stability studies at 5°C, 25°C, and 40°C. The vials were stored upside down during the stability study to minimize contact with the stopper. When visible particles were found in previous studies, 6 weeks of data had been collected for this stability study. In addition, particles were observed in the NaCl formulation between the 3rd and 6th month time points in this study (data not shown), resulting in the NaCl formulation being ruled out and the arginine formulation being supported.

[0176] The additional formulation work required to mitigate visible particle formation is described in the examples below and resulted in a reduction in the polysorbate-20 concentration from 0.02% to 0.006%. No other stability issues were observed for this formulation, and no significant effects of container or fill volume were observed. The one-year stability data are summarized in Fig.14 The results of the present study included visible particle fraction, purity loss by SEC, particle count by HIAC, and potency (not all configurations were tested at all time points). Conclusion of Example 1

[0177] Formulation screening was performed using different stress methods, including freeze / thaw, agitation, silicone oil spiking, and accelerated stability. Long-term stability was used to validate the results of the screening studies. The Phase 2b trehalose formulation was successful for high concentration liquids with increased polysorbate concentrations, but showed instability when extended to low concentration liquids, primarily due to the formation of subvisible particles. Reformulation of the low concentration range was performed by increasing the ionic strength using arginine hydrochloride or sodium chloride, resulting in stable solutions.

[0178] To cover the wide range of possible doses (2–100 mg), three potential formulations are presented as follows: 2-20 g / L, 130 mM arginine hydrochloride, 50 mM trehalose dihydrate, 20 mM histidine / histidine hydrochloride, 0.02% PS-20, pH 6.0; 2-20 g / L, 130 mM sodium chloride, 50 mM trehalose dihydrate, 20 mM histidine / histidine hydrochloride, 0.02% PS-20, pH 6.0; and 20-100 g / L, 250 mM trehalose dihydrate, 20 mM histidine / histidine hydrochloride, 0.02% PS-20, pH 6.0.

[0179] Long-term stability up to 24 months indicated that the three formulations were stable with respect to agitation at 2–8°C, relatively insensitive to silicone oil, and compatible with vials and PFS. In addition, minimal degradation was observed at elevated temperatures. Two low concentration formulations were observed consecutively, and the NaCl option was eliminated due to increased visible particle formation compared to the arginine option, resulting in two formulation intervals. The data indicate that pre-filled syringes are acceptable primary containers for 1 mL or 1 / 2 mL fill volumes. Visible particle formation remains an issue for these formulations, which is addressed as in the following examples. Example 2 - Particle Formation in Anti-IL5R Formulations

[0180] In previous long-term stability studies, visible particles were detected in aqueous anti-IL5R formulations in vials and PFS, with the first detection occurring at the 6-month time point (data not shown). A study was conducted to mitigate visible particle formation in anti-IL5R antibody formulations containing polysorbate 20 (PS-20) stored for long periods of time. The following example describes a long-term study of formulations containing different PS-20 and protein concentrations to verify the importance of PS-20 concentration in mitigating particle formation. This study yielded an acceptable range of 0.002%–0.01% PS-20. Abbreviations and definitions Preliminary research

[0181] Particle formation was detected in the long-term stability study described in Example 1. However, the particles were extremely small and appeared more like a cloud than individual particles. To see the particles, the samples were examined close to the light source. Because these particles were different from those in the vials and PFS standard set, the samples were compared to each other and the standards at each time point.

[0182] Particles were initially tested at the 6 month time point in 100 g / L Tre vials with 0.02% PS-20.

[0183] Six-month and 11-month samples were compared in (1) vials and (2) prefilled syringes (PFS). Both studies showed that particle formation was more severe in the vials than in the PFS, but PFS standards were not available at the time so multiple comparisons could not be made. Samples in the PFS were decanted and injected into vials for appearance testing. No more particles were observed in these vials, confirming that the difference was not a process length effect. This vial-PFS difference was more pronounced in the trehalose formulation than in the arginine formulation.

[0184] The different PS-20 concentrations (0, 0.01%, 0.02%, and 0.03%) in the 100 g / L Tre formulation in the vials and PFS after shipping were compared. At the 11-month time point, the lower PS-20 concentrations of 0 and 0.01% PS-20 were completely free of particles. In both the 0.02% and 0.03% PS-20 vials and PFS, particles were visible. At 20 months, the 0 and 0.01% PS-20 PFS and 0% PS-20 vials were still clear, but the 0.01% vial had very small amounts of rolled particles.

[0185] In the directed study, particles were clearly visible in the vials and PFS at 100 g / L Tre, 0.02% PS-20, in some cases as early as 3-4 months when held close to a light source. At the low concentration (20 g / L) end of the Tre interval, particle formation was much slower and was only observed in vials at month 21. No particles were observed in the 20 g / L Tre PFS at any PS-20 concentration, with data available up to 21 months. Particle formation in the arginine formulations was somewhat slower, with particles observed at the high concentration (20 g / L) end of the interval at months 6-9 with 0.02% PS-20. No particles were observed in the 2 g / L Arg formulation at any PS-20 concentration in either container, with data available up to 16 months.

[0186] Accelerated and stress temperature studies provide no insight into particle formation. At 40°C, there was no particle formation at all over the 3 month test period. At 25°C, particle formation was similar or slightly reduced compared to 5°C.

[0187] This data indicates that the high protein concentration end of the interval has a high risk of visible particles, while the low end is potentially risky in the long term. It was investigated that reducing the PS-20 concentration mitigated particle formation. The primary container is still the platform PFS, but the vials are used as early particle formation readouts because particle formation in the vials is more severe and easier to see. Investigation of different PS-20 concentrations

[0188] An extended stability study was designed to investigate particle formation in different formulations, different PS-20 concentrations, and different antibody concentrations. All samples were filled into PFS and vials, shipped twice to a separate location to simulate the distribution process, and placed in a 5°C stability study. Appearance testing was performed once a month. SEC, HIAC, and MFI testing were also performed on a subset of samples at time zero, 3 months, 6 months, and 9 months. Fig.15 Shown are samples prepared as part of this study.

[0189] like Fig.16 As shown in the MFI data for PFS at time zero in , sub-visible particles were formed after delivery in the absence of any PS-20. The lowest PS-20 tested level (0.002%) or more was sufficient to protect against delivery stress. In addition, it was verified that the lowest PS-20 concentration (0.002%) was sufficient to protect against DS delivery stress in the tank using the scaled-down model (data not shown).

[0190] The particle appearance results showed significant particle formation in the vials above 0.01% PS-20, while some particles were present in both formulation intervals in the PFS at 0.02% PS-20, with fewer particles observed at the lowest concentration endpoints of both intervals. Fig.17 For 100 g / L Tre, particles were first observed at month 3 and for 20 g / L Arg at month 6. The accompanying figures show the effect of protein concentration and PS-20 concentration on particle formation at the latest time point, month 9. It should be noted that particle formation observations were all performed close to the light source.

[0191] The appearance data set an upper limit of 0.01% for PS-20 concentration, while the sub-visible particle count set a lower limit of 0.002% (the actual limit may be lower depending on the available data). The midpoint of this acceptable range (0.006%) was set as the new PS-20 concentration.

[0192] The overall stability of PFS was demonstrated for the interval endpoint samples, including data up to 9 months for 2 g / L Arg and 20 g / L Arg, and up to 18 months for 20 g / L Tre and 100 g / L Tre. The following assays were tested: A. 0.002, 0.006 and 0.01% PS-20: Appearance, HIAC, MFI and SEC. B. 0.006% PS-20 only: Instron, Bioassay, Bioanalyzer, RP, and cIEF. All data indicate that the new formulation is stable and low risk. A summary of the data is shown in Table 2. Table 2

[0193] Data from Example 2 indicate that reducing the PS-20 concentration to 0.006% mitigated particle formation and produced an antibody formulation product that was stable for at least 9 months (arginine formulation) or 18 months (trehalose formulation) with no signs indicating impending deterioration. Example 3 Orthogonal method for particle detection

[0194] The primary method for particle detection and quantification is the appearance test by visual inspection as illustrated in Example 2. Visual inspection is variable for a variety of reasons. In general, due to the inherent variability of human senses, visual inspection is variable, resulting in different results for different individuals. Due to the very small size of these particles, their visibility is highly dependent on the amount of light, and they are different from the standards of vials and PFS. These factors increase the variability of results between time points and between analysts.

[0195] Orthogonal methods were investigated to validate the appearance results. The particles formed in the trehalose formulation were too small to be seen individually, so a sub-visible particle method was investigated. Worst case samples (100 g / L, Tre, 0.02% PS-20, vials) from different batches at week 2, and months 2, 5, and 9 were compared by DLS, FC, HIAC, and MFI.

[0196] DLS was performed at 100 g / L, resulting in an underestimation of the main peak as expected and unreliability of all peak sizes. Large peaks were detected for the 2-, 5-, and 9-month-old samples with sizes of 1.4–2.2 μm. Even though the reported sizes were not reliable, the presence of the peaks correlated with visible particles. However, neither the reported particle size nor the intensity of the particle peaks correlated with the visual appearance results.

[0197] HIAC results were similar for all samples; no particles were detected.

[0198] The MFI counts for particles >10 μm and >25 μm were similar for all samples. However, the MFI counts for particles >1 μm and >2 μm, as well as the FC counts, trended toward visual appearance results and increased with sample age. The results for these samples are shown in Fig.18 middle.

[0199] Further experiments indicated that particles >1 μm as determined by MFI provided a more reliable trend with visual appearance than larger particles or FC counts (data not shown).

[0200] Similar experiments were performed on the 20 g / L Arg sample and showed visible particles, but none of these orthogonal methods successfully detected these particles. The particles formed in the arginine formulation appeared larger than those formed in trehalose and were seen as separate particles, which may be why they were not detected by the sub-visible particle method.

[0201] The MFI was used to verify the effect of PS-20 concentration in Example 2. A comparison of the MFI and appearance scores at the 9-month time point for Example 2 is shown in Fig.19Together with additional measurements (not shown), this comparison indicates that particles are visible when the MFI count exceeds approximately 100,000 particles (>1 μm) / mL. The MFI results provide additional support for the conclusion that particle formation is mitigated by 0.002%-0.01% PS-20, especially in PFS. The vial data was considered a worst-case scenario and was also stable at the target concentration of 0.006% PS-20. Example 4 Stability studies

[0202] Additional stability studies were performed to investigate all configurations previously studied with a new target PS-20 concentration (0.006%) and to add confidence that these formulations are stable. The intervals include protein concentration intervals as well as fill volume intervals and are shown in Figure 1. Fig. 20 middle.

[0203] The increase in fill volume intervals increases the dosage range covered by the trehalose formulations. These additional configurations reduce the risk associated with the arginine formulations, which are more risky because particle formation is slower and cannot be detected by orthogonal methods. The samples included in this stability study were: 0, 2, and 20 g / L, 1 mL fill, arginine formulation, 0.006% PS-20; 0 and 2 g / L, 0.3 mL fill, arginine formulation, 0.006% PS-20; 0, 20, 50 and 100 g / L, 1 mL fill, trehalose formulation, 0.006% PS-20; and 0 and 20 g / L, 0.3 mL fill, trehalose formulation, 0.006% PS-20.

[0204] Samples were shipped twice to an off-site location to simulate the distribution process and then placed in a 5°C, 25°C, and 40°C stability study.

[0205] Nine months of data were collected for the arginine formulation, and twelve months of data have been collected for the trehalose formulation. These results are consistent with historical data, and no particles were observed in these samples until this time point. In addition, no trend of sub-visible particles has been observed over time, although some moderately high super-biased values ​​have occurred. The range of results for all analyses is shown in Table 3. Table 3 43 of the measured values ​​fell within this range, but one outlier value of 347 particles / mL was also measured.

[0206] The data support that this formulation interval is recommended for the full dose range. The trehalose interval is less risky than the arginine interval and is recommended for doses as low as 6 mg. Conclusion of Example 2-4

[0207] Particle observations in previous long-term stability studies led to an investigation of formulation variables that could be changed to improve stability. Key variables, namely protein concentration and polysorbate concentration, were tested and analyzed. Based on the data presented above, an allowable polysorbate range from 0.002% to 0.01% and a target of 0.006% were determined as the optimal formulation for the anti-IL5R antibody formulation. This observation was supported by two stability studies with available data at 18 and 12 months, primarily visual appearance testing.

[0208] It was also observed that by using a volume interval of 0.3-1.0 mL at 20 mg / mL, trehalose formulations in the range of 6-20 mg could be used instead of arginine formulations. Trehalose formulations were found to be more predictable than arginine formulations due to a larger data set and better detection. Example 5

[0209] Additional purification development focused on reducing host cell proteins (HCPs) was performed to determine the impact of HCPs on particle formation. Fig.21 Anti-IL5R was purified as outlined.

[0210] 2D gel analysis of the flow-through of the Protein A column indicated the presence of several protein species. See, e.g. Fig. 22 As determined by reverse phase mass spectrometry (RP-MS), the major impurities included about 60% of Fab fragments, about 5% of light chains (LC) and fragments, and about 40% of non-anti-IL5R related host cell proteins (HCPs). Non-anti-IL5R related HCPs included glutathione S-transferase (GST), fructose bisphosphate aldolase, and dinulopyrrolidase.

[0211] The flow-through from the Protein A column was further passed through a Protein L column to further separate (1) Fab fragments from (2) non-anti-IL5R HCPs. It was found that some material in the non-anti-IL5R HCPs contributed to particle formation and was not a Fab fragment (data not shown).

[0212] To determine which major HCP was responsible for particle formation, GST was first investigated. The effect of GST on particle formation was investigated by: (1) selectively removing GST to determine the effect on particle formation; (2) analyzing the particle pellet to determine the presence of GST in the pellet; and (3) adding GST to the anti-IL5R formulation to determine the effect on particle formation.

[0213] Preliminary evidence for specific removal of GST using an affinity matrix made of glutathione-cross-linked agarose conjugate showed that removal of GST resulted in reduced particle formation (data not shown). Analysis of the pellet formed by the protein A flow-through indicated the presence of high concentrations of GST in the pellet (data not shown).

[0214] GST was added to a purified anti-IL5R formulation lacking particles to determine the effect of GST on the particle formulation. GST was obtained from a commercial source (Prospec). GST was added to a formulation containing 50 mg / mL anti-IL5R, 20 mM histidine hydrochloride buffer, 9% (w / v) trehalose, 0.02% PS-20, pH 6.0, resulting in GST concentrations of 3.8 μg / mg and 7.6 μg / mg. The samples were incubated at 38°C-42°C. The particles were observed by placing the samples under a light box. The incubation time required to observe particle formation depends on the level of GST present. For both 3.8 μg / mg and 7.6 μg / mg GST-spiked samples, particles were observed after 24 hours at 38°C-42°C (data not shown).

[0215] These results indicate that spiking of GST into anti-IL5R formulations resulted in particle formation in the anti-IL5R formulations. Example 6

[0216] The GST activity in different anti-IL5R formulations purified by different means was investigated. As a preliminary, the GST activity assay (BioVision) was used to determine the GST concentration to form a standard curve. GST catalyzes the formation of thiol groups of glutathione (GSH) to electrophilic compounds such as 1-chloro-2,4-dinitrobenzene (CDNB) to form a GS-DNB conjugate, which was detected at 340nM. Therefore, the increase in absorption at 340nM is directly proportional to the GST activity. Using this assay, the GST concentration was determined for different anti-IL5R formulations (samples AJ) purified using different procedures. Note the correlation between GST concentration and particle formation. The results are provided in Table 4. Table 4 sample GST concentration Particle Description A 188.4 A large number of particles (most) B 0.422 Virtually particle-free* C 2.074 Virtually particle-free* D 1930.5 A large number of particles E 1950.6 A large number of particles F 1774.2 A large number of particles G 40.6 Particle formation H 1.073 Some particles I <LLOQ No particles J 5.416 Particle formation *Some particles, but not many LLOQ = Lower Limit of Quantitation.

[0217] This evidence confirms that the presence of GST is correlated with particle formation. Example 7

[0218] Different purification columns were investigated to identify the most effective method to reduce GST concentration in anti-IL5R formulations. See Table 5. GST concentration was determined as outlined in Example 7. Table 5 describe GST concentration (μg / ml) CM Products 5.935 HA Products 1.255 MabSelect Sure Elution Products <LLOQ CaptoAdhere Products <LLOQ CEX Products* 9.228 CEX Products* 21.081 *GST concentrations of two separate lots of anti-IL5R antibody were determined.

[0219] Table 5 shows that both the Protein A column (MabSelect Sure) and the mixed mode chromatography column (CaptoAdhere) were successful in reducing the GST concentration to below detectable levels during the purification of the anti-IL5R antibody. Example 8

[0220] The presence of GST in other antibody preparations was investigated. The presence of particles in these antibody preparations was also investigated. The results are presented in Table 6 along with the evaluation. Table 6

[0221] All the various embodiments or options described herein may be combined in any or all variants. Although the present invention has been specifically shown and described with reference to some of its embodiments, it should be understood by those of ordinary skill in the art that they are presented by way of example only, are not restrictive, and various changes in form and detail may be made therein without departing from the spirit and scope of the present invention. Therefore, the breadth and scope of the present invention should not be limited to any of the exemplary embodiments described above, but should only be limited according to the following claims and their equivalents.

[0222] All documents cited herein (including journal articles or abstracts, published or corresponding U.S. or foreign patent applications, issued or foreign patents, or any other documents) are each incorporated herein by reference in their entirety, including all data, tables, figures, and text presented in the cited documents.

Claims

1. A stable aqueous antibody formulation comprising: a. about 2 mg / mL to about 100 mg / mL of an antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 5-7, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 8-10; and b. From about 0.002% to about 0.01% polysorbate-20.

2. The antibody formulation of claim 1, further comprising an uncharged excipient.

3. The antibody formulation of claim 2, wherein the uncharged excipient is trehalose.

4. The antibody formulation of any one of claims 1 to 3, comprising about 2 to about 20 mg / ml of the antibody.

5. The antibody formulation of any one of claims 1 to 3, comprising about 20 to about 100 mg / ml of the antibody.

6. The antibody formulation of claim 4, wherein the concentration of the uncharged excipient is about 20 mM to about 80 mM.

7. The antibody formulation of claim 5, wherein the concentration of the uncharged excipient is about 200 mM to about 400 mM.

8. The antibody formulation of claim 4, further comprising arginine.

9. The antibody formulation of claim 8, wherein the arginine is L-arginine.

10. The antibody formulation of claim 8, comprising about 100 mM to about 200 mM L-arginine.

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