Stable formulations of igg4 binding agents

Abstract

The present invention provides stable pharmaceutical formulations of antibodies, including liquid and lyophilized formulations, comprising an IgG4 binding agent and a citrate buffer. The pH of the formulation is equal to or less than 6 and at the isoelectric point (pI) of the binding agent. These formulations can be used in the treatment of chronic intestinal diseases or rheumatoid arthritis.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application Serial No. 61 / 615,539, filed March 26, 2012.BACKGROUND OF THE INVENTION

[0002] The human LIGHT antigen is one potential cytokine target that has been implicated in the processes of chronic inflammatory autoimmune disease. As a member of the TNF superfamily (TNFSF) of ligands, LIGHT is also known as TNFSF14 or CD258. LIGHT is expressed on the surface of T cells upon activation in a tightly regulated manner. However, LIGHT is also present at detectable levels constitutively on the surface of immature dendritic cells and on T cells and natural killer (NK) cells of the gut. LIGHT mediates its biologic effects by binding three TNF superfamily receptors, including the lymphotoxin β receptor (LTβR), the herpes virus entry mediator (HVEM), and decoy receptor 3 (DcR3). LIGHT-expressing lymphocytes can induce IBD-like symptoms in humans, and increases of LIGHT expression have been observed in patients with active Crohn's disease and other inflammatory disorders such as Graft-vs.-Host Disease.

[0003] CXCR5, also known as Burkitt lymphoma receptor (BLR1), CD185, MDR15, and MGC117347, is a G protein-coupled receptor that is a member of the CXC chemokine receptor family. The unprocessed CXCR5 precursor is 372 amino acids in length with a molecular weight of 42 K D . CXCR5 has a role in B cell migration and localization within particular anatomic compartments. Knockout mice lack peripheral lymph nodes, have fewer Peyer's patches and have decreased B cell levels. CXCL13, also known as BLC, is a ligand for CXCR5. CXCL13 is a B cell chemoattractant.

[0004] WO 2004 / 071439 discloses a lyophilized formulation with 20-200 mg / mL of an IgG4 antibody (natalizumab; anti-4α-integrin) that can be obtained by using: i) a buffering agent chosen from the group consisting of phosphate, histidine, citrate, acetate or succinate in the concentration range 2-50 mM to provide a pH preferably of 6 + / - 0.5 and ii) a surfactant such as a polysorbate at a concentration of 0.001% to 2.0% to improve stability and decrease reconstitution time.

[0005] Anti-LIGHT binding agents and anti-CXCR5 binding agents are each therapeutically relevant, and a need exists to formulate each of these binding agents into drug products that may be administered to subjects, particularly human subjects, for the treatment of inflammatory diseases.

[0006] In order to develop a pharmaceutical formulation containing an anti-LIGHT binding agent suitable for intravenous or subcutaneous administration, the binding agent must be concentrated to about 20 mg / mL or greater, usually about 100-150 mg / mL, and even up to 250 mg / mL. Many complications can arise at such high concentrations, including an increase in viscosity, a shift of pH, a change of the color of the solution, and the formation of visible and sub-visible particles.

[0007] The formulation of these binding agents is further complicated by the fact that these agents are highly prone to aggregation at such high concentrations.

[0008] The formulation of IgG4 antibodies is even further complicated by the fact that IgG4 antibodies tend to form half-molecules at high concentrations in solution. However, IgG4 antibodies are of therapeutic interest because they have reduced effector function.SUMMARY OF THE INVENTION

[0009] The technical information set out below may in some respects go beyond the disclosure of the claimed invention, which is defined exclusively by the appended claims. The additional technical information is provided to place the actual disclosure in a broader technical context and to illustrate possible related technical developments. Such additional technical information which does not fall within the scope of the appended claims, is not part of the disclosure.

[0010] To meet these and other needs, provided herein are highly stable IgG4 binding agent formulations. Highly stable IgG4 binding agent formulations have surprisingly been found in the form of liquids and lyophilized powders that comprise an IgG4 binding agent and a citrate buffer, wherein the pH of the formulation is at or below both about pH 6 and the isoelectric point (pI) of the binding agent. These formulations improve upon conventional formulations, which often lead to dimerization of the binding agent, such as an antibody, upon increasing the concentration of the binding agent, such as an antibody, in the formulation. In particular, the formulations of the invention reduce the amount of unwanted byproducts, including aggregates, half-molecules, degradation products, low molecular weight proteins (LMWPs), high molecular weight proteins (HMWPs), and rearrangements of acid, basic, and neutral isoforms of the binding agent, such as an antibody, component in the formulation.

[0011] In certain aspects, the invention provides a stable formulation comprising: (a) 150 mg / mL of a fully human IgG4 anti-LIGHT antibody comprising a heavy chain comprising the amount acid sequence of SEQ ID NO: 7 and a light chain comprising the amino acid sequence of SEQ ID NO: 8; (b) 10 mM citrate buffer; (c) 0.005% w / v polysorbate 20; and (d) 4% w / v mannitol, wherein the pH of the formulation is pH 5.5, and wherein 5% or less of antibodies in the formulation form aggregates after storage for at least 6 months at +5°C.

[0012] The antibody binds to lymphotoxin-like, exhibits inducible expression and competes with herpes virus glycoprotein D for herpes virus entry mediator, a receptor expressed on lymphocytes (LIGHT). According tothe invention, the anti-LIGHT binding agent or antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising complementary determining regions (CDRs) comprising the amino acid sequences of SEQ ID NOS: 1, 2, and 3, and the light chain variable region comprising CDRs comprising the amino acid sequences of SEQ ID NOS: 4, 5, and 6. The antibody is a fully human IgG4 anti-LIGHT antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 7 and a light chain comprising the amino acid sequence of SEQ ID NO: 8.

[0013] The antibody concentration is 150 mg / mL.

[0014] The citrate concentration is 10 mM. In some embodiments of the invention, the citrate buffer is sodium citrate dihydrate.

[0015] The pH of the formulation is pH 5.0.

[0016] In certain specific embodiment of the invention, the pI of the binding agent or antibody is from about 6.8 and about 7.2. In alternative specific embodiments of the invention, the pI of the binding agent or antibody is from about 7.6 and about 8.4.

[0017] The formulation comprises a surfactant, wherein the surfactantis polysorbate 20. In some specific embodiments of the invention, the concentration of polysorbate 20 is about 0.005% w / v.

[0018] The formulation comprises a tonicity agent. In certain specific embodiments of the invention, the tonicity agent is a saccharide. In some specific embodiments of the invention, the saccharide is mannitol. According to the invention, the concentration of mannitol is 4%. In alternative specific embodiments described herein, the saccharide is sucrose. In some specific embodiments described herein, the concentration of sucrose is between 1% and 10% w / v. In some specific embodiments described herein, the concentration of sucrose is 5% w / v. In alternative specific embodiments described herein, the concentration of sucrose is 6% w / v. In yet other specific embodiments described herein, the concentration of sucrose is 4.5% w / v. In further specific alternative embodiments described herein, the tonicity agent is sodium chloride In some specific embodiments described herein, the concentration of sodium chloride is 0.2%. In other specific embodiments described herein, the tonicity agent is a combination of sucrose and sodium chloride. In specific embodiments described herein, the concentration of sucrose is between 1% and 10% w / v. In alternative specific embodiments described herein, the concentration of sucrose is 6% w / v and the concentration of sodium chloride is 0.2%. In yet further alternative specific embodiments described herein, the concentration of sucrose is 4.5% w / v and the concentration of sodium chloride is 0.2%.

[0019] In certain embodiments of the invention, the formulation further comprises an amino acid. In certain specific embodiments of the invention, the amino acid concentration is between 0.1% and about 5% w / v. In certain specific embodiments of the invention, the amino acid is proline or arginine. In specific embodiments of the invention, the proline or arginine concentration is between 1% and 2% w / v. In other specific embodiments of the invention, the proline concentration is 1.5% w / v. In alternative specific embodiments of the invention, the arginine concentration is 1% w / v.

[0020] In certain embodiments of the invention, the formulation is a liquid formulation. In other specific embodiments of the invention, the formulation is a lyophilized formulation.

[0021] The formulation is stable for at least 6 months at +5°C, wherein 5% or less of antibodies in the formulation form aggregates after storage for at least 6 months at +5°C. In alterntative embodiments of the invention, the formulation is stable for at least 9 months at +5°C, wherein 5% or less of antibodies in the formulation form aggregates after storage for at least 9 months at +5°C.

[0022] In certain embodiments of the invention, the formulation exhibits a reduced amount of at least one byproduct selected from the group consisting of aggregates, half-molecules, degradation products, low molecular weight proteins, high molecular weight proteins, and and rearrangements of acidic / basic / neutral isoforms of the antibody as compared to either a reference anti-LIGHT formulation comprising an anti-LIGHT antibody in phosphate buffered saline at pH 7.3.

[0023] In certain specific embodiments of the invention, the invention provides a stable liquid antibody formulation suitable for subcutaneous administration, the formulation comprising: a) 150 mg / mL of a fully human IgG4 anti-LIGHT (lymphotoxin-like, exhibits inducible expression and competes with HSV glycoprotein D for HVEM, a receptor expressed by T lymphocytes) antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 7 and a light chain comprising the amino acid sequence of SEQ ID NO: 8; b) 10 mM citrate buffer; c) 0.005% w / v polysorbate 20; and d) 4% w / v mannitol; wherein the pH of the formulation is about pH 5.5.

[0024] In yet other specific embodiments described herein, the invention provides a stable lyophilized antibody formulation suitable for intravenous administration, the formulation comprising: a) 50 mg / mL of a fully human IgG4 anti-LIGHT (lymphotoxin-like, exhibits inducible expression and competes with HSV glycoprotein D for HVEM, a receptor expressed by T lymphocytes) antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 7 and a light chain comprising the amino acid sequence of SEQ ID NO: 8; b) 10 mM citrate buffer; c) 0.01% w / v polysorbate 20; d) 5% w / v sucrose; and e) 1.5% w / v proline; wherein the pH of the formulation is pH 5.5.

[0025] In certain embodiments of the invention, the invention provides a kit comprising a container comprising: 1) the formulation of any one of the previous claims, and 2) a lable or instructions for the administration and use of the formulation. In certain embodiments of the invention, the label comprises one or more of the following: instructions for the administration of the formulation, instructions for use of the formulation, instructions concerning the storage conditions of the formulation, information concerning lot and batch number of the formulation and / or kit, information concerning the composition of the formulation, safety information, information concerning possible adverse reactions, secondary effects, and / or side effects in connection with the administration of the formulation, or information concerning possible indications and / or contra-indications of the formulation.

[0026] In certain embodiments of the invention, the invention provides a pre-filled device or pre-filled container, such as a syringe, cartridge, vial, ampoule, or autoinjector comprising the formulation of the invention. In certain other embodiments, the invention provides a kit comprising such pre-filled syringe, cartridge, vial, ampoule, or autoinjector.BRIEF DESCRIPTION OF THE FIGURES

[0027] Figure 1 is a picture of a gel showing the results of denatured isoelectric focusing experiments that were used to determine the isoelectric point (pI) of the fully human IgG4 anti-LIGHT antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 7 and a light chain comprising the amino acid sequence of SEQ ID NO: 8 formulated in phosphate buffered saline at pH 7.3 at a concentration of 5.5 mg / mL (the "Original Formulation", "PBS Formulation", or "Reference Lot"). Lanes 1 & 5: IEF Calibration Kit High Range pI 5-10.5; lanes 2 & 4: a first batch of Reference Lot; lanes 3 & 4: a second batch of Reference Lot. The pI values are indicated by numbers. Figure 2 is a picture of an SDS-PAGE gel that compared different Reference Lot batches under reducing and non-reducing conditions. Lanes 1 & 10: Biorad Precision Plus Protein Standard; lane 5: empty; lane 2: a first batch of Reference Lot under non-reduced conditions; lanes 3 & 4: a second batch of Reference Lot under non-reduced conditions; lane 6: a first batch of Reference Lot under reduced conditions; lanes 7 & 8: a second batch of Reference Lot under reduced conditions; and lane 9: system control. The sizes are indicated by numbers within the rows. Figure 3 shows an ELISA graph that was used to determine the antigen binding activity of the first and second batches of Reference Lot. Figure 4 shows a size exclusion chromatography (SEC) chromatogram of the first batch of Reference Lot. As shown in Figure 4, SEC detected high molecular weight proteins (HMWP), e.g., di- / oligomers (RRT0.8) or aggregates, and low molecular weight proteins (LMWPs) or degradation products. The first batch of Reference Lot batch had a purity of 97% monomer content. Figure 5 shows a weak cation exchange chromatogram for the first batch of Reference Lot. As shown in Figure 5, rearrangements of acidic, neutral, and basic isoforms occured during stability studies. The first batch of Reference Lot had a distribution of acidic / neutral / basic isoforms of 42.3 / 55.6 / 1.9%. Figure 6 shows a differencial scanning calorimetry thermogram of the first batch of Reference Lot. As shown in Figure 6, the three domains of the antibody unfold at 68°C, 75°C, and 78°C. Figure 7 shows a dynamic light scattering pattern of the first batch of Reference Lot, which was unfiltered. DLS was used to determine the hydrodynamic diameter of the first batch of Reference Lot antibody monomer and potential soluble aggregates. Figure 8 shows a dynamic light scattering pattern of the first batch of Reference Lot, which was filetered. DLS was used to determine the hydrodynamic diameter of the first batch of Reference Lot antibody monomer and potential soluble aggregates. Figure 9 is a flow diagram of the drug product manufacturing process for the high antibody concentration formulation. Figure 10 shows a dynamic light scattering pattern of Formulation 14. DLS was used to determine the hydrodynamic diameter of the antibody monomer and potential soluble aggregates. Figure 11 is a picture of a gel showing the results of isoelectric focusing to determine the pI (isoelectric point) of the Lead CXCR5 Antibody. Lanes 1,6: IEF Calibration High Range pI Kit; Lanes 2,4: Reference Standard Lead Antibody LP08031; and Lanes 3,5: Lead Antibody Drug Substance, RSN0151 (not part of the invention). Figure 12 is a picture of an SDS-PAGE gel that compared different drug substance batches under reducing and non-reducing conditions. The gel was also used to determine the molecular weight of the Lead CXCR5 Antibody, and the presence of any aggregates (not part of the invention). Figure 13 is an ELISA graph that was used to determine antigen binding activity of the Lead CXCR5 Antibody to a 28mer peptide of the CXCR5 antigen (not part of the invention). Figure 14 is a SEC chromatogram of stressed Lead CXCR5 Antibody. SEC could detect high molecular weight proteins (HMWP), e.g., di- / oligomers or aggregatres and low molecular weight proteins (LMWP) or degradation products. The Lead CXCR5 Antibody had a purity of 99% monomer content (not part of the invention). Figure 15 is a WCX chromatogram that was used to determine acidic, neutral, and basic isoforms of the Lead CXCR5 Antibody. The Lead CXCR5 Antibody had a distribution of acidic / neutral / basic isoforms of 14 / 85 / 1% (not part of the invention). Figure 16 is a DLS measurement that was used to determine the hydrodynamic diameter of the antibody monomer and potential soluble aggregates (not part of the invention). Figure 17 is a picture of the Lead CXCR5 Antibody in acetate buffer pH 5.0 (left) and pH 5.5 (right); each v. WFI (water for injection) and after thermal stress. This figure shows that acetate is a suitable buffer system (not part of the invention). Figure 18 is a picture of the Lead CXCR5 Antibody in histidine buffer pH 6.0 (left), pH 5.5 (middle), and pH 5.0 (right); each v. WFI (water for injection) and after thermal stress. This figure shows that histidine is a suitable buffer (not part of the invention). Figure 19 is a picture of the Lead CXCR5 Antibody in TRIS buffer pH 7.5 after UF / DF (left) and after filtration (right); each v. WFI (water for injection) and after thermal stress. This figure shows that TRIS is an incompatible buffer system (not part of the invention). Figure 20 is a picture of the Lead CXCR5 Antibody in citrate buffer pH 6.0 after UF / DF and filtration (not part of the invention). Figure 21 is a picture of the Lead CXCR5 Antibody in acetate buffer pH 5.5 after UF / DF and filtration (not part of the invention). Figure 22 is a picture of the Lead CXCR5 Antibody in succinate buffer pH 5.0 after UF / DF and filtration (not part of the invention). Figure 23 is a picture of the Lead CXCR5 Antibody in histidine buffer pH 5.0 after UF / DF and filtration (not part of the invention). Figure 24 is a picture of the Lead CXCR5 Antibody in arginine buffer pH 6.0 after UF / DF and filtration (not part of the invention). Figure 25 is a picture of the appearance of Lead CXCR5 Antibody LA_09_016 solutions with different surfactants (without surfactant, polysorbate 20, polysorbate 80, Lutrol F68, Cremophor RH40, Solutol HS15, and SDS) after mechanical stress (350 rpm, 2.5 h, RT) (not part of the invention). Figure 26 is a graph that shows an increase of dimers under accelerated conditions, as analyzed by SEC. An increase of dimer formation up to 10% after three months of storage in all four histidine formulation can be seen. Acetate formulations showed an increase of dimer content up to 6%. In all four citrate formulations, the dimer concentration was below 2%, even after three months at +40°C (not part of the invention). Figure 27 is a graph showing an increase of basic isoforms under accelerated conditions, as analyzed by WCX. Histidine is worse for Lead CXCR5 Antibody stability under accelerated conditions. A slight increase of basic isoforms can be noticed for all four acetate formultions. Interestingly, it was not possible to discriminate between the four citrate formulations (not part of the invention). Figure 28 is a graph showing a decrease of neutral isoforms under accelerated conditions, as analyzed by WCX. This figure shows a strong decrease in neutral isoforms for the histidine formulations. A slight decrease was seen in acetate. Citrate was affected the least (not part of the invention). Figure 29 shows the delta pH of all four formulations (A-D) in citrate buffer at accelerated conditions. The most pH stabilizing formulations are the citrate buffered, and especially formulation B and D (not part of the invention). Figure 30 shows the delta pH of all four formulations (A-D) in acetate buffer at accelerated conditions. In acetate buffered solutions of the Lead CXCR5 Antibody, the pH was shifted towards higher value(not part of the invention). Figure 31 shows the delta pH of all four formulations (A-D) in histidine buffer at accelerated conditions. In histidine buffered solutions of the Lead CXCR5 Antibody, the pH was slightly decreasing (not part of the invention). Figure 32 is a graph showing the hydrodynamic diameter of CXCR5 LA_09_027 A-D after 3 months storage at 40°C. Citrate buffered formulations showed only slight aggregates after three weeks in formulation C, and after six weeks of storage in formulation A. Some aggregates could be detected after three months in formulation B as well. But, compared to acetate buffered formulations, the amount was very little (not part of the invention). Figure 33 is a graph showing the hydrodynamic diameter of CXCR5 LA_09_028 A-D after 3 months storage at 40°C. The acetate buffered formulation C showed some aggregates <200 nm after three weeks. Formulation A showed some aggregates after three months (not part of the invention). Figure 34 is a chart showing the effect of increasing Lead CXCR5 Antibody concentration on the Z-average. The Lead CXCR5 Antibody showed a significant increase in the hydrodynamic diameter (Z-Average) by increasing the concentration of the antibody (not part of the invention). Figure 35 is a chart showing the effect of different stabilizers (excipients) on the Z-Average at 100 mg / mL of Lead CXCR5 Antibody after thermal stress. Z-Average was measured before and after thermal stress. The stabilizing effect was simiar to all tested excipients, but the increase in Z-average was generally reduced by using amino acids as stabilizers (argining, lysing, or glycine). Lysine was excluded due to a higher content of aggregates after stress. Arginine showed a better effect than glycine (not part of the invention). Figure 36 is a chart showing the effect of different stabilizers on the Z-Average at 100 mg / mL Lead CXCR5 Antibody after mechanical stress. Z-Average was measured before and after mechanical stress. The same reduction in Z-average was noticed in the presence of amino acids. Sucrose had a better protective effect than trehalose against mechanical stress. Arginine and glycine performed better in combination with NaCl (not part of the invention). Figure 37 is a set of graphs showing particle size distribution, as measured by DLS, of Lead CXCR5 Antibody formulated in 10 mM citrate buffer at pH 6 before mechanical stress (A) and after mechanical stress (B). A higher molecular weight species was measured by DLS after mechanical stress of DS (not part of the invention). Figure 38 is a set of graphs showing particle size distribution, as measured by DLS, of Lead CXCR5 Antibody drug product prototype formulations (A-D; Table 111) before (A) and after (B) mechanical stress (not part of the invention). DETAILED DESCRIPTION A. Definitions

[0028] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of ordinary skill in the art.

[0029] It is noted here that as used in this specification and the appended claims, the singular forms "a", "an", and "the" also include plural reference, unless the context clearly dictates otherwise.

[0030] The term "about" or "approximately" means within 10%, such as within 5% (or 1% or less) of a given value or range.

[0031] The terms "administer" or "administration" refers to the act of injecting or otherwise physically delivering a substance as it exists outside the body (e.g., a formulation of the invention) into a patient, such as by mucosal, intradermal, intravenous, subcutaneous, intramuscular delivery and / or any other method of physical delivery described herein or known in the art. When a disease, or a symptom thereof, is being treated, administration of the substance typically occurs after the onset of the disease or symptoms thereof. When a disease or its symptoms are being prevented, administration of the substance typically occurs before the onset of the disease or symptoms thereof.

[0032] In the context of a polypeptide, the term "analog" refers to a polypeptide that possesses a similar or identical function as a LIGHTpolypeptide, a fragment of a LIGHT, a LIGHT, or an anti-LIGHT antibody, but does not necessarily comprise a similar or identical amino acid sequence of a LIGHT polypeptide, a fragment of a LIGHT polypeptide, a LIGHT epitope, or an anti-LIGHT antibody, or possess a similar or identical structure of a LIGHT polypeptide, a fragment of a LIGHT polypeptide, a LIGHT epitope, or an anti-LIGHT antibody. A polypeptide that has a similar amino acid sequence refers to a polypeptide that satisfies at least one of the following: (a) a polypeptide 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 a LIGHT polypeptide (e.g., SEQ ID NO: 9), a fragment of a LIGHT polypeptide, a LIGHT epitope, or an anti-LIGHT antibody described herein; (b) a polypeptide encoded by a nucleotide sequence that hybridizes under stringent conditions to a nucleotide sequence encoding a LIGHT polypeptide, a fragment of a LIGHT polypeptide, a LIGHT epitope, or an anti-LIGHT antibody (or VH or VL region thereof) described herein of at least 5 amino acid residues, at least 10 amino acid residues, at least 15 amino acid residues, at least 20 amino acid residues, at least 25 amino acid residues, at least 40 amino acid residues, at least 50 amino acid residues, at least 60 amino residues, at least 70 amino acid residues, at least 80 amino acid residues, at least 90 amino acid residues, at least 100 amino acid residues, at least 125 amino acid residues, or at least 150 amino acid residues (see, e.g., Sambrook et al. (2001) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.; Maniatis et al. (1982) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press, Cold Spring Harbor, N.Y.); and (c) a polypeptide encoded by a nucleotide 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 nucleotide sequence encoding a LIGHT polypeptide, a fragment of a LIGHT polypeptide, a LIGHT epitope, or an anti-LIGHT antibody (or VH or VL region thereof) described herein. A polypeptide with similar structure to a LIGHT polypeptide, a fragment of a LIGHT polypeptide, a LIGHT epitope, or an anti-LIGHT antibody refers to a polypeptide that has a similar secondary, tertiary or quaternary structure of a LIGHT polypeptide, a fragment of a LIGHT polypeptide, a LIGHT epitope, or a LIGHT antibody. The structure of a polypeptide can determined by methods known to those skilled in the art, including but not limited to, X-ray crystallography, nuclear magnetic resonance, and crystallographic electron microscopy.

[0033] To determine the percent identity of two amino acid sequences or of two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in the sequence of a first amino acid or nucleic acid sequence for optimal alignment with a 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, then the molecules are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences (i.e., % identity=number of identical overlapping positions / total number of positions X 100%). In one embodiment, the two sequences are the same length.

[0034] The determination of percent identity between two sequences (e.g., amino acid sequences or nucleic acid sequences) can also be accomplished using a mathematical algorithm. A non-limiting example of a mathematical algorithm utilized for the comparison of two sequences is the algorithm of Karlin and Altschul, 1990, Proc. Natl. Acad. Sci. U.S.A. 87:2264 2268, modified as in Karlin and Altschul, 1993, Proc. Natl. Acad. Sci. U.S.A. 90:5873 5877. Such an algorithm is incorporated into the NBLAST and XBLAST programs of Altschul et al., 1990, J. Mol. Biol. 215:403. BLAST nucleotide searches can be performed with the NBLAST nucleotide program parameters set, e.g., for score=100, wordlength=12 to obtain nucleotide sequences homologous to nucleic acid molecules of interest. BLAST protein searches can be performed with the XBLAST program parameters set, e.g., to score 50, wordlength=3 to obtain amino acid sequences homologous to a protein molecule of interest. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al., 1997, Nucleic Acids Res. 25:3389 3402. Alternatively, PSI BLAST can be used to perform an iterated search which detects distant relationships between molecules (Id.). When utilizing BLAST, Gapped BLAST, and PSI Blast programs, the default parameters of the respective programs (e.g., of XBLAST and NBLAST) can be used (see, e.g., National Center for Biotechnology Information (NCBI) on the worldwide web at ncbi dot nlm dot nih dot gov). Another non limiting example of a mathematical algorithm utilized for the comparison of sequences is the algorithm of Myers and Miller, 1988, CABIOS 4:11 17. Such an algorithm is incorporated in the ALIGN program (version 2.0), which is part of the GCG sequence alignment software package. When utilizing the ALIGN program for comparing amino acid sequences, a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4 can be used.

[0035] The percent identity between two sequences can be determined using techniques similar to those described above, with or without allowing gaps. In calculating percent identity, typically only exact matches are counted.

[0036] An "antagonist" or "inhibitor" refers to a molecule capable of inhibiting one or more biological activities of a target molecule. Antagonists may interfere with the binding of a receptor to a ligand and vice versa, by incapacitating or killing cells activated by a ligand, and / or by interfering with receptor or ligand activation (e.g., tyrosine kinase activation) or signal transduction after ligand binding to a receptor. The antagonist may completely block receptor-ligand interactions or may substantially reduce such interactions. All such points of intervention by an antagonist shall be considered equivalent for purposes of the instant invention.

[0037] For example, an "antagonist" or "inhibitor" of LIGHT refers to a molecule that is capable of inhibiting or otherwise decreasing one or more of the biological activities of LIGHT, such as in a cell expressing LIGHT or in a cell expressing a LIGHT ligand, such as a LIGHT receptor. For example, in certain embodiments, antibodies of the invention are antagonist antibodies that inhibit or otherwise decrease secretion of CCL20, IL-8, and / or RANTES from a cell having a cell surface-expressed LIGHT receptor (e.g., HVEM, LTβR and / or DcR3) when said antibody is contacted with said cell. In some embodiments, an antagonist of LIGHT (e.g., an antagonistic antibody of the invention) may, for example, act by inhibiting or otherwise decreasing the activation and / or cell signaling pathways of the cell expressing a LIGHT receptor, thereby inhibiting a LIGHT-mediated biological activity of the cell relative to the LIGHT-mediated biological activity in the absence of antagonist. In certain embodiments of the invention, the anti-LIGHT antibodies are fully human, antagonistic anti-LIGHT antibodies, such as fully human, monoclonal, antagonistic anti-LIGHT antibodies.

[0038] The terms "antibody", "immunoglobulin", or "Ig" may be used interchangeably herein. The term antibody includes, but is not limited to, synthetic antibodies, monoclonal antibodies, recombinantly produced antibodies, multispecific antibodies (including bi-specific antibodies), human antibodies, humanized antibodies, chimeric antibodies, intrabodies, single-chain Fvs (scFv) (e.g., including monospecific, bispecific, etc.), camelized antibodies, Fab fragments, F(ab') fragments, disulfide-linked Fvs (sdFv), anti-idiotypic (anti-Id) antibodies, and epitope-binding fragments of any of the above. In particular, antibodies include immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, i.e., antigen binding domains or molecules that contain an antigen-binding site that specifically binds to a LIGHT antigen (e.g., one or more complementarity determining regions (CDRs) of an anti-LIGHT antibody). The anti-LIGHT antibodies can be of any type (e.g., IgG, IgE, IgM, IgD, IgA and IgY), any class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2), or any subclass (e.g., IgG2a and IgG2b) of immunoglobulin molecule. In some embodiments, the anti-LIGHT antibodies are fully human, such as fully human monoclonal anti-LIGHT antibodies. In certain embodiments, the anti-LIGHT antibodies are IgG antibodies, human IgG4 antibodies.

[0039] As used herein, the term "anti-LIGHT antibody" means an antibody or polypeptide derived therefrom (a derivative) that binds specifically to human LIGHT as defined herein, including, but not limited to, molecules that inhibit or substantially reduce the binding of LIGHT to its ligands or inhibit LIGHT activity.

[0040] The term "B cell activity" means higher than normal B cell levels, which can be local, or evidence of a biological manifestation or function of a B cell, such as antibody expression, Bruton's tyrosine kinase presence or activity, expression or presence of CD19, expression or presence of B cell activating factor and so on.

[0041] The term "binding agent" means any molecule, such as an antibody, a siRNA, a nucleic acid, an aptamer, a protein, or a small molecule organic compound, that binds or specifically binds to LIGHT, or a variant or a fragment thereof.

[0042] The term "by-product" includes undesired products, which detract or diminish the proportion of therapeutic / prophylactic binding agent, such as an antibody, in a given formulation. For example, typical by-products include aggregates of the antibody, fragments of the antibody, e.g. produced by degradation of the antibody by deamidation or hydrolysis, or mixtures thereof. Typically, aggregates are complexes that have a molecular weight greater than the monomer antibody. Antibody degradation products may include, for example, fragments of the antibody, for example, brought about by deamidation or hydrolysis. Typically, degradation products are complexes that have a molecular weight less than the monomer antibody. In the case of an IgG antibody, such degradation products are less than about 150 kD.

[0043] The terms "composition" and "formulation" are intended to encompass a product containing the specified ingredients (e.g., an anti-LIGHT antibody or an anti-CXCR5 antibody) in, optionally, the specified amounts, as well as any product that results, directly or indirectly, from the combination of the specified ingredients in, optionally, the specified amounts.

[0044] The terms "constant region" or "constant domain" refer to a carboxy terminal portion of the light and heavy chain which is not directly involved in binding of the antibody to antigen but exhibits various effector functions, such as interaction with the Fc receptor. The terms refer to the portion of an immunoglobulin molecule having a more conserved amino acid sequence relative to the other portion of the immunoglobulin, the variable domain, which contains the antigen binding site. The constant domain contains the CH1, CH2 and CH3 domains of the heavy chain and the CHL domain of the light chain.

[0045] The term "CXCR5" relates to the naturally occurring, known molecule found on lymphocytes, particularly B cells, and particularly naive B cells; to such a molecule isolated from such cells; to such a molecule manufactured recombinantly using known materials and means, and using a nucleic acid encoding a CXCR5; as well as to portions of CXCR5, such as the extracellular (EC) domain, that retain the characteristics and propertiessuch as CXCL13 binding. A soluble CXCR5 molecule can consist essentially of the EC domain of CXCR5, which includes, generally, about the first sixty amino acids of the molecule, that is, the amino terminal portion of CXCR5.

[0046] CXCR5 is a non-promiscuous receptor. CXCL13 is a ligand of CXCR5 and is expressed constitutively on stromal cells, such as follicular dendritic cells, and in lymphoid tissues. CXCL13 specifically attracts B cells and a small subset of T cells called B helper follicular T cells, TFH. This may not be unexpected given the many interactions between T cell and B cell populations in the immune system. Moreover, activated T cells induce or upregulate CXCR5 expression. Infiltration of lymphocytes into tertiary, ectopic germinal centers (GCs) has been found to correlate well with increased disease severity and tolerance breakdown in certain disorders that present with such atypical lymph node-like structures. Using in vivo murine models, such as CXCR5- / - and CXCL13- / - mice, the absence of either the receptor or the ligand results in an altered GC fine architecture due to altered T and B cell localization, and possibly interaction. These mice are also protected against developing severe collagen-induced arthritis (CIA). As CXCR5 is selectively expressed on mature B cells, which are linked to the pathogenesis of RA, blocking this receptor will modulate the arthritogenic response in affected individuals. Rheumatoid arthritis treatment with biologics (i.e., anti-TNFα and anti-CD20 antibodies, Rituximab) has shown to be clinically effective; in particular, patients on B cell-directed therapy have shown long-lasting improvements in clinical signs and symptoms. Selective targeting of CXCR5, which is only expressed on mature B cells and B helper T cells, will not affect B cell development or immunocompromise the patient. Unlike Rituximab, the disclosed anti-CXCR5 antibody is a neutralizing antibody that does not mediate cell cytotoxicity.

[0047] A "CXCR5 disease" is a malady, disorder, disease, condition, abnormality and so on, that is characterized by or caused by overexpression or increased levels of CXCL13 or other CXCR5 ligand, increased levels of B cells, increased levels of B cell activity, increased levels of CXCR5, or improper metabolism and activity of CXCR5.

[0048] The term "epitope" refers to a localized region on the surface of an antigen, such as a LIGHT polypeptide, or LIGHT polypeptide fragment, that is capable of being bound to one or more antigen binding regions of a binding agent, such as an antibody, and that has antigenic or immunogenic activity in an animal, such a mammal, such as in a human, that is capable of eliciting an immune response. An epitope having immunogenic activity is a portion of a polypeptide that elicits an antibody response in an animal. An epitope having antigenic activity is a portion of a polypeptide to which an antibody specifically binds, as determined by any method well known in the art, for example, such as an immunoassay. Antigenic epitopes need not necessarily be immunogenic. Epitopes usually consist of chemically active surface groupings of molecules, such as amino acids or sugar side chains, and have specific three dimensional structural characteristics, as well as specific charge characteristics. A region of a polypeptide contributing to an epitope may be contiguous amino acids of the polypeptide or the epitope may come together from two or more non-contiguous regions of the polypeptide. The epitope may or may not be a three-dimensional surface feature of the antigen. In certain embodiments, a LIGHT epitope is a three-dimensional surface feature of a LIGHT polypeptide (e.g., in a trimeric form of a LIGHT polypeptide). In other embodiments, a LIGHT epitope is a linear feature of a LIGHT polypeptide (e.g., in a trimeric form or monomeric form of the LIGHT polypeptide). Anti-LIGHT antibodies may specifically bind to an epitope of the monomeric (denatured) form of LIGHT, an epitope of the trimeric (native) form of LIGHT, or both the monomeric (denatured) form and the trimeric (native) form of LIGHT. In specific embodiments, the anti-LIGHT antibodies specifically bind to an epitope of the trimeric form of LIGHT but do not specifically bind the monomeric form of LIGHT.

[0049] The term "excipients" refers to inert substances that are commonly used as a diluent, vehicle, preservative, binder, stabilizing agent, etc. for drugs and includes, but is not limited to, proteins (e.g., serum albumin, etc.), amino acids (e.g., aspartic acid, glutamic acid, lysine, arginine, glycine, histidine, etc.), fatty acids and phospholipids (e.g., alkyl sulfonates, caprylate, etc.), surfactants (e.g., SDS, polysorbate, nonionic surfactant, etc.), saccharides (e.g., sucrose, maltose, trehalose, etc.) and polyols (e.g., mannitol, sorbitol, etc.). See, also, Remington's Pharmaceutical Sciences (1990) Mack Publishing Co., Easton, Pa..

[0050] In the context of a peptide or polypeptide, the term "fragment" refers to a peptide or polypeptide that comprises less than the full length amino acid sequence. Such a fragment may arise, for example, from a truncation at the amino terminus, a truncation at the carboxy terminus, and / or an internal deletion of a residue(s) from the amino acid sequence. Fragments may, for example, result from alternative RNA splicing or from in vivo protease activity. In certain embodiments, hLIGHT fragments include polypeptides comprising an amino acid sequence of at least 5 contiguous amino acid residues, at least 10 contiguous amino acid residues, at least 15 contiguous amino acid residues, at least 20 contiguous amino acid residues, at least 25 contiguous amino acid residues, at least 40 contiguous amino acid residues, at least 50 contiguous amino acid residues, at least 60 contiguous amino residues, at least 70 contiguous amino acid residues, at least 80 contiguous amino acid residues, at least 90 contiguous amino acid residues, at least contiguous 100 amino acid residues, at least 125 contiguous amino acid residues, at least 150 contiguous amino acid residues, at least 175 contiguous amino acid residues, at least 200 contiguous amino acid residues, or at least 250 contiguous amino acid residues of the amino acid sequence of a LIGHT polypeptide or an antibody that specifically binds to a LIGHT polypeptide. In a specific embodiment, a fragment of a LIGHT polypeptide or an antibody that specifically binds to a LIGHT antigen retains at least 1, at least 2, or at least 3 functions of the polypeptide or antibody.

[0051] The terms "fully human antibody" or "human antibody" are used interchangeably herein and refer to an antibody that comprises a human variable region and, possibly a human constant region. In specific embodiments, the terms refer to an antibody that comprises a variable region and constant region of human origin. "Fully human" anti-LIGHT antibodies, in certain embodiments, can also encompass antibodies that bind LIGHT polypeptides and are encoded by nucleic acid sequences that are naturally occurring somatic variants of a human germline immunoglobulin nucleic acid sequence. In a specific embodiment, the anti-LIGHT antibodies are fully human antibodies. The term "fully human antibody" includes antibodies having variable and constant regions corresponding to human germline immunoglobulin sequences as described by Kabat et al. (See Kabat et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242). Methods of producing fully human antibodies are known in the art.

[0052] The phrase "recombinant human antibody" includes human antibodies that are prepared, expressed, created, or isolated by recombinant means, such as antibodies expressed using a recombinant expression vector transfected into a host cell, antibodies isolated from a recombinant, combinatorial human antibody library, antibodies isolated from an animal (e.g., a mouse or cow) that is transgenic and / or transchromosomal for human immunoglobulin genes (see, e.g., Taylor, L. D. et al. (1992) Nucl. Acids Res. 20:6287-6295) or antibodies prepared, expressed, created, or isolated by any other means that involves splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies can have variable and constant regions derived from human germline immunoglobulin sequences (See Kabat, E. A. et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242). In certain embodiments, however, such recombinant human antibodies are subjected to in vitro mutagenesis (or, when an animal transgenic for human Ig sequences is used, in vivo somatic mutagenesis) and thus the amino acid sequences of the VH and VL regions of the recombinant antibodies are sequences that, while derived from and related to human germline VH and VL sequences, may not naturally exist within the human antibody germline repertoire in vivo.

[0053] An "IgG4 binding agent" or a "binding agent comprising at least a portion of an IgG4 Fc region" both refer to binding agents described herein that include at least a fragment of IgG4 Fc. In certain embodiments, the fragment comprises 10, 20, 30, 40, 50, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210 or 220 amino acids of the IgG4 Fc region. In other embodiments, the fragment includes 10-50, 50-100, 100-150, or 150-200 amino acids of the IgG4 Fc region. In other embodiments, the portion of the IgG4 Fc region can have a certain homology to the IgG4 Fc region. For example, the IgG4 binding agent may include a portion of a protein with greater than 50, 60, 70, 80, 90, 93, 95, 96, 97, 98, 99, or 100% homology to the IgG4 Fc region. Exemplary Fc regions of IgG4 are described throughout the specification.

[0054] The term "heavy chain", when used in reference to an antibody, refers to five distinct types, called alpha (α), delta (Δ), epsilon (ε), gamma (γ), and mu (µ), based on the amino acid sequence of the heavy chain constant domain. These distinct types of heavy chains are well known in the art and give rise to five classes of antibodies, IgA, IgD, IgE, IgG, and IgM, respectively, including four subclasses of IgG, namely IgG1, IgG1, IgG3, and IgG4. In some embodiments, the heavy chain is a human heavy chain.

[0055] "Humanized" forms of non-human (e.g., murine) antibodies are chimeric immunoglobulins, immunoglobulin chains or fragments thereof (such as F v , F ab , F ab' , F (ab')2 or other target-binding subsequences of antibodies) that contain sequences derived from non-human immunoglobulin, as compared to a human antibody. In general, the humanized antibody will comprise substantially all of one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin template sequence. The humanized antibody may also comprise at least a portion of an immunoglobulin constant region (F c ), typically that of the human immunoglobulin template chosen. In general, the goal is to have an antibody molecule that is minimally immunogenic in a human. Thus, it is possible that one or more amino acids in one or more CDRs also can be changed to one that is less immunogenic to a human host, without substantially minimizing the specific binding function of the one or more CDRs. Alternatively, the FR can be non-human but those amino acids most immunogenic are replaced with ones less immunogenic. Nevertheless, CDR grafting, as discussed above, is not the only way to obtain a humanized antibody. For example, modifying just the CDR regions may be insufficient as it is not uncommon for framework residues to have a role in determining the three-dimensional structure of the CDR loops and the overall affinity of the antibody for its ligand. Hence, any means can be practiced so that the non-human parent antibody molecule is modified to be one that is less immunogenic to a human, and global sequence identity with a human antibody is not always a necessity. So, humanization also can be achieved, for example, by the mere substitution of just a few residues, particularly those which are exposed on the antibody molecule and not buried within the molecule, and hence, not readily accessible to the host immune system. Such a method is taught herein with respect to substituting "mobile" or "flexible" residues on the antibody molecule, the goal being to reduce or dampen the immunogenicity of the resultant molecule without comprising the specificity of the antibody for its epitope or determinant. See, for example, Studnicka et al., Prot Eng 7(6)805-814, 1994; Mol Imm 44:1986-1988, 2007; Sims et al., J Immunol 151:2296 (1993); Chothia et al., J Mol Biol 196:901 (1987); Carter et al., Proc Natl Acad Sci USA 89:4285 (1992); Presta et al., J Immunol 151:2623 (1993), WO 2006 / 042333 and U.S. Pat. No. 5,869,619.

[0056] An "isolated" or "purified" binding agent, such as an antibody, is substantially free of cellular material or other contaminating proteins from the cell or tissue source from which the binding agent is derived, or substantially free of chemical precursors or other chemicals when chemically synthesized. For example, the language "substantially free of cellular material" includes preparations of an antibody in which the antibody is separated from cellular components of the cells from which it is isolated or recombinantly produced. Thus, an antibody that is substantially free of cellular material includes preparations of antibody having less than about 30%, 20%, 10%, or 5% (by dry weight) of heterologous protein (also referred to herein as a "contaminating protein"). When the antibody is recombinantly produced, it is also desirable to be substantially free of culture medium, i.e., culture medium represents less than about 20%, 10%, or 5% of the volume of the protein preparation. When the antibody is produced by chemical synthesis, in some embodiments it is substantially free of chemical precursors or other chemicals, i.e., it is separated from chemical precursors or other chemicals that are involved in the synthesis of the protein. Accordingly, such preparations of the antibody have less than about 30%, 20%, 10%, 5% (by dry weight) of chemical precursors or compounds other than the antibody of interest. In some embodiments, anti-LIGHT or anti-CXCR5 antibodies are isolated or purified.

[0057] The term "human LIGHT," "hLIGHT" or "hLIGHT polypeptide" and similar terms refer to the polypeptides ("polypeptides," "peptides" and "proteins" are used interchangeably herein) comprising the amino acid sequence of SEQ ID NO: 9 and related polypeptides, including SNP variants thereof. Related polypeptides include allelic variants (e.g., SNP variants); splice variants; fragments; derivatives; substitution, deletion, and insertion variants; fusion polypeptides; and interspecies homologs, in some embodiments, which retain LIGHT activity and / or are sufficient to generate an anti-LIGHT immune response. Also encompassed are soluble forms of LIGHT that are sufficient to generate an anti-LIGHT immunological response. As those skilled in the art will appreciate, an anti-LIGHT binding agent, such as an antibody, can bind to a LIGHT polypeptide, polypeptide fragment, antigen, and / or epitope, as an epitope is part of the larger antigen, which is part of the larger polypeptide fragment, which, in turn, is part of the larger polypeptide. hLIGHT can exist in a trimeric (native) or monomeric (denatured) form.

[0058] The term "Kabat numbering," and like terms are recognized in the art and refer to a system of numbering amino acid residues that are more variable (i.e. hypervariable) than other amino acid residues in the heavy and light chain variable regions of an antibody, or an antigen binding portion thereof (Kabat et al. (1971) Ann. NY Acad. Sci. 190:382-391 and, Kabat et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242). For the heavy chain variable region, the hypervariable region typically ranges from amino acid positions 31 to 35 for CDR1, amino acid positions 50 to 65 for CDR2, and amino acid positions 95 to 102 for CDR3. For the light chain variable region, the hypervariable region typically ranges from amino acid positions 24 to 34 for CDR1, amino acid positions 50 to 56 for CDR2, and amino acid positions 89 to 97 for CDR3.

[0059] The term "light chain" when used in reference to an antibody refers to two distinct types, called kappa (κ) of lambda (λ) based on the amino acid sequence of the constant domains. Light chain amino acid sequences are well known in the art. In some embodiments, the light chain is a human light chain.

[0060] The terms "manage", "managing", and "management" refer to the beneficial effects that a subject derives from a therapy (e.g., a prophylactic or therapeutic agent), which does not result in a cure of the infection. In certain embodiments, a subject is administered one or more therapies (e.g., prophylactic or therapeutic agents, such as a formulation of the invention) to "manage" a LIGHT-mediated disease (e.g., chronic bowel disease, IBD, Crohn's disease, ulcerative colitis, or GVHD), one or more symptoms thereof, so as to prevent the progression or worsening of the disease.

[0061] The term "monoclonal antibody" refers to an antibody obtained from a population of homogenous or substantially homogeneous antibodies, and each monoclonal antibody will typically recognize a single epitope on the antigen. In some embodiments, a "monoclonal antibody" is an antibody produced by a single hybridoma or other cell. The term "monoclonal" is not limited to any particular method for making the antibody. For example, monoclonal antibodies may be made by the hybridoma method as described in Kohler et al.; Nature, 256:495 (1975) or may be isolated from phage libraries. Other methods for the preparation of clonal cell lines and of monoclonal antibodies expressed thereby are well known in the art (see, for example, Chapter 11 in: Short Protocols in Molecular Biology, (2002) 5th Ed.; Ausubel et al., eds., John Wiley and Sons, New York).

[0062] The term "pharmaceutically acceptable" means being approved by a regulatory agency of the Federal or a state government, or listed in the U.S. Pharmacopeia, European Pharmacopeia or other generally recognized Pharmacopeia for use in animals, and more particularly in humans.

[0063] By "pharmaceutically acceptable excipient" is meant any inert substance that is combined with an active molecule, such as a monoclonal antibody, for preparing an agreeable or convenient dosage form. The "pharmaceutically acceptable excipient" is an excipient that is non-toxic to recipients at the dosages and concentrations employed, and is compatible with other ingredients of the formulation comprising the monoclonal antibody.

[0064] The terms "prevent", "preventing", and "prevention" refer to the total or partial inhibition of the development, recurrence, onset or spread of a LIGHT-mediated disease and / or symptom related thereto, resulting from the administration of a therapy or combination of therapies provided herein (e.g., a combination of prophylactic or therapeutic agents, such as a formulation of the invention).

[0065] The term "prophylactic agent" refers to any agent that can totally or partially inhibit the development, recurrence, onset or spread of a LIGHT-mediated disease and / or symptom related thereto in a subject. In certain embodiments, the term "prophylactic agent" refers to a formulation of the invention. In certain other embodiments, the term "prophylactic agent" refers to an agent other than a formulation of the invention. In some embodiments, a prophylactic agent is an agent that is known to be useful to or has been or is currently being used to prevent a LIGHT-mediated disease and / or a symptom related thereto, or impede the onset, development, progression and / or severity of a LIGHT-mediated disease and / or a symptom related thereto. In specific embodiments, the prophylactic agent is a fully human anti-LIGHT antibody, such as a fully human anti-LIGHT monoclonal antibody.

[0066] The term "LIGHT antigen" refers to that portion of a LIGHT polypeptide to which a binding agent, such as an antibody, specifically binds. A LIGHT antigen also refers to an analog or derivative of a LIGHT polypeptide or fragment thereof to which a binding agent, such as an antibody, specifically binds. In some embodiments, a LIGHT antigen is a monomeric LIGHT antigen or a trimeric LIGHT antigen. A region of a LIGHT polypeptide contributing to an epitope may be contiguous amino acids of the polypeptide, or the epitope may come together from two or more non-contiguous regions of the polypeptide. The epitope may or may not be a three-dimensional surface feature of the antigen. A localized region on the surface of a LIGHT antigen that is capable of eliciting an immune response is a LIGHT epitope. The epitope may or may not be a three-dimensional surface feature of the antigen.

[0067] The terms "LIGHT-mediated disease" and "LIGHT-mediated disorder" are used interchangeably and refer to any disease that is completely or partially caused by or is the result of LIGHT. In certain embodiments, LIGHT is aberrantly (e.g., highly) expressed on the surface of a cell. In some embodiments, LIGHT may be aberrantly upregulated on a particular cell type. In other embodiments, normal, aberrant, or excessive cell signaling is caused by binding of LIGHT to a LIGHT ligand. In certain embodiments, the LIGHT ligand is a LIGHT receptor (e.g., HVEM, LTβR, or DCR3), for example, that is expressed on the surface of a cell, such as a colonic epithelial cell. In certain embodiments, the LIGHT-mediated disease is a chronic bowel disease, an inflammatory bowel disease (IBD), such as Crohn's disease (CD) or ulcerative colitis (UC). In other embodiments, the LIGHT-mediated disease is graft-versus-host disease (GVHD).

[0068] The term "saccharide" refers to a class of molecules that are derivatives of polyhydric alcohols. Saccharides are commonly referred to as carbohydrates and may contain different amounts of sugar (saccharide) units, e.g., monosaccharides, disaccharides, and polysaccharides.

[0069] The terms "specifically binds" or "specifically binding" mean specifically binding to an antigen or a fragment thereof and not specifically binding to other antigens. For example, an antibody that specifically binds to an antigen may bind to other peptides or polypeptides with lower affinity, as determined by, e.g., radioimmunoassays (RIA), enzyme-linked immunosorbent assays (ELISA), BIACORE, or other assays known in the art. Antibodies or variants or fragments thereof that specifically bind to an antigen may be cross-reactive with related antigens. In some embodiments, antibodies or variants or fragments thereof that specifically bind to an antigen do not cross-react with other antigens. An antibody or a variant or a fragment thereof that specifically binds to a LIGHT antigen can be identified, for example, by immunoassays, BIAcore, or other techniques known to those of skill in the art. Typically a specific or selective reaction will be at least twice background signal or noise, and more typically more than 10 times background. See, e.g., Paul, ed., 1989, Fundamental Immunology Second Edition, Raven Press, New York at pages 332-336 for a discussion regarding antibody specificity.

[0070] A "stable" or "stabilized" formulation is one in which the binding agent, such as an antibody, therein essentially retains its physical stability, identity, integrity, and / or chemical stability, identity, integrity, and / or biological activity upon storage. Various analytical techniques for measuring protein stability are available in the art and are reviewed in Peptide and Protein Drug Delivery, 247-301, Vincent Lee Ed., Marcel Dekker, Inc., New York, N.Y., Pubs. (1991) and Jones, A. Adv. Drug Delivery Rev. 10:29-90 (1993), for example. Stability can be measured at a selected temperature and other storage conditions for a selected time period. The stability may be determined by at least one of the methods selected from the group consisting of visual inspection, SDS-PAGE, IEF, HPSEC, RFFIT, and kappa / lambda ELISA. For example, an antibody "retains its physical stability" in a pharmaceutical formulation, if it shows no signs of aggregation, precipitation, and / or denaturation upon visual examination of color and / or clarity, or as measured by UV light scattering, SDS-PAGE, or by (high pressure) size exclusion chromatography (HPSEC). In some embodiments, when using the formulations of the invention, 5% or less, typically 4% or less, typically 3% or less, more typically 2% or less, and particularly 1% or less of the antibodies forms aggregates, as measured by HPSEC or any other suitable method for measuring aggregation formation. For example, an antibody is considered stable in a particular formulation if the antibody monomer has a purity of about 90%, typically about 95%, in particular about 98% after a certain predetermined period of time under certain storage conditions in a particular formulation. Chemical stability can be assessed by detecting and quantifying chemically altered forms of the protein. Chemical alteration may involve size modification (e.g., clipping), which can be evaluated using (HP)SEC, SDS-PAGE, and / or matrix-assisted laser desorption ionization / time-of-flight mass spectrometry (MALDI / TOF MS), for example. Other types of chemical alteration include charge alteration (e.g., occurring as a result of deamidation), which can be evaluated by ion-exchange chromatography, for example. An antibody "retains its biological activity" in a pharmaceutical formulation at a given time, if the biological activity of the antibody at a given time is at least about 90% (within the errors of the assay) of the biological activity exhibited at the time the pharmaceutical formulation was prepared, as determined in an antigen binding assay or virus neutralizing assay, for example.

[0071] The terms "subject" and "patient" are used interchangeably. As used herein, a subject is typically a mammal, such as a non-primate (e.g., cows, pigs, horses, cats, dogs, rats, etc.) or a primate (e.g., monkey and human), and in some embodiments a human. In one embodiment, the subject is a mammal, such as a human, having a LIGHT-mediated disease. In another embodiment, the subject is a mammal, such as a human, at risk of developing a LIGHT-mediated disease.

[0072] The term "therapeutically effective amount" refers to the amount of a therapy (e.g., a formulation of the invention) that is sufficient to reduce and / or ameliorate the severity and / or duration of a given disease and / or a symptom related thereto. This term also encompasses an amount necessary for the reduction or amelioration of the advancement or progression of a given disease, reduction or amelioration of the recurrence, development or onset of a given disease, and / or to improve or enhance the prophylactic or therapeutic effect(s) of another therapy (e.g., a therapy other than a formulation of the invention). In some embodiments, the therapeutically effective amount of an antibody of the invention is from about 0.1 mg / kg (mg of antibody per kg weight of the subject) to about 100 mg / kg. In certain embodiments, a therapeutically effective amount of an antibody provided therein is about 0.1 mg / kg, about 0.5 mg / kg, about 1 mg / kg, 3 mg / kg, 5 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 25 mg / kg, about 30 mg / kg, about 35 mg / kg, about 40 mg / kg, about 45 mg / kg, about 50 mg / kg, about 60 mg / kg, about 70 mg / kg, about 80 mg / kg about 90 mg / kg or about 100 mg / kg (or a range therein). In some embodiments, "therapeutically effective amount" as used herein also refers to the amount of an antibody of the invention to achieve a specified result (e.g., inhibition of a LIGHT biological activity of a cell, such as inhibition of secretion of CCL20, IL-8, or RANTES from the cell).

[0073] The term "therapeutic agent" refers to any agent that can be used in the treatment, management or amelioration of a LIGHT-mediated disease and / or a symptom related thereto. In certain embodiments, the term "therapeutic agent" refers to a formulation of the invention. In certain other embodiments, the term "therapeutic agent" refers to an agent other than a formulation of the invention. In some embodiments, a therapeutic agent is an agent that is known to be useful for, or has been or is currently being used for the treatment, management or amelioration of a LIGHT-mediated disease or one or more symptoms related thereto.

[0074] The term "therapy" refers to any protocol, method, and / or agent that can be used in the prevention, management, treatment, and / or amelioration of a LIGHT-mediated disease (e.g., IBD or GVHD). In certain embodiments, the terms "therapies" and "therapy" refer to a biological therapy, supportive therapy, and / or other therapies useful in the prevention, management, treatment, and / or amelioration of a LIGHT-mediated disease known to one of skill in the art, such as medical personnel.

[0075] The terms "treat", "treatment", and "treating" refer to the reduction or amelioration of the progression, severity, and / or duration of a LIGHT-mediated disease (e.g., chronic bowel disease, IBD, or GVHD) resulting from the administration of one or more therapies (including, but not limited to, the administration of one or more prophylactic or therapeutic agents, such as a formulation of the invention). In specific embodiments for LIGHT, such terms refer to the reduction or inhibition of the binding of LIGHT to HVEM, the reduction or inhibition of the binding of LIGHT to LTβR, the reduction or inhibition of the binding of LIGHT to DcR3, the reduction or inhibition of the production or secretion of CCL20 from a cell expressing a LIGHT receptor of a subject, the reduction or inhibition of the production or secretion of IL-8 from a cell expressing a LIGHT receptor of a subject, the reduction or inhibition of the production or secretion of RANTES from a cell expressing a LIGHT receptor of a subject, and / or the inhibition or reduction of one or more symptoms associated with a LIGHT-mediated disease, such as a chronic bowel disease, IBD, or GVHD.

[0076] The terms "variable region" or "variable domain" refer to a portion of the light and heavy chains, typically about the amino-terminal 120 to 130 amino acids in the heavy chain and about 100 to 110 amino acids in the light chain, which differ extensively in sequence among antibodies and are used in the binding and specificity of each particular antibody for its particular antigen. The variability in sequence is concentrated in those regions called complementarity determining regions (CDRs), while the more highly conserved regions in the variable domain are called framework regions (FR). The CDRs of the light and heavy chains are primarily responsible for the interaction of the antibody with antigen. Numbering of amino acid positions is according to the EU Index, as in Kabat et al. (1991) Sequences of proteins of immunological interest. (U. S. Department of Health and Human Services, Washington, D.C.) 5th ed. ("Kabat et al."). In some embodiments, the variable region is a human variable region.B. Formulations and Formulation Components

[0077] As stated previously, the formulations of the invention have surprisingly been found in the form of liquids and lyophilized powders that comprise an IgG4 binding agent and a citrate buffer, wherein the pH of the formulation is at or below both about pH 6 and the isoelectric point (pI) of the binding agent. The formulations of the invention provide significant improvements over conventional IgG4 binding agent formulations (e.g., phosphate buffered saline (PBS) formulations), which form unwanted byproducts upon increasing the concentration of the binding agent in the formulation. In particular, the formulations of the invention have a reduced amount of aggregates, half-molecules, degradation products, low molecular weight proteins (LMWPs), high molecular weight proteins (HMWPs), and rearrangements of acid, basic, and neutral isoforms of the binding agent in the formulations.i. Anti-LIGHT Binding Agents, and variants and fragments thereof

[0078] In certain embodiments, the formulations of the invention include an anti-LIGHT binding agent. The binding agents such as an antibody, that binds or specifically binds to LIGHT, or a variant or a fragment thereof. In some embodiments, the binding agent is an anti-LIGHT antibody, or a variant thereof, or an antigen binding fragment thereof. Anti-LIGHT antibodies specifically bind to a LIGHT (lymphotoxin-like, exhibits inducible expression and competes with HSV glycoprotein D for HVEM, a receptor expressed by T lymphocytes) protein, polypeptide fragment, or epitope. The LIGHT molecule may be from any species. In some embodiments, the LIGHT molecule is from a human, known herein as "hLIGHT". hLIGHT has the following amino acid sequence, which is identified as SEQ ID NO: 9:

[0079] In certain specific embodiments, the anti-LIGHT antibody comprises a heavy chain variable region (VH) comprising the amino acid sequence of any one or more of the following complementary determining regions (CDRs): HCDR1 - GYNWH (SEQ ID NO: 1 ); HCDR2 - EITHSGSTNYNPSLKS (SEQ ID NO: 2 ); or HCDR3 - EIAVAGTGYYGMDV (SEQ ID NO: 3).

[0080] In other specific embodiments, the anti-LIGHT antibody comprises a light chain variable region (VL) comprising the amino acid sequence of any one or more of the following complementary determining regions (CDRs): LCDR1 - RASQGINSAFA (SEQ ID NO: 4 ); LCDR2 - DASSLES (SEQ ID NO: 5); or LCDR3 - QQFNSYPLT (SEQ ID NO: 6).

[0081] In one specific embodiment, the anti-LIGHT antibody comprises a heavy chain variable region (VH) comprising the amino acid sequences of SEQ ID NOs: 1, 2, and 3.

[0082] In another specific embodiment, the anti-LIGHT antibody comprises a light chain variable region (VL) comprising the amino acid sequences of SEQ ID NOs: 4, 5, and 6.

[0083] In more specific embodiments, the anti-LIGHT antibody comprises a heavy chain variable region comprising the amino acid sequences of SEQ ID NOs: 1, 2, and 3; and a light chain variable region comprising the amino acid sequences of SEQ ID NOs: 4, 5, and 6.

[0084] The anti-LIGHT antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 7: Positions 1-122:variable region of the heavy chain (VH). The CDRs (complementary determining regions, according to Kabat definition) are underlined.Positions 123-448:constant region of human IgG4 (SwissProt IGHG4 _HUMAN with the two mutations S241P and L248E, according to Kabat numbering).

[0085] The anti-LIGHT antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 8: Positions 1-107:variable region of the light chain (VL). The CDRs (complementary determining regions, according to Kabat definition) are underlined.Positions 108-214:constant region of human Cκ.

[0086] In further embodiments, the anti-LIGHT antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 7, and a light chain comprising the amino acid sequence of SEQ ID NO: 8.

[0087] In certain embodiments, the anti-LIGHT antibody comprises a heavy chain derived from the amino acid sequence of SEQ ID NO: 10, which may be encoded by the nucleic acid sequence of SEQ ID NO: 11. Amino acids 1-19: signal peptide Amino acids 20-141: 124F19k2 variable region (VH) Amino acids 142-end: hIgG4 PE constant region Nucleic acids 1-57: nucleic acids encoding the signal peptide Nucleic acids 58-423: nucleic acids encoding the 124F19k2 variable region (VH) Nucleic acids 424-end: nucleic acids encoding the hIgG4 PE constant region

[0088] In alternative specific embodiments, the anti-LIGHT antibody comprises a light chain derived from the amino acid sequence of SEQ ID NO: 12 , which may be encoded by the nucleic acid sequence of SEQ ID NO: 13. Amino acids 1-22: signal peptide Amino acids 23-129: 124F19k2 variable region (VL) Amino acids 130-end: hKappa constant region Nucleic acids 1-66: nucleic acids encoding the signal peptide Nucleic acids 67-387: nucleic acids encoding the 124F19k2 variable region (VL) Nucleic acids 388-end: nucleic acids encoding the hKappa constant region

[0089] In an embodiment of the invention, the anti-LIGHT antibody is a fully human antibody. Examples of fully human antibody isotypes include IgA, IgD, IgE, IgG, and IgM. In some embodiments, the anti-LIGHT antibody is an IgG antibody. There are four forms of IgG. In some embodiments, the anti-LIGHT antibody is an IgG4 antibody. In some embodiments of the invention, the anti-LIGHT antibody is a fully human IgG4 antibody.

[0090] In some embodiments, the anti-LIGHT antibody further comprises a constant region, e.g., a human IgG constant region. In some embodiments, the constant region is a human IgG4 constant region. In additional embodiments, the constant region is a modified human IgG4 constant region. In other embodiments, the constant region is a human Cκ constant region.

[0091] The anti-LIGHT antibody is a fully human IgG4 anti-LIGHT antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 7 and a light chain comprising the amino acid sequence of SEQ ID NO: 8 (the "Lead LIGHT Antibody"). In alternative embodiments of the invention, the anti-LIGHT antibody is a fully human IgG4 anti-LIGHT antibody comprising a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising 3 complementary determining regions (CDRs) comprising the amino acid sequences of SEQ ID NOs: 1, 2, and 3, and the light chain variable region comprising 3 CDRs comprising the amino acid sequences of SEQ ID NOs: 4, 5, and 6. Identification, isolation, preparation, and characterization of anti-LIGHT antibodies, including the anti-LIGHT antibody comprising a heavy chain amino acid sequence comprising SEQ ID NO: 7 and a light chain amino acid sequence comprising SEQ ID NO: 8, have been described in detail in U.S. Patent No. 8,058,402, corresponding to PCT Publication WO 2008 / 027338.

[0092] Embodiments described herein also include antigen binding fragments of the anti-LIGHT binding agents, such as antibodies. The term "antigen binding domain," "antigen binding region," "antigen binding fragment," and similar terms refer to that portion of an antibody which comprises the amino acid residues that interact with an antigen and confer on the binding agent its specificity and affinity for the antigen (e.g., the complementary determining regions (CDR)). The antigen binding region can be derived from any animal species, such as rodents (e.g., rabbit, rat or hamster) and humans. In some embodiments, the antigen binding region will be of human origin. Non-limiting examples of antigen binding fragments include: Fab fragments, F(ab')2 fragments, Fd fragments, Fv fragments, single chain Fv (scFv) molecules, dAb fragments, and minimal recognition units consisting of the amino acid residues that mimic the hypervariable region of the antibody.

[0093] In some embodiments of the invention, the anti-LIGHT binding agents (or a variant thereof) will ameliorate, neutralize, or otherwise inhibit LIGHT biological activity in vivo (e.g., the LIGHT-mediated production or secretion of CCL20, IL-8, or RANTES from a cell expressing a LIGHT receptor, e.g., HVEM, LTβR, and / or DcR3).

[0094] In some embodiments of the invention, the anti-LIGHT binding agents (or a variant thereof) are antagonist binding agents that ameliorate, neutralize, or otherwise inhibit LIGHT biological activity in vivo (e.g., the LIGHT-mediated production or secretion of CCL20, IL-8, or RANTES from cells expressing a LIGHT ligand, such as a LIGHT receptor, (e.g., HVEM, LTβR, and / or DcR3).

[0095] The anti-LIGHT binding agent (or a variant thereof thereof) is present in the formulations in an amount of 150 mg / mL.

[0096] In an embodiment, the anti-LIGHT binding agent is present in the formulation in an amount of 150 mg / mL. In another exemplary embodiment, a fully human IgG4 anti-LIGHT antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 7 and a light chain comprising the amino acid sequence of SEQ ID NO: 8 is present in the formulation in an amount of 150 mg / mL.iii . Buffering Agents

[0097] The formulations of the invention comprise a citrate buffer as a buffering agent. A buffering agent maintains a physiologically suitable pH. In addition, a buffering agent enhances isotonicity and chemical stability of the formulation. The citrate buffer is present in the formulations at a concentration of 10 mM.

[0098] The formulations of the invention have a pH of 5.5. The pH of the formulation may be measured by any means known to those of skill in the art. A means for measuring pH is using a pH meter with a micro-electrode. The pH of the formulation may be adjusted using any means known in the art. Exemplary chemicals for altering the pH of the formulations are hydrochloric acid (HCl) and sodium hydroxide (NaOH).

[0099] In certain embodiments, the formulations of the invention have a pH at or below the isoelectric point (pI) of the binding agent, such as an antibody. The isoelectric point is the pH at which a particular molecule or surface carries no net electrical charge. The pI of an anti-LIGHT binding agent may be determined by any means known to those of skill in the art. In some embodiments, the pI of an anti-LIGHT antibody is determined by denaturated isoelectric focusing. As shown in Figure 1, the pI of a fully human IgG4 anti-LIGHT antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 7 and a light chain comprising the amino acid sequence of SEQ ID NO: 8 is 6.8-7.2.iv. Surfactants

[0100] The formulations of the invention comprise a surfactant, wherein the surfactant is polysorbate 20.

[0101] The surfactant is present in the formulations in an amount of 0.005% (w / v).v. Tonicity Agents

[0102] The formulations of the invention comprise a tonicity agent. Typically, tonicity agents are used to adjust or maintain the osmolality of the formulations in order to bring it closer to the osmotic pressure of body fluids, such as blood or plasma. Tonicity agents may also maintain the binding agent levels in a formulation. In part, the tonicity agent contributes to preserving the level, ratio, or proportion of the therapeutically active binding agent present in the formulation. As used herein, the term "tonicity" refers to the behavior of biologic components in a fluid enviornment or solution. Isotonic solutions possess the same osmotic pressure as blood plasma, and can be intravenously infused into a subject without changing the osmotic pressure of the subject's blood plasma. Indeed, in certain embodiments of the invention, the tonicity agent is present in an amount sufficient to render the formulation suitable for intravenous infusion. Often, the tonicity agent serves as a bulking agent or a stabilizing agent as well. As such, the tonicity agent may allow the binding agent to overcome various stresses, such as freezing and shear. Tonicity agents may include, but are not limited to, saccharides, sugars, glycerol, sorbitol, mannitol, sodium chloride, potassium chloride, magnesium chloride, and other inorganic salts. Those skilled in the art are aware that other tonicity agents can be used as long as they are pharmaceutically acceptable, i.e. suitable for administration to subjects.

[0103] The tonicity agent is present in the formulations in an amount from 0.1% to 10% (w / v). For example, the tonicity agent may be present in the formulation in an amount of 0.1% (w / v), 0.2% (w / v), 0.3% (w / v), 0.4% (w / v), 0.5% (w / v), 0.6% (w / v), 0.7% (w / v), 0.8% (w / v), 0.9% (w / v), 1% (w / v), 2% (w / v), 3% (w / v), 4% (w / v), 4.5% (w / v), 5% (w / v), 5.5% (w / v), 6% (w / v), 7% (w / v), 8% (w / v), 9% (w / v), and 10% (w / v). Alternatively, the tonicity agent may be present in the formulation in an amount from 2% to 8% (w / v), from 3% to 7% (w / v), and from 4% to 6% (w / v). In further alternative embodiments, the tonicity agent may be present in the formulation in an amount from 0.1% to 1%, from 0.1% to 0.5%, from 0.1 to 0.3%, and 0.2%.

[0104] In certain exemplary embodiments, the tonicity agent is a saccharide. Examples of saccharides include glucose, sucrose (which is also known as saccharose), maltose, trehalose, dextrose, xylitol, fructose and mannitol. For example, mannitol is present in an amount of 4% (w / v). Alternatively, sucrose (which is also known as saccharose) may be present in an amount of 1% to 10% (w / v), 3% to 8% (w / v), or 4% to 6% (w / v), e.g., 4.5, 5, 5.5, or 6% (w / v).

[0105] In certain other exemplary embodiments, the tonicity agent is sodium chloride. For example, sodium chloride may be present in an amount of 0.1% (w / v), 0.2% (w / v), 0.3% (w / v), 0.4% (w / v), 0.5% (w / v), 0.6% (w / v), 0.7% (w / v), 0.8% (w / v), 0.9% (w / v), and 1% (w / v). Alternatively, sodium chloride may be present in the formulation in an amount from 0.1% to 1%, from 0.1% to 0.5%, from 0.1 to 0.3%, and 0.2%.

[0106] In further exemplary embodiments, the formulations may comprise one or more tonicity agents. For example, the formulations may comprise one or more of the above tonicity agents in the above concentrations. In certain specific embodiments, the formulations may comprise sucrose and sodium chloride, wherein each of the sucrose and sodium chloride concentrations is between 0.1% to 10% (w / v). In some embodiments, the sucrose concentration is 6% and the sodium chloride concentration is 0.2%. Alternatively, the sucrose concentration is 4.5% and the sodium chloride concentration is 0.2%.

[0107] In certain embodiments of the invention, the osmolality of the formulations range from 200 mOsm / kg to 350 mOsm / kg, 270 mOsm / kg to 330 mOsm / kg, 280 mOsm / kg to 320 mOsm / kg, or 290 mOsm / kg to 310 mOsm / kg, e.g., 300 mOsm / kg. In other words, the formulations of the invention are, in some embodiments, substantially isotonic, i.e. having substantially the same osmotic pressure as human blood. Osmolality can be measured by any means known to those of skill in the art, such as using vapor pressure or ice-freezing type osmometers. The osmolality of the formulations of the invention can, for instance, be regulated by the one or more tonicity agents described herein.vi. Amino Acids

[0108] The formulations of the invention may, optionally, further comprise an amino acid. Examples of amino acids include, but are not limited to, glycine, alanine, aspartic acid, lysine, serine, tyrosine, cysteine, glutamine, methionine, arginine, and proline. In exemplary embodiments, the amino acid is present in the formulations in an amount from 0.1% to 5% (w / v). For example, the amino acid may be present in the formulation in an amount of 0.1% (w / v), 0.2% (w / v), 0.3% (w / v), 0.4% (w / v), 0.5% (w / v), 0.6% (w / v), 0.7% (w / v), 0.8% (w / v), 0.9% (w / v), 1.0% (w / v), 1.1% (w / v), 1.2% (w / v), 1.3% (w / v), 1.4% (w / v), 1.5% (w / v), 1.6% (w / v), 1.7% (w / v), 1.8% (w / v), 1.9% (w / v), 2.0% (w / v), 3% (w / v), 4% (w / v), and 5% (w / v). Alternatively, the amino acid is present in the formulation in an amount from 1.3% to 1.8% (w / v), or 1.4% to 1.6% (w / v), e.g., 1.5% (w / v). In further alternative embodiments, the amino acid is present in the formulation in an amount from 0.5% to 1.5% (w / v), or 0.8% to 1.2% (w / v), e.g., 1.0% (w / v). An exemplary amino acid is proline or arginine. For example, proline may be present in the formulation in an amount from 1% to 2%, (w / v) 1.3% to 1.8% (w / v), 1.4% to 1.6% (w / v), e.g., 1.5% (w / v). Alternatively, arginine may be present in the formulation in an amount from 0.5% to 1.5% (w / v), or 0.8% to 1.2% (w / v), e.g., 1.0% (w / v).vii. Other excipients

[0109] Furthermore, the formulations of the invention may comprise other excipients including, but not limited to, water for injection, diluents, solubilizing agents, soothing agents, additional buffers, inorganic or organic salts, antioxidants, or the like. In some embodiments, however, the formulations of the invention comprise no other excipients, except those described above. Other pharmaceutically acceptable carriers, excipients, or stabilizers, such as those described in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980) may be included in the formulation provided that they do not adversely affect the desired characteristics of the formulation. In a particular embodiment, the formulation is substantially free of preservatives, although, in alternative embodiments, preservatives may be added as necessary. For example, cryoprotectants or lyoprotectants may be included in lyophilized formulations.viii. Liquid or lyophilized formulations

[0110] The formulations of the invention may either be liquid formulations or lyophilized formulations. In some embodiments, the formulations are liquid formulations. In some embodiments, the liquid formulations are ready for injection. Alternatively, the formulations may be lyophilized powders. In some embodiments, the lyophilized powders are ready to be combined with a solvent just prior to administration.ix. Exemplary formulations

[0111] In one exemplary embodiment of the invention, the invention provides a stable liquid antibody formulation suitable for subcutaneous administration, the formulation comprising: a) 150 mg / ml, of a fully human IgG4 anti-LIGHT (lymphotoxin-like, exhibits inducible expression and competes with HSV glycoprotein D for HVEM, a receptor expressed by T lymphocytes) antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 7 and a light chain comprising the amino acid sequence of SEQ ID NO: 8; b) 10 mM citrate buffer; c) 0.005% (w / v) polysorbate 20; and d) 4% (w / v) mannitol; wherein the pH of the formulation is pH 5.5

[0112] In certain exemplary embodiments, this formulation may be manufactured by: a) dissolving 10 mM sodium citrate dihydrate in water for injection and adjusting the pH of the buffered solution to pH 5.5, e.g., using either hydrochloric acid or sodium hydroxide; b) adding greater than 80 mg / ml, e.g., 150 mg / ml, of a fully human IgG4 anti-LIGHT (lymphotoxin-like, exhibits inducible expression and competes with HSV glycoprotein D for HVEM, a receptor expressed by T lymphocytes) antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 7 and a light chain comprising the amino acid sequence of SEQ ID NO: 8, 4% (w / v) mannitol, and 0.005% (w / v) polysorbate 20 to the buffered solution from step a) while stirring in a vessel made of inert material until completely dissolved, and then adjusting the pH of the resulting formulation to pH 5.5 using either hydrochloric acid or sodium hydroxide, and then adding buffered solution from step a) to adjust the final weight of the resulting formulation; c) filtering the formulation from step b) under aseptic conditions using a sterilized, compatible membrane filter having a nominal pore size of 0.2 µM, and then sterilizing the formulation by filtration under aseptic conditions into sterilized containers made out of inert material using a sterilized, compatible membrane filter having a nominal pore size of 0.2 µM; d) filling the formulation from step c) under aseptic conditions into sterilized vials that are closed with stoppers and flip-off caps with a flange; and, optionally, e) inspecting the containers from step d) for coarse contaminants, intact sealing, and visible particles.

[0113] A stable lyophilized antibody formulation suitable for intravenous administration is also described herein, the formulation comprising: a) 5 to about 80 mg / mL, e.g., 50 mg / mL, of a fully human IgG4 anti-LIGHT (lymphotoxin-like, exhibits inducible expression and competes with HSV glycoprotein D for HVEM a receptor expressed by T lymphocytes) antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 7 and a light chain comprising the amino acid sequence of SEQ ID NO: 8; b) 10 mM citrate buffer; c) 0.01% (w / v) polysorbate 20; d) 5% (w / v) sucrose; and e) 1.5% (w / v) proline; wherein the pH of the formulation is pH 5.5.x. Stability

[0114] The formulations of the invention are stable at 5°C for at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months or more, and typically at least about 12, 18 or 24 months or more. In exemplary embodiments, they are stable at 5°C for at least about 6 months or more. In other exemplary embodiments, they are stable at 5°C for at least about 9 months. In further exemplary embodiments, they are stable at 5°C for at least about 1 year or more, and typically about 2 years.C. Modes of Administration

[0115] In certain embodiments of the invention, the formulations are suitable for administration parenterally, intravenously, intramuscularly, intradermally, subcutaneously, or a combination thereof. The formulations of the invention are suitable for delivery by a variety of techniques.

[0116] In some embodiments of the invention, the formulation is administered intravenously. For example, it is desirable that formulations containing 80 mg / mL of IgG4 binding agent, such as an antibody, or less are administered intravenously. Therefore, the formulations are typically sterile. Methods for making formulations sterile are well known in the art and include, for example, filtration through sterile filtration membranes or autoclaving the ingredients of the formulation, with the exception of the antibodies, at about 120°C for about 30 minutes.

[0117] In some embodiments of the invention, the formulation is administered subcutaneously. For example, it is desirable that formulations containing more than 80 mg / mL of IgG4 binding agent, such as an antibody, are administered subcutaneously. In a specific embodiment, it is desirable to administer subcutaneously to subjects a stable liquid antibody formulation comprising: a) about 150 mg / mL of a fully human IgG4 anti-LIGHT (lymphotoxin-like, exhibits inducible expression and competes with HSV glycoprotein D for HVEM, a receptor expressed by T lymphocytes) antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 7 and a light chain comprising the amino acid sequence of SEQ ID NO: 8; b) 10 mM citrate buffer; c) 0.005% (w / v) polysorbate 20; d) a4% (w / v) mannitol; and wherein the pH of the formulation is about pH 5.5.D. Dosages and Dosage Forms

[0118] Effective doses of the formulations of the invention vary depending upon many different factors, including means of administration, target site, physiological state of the subject, whether the subject is human or an animal, other medications administered, and whether treatment is prophylactic or therapeutic. Usually, the subject is a human, but non-human mammals including transgenic mammals can also be treated. Treatment dosages may need to be titrated to optimize safety and efficacy.

[0119] The formulations of the invention may be administered on multiple occasions. Intervals between single dosages can be daily, weekly, biweekly, monthly or yearly. Intervals can also be irregular. In some methods, the dosage is adjusted to achieve a certain plasma binding agent, such as an antibody, concentration. Dosage and frequency will vary depending on the half-life of the anti-LIGHT binding agent, such as an antibody, in the subject. In general, human antibodies show the longest half-life, followed by humanized antibodies, chimeric antibodies, and nonhuman antibodies.

[0120] In further embodiments, the invention provides a pharmaceutical unit dosage form comprising a therapeutically effective amount of a formulation of the invention for the treatment of one or more diseases in a subject through administration of the dosage form to the subject. In some embodiments, the subject is a human. The human may be an adult or may be an infant. The term "pharmaceutical unit dosage form" refers to a physically discrete unit suitable as unitary dosages for the subjects to be treated, each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic / prophylactic effect in association with the required citrate buffer and pH.

[0121] The unit dosage form may be a container comprising the formulation. Suitable containers include, but are not limited to, sealed ampoules, vials, bottles, syringes, and test tubes. The containers may be formed from a variety of materials, such as glass or plastic, and may have a sterile access port (for example, the container may be a vial having a stopper pierceable by a hypodermic injection needle). In some embodiments, the container is a vial. Generally, the container should maintain the sterility and stability of the formulation.

[0122] In specific embodiments, the formulations are packaged in 2 mL vials that are made of clear, colorless type I glass, and closed with a stopper (fluoropolymer-coated bromobutyl) sealed with flip-of caps with flange (polypropylene). The vials are, in some embodiments, filled with 1.2 mL of the formulations so that the vial has an overfill volume of about 0.2 mL per vial, and an extractable volume of 1.0 mL. For example, this means that the dosage strength of anti-LIGHT antibody (e.g., 150 mg / mL) will be contained within 1 mL of solution.

[0123] In specific embodiment, the formulations are secondarily packaged in a container, such as a cardboard box, that protects the vials from light.F. Kits

[0124] Certain embodiments of the invention include a kit comprising a formulation of the invention. The kit may further comprise one or more containers comprising pharmaceutically acceptable excipients, and include other materials desirable from a commercial and user standpoint, including filters, needles and syringes. Associated with the kits can be instructions customarily included in commercial packages of therapeutic, prophylactic or diagnostic products, that contain information about, for example, the indications, usage, dosage, manufacture, administration, contra-indications, and / or warnings concerning the use of such therapeutic, prophylactic or diagnostic products. The kit can also be associated with a label that can be any kind of data carrier (e.g., a leaflet, sticker, chip, print or bar code) comprising information. In certain embodiments, the instructions etc. as listed above can be comprised in or on the label. The kit can further comprise a device for administration of the formulation, and particularly a device that contains the formulation, i.e., a pre-filled device such as, but not limited to, a pre-filled syringe or a pre-filled autoinjector. The kit can also comprise a container comprising the formulation, i.e., a pre-filled container, such as a pre-filled vial, cartouche, sachet, or ampoule.G. Combination of different embodiments

[0125] In the context of the present invention, any of the herein described embodiments can be combined with one or more of the other herein described embodiments unless explicitly stated to the contrary. Particularly, any of the herein described binding agents and antibodies and the herein described formulations thereof can be used in combination with any of the kits, pre-filled devices or pre-filled containers, or can be used in the methods of treatment or medical uses as described herein in connection with the respective antibody (e.g., the stable formulations comprising the anti-LIGHT antibodies can be combined with any of the herein described kits, containers or devices). Any of the herein described binding agents specifically binding an antigen (e.g., a binding agent specifically binding LIGHT) can also be used in any of the methods of treatment that are described herein in connection with the respective antibodies (i.e., anti-LIGHT) and vice versa.EXAMPLES

[0126] To help illustrate the invention, the following examples are provided. In general, the practice of the present invention employs, unless otherwise indicated, conventional techniques of pharmaceutical formulation, chemistry, molecular biology, recombinant DNA technology, immunology such as antibody technology, and standard techniques of polypeptide preparation as described, for example, in Sambrook, Fritsch and Maniatis, Molecular Cloning: Cold Spring Harbor Laboratory Press (1989); Antibody Engineering Protocols (Methods in Molecular Biology), volume 51, Ed.: Paul S., Humana Press (1996); Antibody Engineering: A Practical Approach (Practical Approach Series, 169), Eds.: McCafferty J. et al., Humana Press (1996); Antibodies: A Laboratory Manual, Harlow and Lane, Cold Spring Harbor Laboratory Press (1999); and Current Protocols in Molecular Biology, Eds. Ausubel et al., John Wiley & Sons (1992).Anti-LIGHT

[0127] A fully human IgG4 anti-LIGHT antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 7 and a light chain comprising the amino acid sequence of SEQ ID NO: 8 (the "Lead LIGHT Antibody") was used in Examples 1-9 in order to determine optimal formulation conditions.MATERIALS Drug substance batch

[0128] The Lead Antibody, formulated in phosphate buffered saline (PBS) at a concentration of 5.5 mg / mL and at a pH of 7.3 (the "Original Formulation", "PBS Formulation", or "Reference Lot"), was used in the following examples.Excipients

[0129] Table 1 shows the excipients that were used in the following examples, which were chosen according to their acceptability / suitability for use in parenteral products. Table 1 - Excipients used in this study Excipients Art. No. / Charge Supplier Arginine1.01587MerckCitric acid100241MerckHCl114027MerckSodium acetate1.06265MerckSodium chloride10158Riedel de HaenSodium hydroxide114076MerckSodium citrate114196Boehringer Ingelheim KGPolysorbate 20139850FlukaTrehalose-dihydratT9531Sigma-Aldrich METHODS

[0130] The following methods were used to manufacture the experimental formulations and the formulations of the invention containing the Lead LIGHT Antibody.Manufacturing & composition of buffers

[0131] All buffers were manufactured under stirring to dissolve the respective excipients. pH was adjusted using 0.1 M HCl or 0.1 M NaOH. The general concentration of all buffers was 10 mM.Manufacturing & composition of excipient stock solutions

[0132] All stock solutions were manufactured under stirring to dissolve the excipients. Concentration was given as weight / weight (w / w).Sterile filtration of samples

[0133] All samples, solutions, buffers, etc. were sterile filtered (0.22 µm) using a Sartopore-2 membrane. The samples were filtered into sterilized bottles or vials and closed under aseptic conditions inside a clean-bench to prevent microbiological contamination.Mechanical stress test

[0134] Mechanical stress with an agitation speed of 350 / minute for 2.5 hours at room temperature was performed using a horizontal laboratory shaker with a 26 mm distance (shaker & incubation hood from Bühler Company). 2R vials were filled with 1 mL solution with a head space of about 2.5 mL. The mechanical stress test was planned and performed during the first pre-formulation studies and during relevant studies for surfactant selection.Thermal stress test

[0135] Thermal stress was used as a stress test during all steps of the pre-formulation program. The samples were stored at +40°C for either 24 hours or 7 days, depending on the study.Analytical Methods in Formulation Fill and Finish

[0136] The following analytical methods were used in the formulation fill and finish in the following examples.Appearance

[0137] Appearance of the antibody solutions were checked visually, and additionally documented by taking a picture with a digital camera.pH

[0138] All pH measurements were performed using a pH-meter with a micro-electrode.Concentration using UV

[0139] The protein concentrations of all antibody solutions were measured against buffer using a NanoDrop ND1000. Proteins concentrations near or below 5 mg / mL were diluted 1:3, while higher protein concentrations near 20 mg / mL were diluted 1:20, and the absorption was measured at 215 nm and 280 nm.Dynamic light scattering (DLS)

[0140] The hydrodynamic diameter of the molecule was measured using laser light scattering. The samples were sterile filtered prior to the analytics if turbidity was observed, thus only soluble aggregates could be detected.Differencial scanning calorimetry (DSC)

[0141] Aliquots of most pre-formulation samples were examined by DSC using a VPCapillary DSC from Microcal and scanned in the autosampling instrument at 90°C / hour with a filter time of 2 seconds. 400 µl samples were placed into 96-well plates and analyzed for the unfolding temperature Tm.Osmolarity

[0142] Osmolarity was measured using an automated Knaur Osmometer.Density

[0143] Density of the formulations was measured using a falling sphere viscosimeter DMA4500 Anton Paar.Analytical methods in Bioanalytics FF

[0144] The following analytical methods were used in the bioanalytics fill and finish in the following examples.Size Exclusion Chromatography (SEC)

[0145] Aggregates, as well as degradation products of the Lead Antibody, were quantified using size exclusion GL chromatography. The test was carried out by isocratic HPLC with a SUPERDEX 200 10 / 300 column.SDS -PAGE, reducing and non-reducing

[0146] Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was used to analyze the molecular integrity (e.g., half molecules) and the purity. This electrophoretic analysis was performed with 4-12% gradient gels under reducing and non-reducing conditions. The proteins were visualized with Coomassie staining after electrophoretic separation.Weak Cation Exchange (WCX)

[0147] Weak cation exchange chromatography was used to monitor the charge heterogeneity of the antibody. The percentages of basic, neutral, and acidic isoforms were reported. The test was carried out by discontinuous high performance liquid chromatography (HPLC) with a ProPac WCX10 column.Antigen-Enzyme Linked Immunosorbent Assay (Antigen-ELISA)

[0148] Antigen-ELISA was performed to determine the functionality of the antibody. The binding properties to native LIGHT protein were monitored in comparison to the current standard of the antibody. This potency was reported as the relative EC 50 .Isoelectric focusing (IEF)

[0149] IEF was performed. The isoelectric pattern was specific for the Lead Antibody and served as an identification test. Degradation could be seen by a different charge pattern.Storage

[0150] All buffer solutions, excipient solutions, and samples were stored at 5°C (± 3°C), if not otherwise mentioned.SUMMARY OF ALL FORMULATIONS PREPARED & ANALYZED

[0151] Table 2 below shows a summary of all of the formulations that were prepared and analyzed in the following examples. Each of the formulations contained the Lead LIGHT Antibody at the concentration listed. Table 2 - Summary of all formulations prepared and analyzed Sample number Buffer pH Concentration Comment Formulation 1.1Citrate 10 mM5.05.5 mg / mLFormulation 1.2Citrate 10 mM5.55.5 mg / mLFormulation 1.3Citrate 10 mM6.05.5 mg / mLFormulation 2PBS7.3< 80 5 mg / mLmg / mLVery turbidCitrate 10 mM5.0Formulation 3.1PS20 0.01 %5 mg / mLCitrate 10 mM5.5Formulation 3.2PS20 0.01 %5 mg / mLCitrate 10 mM5.5Formulation 4PS20 0.01 %80 mg / mLclearCitrate 10 mM5.0Formulation 5PS20 0.01 %5 mg / mLCitrate 10 mM5.5PS20 0.01 %Proline 1.5 %Formulation 6.1Sucrose 5 %50 mg / mLLyoCitrate 10 mM5.5PS20 0.01 %Formulation 6.2Sucrose 5 %50 mg / mLLyoFormulation 7Histidine 10 mM5.550 mg / mLHistidine 10 mM5.5Formulation 8PS20 0.01 %50 mg / mLFormulation 9Citrate 10 mM5.550 mg / mLCitrate 10 mM5.5Formulation 10PS20 0.01 %50 mg / mLCitrate 10 mM5.5Formulation 11Sucrose 5 %50 mg / mLlyoFormulation 12Citrate 10 mM7.05 mg / mLµDSCFormulation 13PBS5.05 mg / mLµDSC EXAMPLE 1 - Characterization of a phosphate buffered saline (PBS) formulation and disadvantages associated therewith

[0152] In this example, the Reference Lot was characterized. As stated in the Materials section above, the Reference Lot contains the Lead LIGHT Antibody formulated in phosphate buffered saline (PBS) at a concentration of 5.5 mg / mL and at a pH of 7.3, and produced in research solutions Vitry (BioSCP).

[0153] Isoelectric focusing (IEF) was used to determine the isoelectric point (pI) of the Lead Antibody. The pI of the Lead LIGHT Antibody was theoretically calculated as 6.28, and then measured by denaturated isoelectric focusing using standard methods known in the art. As shown in Figure 1, the main bands show that the pI of the Lead LIGHT Antibody was 6.8-7.2.

[0154] SDS-PAGE was used to identify the molecular weight of the antibody monomer, potential aggregates, or the presence of half-molecules. Figure 2 shows an SDS-PAGE gel that compared different Reference Lot batches under reducing and non-reducing conditions. An ELISA was used to determine the antigen binding activity of the Lead LIGHT Antibody. Figure 3 shows an ELISA graph that was used to determine the antigen binding activity of the first and second batches of Reference Lot.

[0155] SEC was used to determine the presence of aggregates, as well as degradation products of the first batch of Reference Lot. As shown in Figure 4, size exclusion chromatography detected high molecular weight proteins (HMWP), e.g., di- / oligomers (RRT0.8) or aggregates, and low molecular weight proteins (LMWPs) or degradation products. The first batch of Reference Lot had a purity of 97% monomer content.

[0156] WCX was used to monitor the charge heterogeneity of the first batch of Reference Lot. As shown in Figure 5, rearrangements of acidic, neutral, and basic isoforms occured during stability studies. The first batch of Reference Lot had a distribution of acidic / neutral / basic isoforms of 42.3 / 55.6 / 1.9%.

[0157] DSC was used to analyze the unfolding temperature Tm of the first batch of Reference Lot. As shown in Figure 6, the three domains of the antibody unfold at 68°C, 75°C, and 78°C.

[0158] DLS was used to determine the hydrodynamic diameter of the antibody monomer and potential soluble aggregates. As shown in Figures 7 & 8, a hydrodynamic diameter of about 10 nm was detected, but aggregates were seen in PBS. However, aggregates were not seen in citrate buffer (Figure 10).EXAMPLE 2 - Development of Citrate-Buffered Formulations, and advantages associated therewith

[0159] The original buffer, phosphate buffered saline (PBS) at a pH of 7.3, was, in terms of pH, very close to the isoelectric point (pI) of the Lead Antibody (see Example 1). In addition, the Original Formulation exhibited aggregates; half-molecules; degradation products; low molecular weight proteins (LMWPs); high molecular weight proteins (HMWPs); and rearrangements of acidic, basic, and neutral antibody isoforms (see Example 1). Thus, there was a need for an improved formulation that does not suffer from these disadvantages.

[0160] Formulations of the Lead LIGHT Antibody (a fully human IgG4 anti-LIGHT antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 7 and a light chain comprising the amino acid sequence of SEQ ID NO: 8) containing 10mM citrate buffer at a pH of 5, 5.5, and 6, with and without polysorbate 20 were tested. Table 3 shows the analytical results of the first batch of Reference Lot, and the various experimental formulations of the Lead LIGHT Antibody formulated into citrate, at a pH of 5.0 and 5.5 and 6.0, with and without polysorbate 20. Aggregates were found in dynamic light scattering (DLS) measurements for the Reference Lot, but not in all other tested formulations. Tm, as measured by differencial scanning calorimetry (µDSC), indicated that the higher the pH, the higher the thermodynamic stability could be assumed. But for high antibody concentrated formulations, the pH had to be chosen below the pI of the antibody.

[0161] As shown in Table 4, size exclusion chromatography (SEC) data showed a significantly reduced amount of high molecular weight proteins (HMWPs) for the Lead LIGHT Antibody in citrate buffer as compared to the Reference Lot (phosphate buffer at pH 7.3). In contrast, no differences could be detected with SDS-PAGE (Table 5). Table 3 - Analytical Results of Formulations Sample number Tm1 [°C] Tm2 [°C] Tm3 [°C] pH ZAve [nm] Concentration [mg / mL] Buffer Reference Lot67.9475.0077.377.3179.855.5PBSFormulation 1.158.3969.9875.755.010.975.0Citrate 10 mMFormulation 1.262.0272.2676.595.510.715.0Citrate 10 mMFormulation 1.365.4673.7477.026.010.815.0Citrate 10 mMFormulation 3.158.3369.9375.745.013.145.0Citrate 10 mMPS20 0.01 %Formulation 3.261.4271.9776.455.512.795.0Citrate 10 mMPS20 0.01 % Table 4 - SEC data of Formulations ANTIBODY RRTO.8 LMWP HMWPs Sample NameAreaRel.Area AreaRel.Area AreaRel.Area AreaRel.Area Monomer ContentmAU*min% mAU*min% mAU*min% mAU*min% [mg / mL]Ref. Lot255.6198.00 3.981.52 1.500.57 0.590.23 Formulation 3.1223.2398.07 3.221.42 1.010.44 0.160.07 45.49Formulation 3.2257.0998.24 3.741.43 0.790.30 0.090.03 48.92 Table 5 - SDS-PAGE data of Formulations Sample NamesizeRel.QTYsizeRel.QTYsizeRel.QTYsizeRel.QTYcommentkDa%kDa%kDa%kDa%Ref. Lot172.598.4150.11.468.40.2Additional bands <0.5%Formulation 3.1166.197.7147.8271.50.3Identical pattern to Ref. LotFormulation 3.2166.296.2147.23.471.40.4Identical pattern to Ref. Lot EXAMPLE 3 - Development of High-Concentration Antibody Formulations

[0162] In view of the improvements provided by the Citrate-Buffered Antibody Formulation of Example 2, the citrate buffer components were optimized for increased concentrations of Lead LIGHT Antibody. Table 6 shows the analytical results of the first batch of high concentration (about 40 mg / ml) antibody formulations: high phosphate buffered saline (PBS) at a pH of 7.3 (Formulation 2) or citrate at a pH of 5.5 with polysorbate 20 (Formulation 4). Table 6 - Analytical results of Formulations 2 & 4 Sample number Tm1 [°C] Tm2 [°C] Tm3 [°C] pH ZAve [nm] Concentration [mg / mL] Buffer Reference Lot67.9475.0077.377.310.055.5PBSFormulation 267.8774.8777.287.312.8942.1PBSFormulation 461.5572.0076.485.516.7139.97Citrate 10 mMPS20 0.01 %

[0163] Slightly reduced monomer content was observed after concentrating the protein solution in citrate buffer. Moreover, dimer concentration was reduced and high molecular weight proteins (HMWPs) could be significantly reduced as well (see Table 7). In contrast, these impurities and byproducts were increased by increasing the concentration in phosphate buffer. No differences could be detected with SDS-PAGE analysis (Table 8). Table 7 - SEC data of Formulations 2 & 4 SEC Analysis ANTIBODY RRT0.8 LMWP HMWPs Sample NameAreaRel.AreaAreaRel.AreaAreaRel.AreaAreaRel.AreaMonomer GehaltmAU*min%mAU*min%mAU*min%mAU*min%[mg / mL]Ref. Lot255.6198.00 3.981.52 1.500.57 0.590.23 Formulation 2121.4297.39 2.131.71 0.980.79 0.150.12 44.08Formulation 4141.9097.65 2.171.49 1.160.80 0.090.06 45.83 Table 8 - SDS-PAGE data of Formulations 2 & 4 SDS-PAGE Analysis Antibody Main 2. band Half molecules Additional bands Sample NamesizeRel.QTYsizeRel.QTYsizeRel.QTYsizeRel.QTYcommentkDa%kDa%kDa%kDa%Ref. Lot172.598.4150.11.468.40.2Additional bands <0.5%Formulation 2170.697.9147.61.972.20.2Identical pattern to Ref. LotFormulation 417197.21492.570.50.3Identical pattern to Ref. Lot EXAMPLE 4 - Development of Lyophilized Antibody Formulations

[0164] To test the feasibility of lyophilization, different lyophilized experimental formulations were manufactured and subjected to stability analysis. The concentration of the Lead LIGHT Antibody was increased to 50 mg / mL.

[0165] Table 9 shows the freeze drying program that was used in this example. Table 9 - Freeze drying program Lyo program (vacuum) N° 8 Chamber loading5 min / RT / 100%Freezing2 h / -45°C / 100%Main drying I30 min / -45°C / 30%Main drying II5h / -20°C / 30 %Main drying III8h / +20°C / 30%Final drying2 h / + 20°C / 3%

[0166] Table 10 shows the analytical results of the first batch of Reference Lot, and the various experimental lyophilized formulations of the Lead LIGHT Antibody formulated into various combinations of citrate buffer, sucrose, polysorbate 20, and proline.

[0167] As shown in Table 11, high molecular weight proteins (HMWPs) could clearly be reduced by using citrate buffer. No differences in dimer content were seen over the time of storage at 40°C. An increase of low molecular weight proteins (LMWPs) after freeze drying was observed. As before, these differences could not be detected with SDS-PAGE analysis (Table 12). Table 10 - Analytical data of Formulations 6-6.2 & 11 Sample number Tm1 Tm2 Tm3 Time / Temp. pH ZAve [nm] Concentration Buffer Reference Lot67.9475.0077.377.310.055.5 mg / mLPBSFormulation 6NdNdNdN / A5.717.4657.32Citrate 10 mMPS20 0.01 %Formulation 6.164.3072.6177.02T05.759.66NdCitrate 10 mMT1 / 5°C5.718.85NdPS20 0.01 %T1 / 40°C5.719.12NdProlin 1.5 %T2 / 5°C5.7NdNdSucrose 5%T2 / 40°C5.7NdNdFormulation 6.265.4575.0879.37T05.719.58NdCitrate 10 mMT1 / 5°C5.731.34NdPS20 0.01 %T1 / 40°C5.718.1NdSucrose 5%T2 / 5°C5.7NdNdT2 / 40°C5.7NdNdFormulation 1168.8475.6177.91T07.098.6056.49PBST1 / 5°C5.720.22NdSucrose 5%T1 / 40°C5.722.68NdT2 / 5°C5.7NdNdT2 / 40°C5.7NdNd Table 11 - SEC data of Formulations 6.1-6.2 & 11 SEC Analysis ANTIBODY RRT0.8 LMWP HMWPs Sample NameTimeArea [mAU*min]Rel.Area [%]Area [mAU*min]Rel.Area [%]Area [mAU*min]Rel.Area [%]Area [mAU*min]Rel.Area [%]Ref. Lot255.6198.00 3.981.52 1.500.57 0.590.23 Formulation 6.1T0222.9497.43 3.891.70 1.910.83 0.090.04 Formulation 6.1T1 5°C369.7297.57 6.311.66 2.750.73 0.180.05 Formulation 6.1T1 40°C405.4997.35 7.331.76 3.600.86 0.120.03 Formulation 6.1T2 5°C422.4697.59 7.011.62 2.740.63 0.680.16 Formulation 6.1T2 40°C289.6597.28 5.501.85 2.130.72 0.480.16 Formulation 6.2T0230.0697.61 3.931.67 1.640.70 0.070.03 Formulation 6.2T1 5°C407.1797.56 6.811.63 3.230.77 0.170.04 Formulation 6.2T1 40°C468.7497.36 8.791.83 3.780.78 0.160.03 Formulation 6.2T2 5°C552.3197.64 9.801.73 2.960.52 0.610.11 Formulation 6.2T2 40°C249.9596.78 5.392.09 2.390.93 0.520.20 Formulation 11T0211.4597.49 3.641.68 1.470.68 0.350.16 Formulation 11T1 5°C339.0897.71 5.451.57 2.280.66 0.230.07 Formulation 11T1 40°C700.9197.30 12.691.76 5.190.72 1.600.22 Formulation 11T2 5°C325.8097.17 5.801.73 2.170.65 1.520.45 Formulation 11T2 40°C229.2996.96 4.331.83 1.780.75 1090.46 Table 12 - SDS-PAGE data of Formulations 6.1-6.2 & 11 SDS-PAGE Analysis ANTIBODY Main 2.Band HM Additional bands Sample NameSample IDsizeRel.QTYsizeRel.QTYsizeRel.QTYsizeRel.QTYcommentRef. Lot182.995.6161.22.373.80.5Formulation 6.1T0175.694.7156.12.773.50.5Formulation 6.1T1 5°C180.286.9159.911.475.90.1Formulation 6.1T1 40°C179.290.4158.57.576.10.4Formulation 6.1T2 5°C177.395.6157.92.174.90.3Formulation 6.1T2 40°C179.894.7159.82.975.40.3Formulation 6.2T0176.694.9156.32.673.60.5Formulation 6.2T1 5°C180.289.8159.37.976.30.4Formulation 6.2T1 40°C182.188.7160.99.476.30.1Formulation 6.2T2 5°C177.595.5160.22.975.40.2Formulation 6.2T2 40°C180.995.5161.52.475.70.3Formulation 11T0178.795.1156.52.373.70.4Formulation 11T1 5°C181.070.0154.725.774.50.3Formulation 11T1 40°C181.366.2154.228.974.50.3Formulation 11T2 5°C177.787.5155.910.975.20.3Formulation 11T2 40°C176.886.2155.212.074.50.3 EXAMPLE 5 - Accelerated Stability Study

[0168] An accelerated stability study was performed with citrate and histidine buffers. Table 13 shows the analytical results of the first batch of Reference Lot, and the various xperimental formulations of the Lead LIGHT Antibody formulated into various combinations of citrate buffer or histidine buffer. Notably, the citrate formulation of the invention appeared in all experiments to perform better than histidine. In particular, citrate formulations had a higher monomer content compared to the both the Reference Lot batch and the histidine (Table 13) and the content or low molecular weight proteins (LMWPs) and high molecular weight proteins (HMWPs) were also significantly lower (Table 14). As before, these differences could not be detected with SDS-PAGE analysis (Table 15). Table 13 - Analytical data of Formulations 7, 8, 9 & 10 Sample number Tm1 [°C] Tm2 [°C] Tm3 [°C] pH ZAve [nm] Concentration [mg / mL] Buffer Ref. Lot67.9475.0077.377.310.055.5PBSFormulation 758.9568.5176.205.512.9753.65Histidine 10 mMFormulation 858.6968.2376.125.413.2958.72Histidine 10 mMPS20 0.01 %Formulation 961.6772.0176.535.659.2655.01Citrate 10 mMFormulation 1062.2472.3276.615.617.355.8Citrate 10 mMPS20 0.01 % Table 14 - SEC Analysis of Formulations 7 & 8 & 9 & 10 ANTIBODY RRT0.8 LMWP HMWPs Sample NameTimeArea [mAU*min]Rel.Area [%]Area [mAU*min]Rel.Area [%]Area [mAU*min]Rel.Area [%]Area [mAU*min]Rel.Area [%]Monomer Content [mg / mL]Ref. Lot282.4297.40 4.551.57 2.150.74 0.850.29 Formulation 7T0184.495.1 163.81.6 72.81 Formulation 7T1 5°C390.8497.85 5.721.43 2.800.70 0.060.01 Formulation 7T1 40°C379.8196.74 7.041.79 5.771.47 00 Formulation 7T2 5°C863.0197.75 14.191.61 4.140.47 1.540.18 164.88Formulation 7T2 40°C1085.9195.22 29.592.60 23.252.04 1.690.15 207.47Formulation 8T0184.694.9 165.92.1 72.70.8 Formulation 8T1 5°C507.6497.74 7.521.45 4.190.81 0.050.01 Formulation 8T1 40°C461.4496.98 8.051.69 6.191.30 00 Formulation 8T2 5°C416.5497.46 6.591.54 3.490.82 0.790.18 79.58Formulation 8T2 40°C422.2193.23 11.172.47 18.404.06 1.110.25 80.66Formulation 9T0229.0197.63 3.751.60 1.630.70 0.190.08 45.28Formulation 9T1 5°C307.9497.96 4.21.34 2.200.7 00 Formulation 9T1 40°C319.1097.54 5.241.60 2.590.79 0.230.07 Formulation 9T2 5°C337.1597.48 5.411.56 2.840.82 0.490.14 64.41Formulation 9T2 40°C325.5496.26 7.782.30 3.661.08 1.200.36 62.20Formulation 10T0233.1197.54 3.841.61 1.970.82 0.080.03 46.09Formulation 10T1 5°C343.3897.77 5.211.48 2.580.73 0.040.01 Formulation 10T1 40°C329.5697.21 5.061.49 4.291.26 0.130.04 Formulation 10T2 5°C343.3397.43 5.471.55 3.060.87 0.530.15 65.59Formulation 10T2 40°C257.2094.59 5.592.06 8.983.30 0.150.05 49.14 Table 15 - SDS-PAGE data of Formulations 7 & 8 & 9 & 10 SDS-PAGE Analysis ANTIBODY Main 2. band Half molecules Additional bands Sample NameTime / Tempsize [kDa]Rel.QTY [%]size [kDa]Rel.QTY [%]size [kDa]Rel.QTY[ %]size [kDa]Rel.QTY [%]commentRef. Lot173.696.3155.82.2740.4Formulation 7T0184.495.1163.81.672.81Formulation 7T1 5°C183.091.1159.97.276.10.4Formulation 7T1 40°C182.283.1158.413.874.00.4Formulation 7T2 5°C181.595.7160.32.775.60.3Formulation 7T2 40°C173.084.6151.110.373.90.712.10.9more LMWPsFormulation 8T0184.694.9165.92.172.70.8Formulation 8T1 5°C180.186.2158.311.473.90.4Formulation 8T1 40°C180.979.4158.216.974.00.3Formulation 8T2 5°C175.195.2154.93.174.40.3Formulation 8T2 40°C174.884.7150.59.274.00.912.11.5more LMWPsFormulation 9T0187.795.5163.11.172.90.9Formulation 9T1 5°C178.965.8160.429.673.70.9Formulation 9T1 40°C184.782.9160.314.874.40.3Formulation 9T2 5°C176.295.6155.62.673.60.3Formulation 9T2 40°C174.391.5153.93.173.10.312.10.2more LMWPsFormulation 10T0182.595.2161.31.672.30.8Formulation 10T1 5°C184.568.4156.426.675.10.3Formulation 10T1 40°C180.865.4153.828.874.80.3Formulation 10T2 5°C188.788.6165.09.673.50.2Formulation 10T2 40°C181.778.9158.215.875.70.812.61.3more LMWPs EXAMPLE 6 - Development of High Antibody Concentration Formulation for Subcutaneous Administration

[0169] Based on the successful results of the citrate-buffered formulations of Examples 2-5, a high-concentration (150 mg / ml) antibody formulation suitable for subcutaneous administration was developed. Formulation development was performed on the Lead LIGHT Antibody with the goal of developing a liquid dosage form with an acceptable shelf life when stored at +2 to +8°C. Preliminary stress studies showed the formation of subvisible and visible particles, high molecular weight species and more basic species. Therefore, these parameters were monitored during the screening of formulation candidates using visual assessment, dynamic light scattering, light obscuration, size exclusion chromatography, sodium dodocyl sulphate polyacrylamide gel electrophoresis, and weak cationic exchange chromatography. Different liquid formulations were used in the pre-formulation and formulation trials prior to selection of the clinical formulation. According to the findings, a formulation in 10 mM citrate buffer adjusted to pH 5.5 (Formulation 14) was selected for further development. The pH of the formulation is in the region of optimal physical and chemical stability of the drug substance and acceptable physiological tolerability (e.g., osmolarity).

[0170] As shown in Table 16, Formulation 14 is a solution for injection and is an aqueous, sterile, and clear solution containing the Lead LIGHT Antibody, sodium citrate dihydrate (buffering agent), polysorbate 20 (stabilizing agent), and mannitol (tonicity agent). Sodium hydroxide solution and hydrochloric acid were used to adjust the pH to 5.5.

[0171] All excipients were soluble and well tolerated pharmacopoeial standard excipients for parenterals and listed in Ph. Eur. and USP. Table 16 - Composition Components a< Composition Function Reference to standards b< per mL (mg) per vial (1.2 mL) (mg) Lead Antibody150.00180.00Drug substanceIn-houseSodium citrate dehydrate2.943.53Buffering agentPh. Eur., USPMannitol40.0048.00Tonicity agentPh. Eur., USPPolysorbate 200.050.06Stabilizing agentPh. Eur., NF, JPHydrochloric acid, concentrated [Hydrochloricq.s pH 5.5q.s. pH 5.5Acidifying agentPh. Eur., NFacid]Sodium hydroxideq.s.pH 5.5q.s. pH 5.5Alkalizing agentPh. Eur., NFWater for injectionq.s. 1 mLq.s. 1.2 mLSolventPh. Eur., USPNitrogenProcess aid for filtrationPh. Eur., NF a< Components are listed according to their pharmacopoeial names. If more than one monograph exists, other names are given in brackets, along with the compendial origin. b< Reference is made to the current edition of the Pharmacopoeia. EXAMPLE 7 - Manufacturing Process for Subcutaneous Antibody Formulation

[0172] A GMP-compliant manufacturing process was developed for the subcutaneous, high-concentration antibody formulation (Formulation 14) of Example 6. The manufacturing procedure consisted of dissolving, pH adjustment, sterile filtration, filling, and packaging steps.

[0173] Drug substance (the Lead LIGHT Antibody) is provided in a liquid form in the formulation buffer (10 mM citrate buffer at pH 5.5). The excipients were all water-soluble and dissolved in the initial aqueous portion of the formulation buffer during manufacture. The bulk drug substance solution was further diluted with the same formulation buffer to reach the concentration of 150 mg / mL of Lead LIGHT Antibody. The bulk solution was well mixed to facilitate the dissolution process and to ensure homogeneity.

[0174] Sterilization by filtration was carried out (according to Ph. Eur. and USP) using bacteria retentive filters having a nominal pore size of 0.2 µm. An additional filtration procedure before "sterilization by filtration" was performed to ensure a low bioburden. Bioburden sampling was done before the pre-filtration step as well as the sterile filtration step.

[0175] Preparation and filling of the sterilized solution into the suitable containers was performed under aseptic conditions. This was in accordance with pharmacopoeial monographs and related guidelines, which required sterilization by filtration and subsequent aseptic processing. The equipment, which comes into contact with the product after the step "sterilization by filtration", was sterilized by heat or steam using a validated method (according to Ph. Eur. / USP).

[0176] Vials were filled to about 1.2 mL to ensure an extractable volume of 1.0 mL. The 2 mL vials were made of clear, colorless type I glass, and closed with a stopper (fluoropolymer-coated bromobutyl) sealed with flip-off caps with a flange (polypropylene). The primary packaging materials met the requirements of the Ph. Eur. and USP. The suitability of the primary packaging materials was substantiated by the results of the stability tests. Incompatibilities with the primary packaging material used were not observed. Secondary packaging which provides protection of the product from light.

[0177] Compatibility with application syringes was assessed using 3 different syringe sizes and needles diameters (between 25 and 28 gauges) on the drug product solution. No differences in terms of product quality were obtained. Compatibility was proven for a time period of 8 hours.

[0178] Formulation 14 was made in 5L batches, the composition of which is shown in Table 17. However, the batch size may be adjusted according to clinical requirements. Table 17 - Batch formula Components Batch size 5 Liter a< [g]Lead Antibody b< 750.00Mannitol200.00Polysorbate 200.25Sodium citrate dihydrate14.71Hydrochloric acid, concentrated c< q.s. pH 5.5Sodium hydroxide c< q.s. pH 5.5Water for injectionAd 5285.25 d< NitrogenProcess aid for filtration a< The vials were filled with a volume of 1.2 mL to ensure an extractable volume of 1.0 mL. A 6.0 L batch size therefore results in a theoretical yield of 5000 vials. b< For pH adjustment. c< This was calculated according to the density of the final drug product solution (1.05705 mg / mL)

[0179] The manufacturing process and process controls for Formulation 14 are shown in the flow diagram in Figure 9. Batch manufacturing included the following steps: I. Sodium citrate dihydrate was dissolved in water for injection while stirring in a vessel made of inert material (e.g., stainless steel or glass), until completely dissolved. The pH value was adjusted to 5.5 using hydrochloric acid, diluted (e.g., 0.1 M hydrochloric acid) and / or sodium hydroxide solution (e.g., 0.1 M sodium hydroxide), if necessary. II. Lead Antibody, mannitol, and polysorbate 20 were diluted in the buffer solution from step 1 while stirring in a vessel made of inert material (e.g., stainless steel or glass) until completely dissolved. If necessary, the pH value was adjusted to 5.5 using hydrochloric acid, diluted (e.g., 1 M hydrochloric acid) or sodium hydroxide solution (e.g., 1 M sodium hydroxide). Buffer solution from step 1 (remainder) was added to adjust the final weight. III. a) Pre-filtration: Solution from step II was filtered under aseptic conditions using a sterilized, compatible membrane filter (e.g., polyether sulfone or polyvinylidene difluoride) having a nominal pore size of 0.2 µm. b) Sterilization by filtration: Solution from step IIII.a was sterilized by filtration under aseptic conditions into sterilized containers made out of inert material (e.g., stainless steel or glass) using a sterilized, compatible membrane filter (e.g., polyether sulfone or polyvinylidene difluoride) having a nominal pore size of 0.2 µm. IV. Solution from step III.b was filled under aseptic conditions into sterilized vials, which were closed with stoppers and flip-off caps with a flange. V. The containers from step IV were inspected for coarse contaminants, intact sealing, and visible particles. VI. The inspected containers from step V were additionally packaged in suitable containers (e.g., cardboard boxes).

[0180] In addition, DLS was used to determine the hydrodynamic diameter of the antibody monomer and potential soluble aggregates. As shown in Figure 10, aggregates were not seen in citrate buffer. However, as shown in Figures 7 & 8, aggregates were seen in PBS. Due to the higher concentration of antibody, an increase in ZAve to 28 nm was observed, compared to the sample in PBS.EXAMPLE 8 - Stability Profile for Subcutaneous Antibody Formulation

[0181] The stability profile of the clinical batch (batch 2) of Example 7 was assessed for storage under long term and accelerated testing conditions according to ICH guidelines. Samples were packed and stored in 2 mL clear and colorless vials (glass type I) closed with stoppers (fluoropolymer-coated bromobutyl) and flip-off caps with a flange (polypropylene). The following tests were performed during stability: appearance (clarity, color), assay (antigen ELISA, UV), purity (SEC, SDS-PAGE under reducing and non-reducing conditions), molecular integrity (SDS-PAGE under non-reducing conditions), charge heterogeneity (weak cation exchange chromatography, isoelectric focusing), pH, sterility, bacterial endotoxins, particulate matter (visible and subvisible particles), and closure integrity.

[0182] The samples were stored in inverted and upright positions. The results of the inverted storage were presented as the more stringent condition. Stability data at -20°C, +5°C and +25°C are presented in Tables 18-20, respectively. The investigations on physical, chemical, and biological properties of storage under long term testing conditions confirmed a good stability of the drug product at 5°C (see Table 19). Under accelerated testing conditions (+25°C), only a slight decrease in the purity was detected by size exclusion chromatography (see Table 20). Therefore, it was concluded that the drug product should be stored at +2°C to +8°C protected from light. EXAMPLE 9 - Development of Ultra-High Antibody Concentration Formulation for Subcutaneous Administration

[0183] Based upon the successful results of the citrate-buffered formulations for antibody concentrations up to 150 mg / mL in Example 7, higher concentrated (up to 250 mg / ml) antibody formulations suitable for subcutaneous administration were developed.

[0184] Preliminary data showed that the formulation of antibody concentrations above 150 mg / mL may lead to higher viscosities affecting usage of the formulation. Table 21 - Ultra high concentrations with formulation 14 Sample Concentration [mg / mL] Density [kg / m-3] Viscosity [mPa] DLS Size exclusion chromatography z-average [nm]HMWPsMonomerLMWPsLead LIGHT Antibody_11_30A2371.06642.29301.398.60.0Lead LIGHT Antibody_11_30B2121.05922.58391.398.70.0Lead LIGHT Antibody_11_30C1811.05213.57281.398.70.1Lead LIGHT Antibody_11_30D1731.0468.8271.298.80.0Lead LIGHT Antibody_11_30E1431.0396.16251.198.80.1

[0185] As can be seen in Table 21, the viscosity decreases with lower antibody concentrations, yet still being in an acceptable range at the higher concentration formulated with formulation 14. All other parameters seemed to be unaffected or just slightly affected by the ultra-high concentrations.

[0186] As shown in Table 22, the antibody concentrations did not affect the stability of the formulations, which was indicated by identical 1 month stability data at long term and stress conditions. Table 22 - 1 month stability data of ultra high concentrated Lead Antibody formulations Concentration [mg / mL] HMWPs Monomer LMWPs Lead LIGHT Antibody_11 30A 40°C2374.795.20.2Lead LIGHT Antibody _11_30B 40°C2124.495.40.2Lead LIGHT Antibody _11_30C 40°C1815.891.72.6Lead LIGHT Antibody _11_30D 40°C1733.996.00.2Lead LIGHT Antibody _11_30E 40°C1434.294.71.1Lead LIGHT Antibody _11_30A 5°C2371.498.60.0Lead LIGHT Antibody _11_30B 5°C2121.398.70.0Lead LIGHT Antibody _11_30C 5°C1811.398.70.0Lead LIGHT Antibody _11_30D 5°C1731.298.80.0Lead LIGHT Antibody _11_30E 5°C1431.198.90.0

Claims

1. A formulation comprising: a) 150 mg / mL of a fully human IgG4 anti-LIGHT antibody comprising a heavy chain comprising the amount acid sequence of SEQ ID NO: 7 and a light chain comprising the amino acid sequence of SEQ ID NO: 8; b) 10 mM citrate buffer; c) 0.005% w / v polysorbate 20; and d) 4% w / v mannitol, wherein the pH of the formulation is pH 5.5, and wherein 5% or less of antibodies in the formulation form aggregates after storage for at least 6 months at +5°C.

2. The formulation of claim 1, wherein the formulation exhibits a reduced amount of at least one byproduct selected from the group consisting of aggregates, half-molecules, degradation products, low molecular weight proteins, high molecular weight proteins, and rearrangement of acidic / basic / neutral isoforms of the antibody as compared to a reference anti-LIGHT formulation comprising an anti-LIGHT antibody in phosphate buffered saline at pH 7.3.

3. The formulation of any of the preceding claims, wherein 5% or less of antibodies in the formulation form aggregates after storage for at least 9 months at +5°C.

4. The formulation of any of the preceding claims, wherein the formulation is a liquid formulation or a lyophilized formulation.

5. A kit comprising a container comprising: 1) the formulation of any of the preceding claims, and 2) a label or instructions for the administration and use of the formulation.

6. A pre-filled device or pre-filled container, such as a syringe, cartridge, vial, ampoule, or autoinjector comprising the formulation of any of claims 1 to 4.

Images