Formulations for dual vegf / il-6 inhibitors, Anti-vegf antibodies, and conjugates thereof

Abstract

The present disclosure provides formulations comprising antagonizing antibodies that bind to IL-6, fusion proteins thereof with VEGF Trap, and conjugates of either thereof, and methods of making and using same. The anti-IL-6 antibodies, fusion proteins, and conjugates thereof, and formulations thereof can be used therapeutically alone or in combination with other therapeutics to treat diseases. Also provided are formulations comprising anti-VEGF antibodies and conjugates thereof.

Description

FORMULATIONS FOR DUAL VEGF / IL-6 INHIBITORS, ANTI-VEGF ANTIBODIES, AND CONJUGATES THEREOFINCORPORATION BY REFERENCE TO ANY PRIORITY APPLICATIONS

[0001] Any and all applications for which a foreign or domestic priority claim is identified in the Application Data Sheet as filed with the present application are hereby incorporated by reference under 37 CFR §1.57. The present Application claims priority to U.S. Provisional Application Nos. 63 / 606284 and 63 / 656917, filed December 5, 2023 and June 6, 2024, respectively. The contents of each of these related applications are hereby incorporated by reference.SEQUENCE LISTING

[0002] The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is provided as a file entitled KDIAK222WOseqlist.xml, created December 3, 2024, which is 381,304 bytes in size. The information in the electronic format of the Sequence Listing is incorporated herein by reference in its entirety.BIOLOGICAL SAMPLE DEPOSIT STATEMENT

[0003] In some embodiments, anti-IL-6 antibodies, fusions, and conjugates thereof are provided, which were deposited in the American Type Culture Collection (ATCC), in accordance with the Budapest Treaty, under the numbers PTA-125807 and PTA-125808, on March 13, 2019.

[0004] These deposits are made under the provisions of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure and the Regulations thereunder (Budapest Treaty). This assures maintenance of the deposit for 30 years from date of deposit. The deposit will be made available by the ATCC under the terms of the Budapest Treaty, and subject to an agreement between Applicant and the ATCC, which assures permanent and unrestricted availability of the deposit to the public upon issuance of the pertinent U.S. patent or upon laying open to the public of any U.S. or foreign patent application, whichever comes first, and assures availability of the deposit to one determined by the U.S. Commissioner of Patents and Trademarks to be entitled thereto according to 35 U.S.C. § 122 and theCommissioner’s Rules pursuant thereto (including 37 C.F.R. § 1.14). Availability of the deposited biological material is not to be construed as a license to practice the invention in contravention of the rights granted under the authority of any Government in accordance with its Patent Laws.FIELD

[0005] Provided herein are formulations comprising fusion constructs that bind to IL- 6 and / or VEGF, or comprising anti-VEGF-A antibodies, and conjugates thereof.BACKGROUND

[0006] Vascular endothelial growth factor A (VEGF-A) is a signal protein that mediates pro-angiogenic functions such as endothelial cell survival, proliferation, migration, and cell-cell permeability. Its activity has been shown to promote progression of retinal diseases such as choroidal neovascularization (CNV) in Age Related Macular Degeneration (AMD) and Diabetic Macular Edema (DME). Inhibition of VEGF-A signaling has been proven to be an effective means to stop progression of neovascular retinal diseases (Ferrara et al, Retina, 2006). Various therapeutic molecules have been developed to inhibit VEGF function. Among these, anti- VEGF monoclonal antibodies such as Ranibizumab and Bevacizumab have been shown to be safe and effective treatments against pathological angiogenesis. More recently, recombinantly made VEGFR fusion proteins such as Aflibercept (Eylea) and Conbercept (China), which act as VEGF “traps,” arc proving to be more effective and longer lasting than their antibody competitors.

[0007] Inflammation has been implicated in the pathogenesis of retinal diseases, and anti-inflammatory therapies such as steroids have been effective in treating uveitis and diabetic macular edema (DME). Detailed studies looking at inflammation and infection in the eye have shown that the pro-inflammatory cytokine, interleukin-6 (IL-6) is significantly elevated in the ocular fluids of refractory / chronic uveitis patients, and inhibition of IL-6 in animal models inhibits the onset of uveitis. High ocular fluid levels of IL-6 are also found in patients with DME and retinal vein occlusion. Additionally, chronic inflammatory cells have been seen on the surface of the Bruch’s membrane in eyes with neovascular AMD, and patients with AMD have been reported to have increased serum levels of IL-6. Interestingly, IL-6 has also been observed to stimulate defective angiogenesis. In addition to autoimmune disorders such as rheumatoid arthritis, anti-IL- 6 treatment has been shown to effectively treat uveitis and uveitic macular edema.SUMMARY

[0008] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein, a buffer; and a surfactant, and optionally a tonicity agent. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap.

[0009] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of a fusion protein; a buffer solution, the buffer solution comprising sodium acetate, a surfactant, and optionally a tonicity agent. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap.

[0010] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising sodium acetate mixed with acetic acid; a surfactant, wherein the surfactant comprises polysorbate 20 or polysorbate 80; and optionally a tonicity agent. In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate. In some embodiments, the fusion protein comprises: an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114. In some embodiments, the fusion protein comprises the following structure:Formula (17)

[0011] In some embodiments, each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%, wherein if the conjugatecomprises a VEGF Trap, the VEGF Trap is fused: to the N-terminal end of the heavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain.

[0012] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution; a surfactant, and optionally a tonicity agent. In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate. In some embodiments, the unconjugated antibody is present in the formulation at about 10-60% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114. In some embodiments, the fusion protein comprises the following structure:Formula (17)

[0013] In some embodiments, each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-terminal end of theheavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain.

[0014] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising sodium acetate, and a surfactant, wherein the surfactant comprises polysorbate 20 or polysorbate 80, and optionally a tonicity agent. In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate. In some embodiments, the unconjugated fusion protein is present in the formulation at about 10-60% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein. In some embodiments, the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114, wherein the fusion protein has a light chain with at least 80% identity to the sequence of SEQ ID NO: 169, wherein the fusion protein has a heavy chain with at least 80% identity to the sequence of SEQ ID NO: 170. In some embodiments, the fusion protein comprises the following structure:Formula (17)

[0015] In some embodiments, each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-terminal end of theheavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain.

[0016] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of a fusion protein; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising sodium acetate, a tonicity agent; and a surfactant, wherein the surfactant comprises polysorbate 20 or polysorbate 80. In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate, and, wherein the unconjugated fusion protein is present in the formulation at about 30% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114, wherein the fusion protein has a light chain with at least 80% identity to the sequence of SEQ ID NO: 169, wherein the fusion protein has a heavy chain with at least 80% identity to the sequence of SEQ ID NO: 170. In some embodiments, the fusion protein comprises the following structure:Formula (17 A)

[0017] In some embodiments, part of each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), whichbond is depicted on one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-terminalend of the heavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain.

[0018] In some embodiments, pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein, a buffer, wherein the buffer comprises histidine; a surfactant, and optionally a tonicity agent. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap.

[0019] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of a fusion protein; a buffer solution, the buffer solution comprising histidine acetate, a surfactant, and optionally a tonicity agent. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap.

[0020] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising histidine mixed with acetic acid; a tonicity agent; and a surfactant, wherein the surfactant polysorbate 20 or polysorbate 80. In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate. In some embodiments, the fusion protein comprises: an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114. In some embodiments, the fusion protein comprises the following structure:Formula (17)

[0021] In some embodiments, each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-terminal end of theheavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain.

[0022] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising histidine acetate; a tonicity agent, and a surfactant, wherein the surfactant comprises polysorbate 20 or polysorbate 80, and a tonicity agent. In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate, and wherein the unconjugated fusion protein is present in the formulation at about 10-60% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114. In some embodiments, the fusion protein comprises the following structure:Formula (17)

[0023] In some embodiments, each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, 116, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-terminal end of theheavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain.

[0024] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein, a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising histidine acetate, a tonicity agent; and a surfactant. In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate, and wherein the unconjugated fusion protein is present in the formulation at about 10-60% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114, wherein the fusion protein has a light chain with at least 80% identity to the sequence of SEQ ID NO: 169, wherein the fusion protein has a heavy chain with at least 80% identity to the sequence of SEQ ID NO: 170. In some embodiments, the fusion protein comprises the following structure:Formula (17)

[0025] In some embodiments, each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-terminal end of theheavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain.

[0026] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of a fusion protein; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising histidine acetate, a tonicity agent; and a surfactant, wherein the surfactant comprises polysorbate 20 or polysorbate 80. In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate, and wherein the unconjugated fusion protein is present in the formulation at about 30% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114, wherein the fusion protein has a light chain with at least 80% identity to the sequence of SEQ ID NO: 169, wherein the fusion protein has a heavy chain with at least 80% identity to the sequence of SEQ ID NO: 170. In some embodiments, the fusion protein comprises the following structure:Formula (17 A)

[0027] In some embodiments, part of each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), whichbond is depicted on one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, 116, n7, 118 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-terminalend of the heavy chain; or between a hinge region and a Fab region (after the CH1 domain) of the heavy chain.

[0028] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of a fusion protein, a buffer solution, the buffer solution comprising sodium acetate, a tonicity agent; a surfactant, and wherein the surfactant comprises polysorbate 20 or polysorbate 80. In some embodiments, the fusion protein comprises an anti-IL-6 antibody and a VEGF trap, wherein the fusion protein comprises SEQ ID NOs: 169 and 170.

[0029] In some embodiments, a pharmaceutical formulation is provided, comprising: a pharmaceutically effective amount of a fusion protein; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising sodium acetate, a tonicity agent; and a surfactant, wherein the surfactant comprises polysorbate 20 or polysorbate 80. In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate, and wherein the unconjugated fusion protein is present in the formulation at about 30% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the fusion protein comprises an anti-IL-6 antibody and a VEGF trap, wherein the fusion protein comprises SEQ ID NOs: 169 and 170. In some embodiments, the fusion protein comprises the following structure:Formula (17Br)

[0030] In some embodiments, part of each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), whichbond is depicted on one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is Br; and n 1 , n2, n3, n4, n5, n6, n7, n8 and n9 arc the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%.

[0031] In some embodiments, a pharmaceutical formulation is provided, comprising: a pharmaceutically effective amount of a fusion protein; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising histidineacetate, a tonicity agent; and a surfactant. In some embodiments, the formulation comprises unconjugatcd fusion protein and fusion protein conjugate, and wherein the unconjugatcd fusion protein is present in the formulation at about 30% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the fusion protein comprises an anti-IL-6 antibody and a VEGF trap. In some embodiments, the fusion protein comprises SEQ ID NOs: 169 and 170, wherein the fusion protein comprises the following structure:Formula (17Br)

[0032] In some embodiments, part of each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), whichbond is depicted on one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is Br; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 arc the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, 118 and n9 is about 3500 plus or minus about 10% to about 20%.

[0033] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein, a buffer, wherein the buffer comprises histidine; and an emulsifier, and a tonicity agent. In some embodiments, the fusion protein comprises an anti-IL-6 antibody and a VEGF trap.

[0034] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of a fusion protein; a buffer solution, the buffer solution comprising sodium acetate, a surfactant, and optionally a tonicity agent. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap.

[0035] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein, a buffer, wherein the buffer comprises sodium acetate, a tonicity agent; and a surfactant. In some embodiments, the fusion protein comprises an anti-IL-6 antibody and a VEGF trap, wherein the fusion protein comprises SEQ ID NOs: 169 and 170 (with or without the C-terminal lysine in SEQ ID NO: 170).

[0036] Also provided herein is a formulation comprising about 40 to about 60 mM sodium acetate, about 0.01% to about 0.04% polysorbate 20, about 40 to about 60 mg / mL (total protein concentration) of a mixture of OG1950 and OG1953, the mixture containing about 15% to about 25% OG1950 and about 75% to about 85% OG1953 by molar amount, at pH about 4.5 to about 5.5.

[0037] Provided herein is formulation comprising, consisting of, or consisting essentially of, about 50 mM sodium acetate, about 0.025% polysorbate 20, about 50 mg / mL (total protein concentration) of a mixture of OG1950 and OG1953, the mixture containing about 20% OG1950 and about 80% OG1953 by molar amount, at about pH 5.

[0038] Provided herein is a pharmaceutical formulation comprising: a fusion protein conjugate comprising a first fusion protein conjugated to a phosphorylcholine-containing polymer;an unconjugated fusion protein comprising a second fusion protein that is not conjugated to a phosphorylcholinc-containing polymer; a buffer, wherein the buffer comprises histidine; and a surfactant, wherein the first fusion protein and the second fusion protein each comprises: an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap, wherein the unconjugated fusion protein is present in the formulation at 10-60% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein.

[0039] Further provided is a pharmaceutical formulation comprising: a first fusion protein comprising: a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 170, with or without the C-terminal lysine; and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 169; and a phosphorylcholine-containing polymer conjugated to the heavy chain, wherein the fusion protein conjugate comprises the following structure:Formula (17 A) wherein: part of each heavy chain of the first fusion protein is denoted by the letter H, and each light chain of the first fusion protein is denoted by the letter L; the polymer is bonded to the heavy chain of the first fusion protein through the sulfhydryl of C443 (EU numbering), which bond is depicted on one of the heavy chains; PC is, wherein the curvy line indicates the point of attachment to the rest of the polymer, wherein X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) -H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, 116, n7, 118 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 2500 plus or minus 15%; an unconjugated fusion protein comprising a second fusion protein that is not conjugated to a phosphorylcholine- containing polymer, the unconjugated fusion protein comprising: a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 170, with or without the C-terminal lysine, or a variant thereof having L449 as numbered according to SEQ ID NO: 170; and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 169, wherein the concentration of the first fusion protein and second fusion protein combined is 40-60 mg / mL, wherein the unconjugated fusion protein is present in the formulation at about 30% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, and wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein; histidine acetate at 1- 60 mM; and a surfactant selected from polysorbate 20, polysorbate 80, and poloxamer 188, wherein the surfactant is present in the formulation at 0.01% (w / w) to 0.1% (w / w), wherein the formulation has a pH from 4.9 to 6.2.

[0040] Also provided is a pharmaceutical formulation, the formulation comprising: a pharmaceutically effective amount of a fusion protein, wherein the concentration of the fusion protein is about 50 mg / mL; a phosphorylcholine-containing polymer, wherein the fusion protein either is conjugated to the polymer or is unconjugated; histidine acetate at about 15 mM; and a surfactant at about 0.025% (weight / weight), wherein the surfactant comprises polysorbate 20, polysorbate 80, and / or poloxamer 188, wherein the formulation has a pH of about 5.6, wherein theunconjugated fusion protein is present in the formulation at about 30% of a total molar amount of the fusion protein conjugate and the unconjugatcd fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein, and wherein the fusion protein comprises: a light chain comprising an amino acid sequence at least 80% identical to SEQ ID NO: 169; and a heavy chain comprising an amino acid sequence at least 80% identical to SEQ ID NO: 170, wherein the fusion protein conjugate comprises the following structure:Formula (17A) wherein: part of each heavy chain of the fusion protein is denoted by the letter H, and each light chain of the fusion protein is denoted by the letter L; the polymer is bonded to the heavy chain through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , wherein the curvy lineindicates the point of attachment to the rest of the polymer, wherein X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) -H, or c) any halogen, including -Br, -Cl, or - I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 2500 plus or minus 15%.

[0041] Also provided is a pharmaceutical formulation comprising; a fusion protein comprising: an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap; and a buffer, wherein the pharmaceutical formulation has a pH in a range of 4.5-6.8, optionally, wherein the formulation comprises a surfactant.

[0042] Provided herein is pharmaceutical formulation comprising: a fusion protein comprising: an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap; sodium acetate or histidine acetate at 10-50 mM; polysorbate 20 at 0-0.1% (w / w); and sucrose or trehalose at 0-8% (w / v), wherein the pharmaceutical formulation has a pH in a range of 4.5-6.8, and the fusion protein is present at 20- 200 mg / mL.

[0043] Further provided herein is a pharmaceutical formulation comprising: a fusion protein comprising: a light chain comprising SEQ ID NO: 169, or a sequence at least 80% identical thereto; and a heavy chain comprising SEQ ID NO: 170 (with or without a C-terminal lysine), or a sequence at least 80% identical thereto; sodium acetate at about 15 mM; polysorbate 20 at about 0.025% (w / w); and sucrose about 5.2% (w / v); wherein the pharmaceutical formulation has a pH of about 5.7, and the fusion protein is present at about 100 mg / mL.

[0044] Also provided is a pharmaceutical formulation comprising: a fusion protein comprising: a light chain comprising SEQ ID NO: 169, or a sequence at least 80% identical thereto; and a heavy chain comprising SEQ ID NO: 170 (with or without a C-terminal lysine), or a sequence at least 80% identical thereto; histidine acetate at about 15 mM; polysorbate 20 at about 0.025% (w / w); and sucrose about 5.2% (w / v); wherein the pharmaceutical formulation has a pH of about 6.3, and the fusion protein is present at about 100 mg / mL.BRIEF DESCRIPTION OF THE DRAWINGS

[0045] FIG. 1 depicts a sequence of IL-6.

[0046] FIG. 2A shows Compound L.

[0047] FIG. 2B shows Compound K.

[0048] FIG. 2C shows the synthesis of OG1802 from R3707.

[0049] FIG. 2D shows OG1786.

[0050] FIG. 2E shows the synthesis of OG1546 from OG1550.

[0051] FIG. 2F shows the synthesis of OG1784 from OG1546 and OG1563.

[0052] FIG. 2G shows the synthesis of OG1405 from OG1784.

[0053] FIG. 2H shows the synthesis of OG 1785 from OG1405.

[0054] FIG. 21 shows the synthesis of OG1786 from OG1785.

[0055] FIG. 2J shows OG1801.

[0056] FIG. 2K shows OG1802.

[0057] FIG. 2L shows Compound E.

[0058] FIG. 3 depicts a flow chart for antibody selection and optimization.

[0059] FIG. 4 depicts ELISA data showing that anti-IL-6 mAb binds to IL-6, but not an IL-6 / IL-6R complex, and that IL-6 / IL-6R complex formation is inhibited by anti-IL-6 mAb.

[0060] FIG. 5 depicts some embodiments of the heavy and light chain variable regions of an IL-6-Ab. Embodiments of CDRs are shown in boxed regions. These sequences can also be employed in an IL-6 Ab-VEGF Trap fusion construct.

[0061] FIG. 6 depicts some embodiments of an IL-6- VEGF Trap fusion protein. The VEGF Trap domains are positioned either at the N-terminus immediately preceding the variable domain (left) or positioned between the Fab region and the hinge region of the antibody (right).

[0062] FIG. 7 depicts sensorgrams and table that demonstrate dual inhibitor molecules VEGFR-AntiIL-6 and AntiIL-6-VEGFR bind with similar affinity to VEGF-A as the anti- VEGF antibody OG1950 and Eylea. Thus, the position of the VEGF trap does not alter its affinity for its target.

[0063] FIG. 8 depicts sensorgrams of independent or combined VEGF-A and IL-6 binding to dual inhibitors. IL-6Sensorgrams of mixed targets were compared to a theoretical curve (sum of individual IL-6 and VEGF-A sensorgrams). Results show theoretical and experimental curves superpose, which qualitatively indicates that both targets can bind to the dual inhibitor molecule without influencing each other’s binding.

[0064] FIG. 9 depicts OD450nm the results from various ELISA IL-6 assays. In the bridging ELISA (top), both dual inhibitors bridged btVEGF to IL-6, indicating both configurations can bind to both targets. EC50 VEGF Trap-antiIL-6 = 0.079 nM and antiIL-6-VEGF Trap = 0.026nM. Eylea and anti-IL-6 served as negative controls. FIG. 9 also shows the results of an IL-6 / IL- 6R complex ELISA (middle). Dual inhibitors and antiIL-6 inhibited IL-6 / IL-6R complex formation to the same degree. IC50 values: anti-IL-6 = 0.36 nM, VEGF Trap-antiIL-6 = 0.47 nM, and antiIL-6-VEGF Trap = 0.32 nM. Eylea served as a negative control. FIG. 9 also shows the results of a VEGF / VEGFR competitive ELISA (bottom). Dual inhibitors, Eylea, and OG1950 inhibited VEGF binding to VEGFR to varying degrees. Eylea and the antiIL-6-VEGF Trap construct are comparable (4.24 nM vs. 4.53 nM), while the VEGF Trap-antiIL-6 construct was ~2 fold better (1.74 nM), and OG1950 showed superior maximal inhibition to other inhibitors (1.55 nM).

[0065] FIG. 10 depicts some embodiments of a method for preparing an antibody conjugate (which can also be applied for an Ab-Trap or Trap-Ab conjugate as well). While depicted as an antibody, one of skill in the art will appreciate, in the present context, that the Ab depiction within FIG. 10 can be swapped with a trap fusion (as shown in FIG. 6). For the sake of simplicity, the generic antibody depicted herein represents both options as an antibody and options as a fusion arrangement in the fusion trap context (unless, of course, it is already depicted as a fusion).

[0066] FIGs. 11 A- 1 IB depict SDS-PAGE bands of SeeBlue®Plus2 standard, Anti-IL- 6, Anti-IL-6-VEGFR, and VEGFR- Anti-IL-6. FIG. 11C depicts the conjugate construct of VEGFR- Anti-IL6, which is a fusion of Anti- VEGF (VEGFR1 / 2) and Anti-IL-6 conjugated with a phosphorylcholine-based polymer.

[0067] FIG. 12 depicts results of transfer to a PVDF membrane, N-terminal Edman Sequencing, which show that the cleaved products share the same N-Terminal sequence (LTHRQT), which shows that the cleavage site is located at the VEGF trap region.

[0068] FIGs. 13A-13B depict SDS-PAGE bands of SeeBlue®Plus2 standard and 19 VEGF trap constructs.

[0069] FIGs. 13C-13E depict the sequence listings of VEGF_trap_variant_l, VEGF_trap_variant_2, and VEGF_trap_variant_3.

[0070] FIGs. 13F-13G depict SDS-PAGE bands of SeeBlue®Plus2 standard and 4 VEGF trap variants, including VEGFR_variant_3, which has double point mutations T94I and H95I.

[0071] FIGs. 14A-14F depict Biacore assay results and measurements of affinity to VEGF-A.

[0072] FIG. 15 depicts a cell-based VEGF stimulated VEGFR reporter assay.

[0073] FIG. 16A depicts an assay of inhibition of VEGF / IL6 mediated human umbilical vein endothelial cells (“HUVEC”) tubule formation.

[0074] FIGs. 16B-16C depict statistics of the tubule formation assays with different parameters.

[0075] FIG. 17 depicts the results of an HUVEC proliferation assay.

[0076] FIG. 18 illustrates embodiments of Anti-IL-6 heavy chain variable region sequences. CDRs are underlined.

[0077] FIG. 19 illustrates various embodiments of VEGF trap sequences. Section that varies between the sequences are in bold and underlined.

[0078] FIG. 20 illustrates some embodiments of linker (GS) sequence embodiments. It can be present as a double repeat Gly-Gly-Gly-Gly-Ser linker (GS).

[0079] FIGs. 21A-21B illustrate some embodiments of heavy chain sequence for Anti- IL-6 molecules. CDRs are underlined.

[0080] FIG. 22A-22B illustrates some embodiments of light chain sequences for Anti- IL-6 molecules. CDRs are underlined.

[0081] FIGS. 23A-23B illustrate some embodiments of heavy chain sequences for Anti-IL-6 molecules. CDRs are underlined.

[0082] FIGs. 24A-24B illustrate some embodiments of combinations of CDRs of FIGs.21A-23B.

[0083] FIG. 25 illustrates some embodiments of VEGFR-Fc sequence variants. Sections that vary between the sequences are in bold and underlined.

[0084] FIGs. 26A-26C illustrate affinity binding data.

[0085] FIG. 27 depicts the sequences of some embodiments of the VEGFR- AntiIL6- sequences. The CDRs (as defined by Kabat) are underlined. The greyed sections indicate the VEGFR constructs. The bolded text indicates the linker section. Mutations L234A, L235A, G237A and L443C (EU numbering) are double underlined. Each of these sections can be exchanged for other corresponding sections provided herein (e.g., alternative linkers or CDRs, etc.)

[0086] FIG. 28 depicts a SDS-PAGE of VEGFR- AntiIL6 reduction (Cys - decapping) reaction products. The lanes arc as follows: 1. VEGFR- AntiIL6; 2. VEGFR- AntiIL6 - fully reduced (TCEP); 3. Novex sharp pre-stained protein standard; 4. VEGFR-AntiIL6 + 30x TCEP, initial point; 5. VEGFR-AntiIL6 + 30x TCEP, after 30 min; 6. VEGFR- AntiIL6 + 30x TCEP, after 60 min; 7. VEGFR- AntiIL6 TCEP treated, buffer exchanged; 8.VEGFR-AntiIL6 + 15x dHAA, initial point; 9. VEGFR-AntiIL6 + 15x dHAA, after 30 min; 10. VEGFR- AntiIL6 + 15x dHAA, after 60 min; 11. VEGFR- AntiIL6 decapped; 12. VEGFR- AntiIL6 decapped, fully reduced (TCEP); Gel: NuPAGE Bis-Tris 4-12% Protein amount: 4 pg I lane; 30x TCEP = 30 times molar excess TCEP 15x dHAA = 15 times molar excess dHAA

[0087] FIG. 29 depicts an SDS-PAGE of VEGFR-AntiIL6-OG1802 conjugate CEX chromatography. The Non-reducing gel: NuPAGE Bis-Tris 4-12%. Buffer A: 20 mM sodium acetate pH 5.5. Buffer B: 20 mM sodium acetate pH 5.5, 500 mM NaCl. The lanes are as follows: 1. VEGFR- AntiIL6; 2. VEGFR-AntiIL6-OG1802 (load); 3. Novex sharp pre-stained protein standard; 4. Flow-through; 5. Chase; 6. 30% buffer B - aliquot 1; 7. 30% buffer B - aliquot 2; 8. 30% buffer B - aliquot 3; 9. 30% buffer B - aliquot; 10. 40% buffer B - aliquot 1; 11. 40% buffer B - aliquot 2; 12. 40% buffer B - aliquot 3; 13. 40% buffer B - aliquot 4; 14. 60% buffer B - aliquot 1; 15. 60% buffer B - aliquot 2; 16. 60% buffer B - aliquot 3; 17. 100% buffer B - strip. Lanes 6-13 show protein conjugated material, conjugate cannot penetrate gel due to large size. Lanes 14-17 show protein mixture possibly containing aggregated, conjugated and non-conjugated protein material

[0088] FIG. 30 depicts a SDS-PAGE of a protein-polymer conjugate vs non-conjugate protein material. Gel analysis demonstrates 57% heavy chain and 96% light chain band intensity ratios when bioconjugate (lane 3) is compared to VEGFR-AntiIL6 reference standard (lane 2), which indicates the presence of one OG1802 polymer per VEGFR- AntiIL6 molecule Reducing gel is: NuPAGE Bis-Tris 4-12% Reducing gel is: NuPAGE Bis-Tris 4-12%.

[0089] FIG. 31 depicts a SEC-MALS of VEGFR-AntiIL6-OG1802. The molecular weight of VEGFR-AntiIL6 conjugate was determined with integrated size exclusion chromatography (Shodex-SB806M-HQ) and light scattering (MALS). Top panel. Chromatogram shows the presence of a single eluting peak. Absence of additional peaks and shoulder suggest no aggregates and degraded material were present after conjugation and subsequent CEX separation steps. Bottom panel. Protein conjugate analysis of selected peak showed an experimentallymeasured average molecular weight (Mw) of 983 kDa for the VEGFR-AntiIL6-OG1802 bioconjugatc. This value results from the conjugation of one VEGFR-AntiIL6 molecule (Mw ~ 189 kDa) and one OG1802 polymer (Mw ~ 794 kDa).

[0090] FIG. 32 depicts the results of a VEGFR-AntiIL6 CEX chromatography. Gel: is Novex 8-16% Tris-Glycine (reducing conditions), M = SeeBlue®Plus2 standard Lanes D12-F11 correspond to samples aliquoted at different buffer B concentrations as indicated on chromatogram Intact (I) and cleaved (C) heavy chains are indicated. In Column Poros pXS Buffer A is 20 mM Sodium Phosphate pH 6 and buffer B is 20 mM Sodium Phosphate pH 6, 1 M NaCl.

[0091] FIG. 33 depicts the results of a VEGFR-AntiIL6 HIC chromatography. Gel: is Novex 8-16% Tris-Glycine / reducing conditions, M = SeeBlue®Plus2 standard. L = VEGFR- AntiIL6 intact and cleaved mixture (load). Remaining lanes correspond to samples aliquoted at different buffer B concentrations as indicated on chromatogram. Intact (I) and cleaved (C) heavy chains are indicated. In Column Hi Trap Butly HP Buffer A is: 20 mM Sodium Phosphate pH 6, 1 M Ammonium sulfate. Buffer B is: 20 mM Sodium Phosphate pH 6.

[0092] FIG. 34 depicts the results of a VEGF / VEGFR competitive ELISA.

[0093] FIG. 35 depicts the results of a IL6 / IL6R complex ELISA.

[0094] FIG. 36 depicts the results of a cell-based VEGF stimulated VEGFR reporter assay.

[0095] FIG. 37 depicts the lipopolysaccharide stimulated tubule formation in HUVECs.

[0096] FIG. 38 depicts the lipopolysaccharide stimulated tubule formation in HUVECs

[0097] FIG. 39 depicts lipopolysaccharide stimulated tubule formation in HUVECs

[0098] FIG. 40A depicts the inhibition of VEGF / IL6 mediated proliferation of HUVECs.

[0099] FIG. 40B depicts the inhibition of HUVEC proliferation with increasing number of cells per well.

[0100] FIG. 41A-41C depicts an SDS-PAGE of antibodies and antibody conjugates treated with various reagents for reduction and re-oxidation. 41A lanes are as follows: Lane 4: antibody (IL-6 Ab-VEGF Trap) starting material, lane 5: 3x TCEP, lane 6: 6x TCEP, lane 7: lOx TCEP, and lane 8: 30x TCEP. 42B lanes are as follows: Lane 2: antibody (IL-6 Ab-VEGF Trap) starting material, lane 3: 3x TCEP (re-oxidized over time), lane 4: 6x TCEP (re-oxidized overtime), lane 5: 3x TCEP after re-oxidation, lane 6: 6x TCEP after re-oxidation, lane 7: lOx TCEP after rcoxidation, lane 8: 30x TCEP after rc-oxidation, lane 9: 3x TCEP after conjugation, lane 10: 6x TCEP after conjugation, lane 11: lOx TCEP after conjugation, lane 12: 30x TCEP after conjugation. FIG. 41C lanes are as follows: Lane 7: antibody (IL-6 Ab-VEGF Trap) starting material, lane 8: 30x TCEP after reoxidation, lane 9: 30x TCEP after conjugation. IEF PAGE gel (right). Lane 7: marker, lane 8: antibody (IL-6 Ab-VEGF Trap) starting material, lane 9: 30x TCEP after re-oxidation, lane 10: 30x TCEP after conjugation.

[0101] FIG. 42 depicts a set of SEC-HPLC chromatograms with varying excess TCEP. Top: SEC-HPLC chromatograms of antibody (IL-6 Ab-VEGF Trap) reduced with different excess of TCEP over antibody (IL-6 Ab-VEGF Trap) and subsequently conjugated with biopolymer OG1802 to yield fusion protein. 3x TCEP after conjugation (blue), 6x TCEP after conjugation (magenta), lOx TCEP after conjugation (brown), and 30x TCEP after conjugation (black). Bottom: Graph of fusion protein conversion versus TCEP excess.

[0102] FIG. 43 depicts an SDS-PAGE. Lanes are as follows: Lane 1: antibody (IL-6 Ab-VEGF Trap) starting material, lane 2: reduction with 30x TCEP, lane 3: re-oxidation with lOx DHAA, lane 4: marker, lane 5: re-oxidation with 15x DHAA, lane 6: re-oxidation with 20x DHAA, lane 7: re-oxidation with 30x DHAA.

[0103] FIGs. 44 and 45 depict flow chart diagrams of the production of Fusion Antibody (IL-6 Ab-VEGF Trap) and a combined dual inhibitor molecule (VEGFR-AntiIL6- OG1802), respectively.

[0104] FIG. 46 depicts a design to screen the impact of pH, sucrose %, NaCl concentration, sodium acetate buffering strength, and protein concentration of the OG2072 formulation by SEC-HPLC, VEGF potency, viscosity, and osmolality. Sample buffer was Na- acetate supplemented with 0.025% polysorbate 20, and samples were incubated at 37 °C.

[0105] FIGs. 47A-47B depict the results of an SEC-HPLC assessment of the main peak of OG2072 for the sample formulations depicted in FIG. 46. Sample formulations were analyzed at days 0, 7, 14, and 28.

[0106] FIGs. 48A-48B depict the results of an VEGF potency assessment for the sample formulations depicted in FIG. 46. Sample formulations were analyzed at days 0, 7, 14, and 28.

[0107] FIGs. 49A-49B depict the results of a viscosity assessment for the sample formulations depicted in FIG. 46. Sample formulations were analyzed at day 0.

[0108] FIGs. 50A-50B depict the results of an osmolarity assessment for the sample formulations depicted in FIG. 46. Sample formulations were analyzed at day 0.

[0109] FIG. 51 depicts a summary of the results of FIGs. 47A-50B, where impact of a formulation parameter on SEC-HPLC, potency, viscosity, or osmolarity, is represented by a ‘+’, or 0, with a more pronounced impact indicated by repeated symbols, i.e. ‘+++’.

[0110] FIG. 52 depicts parameters for OG2072 (unconjugated) and OG2074 (conjugated) formulations, indicating values for sodium acetate or histidine acetate concentration, sucrose %, polysorbate 20 %, pH, protein concentration, and conjugatedmnconjugated ratio.

[0111] FIG. 53 depicts formulation information for six preparations of formulation including both OG2074 (conjugated) and OG2072 (unconjugated) antibody, indicating conjugatedmnconjugated ratio, protein concentration, pH, and presence and absence of sucrose.

[0112] FIG. 54 is a table showing the sample setup for a long-term stability plan for various formulations of OG1953 conjugate containing free proteins from 7.5-20% with a total combined protein concentrations of 50-65mg / ml.

[0113] FIGs. 55 and 56 are a collection of tables and plots showing the results of ELISA assays measuring the potency of various formulations of OG1953 conjugate containing free proteins from 7.5-20% with a total combined protein concentrations of 50-65mg / ml after storage.

[0114] FIG. 57 is a collection of a table and a graph showing protein concentration of formulations measured by SoloVPE OD280nm method.

[0115] FIG. 58 is a data table showing a summary of the results of the stability testing of formulations.

[0116] FIG. 59A is a collection of SEC-HPLC traces of formulation #4 (20% OG1950, 80% OG1953, 50 mM Na-acetate, 0.025% Tween 20, pH 5.0) after 6 months at the temperatures highlighted. FIG. 59B provides a table and graphs showing size exclusion chromatography analysis of formulations for the aggregation and degradation level of the OG1953 conjugate.

[0117] FIG. 60A shows a schematic diagram showing an overview of the Tandem HPLC method, which combines the CEX-HPLC in tandem with a SEC-HPLC column.

[0118] FIG. 60B is a collection of traces, a table, and graphs showing tandem method analysis of the OG1950 free protein and its aggregated forms (Pl and P2).

[0119] FIGs. 60C and 60D are a collection of traces and a table showing tandem method analysis of formulations.

[0120] FIGs. 61A-61C are a collection of plots showing the level of impurities (e.g., aggregation and / or degradation level) in OG1953 conjugate formulations over time.

[0121] FIGs. 62 A and 62B are a collection of plots and tables showing potency comparison of various OG1953 formulations using ELISA or cell-based assay.

[0122] FIGs. 63A-63C are a collection of schematic diagrams showing components of non-limiting examples of formulations of the present disclosure.

[0123] FIG. 64 depicts KSI-501DS Batches 1-3 Lot Release Data.

[0124] FIGs. 65 depicts an injection force comparison of (Panel A) OG1953 (100%) conjugate versus the KSI-301 mix formulation using a 27G or 29G dosing needle; (Panel B) OG2074 (100%) conjugate versus the KSI-501_batch 2 mix formulation using a 27G or 29G dosing needle.

[0125] FIG. 66 depicts a viscosity comparison at ambient temperature of (Panel A) OG1953 (100%) conjugate versus the KSI-301 mix formulation; (Panel B) OG2074 (100%) conjugate versus various batches of the KSL501 mix formulation.

[0126] FIGs. 67A-67D depict continuous 80min tandem method separation with PhotoDiol Array (PDA) detection set at 200-350nm. FIG. 67 A depicts the 2D contour view of elution time versus wavelength; FIG. 67B depicts the extracted wavelength profile at 280nm and the peak identification of the various eluted fractions collected for further characterization using SDS-PAGE analysis followed by Silver Staining, and results were shown in FIG. 67C as a nonreducing gel and FIG. 67D as a reducing gel.

[0127] FIG. 68 depicts some embodiments of the heavy chain amino acid sequence of OG1950.

[0128] FIG. 69 depicts some embodiments of the light chain amino acid sequence of OG1950.

[0129] FIGs. 70A-70B depict some embodiments of a Design of experiment (DoE) analysis to investigate the impact of formulation parameters on stability of OG2074 by CEX- HPLC.

[0130] FIGs. 71 A-71B depict some embodiments of a DoE analysis to investigate the impact of formulation parameters on stability of OG2072 by SEC-HPLC.

[0131] FIGs. 72A-72B depict some embodiments of a DoE analysis to investigate the impact of formulation parameters on stability of OG2072 by Tandem HPLC model.

[0132] FIGs. 73A-73B depict some embodiments of a DoE analysis to investigate the impact of formulation parameters on stability of impurities by Tandem HPLC model.

[0133] FIGs. 74A-74B depict some embodiments of an assessment of VEGF potency (FIG. 74A) and IL-6 potency (FIG. 74B).

[0134] FIGs. 75A-75B depict some embodiments of a DoE analysis, showing exemplary CEX-HPLC and SEC-HPLC profiles for stability at 37°C.

[0135] FIG. 76 depicts some embodiments of a DOE analysis, showing tandem HPLC profiles for stability at day 14.

[0136] FIG. 77 A depicts some embodiments of a DoE analysis, where DoE samples of day 0, day 14 and day 56 were analyzed by tandem HPLC. FIG. 77B depicts a summary table of some embodiments of model analysis from a DoE analysis. FIG. 77C depicts some embodiments of pH after vacuum assisted concentration, actual protein concentration, and actual buffer strength of fractions from the tandem HPLC analysis.

[0137] FIG. 78 depicts a table summarizing some embodiments of a one-factor- at-a- time (OFAT) analysis to investigate the impact of formulation parameters on stability on KSI-501 stability.

[0138] FIG. 79A-79B depict some embodiments of a OFAT analysis, showing improvement of stability as measured by CEX-HPLC (Fig. 79A) and SEC-HPLC (Fig. 79B).

[0139] FIGs. 8OA-8OB depict some embodiments of an experiment for pH fine tuning in the presence versus the absence of sucrose. FIG. 80A depicts some embodiments of a summary table of sample pH and sucrose concentration. FIG. 80B depicts some embodiments of sample turbidity at pH 5.6, 5.9, 6.2, and 6.5.

[0140] FIG. 81 depicts some embodiments of stability data of KSI-501 GMP batches with two different formulations at 3-month time points.

[0141] FIG. 82A depicts some embodiments of a KSI-501 formulation comprising 50 mM sodium acetate, pH 5.0, 4% sucrose, 0.025% polysorbate 20; 30% OG2072, 70% OG2074 at50.0 mg / ml. FIG. 82B depicts some embodiments of a KSI-501 formulation comprising 15 mM histidine acetate, pH 5.6, 0.025% polysorbate 20; 30% OG2072, 70% OG2074 at 50 mg / ml.

[0142] FIGs. 83A-83B depict the results of an SEC-HPLC assessment of the main peak of OG2072 for sample formulations of OG2072 in a sodium acetate buffered system.

[0143] FIGs. 84A-84B depicts the results of an VEGF potency assessment for sample formulations of OG2072 in a sodium acetate buffered system.

[0144] FIG. 85 depicts a summary of the results of FIGs. 83A-84B, where impact of a formulation parameter on SEC-HPLC, potency, viscosity, or osmolarity, is represented by a ‘+’, or 0, with a more pronounced impact indicated by repeated symbols, i.e. ‘+++’.

[0145] FIG. 86 depicts the parameters of each of the 24 experimental formulations.

[0146] FIGs. 87A-87C depict the results of an SEC-HPLC assessment of the main peak of OG2072 for sample formulations of OG2072 in a histidine hydrochloride buffered system. FIG. 87C shows OG2072 stability at day 28 for an incubation at 37°C.

[0147] FIGs. 88A-88C depicts the results of an VEGF potency assessment for sample formulations of OG2072 in a histidine hydrochloride buffered system. FIG. 87C shows VEGF potency at day 28 for an incubation at 37°C.

[0148] FIG. 89 depicts the results of a viscosity assessment for sample formulations of OG2072 in a histidine hydrochloride buffered system.

[0149] FIG. 90 depicts the results of an osmolality assessment for sample formulation of OG2072 in a histidine hydrochloride buffered system.

[0150] FIG. 91 depicts a summary of the results of FIGs. 87A-90, where impact of a formulation parameter on SEC-HPLC, potency, viscosity, or osmolarity, is represented by a ‘+’, or 0, with a more pronounced impact indicated by repeated symbols, i.e. ‘+++’.

[0151] FIGs. 92A-92B provide an overlay of the SEC-HPLC traces summarizing some embodiments of a one-factor-at-a-time (OFAT) analysis to investigate the impact of formulation parameters on KSL101 stability.

[0152] FIGs. 93A-93B show corrected VEGF potency and OG2072 SEC-HPLC main peak area, respectively, at day 28 for an incubation at 37°C.

[0153] FIGs. 94A-94B depict the osmolality and viscosity of KSL101 formulation in a sodium acetate buffer system vs. histidine hydrochloride buffer system.

[0154] FIG. 95 depicts flow chart diagrams of the production of a fusion protein (IL-6 Ab-VEGF Trap).DETAILED DESCRIPTION

[0155] To directly reduce the concurrent inflammation and defective angiogenesis that drive pathogenesis of neovascular retinal pathologies, presented herein are formulations of designed molecules that simultaneously block the functions of the pro -inflammatory cytokine IL- 6 and the pro-angiogenic signal protein VEGF, as well methods of manufacture of formulations thereof. These molecules arc comprised of (1) an anti-IL-6 monoclonal antibody fused to (2) two VEGF binding domains of VEGF receptors (VEGFRs). The anti-IL-6 moiety specifically binds IL-6 and inhibits its interaction with the IL-6 receptor (IL-6R). The VEGF Trap moiety includes of a fusion of two VEGF binding domains (VEGFR1 domain 2, VEGFR2 domain 3) that work as a VEGF trap, preventing VEGF from binding to VEGF receptors. Additionally, in some embodiments, each of these dual inhibitor molecules is equipped with an unpaired cysteine at its C-terminus which can be conjugated with a half-life extending phosphorylcholine based biopolymer. In some embodiments, various formulations of fusion protein conjugates, e.g., KSL 501, were tested, as provided herein. When monovalent buffers were tested e.g. sodium acetate or imidazole, there was turbidity formation at lower pH values. Histidine acetate showed turbidity formation only at higher pH values. Thus, it was hypothesized that monovalent ions of the buffer bind as counterions to the biopolymer of the conjugate and thereby neutralize the charge. Without being bound by theory, trivalent histidine binds to the biopolymer but maintains two extra charges, which may help in repelling molecules from interacting with it, and thereby reduce turbidity.

[0156] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein, a buffer; and a surfactant, and optionally a tonicity agent. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap.

[0157] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of a fusion protein; a buffer solution, the buffer solution comprising sodium acetate, a surfactant, and optionally a tonicity agent. Insome embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap.

[0158] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising sodium acetate mixed with acetic acid; a tonicity agent; a surfactant, wherein the surfactant comprises polysorbate 20 or polysorbate 80 (or poloxamer 188). In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate. In some embodiments, the fusion protein comprises: an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114. In some embodiments, the fusion protein comprises the following structure:Formula (17)

[0159] In some embodiments,: each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%, wherein if the conjugatecomprises a VEGF Trap, the VEGF Trap is fused: to the N-terminal end of the heavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain.

[0160] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution; a surfactant, and optionally a tonicity agent. In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate. In some embodiments, the unconjugated antibody (or fusion protein) is present in the formulation at about 10-60% of a total molar amount of the antibody (or fusion protein) conjugate and the unconjugated antibody (or fusion protein), wherein the total molar amount is the sum of the molar amount of the antibody (or fusion protein) conjugate and the molar amount of the unconjugated antibody (or fusion protein). In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114. In some embodiments, the fusion protein comprises the following structure:Formula (17)

[0161] In some embodiments, each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-terminal end of theheavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain.

[0162] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising sodium acetate, a tonicity agent, and a surfactant, wherein the surfactant comprises polysorbate 20 or polysorbate 80 (or poloxamer 188). In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate. In some embodiments, the unconjugated antibody (or fusion protein) is present in the formulation at about 10-60% of a total molar amount of the antibody (or fusion protein) conjugate and the unconjugated antibody (or fusion protein). In some embodiments, the total molar amount is the sum of the molar amount of the antibody (or fusion protein) conjugate and the molar amount of the unconjugated antibody (or fusion protein). In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114, wherein the fusion protein has a light chain with at least 80% identity to the sequence of SEQ ID NO: 169, wherein the fusion protein has a heavy chain with at least 80% identity to the sequence of SEQ ID NO: 170 (with or without the C-terminal lysine). In some embodiments, the fusion protein conjugate comprises the following structure:Formula (17)

[0163] In some embodiments, each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-terminal end of theheavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain.

[0164] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of a fusion protein; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising sodium acetate, a tonicity agent; and a surfactant, wherein the surfactant comprises polysorbate 20 or polysorbate 80 (or poloxamer 188). In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate, and, wherein the unconjugated antibody (or fusion protein) is present in the formulation at about 30% of a total molar amount of the antibody (or fusion protein) conjugate and the unconjugated antibody (or fusion protein), wherein the total molar amount is the sum of the molar amount of the antibody (or fusion protein) conjugate and the molar amount of the unconjugated antibody (or fusion protein). In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114, wherein the fusion protein has a light chain with at least 80% identity to the sequence of SEQ ID NO: 169, wherein the fusion protein has a heavy chain with at least 80% identity to the sequence of SEQ ID NO: 170. In some embodiments, the fusion protein comprises the following structure:Formula (17 A)

[0165] In some embodiments, part of each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), whichbond is depicted on one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-terminal end of the heavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain.

[0166] In some embodiments, pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein, a buffer, wherein the buffer comprises histidine; a surfactant, and optionally a tonicity agent. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap.

[0167] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of a fusion protein; a buffer solution, the buffer solution comprising histidine acetate, a surfactant, and optionally a tonicity agent. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap.

[0168] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising histidine mixed with acetic acid; a tonicity agent; and a surfactant, wherein the surfactant polysorbate 20 or polysorbate 80 (or poloxamer 188). In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate. In some embodiments, the fusion protein comprises: an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114. In some embodiments, the fusion protein comprises the following structure:Formula (17)

[0169] In some embodiments, each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-terminal end of theheavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain.

[0170] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising histidine acetate; a tonicity agent, and a surfactant, wherein the surfactant comprises polysorbate 20 or polysorbate 80 (or poloxamer 188), and optionally a tonicity agent. In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate, and wherein the unconjugated antibody (or fusion protein) is present in the formulation at about 10-60% of a total molar amount of the antibody (or fusion protein) conjugate and the unconjugated antibody (or fusion protein), wherein the total molar amount is the sum of the molar amount of the antibody (or fusion protein) conjugate and the molar amount of the unconjugated antibody (or fusion protein). In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114. In some embodiments, the fusion protein comprises the following structure:Formula (17)

[0171] In some embodiments, each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-terminal end of theheavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain.

[0172] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein, a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising histidine acetate, a tonicity agent; and a surfactant. In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate, and wherein the unconjugated antibody (or fusion protein) is present in the formulation at about 10-60% of a total molar amount of the antibody (or fusion protein) conjugate and the unconjugated antibody (or fusion protein), wherein the total molar amount is the sum of the molar amount of the antibody (or fusion protein) conjugate and the molar amount of the unconjugated antibody (or fusion protein). In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114, wherein the fusion protein has a light chain with at least 80% identity to the sequence of SEQ ID NO: 169, wherein the fusion protein has a heavy chain with at least 80% identity to the sequence of SEQ ID NO: 170. In some embodiments, the fusion protein comprises the following structure:Formula (17)

[0173] In some embodiments, each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-terminal end of theheavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain.

[0174] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of a fusion protein; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising histidine acetate, a tonicity agent; and a surfactant, wherein the surfactant comprises polysorbate 20 or polysorbate 80 (or poloxamer 188). In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate, and wherein the unconjugated fusion protein is present in the formulation at about 30% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114, wherein the fusion protein has a light chain with at least 80% identity to the sequence of SEQ ID NO: 169, wherein the fusion protein has a heavy chain with at least 80% identity to the sequence of SEQ ID NO: 170. In some embodiments, the fusion protein comprises the following structure:Formula (17 A)

[0175] In some embodiments, part of each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), whichbond is depicted on one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-terminal end of the heavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain.

[0176] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of a fusion protein, a buffer solution, the buffer solution comprising sodium acetate, a tonicity agent; a surfactant, and wherein the surfactant comprises polysorbate 20 or polysorbate 80 (or poloxamer 188). In some embodiments, the fusion protein comprises an anti-IL-6 antibody and a VEGF trap, wherein the fusion protein comprises SEQ ID NOs: 169 and 170. In any formulation or composition herein, in some embodiments, the surfactant can be poloxamer 188 instead of polysorbate 20 or polysorbate 80.

[0177] In some embodiments, a pharmaceutical formulation is provided, comprising: a pharmaceutically effective amount of a fusion protein; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising sodium acetate, a tonicity agent; and a surfactant, wherein the surfactant comprises polysorbate 20 or polysorbate 80 (or poloxamer 188). In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate, and wherein the unconjugated fusion protein is present in the formulation at about 30% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the fusion protein comprises an anti-IL-6 antibody and a VEGF trap, wherein the fusion protein comprises SEQ ID NOs: 169 and 170. In some embodiments, the fusion protein comprises the following structure:Formula (17Br)

[0178] In some embodiments, part of each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), whichbond is depicted on one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is Br; and n 1 , n2, n3, n4, n5, n6, n7, n8 and n9 arc the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%.

[0179] In some embodiments, a pharmaceutical formulation is provided, comprising: a pharmaceutically effective amount of a fusion protein; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising histidineacetate, a tonicity agent; and a surfactant. In some embodiments, the formulation comprises unconjugatcd fusion protein and fusion protein conjugate, and wherein the unconjugatcd fusion protein is present in the formulation at about 30% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the fusion protein comprises an anti-IL-6 antibody and a VEGF trap. In some embodiments, the fusion protein comprises SEQ ID NOs: 169 and 170, wherein the fusion protein comprises the following structure:Formula (17Br)

[0180] In some embodiments, part of each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), whichbond is depicted on one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is Br; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 arc the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, 118 and n9 is about 3500 plus or minus about 10% to about 20%.

[0181] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein, a buffer, wherein the buffer comprises histidine; and a surfactant, and a tonicity agent. In some embodiments, the fusion protein comprises an anti-IL-6 antibody and a VEGF trap.

[0182] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of a fusion protein; a buffer solution, the buffer solution comprising sodium acetate, a surfactant, and a tonicity agent. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap.

[0183] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein, a buffer, wherein the buffer comprises sodium acetate, a tonicity agent; and a surfactant. In some embodiments, the fusion protein comprises an anti-IL-6 antibody and a VEGF trap, wherein the fusion protein comprises SEQ ID NOs: 169 and 170 (with or without the C-terminal lysine in SEQ ID NO: 170).

[0184] In some embodiments, provided herein are pharmaceutical formulations comprising a pharmaceutically effective amount of a fusion protein, a buffer; and an emulsifier. In some embodiments, the fusion protein is that in FIG. 27 and / or includes at least one, two, three, four or more of the components in FIG. 27, and / or includes SEQ ID NO: 169 and / or 170 (but can be any fusion protein provided herein). In some embodiments, the fusion protein is at least 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical in sequence to SEQ ID NOs: 169 and / or 170, and / or includes the 3 heavy and / or 3 light CDRs therein. In some embodiments, the fusion protein is conjugated to a polymer, optionally as shown in Formula 17, 17A, or 17Br. It is noted that Formula 17 and 17A depict the same active ingredient bound to an antibody. In some embodiments, in Formula 17Br, X is Br.

[0185] In some embodiments, the formulation comprises a pharmaceutically effective amount of fusion protein, wherein the concentration of the fusion protein is between about 30 to about 85 mg / mL; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising sodium acetate mixed with acetic acid, wherein the buffer solution is between about 0.1 mM to about 25 mM, wherein the buffer solution pH is about 5.0; and a surfactant, wherein the surfactant comprises between about between about 0.01% to about 0.05% (weight / weight) polysorbate 20 or polysorbate 80. In some embodiments, the fusion protein comprises: an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap, wherein the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114, (or wherein the fusion protein has the sequences in FIG. 27). In some embodiments, the fusion protein comprises the following structure:Formula (17) wherein: each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depicted on one of the heavy chains; PC is, where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n6, n7, n8 and n9 is 2500 plus or minus 15%, wherein if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to theN-terminal end of the heavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain.

[0186] In some embodiments, the fusion protein comprises the following structure:Formula (17A) wherein: part of each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depicted on one of theheavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, 116, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, 116, n7, 118 and n9 is 2500 plus or minus 15%, wherein if the conjugate comprises a VEGF Trap, the VEGFTrap is fused: to the N-terminal end of the heavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain.

[0187] Various embodiments provided herein will employ, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry and immunology, which are within the skill of the art. Such techniques are explained fully in the literature, such as, Molecular Cloning: A Laboratory Manual, second edition (Sambrook et al., 1989) Cold Spring Harbor Press; Oligonucleotide Synthesis (M.J. Gait, ed., 1984); Methods in Molecular Biology, Humana Press; Cell Biology: A Laboratory Notebook (J.E. Cellis, ed., 1998) Academic Press; Animal Cell Culture (R.I. Freshney, ed., 1987); Introduction to Cell and Tissue Culture (J.P. Mather and P.E. Roberts, 1998) Plenum Press; Cell and Tissue Culture: Laboratory Procedures (A. Doyle, J.B. Griffiths, and D.G. Newell, eds., 1993- 1998) J. Wiley and Sons; Methods in Enzymology (Academic Press, Inc.); Handbook of Experimental Immunology (D.M. Weir and C.C. Blackwell, eds.); Gene Transfer Vectors for Mammalian Cells (J.M. Miller and M.P. Calos, eds., 1987); Current Protocols in Molecular Biology (F.M. Ausubel et al., eds., 1987); PCR: The Polymerase Chain Reaction, (Mullis et al., eds., 1994); Current Protocols in Immunology (J.E. Coligan et al., eds., 1991); Short Protocols in Molecular Biology (Wiley and Sons, 1999); Immunobiology (C.A. Janeway and P. Travers, 1997); Antibodies (P. Finch, 1997); Antibodies: a practical approach (D. Catty., ed., IRL Press, 1988- 1989); Monoclonal antibodies: a practical approach (P. Shepherd and C. Dean, eds., Oxford University Press, 2000); Using antibodies: a laboratory manual (E. Harlow and D. Lane (Cold Spring Harbor Laboratory Press, 1999); The Antibodies (M. Zanetti and J.D. Capra, eds., Harwood Academic Publishers, 1995).

[0188] The following terms, unless otherwise indicated, shall be understood to have the following meanings: the term “isolated molecule” as referring to a molecule (where the molecule is, for example, a polypeptide, a polynucleotide, or an antibody) that by virtue of its origin or source of derivation (1) is not associated with naturally associated components that accompany it in its native state, (2) is substantially free of other molecules from the same source, e.g., species, cell from which it is expressed, library, etc., (3) is expressed by a cell from a different species, or (4) does not occur in nature. Thus, a molecule that is chemically synthesized, or expressed in a cellular system different from the system from which it naturally originates, will be “isolated” from its naturally associated components. A molecule also may be renderedsubstantially free of naturally associated components by isolation, using purification techniques well known in the art. Molecule purity or homogeneity may be assayed by a number of means well known in the ail. For example, the purity of a polypeptide sample may be assayed using polyacrylamide gel electrophoresis and staining of the gel to visualize the polypeptide using techniques well known in the art. For certain purposes, higher resolution may be provided by using HPLC or other means well known in the art for purification.

[0189] As used herein, unless designated otherwise, the term “IL-6” or “IL6” refers to human IL-6. In some embodiments, other forms of IL-6 are contemplated, and will be designated by specific reference to the other organisms, e.g., canine, feline, equine, and bovine. One exemplary human IL-6 is found as UniProt Accession NumberP05231.

[0190] Anti-IL-6 antibodies or other biologies described herein are typically provided in isolated form. This means that an antibody is typically at least 50% w / w pure of interfering proteins and other contaminants arising from its production or purification but does not exclude the possibility that the antibody is combined with a pharmaceutically acceptable excipient intended to facilitate its use. Sometimes antibodies are at least 60, 70, 80, 90, 95 or 99% w / w pure of interfering proteins and contaminants from production or purification. Often an antibody (or antibody conjugate) is the predominant macromolecular species remaining after its purification.

[0191] An “antibody” is an immunoglobulin molecule capable of specific binding to a target, such as a carbohydrate, polynucleotide, lipid, polypeptide, etc., through at least one antigen recognition site, located in the variable region of the immunoglobulin molecule. As used herein, the term encompasses not only intact polyclonal or monoclonal antibodies, but also, unless otherwise specified, any antigen binding portion thereof that competes with the intact antibody for specific binding, fusion proteins comprising an antigen binding portion, and any other modified configuration of the immunoglobulin molecule that comprises an antigen recognition site. Antigen binding portions include, for example, Fab, Fab’, F(ab’)2, Fd, Fv, domain antibodies (dAbs, e.g., shark and camelid antibodies), fragments including complementarity determining regions (CDRs), single chain variable fragment antibodies (scFv), maxibodies, minibodies, intrabodies, diabodies, triabodies, tetrabodies, v-NAR and bis-scFv, and polypeptides that contain at least a portion of an immunoglobulin that is sufficient to confer specific antigen binding to the polypeptide. An antibody includes an antibody of any class, such as IgG, IgA, or IgM (or sub-class thereof), and the antibody need not be of any particular class. Depending on the antibody amino acid sequenceof the constant region of its heavy chains, immunoglobulins can be assigned to different classes. There arc five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgGi, IgGa, IgGs, IgG4, IgAi and IgA2. The heavy-chain constant regions that correspond to the different classes of immunoglobulins are called alpha, delta, epsilon, gamma, and mu, respectively. The subunit structures and three- dimensional configurations of different classes of immunoglobulins are well known.

[0192] A “variable region” of an antibody refers to the variable region of the antibody light chain or the variable region of the antibody heavy chain, either alone or in combination. As known in the art, the variable regions of the heavy and light chains each consist of four framework regions (FRs) connected by three complementarity determining regions (CDRs) also known as hypervariable regions, and contribute to the formation of the antigen binding site of antibodies. If variants of a subject variable region are desired, particularly with substitution in amino acid residues outside of a CDR region (i.e., in the framework region), appropriate amino acid substitution, preferably, conservative amino acid substitution, can be identified by comparing the subject variable region to the variable regions of other antibodies which contain CDR1 and CDR2 sequences in the same canonical class as the subject variable region (Chothia and Lesk, J Mol Biol 196(4): 901-917, 1987).

[0193] In certain embodiments, definitive delineation of a CDR and identification of residues comprising the binding site of an antibody is accomplished by solving the structure of the antibody and / or solving the structure of the antibody-ligand complex. In certain embodiments, that can be accomplished by any of a variety of techniques known to those skilled in the art, such as X-ray crystallography. In certain embodiments, various methods of analysis can be employed to identify or approximate the CDR regions. In certain embodiments, various methods of analysis can be employed to identify or approximate the CDR regions. Examples of such methods include, but are not limited to, the Kabat definition, the Chothia definition, the IMGT approach (Lefranc et al., 2003) Dev Comp Immunol. 27:55-77), computational programs such as Paratome (Kunik et al., 2012, Nucl Acids Res. W521-4), the AbM definition, and the conformational definition.

[0194] The Kabat definition is a standard for numbering the residues in an antibody and is typically used to identify CDR regions. See, e.g., Johnson & Wu, 2000, Nucleic Acids Res., 28: 214-8. The Chothia definition is similar to the Kabat definition, but the Chothia definition takes into account positions of certain structural loop regions. See, e.g., Chothia et al., 1986, J. Mol.Biol., 196: 901- 17; Chothia et al., 1989, Nature, 342: 877-83. The AbM definition uses an integrated suite of computer programs produced by Oxford Molecular Group that model antibody structure. See, e.g., Martin et al., 1989, Proc Natl Acad Sci (USA), 86:9268-9272; “AbM™, A Computer Program for Modeling Variable Regions of Antibodies,” Oxford, UK; Oxford Molecular, Ltd. The AbM definition models the tertiary structure of an antibody from primary sequence using a combination of knowledge databases and ab initio methods, such as those described by Samudrala et al., 1999, “Ab Initio Protein Structure Prediction Using a Combined Hierarchical Approach,” in PROTEINS, Structure, Function and Genetics Suppl., 3:194-198. The contact definition is based on an analysis of the available complex crystal structures. See, e.g., MacCallum et al., 1996, I. Mol. Biol., 5:732-45. In another approach, referred to herein as the “conformational definition” of CDRs, the positions of the CDRs may be identified as the residues that make enthalpic contributions to antigen binding. See, e.g., Makabe et al., 2008, Journal of Biological Chemistry, 283:1156-1166. Still other CDR boundary definitions may not strictly follow one of the above approaches, but will nonetheless overlap with at least a portion of the Kabat CDRs, although they may be shortened or lengthened in light of prediction or experimental findings that particular residues or groups of residues do not significantly impact antigen binding. As used herein, a CDR may refer to CDRs defined by any approach known in the art, including combinations of approaches. The methods used herein may utilize CDRs defined according to any of these approaches. For any given embodiment containing more than one CDR, the CDRs may be defined in accordance with any of Kabat, Chothia, extended, IMGT, Paratome, AbM, and / or conformational definitions, or a combination of any of the foregoing.

[0195] As known in the art, a “constant region” of an antibody refers to the constant region of the antibody light chain or the constant region of the antibody heavy chain, either alone or in combination.

[0196] As used herein, “monoclonal antibody” refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally-occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Furthermore, in contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. The modifier “monoclonal”indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies may be made by the hybridoma method first described by Kohler and Milstein, 1975, Nature 256:495, or may be made by recombinant DNA methods such as described in U.S. Pat. No. 4,816,567. The monoclonal antibodies may also be isolated from phage libraries generated using the techniques described in McCafferty et al., 1990, Nature 348:552-554, for example. As used herein, “humanized” antibody refers to forms of non-human (e.g. murine) antibodies that are chimeric immunoglobulins, immunoglobulin chains, or fragments thereof (such as Fv, Fab, Fab’, F(ab’)2 or other antigen-binding subsequences of antibodies) that contain minimal sequence derived from non-human immunoglobulin. Preferably, humanized antibodies are human immunoglobulins (recipient antibody) in which residues from a CDR of the recipient are replaced by residues from a CDR of a non-human species (donor antibody) such as mouse, rat, or rabbit having the desired specificity, affinity, and capacity. The humanized antibody may comprise residues that are found neither in the recipient antibody nor in the imported CDR or framework sequences, but are included to further refine and optimize antibody performance.

[0197] A “human antibody” is one which possesses an amino acid sequence which corresponds to that of an antibody produced by a human and / or has been made using any of the techniques for making human antibodies as disclosed herein. This definition of a human antibody specifically excludes a humanized antibody comprising non-human antigen binding residues.

[0198] The term “chimeric antibody” is intended to refer to antibodies in which the variable region sequences are derived from one species and the constant region sequences are derived from another species, such as an antibody in which the variable region sequences are derived from a mouse antibody and the constant region sequences are derived from a human antibody. The term “epitope” refers to that portion of a molecule capable of being recognized by and bound by an antibody at one or more of the antibody ’ s antigen-binding regions. Epitopes often consist of a surface grouping of molecules such as amino acids or sugar side chains and have specific three-dimensional structural characteristics as well as specific charge characteristics. In some embodiments, the epitope can be a protein epitope. Protein epitopes can be linear or conformational. In a linear epitope, all of the points of interaction between the protein and the interacting molecule (such as an antibody) occur linearly along the primary amino acid sequenceof the protein. A “nonlinear epitope” or “conformational epitope” comprises noncontiguous polypeptides (or amino acids) within the antigenic protein to which an antibody specific to the epitope binds. The term “antigenic epitope” as used herein, is defined as a portion of an antigen to which an antibody can specifically bind as determined by any method well known in the art, for example, by conventional immunoassays. Once a desired epitope on an antigen is determined, it is possible to generate antibodies to that epitope, e.g., using the techniques described in the present specification. Alternatively, during the discovery process, the generation and characterization of antibodies may elucidate information about desirable epitopes. From this information, it is then possible to competitively screen antibodies for binding to the same epitope. An approach to achieve this is to conduct competition and cross-competition studies to find antibodies that compete or cross-compete with one another for binding to IL-6, e.g., the antibodies compete for binding to the antigen.

[0199] The term “compete,” as used herein with regard to an antibody, means that a first antibody, or an antigen-binding portion thereof, binds to an epitope in a manner sufficiently similar to the binding of a second antibody, or an antigen-binding portion thereof, such that the result of binding of the first antibody with its cognate epitope is detectably decreased in the presence of the second antibody compared to the binding of the first antibody in the absence of the second antibody. The alternative, where the binding of the second antibody to its epitope is also detectably decreased in the presence of the first antibody, can, but need not be the case. That is, a first antibody can inhibit the binding of a second antibody to its epitope without that second antibody inhibiting the binding of the first antibody to its respective epitope. However, where each antibody detectably inhibits the binding of the other antibody with its cognate epitope or ligand, whether to the same, greater, or lesser extent, the antibodies are said to “cross-compete” with each other for binding of their respective epitope(s). Both competing and cross -competing antibodies are provided herein. Regardless of the mechanism by which such competition or cross-competition occurs (e.g., steric hindrance, conformational change, or binding to a common epitope, or portion thereof), the skilled artisan would appreciate, based upon the teachings provided herein, that such competing and / or cross-competing antibodies are encompassed and can be useful for the methods disclosed herein.

[0200] As used herein, an antibody “interacts with” IL-6 when the equilibrium dissociation constant is equal to or less than 20 nM, preferably less than about 6 nM, morepreferably less than about 1 nM, most preferably less than about 0.75 nM. In some embodiments, the affinity of the antibody is between 400 and 800 pM, c.g., 450-700, or 500-600 pM.

[0201] An IL-6 antagonist antibody encompasses antibodies that block, antagonize, suppress or reduce (to any degree including significantly) a IL-6 biological activity such as binding to IL-6R, IL-6 / IL-6R complex binding to gpl30, phosphorylation and activation of Stat3, cell proliferation, and stimulation of IL-6 mediated inflammatory or pro-angiogenic pathways. For purpose of the present disclosure, it will be explicitly understood that the term “IL-6 antagonist antibody” encompasses all the previously identified terms, titles, and functional states and characteristics whereby the IL-6 itself, an IL-6 biological activity, or the consequences of the biological activity, are substantially nullified, decreased, or neutralized in any meaningful degree. In some embodiments, an IL-6 antagonist antibody binds IL-6. Examples of IL-6 antagonist antibodies are provided herein.

[0202] An antibody that “preferentially binds” or “specifically binds” (used interchangeably herein) to an epitope is a term well understood in the ait, and methods to determine such specific or preferential binding are also well known in the art. A molecule is said to exhibit “specific binding” or “preferential binding” if it reacts or associates more frequently, and / or more rapidly, and / or with greater duration and / or with greater affinity with a particular cell or substance than it does with alternative cells or substances. An antibody “specifically binds” or “preferentially binds” to a target if it binds with greater affinity, and / or avidity, and / or more readily, and / or with greater duration than it binds to other substances. For example, an antibody that specifically or preferentially binds to an IL-6 epitope is an antibody that binds this epitope with greater affinity, and / or avidity, and / or more readily, and / or with greater duration than it binds to other IL-6 epitopes or non-IL-6 epitopes. It is also understood by reading this definition that, for example, an antibody (or moiety or epitope) that specifically or preferentially binds to a first target may or may not specifically or preferentially bind to a second target. As such, “specific binding” or “preferential binding” does not necessarily require (although it can include) exclusive binding. Generally, but not necessarily, reference to binding means preferential binding.

[0203] As used herein, “substantially pure” refers to material which is at least 50% pure (i.e., free from contaminants), more preferably, at least 90% pure, more preferably, at least 95% pure, yet more preferably, at least 98% pure, and most preferably, at least 99% pure.

[0204] A “host cell” includes an individual cell or cell culture that can be or has been a recipient for vcctor(s) for incorporation of polynucleotide inserts. Host cells include progeny of a single host cell, and the progeny may not necessarily be completely identical (in morphology or in genomic DNA complement) to the original parent cell due to natural, accidental, or deliberate mutation. A host cell includes cells transfected in vivo with a polynucleotide(s) provided herein.

[0205] As known in the art, the term “Fc region” is used to define a C-terminal region of an immunoglobulin heavy chain. The “Fc region” may be a native sequence Fc region or a variant Fc region. Although the boundaries of the Fc region of an immunoglobulin heavy chain might vary, the human IgG heavy chain Fc region is usually defined to stretch from an amino acid residue at position Cys226, or from Pro230, to the carboxyl-terminus thereof. The numbering of the residues in the Fc region is that of the EU index as in Kabat. Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991. The Fc region of an immunoglobulin generally comprises two constant domains, CH2 and CH3. As is known in the art, an Fc region can be present in dimer or monomeric form.

[0206] As used in the art, “Fc receptor” and “FcR” describe a receptor that binds to the Fc region of an antibody. The preferred FcR is a native sequence human FcR. Moreover, a preferred FcR is one which binds an IgG antibody (a gamma receptor) and includes receptors of the FcyRI, FcyRII, and FcyRIII subclasses, including allelic variants and alternatively spliced forms of these receptors. FcyRII receptors include FcyRIIA (an “activating receptor”) and FcyRIIB (an “inhibiting receptor”), which have similar amino acid sequences that differ primarily in the cytoplasmic domains thereof. FcRs are reviewed in Ravetch and Kinet, 1991, Ann. Rev. Immunol., 9:457-92; Capel et al., 1994, Immunomethods, 4:25-34; and de Haas et al., 1995, J. Lab. Clin. Med., 126:330-41. “FcR” also includes the neonatal receptor, FcRn, which is responsible for the transfer of maternal IgGs to the fetus (Guyer et al., 1976, J. Immunol., 117:587 ; and Kim et al., 1994, J. Immunol., 24:249).

[0207] A “functional Fc region” possesses at least one effector function of a native sequence Fc region. Exemplary “effector functions” include Clq binding; complement dependent cytotoxicity; Fc receptor binding; antibody-dependent cell-mediated cytotoxicity; phagocytosis; down-regulation of cell surface receptors (e.g. B cell receptor), etc. Such effector functions generally require the Fc region to be combined with a binding domain (e.g. an antibody variabledomain) and can be assessed using various assays known in the art for evaluating such antibody effector functions.

[0208] A “native sequence Fc region” comprises an amino acid sequence identical to the amino acid sequence of an Fc region found in nature. A “variant Fc region” comprises an amino acid sequence which differs from that of a native sequence Fc region by virtue of at least one amino acid modification, yet retains at least one effector function of the native sequence Fc region. Preferably, the variant Fc region has at least one amino acid substitution compared to a native sequence Fc region or to the Fc region of a parent polypeptide, e.g. from about one to about ten amino acid substitutions, and preferably, from about one to about five amino acid substitutions in a native sequence Fc region or in the Fc region of the parent polypeptide. The variant Fc region herein will preferably possess at least about 80% sequence identity with a native sequence Fc region and / or with an Fc region of a parent polypeptide, and most preferably, at least about 90% sequence identity therewith, more preferably, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% sequence identity therewith.

[0209] As used herein, “treatment” is an approach for obtaining beneficial or desired clinical results.

[0210] As used herein, “IL-6 and / or VEGF related disorders” include, for example, ocular disorders and systemic disorders. Ocular disorders include ocular disorders such as the ophthalmic inflammatory disease scleritis, non-proliferative diabetic retinopathy, proliferative diabetic retinopathy, diabetic macular edema, prevention of diabetic macular edema, prevention of proliferative diabetic retinopathy, wet age-related macular degeneration, prevention of wet age- related macular degeneration, dry age-related macular degeneration, venous, arterial or other blockage of the ocular and or retinal blood vessels with or without retinal edema, anterior and posterior uveitis, uveitic macular edema, and intraocular tumors. IL-6 related disorders also include disorders where there is an elevated level of IL-6 activity due to IL-6 interacting with IL- 6R or soluble IL-6R (sIL-6R). In some embodiments, any one or more of the fusion proteins provided herein and / or any one or more of the conjugates provided herein can be used for treatment or prevention of any one or more of the IL-6 and / or VEGF related disorders. In some embodiments, the disorders include systemic diseases that affect the eye such as Grave’s disease or neuromyelitis optica, or systemic diseases that do not affect the eye such as multiple sclerosis,rheumatoid arthritis. Tn some embodiments, the disorders include cytokine release syndrome following CAR-T or similar immune-oncology therapeutics.

[0211] Additionally, anti-IL6 molecules abrogate the induction of IL-6 expression observed following treatment with anti-PD-l / PD-Ll molecules (Tsukamoto et al, Cancer Res; 2018 78(17); 5011-22). It has also been shown that VEGF signaling blockade can improve anti- PD-L1 treatment (Allen et al, Sci Transl Med 2017 April 12: 9(385)). Thus, these dual inhibitors can be used in combination with PD-l / PDL-1 modulators and / or other immune checkpoint inhibitors, to synergistically treat cancer. Other disorders can include cerebral edema in glioblastoma where anti-IL6 therapy may show additional benefits to anti- VEGF treatments. Other disorders include those with solid tumors.

[0212] As used herein, “Ameliorating” means a lessening or improvement of one or more symptoms as compared to not administering an IL-6 antibody, IL-6 antibody-VEGF Trap fusion, conjugate of IL-6 antibody, and / or conjugate of IL-6 antibody-VEGF Trap fusion. “Ameliorating” also includes shortening or reduction in duration of a symptom.

[0213] As used herein, “VEGF Trap” or similar term denotes the VEGF binding domains (VEGFR1 domain 2, VEGFR2 domain 3). This fragment allows for the protein to work as a VEGF trap, preventing VEGF from binding to cellularly expressed VEGF receptors. An example of this sequence can be found in Table 10. In some embodiments, the VEGF Trap only includes VEGFR1 domain 2, VEGFR2 domain 3. Various embodiments of Trap proteins are known in the art and can be found, for example in U.S. Pub. No. 20150376271, the entirety of which, with respect to various VEGF Trap embodiments (which are VEGFR proteins or fragments thereof) and fusions thereof, is incorporated herein by reference. In some embodiments, the term “VEGF Trap” or similar term refers to a full length extracellular region or any portion thereof, or combination of portions from different VEGF receptors that can antagonize signaling between at least one VEGF and VEGFR.

[0214] As used herein, “IL-6 antibody-VEGF Trap fusion”, “IL-6 antibody-VEGF Trap”, “Ab IL-6-VEGF Trap”, “AntiIL-6-VEGF Trap”, “VEGFR- AntiIL6”, “VEGFR- AntiIL-6”, “VEGF Trap-anti-IL6 Antibody Fusion (TAF)”, “VEGF Trap-IL6”, “VEGFR IL-6”, “IL6- VEGFR” or similar term or inverse terms (e.g. “VEGF Trap-IL-6 Ab,” “VEGF Trap-IL-6 antibody fusion,” etc.) denote the fusion between the IL-6 antibody and the VEGF Trap. Embodiments are depicted in FIG. 6. When used generically, the order of the two terms can be swapped. Whenused specifically, the order of the two terms denotes the relative position of the components in the construct, the term “Ab-Trap”, “IL-6 Ab-VEGF Trap” “Ab IL-6 VEGF Trap” or “Ab IL-6-Trap” or “antiIL-6 VEGF Trap”, “Trap-Ab”, AntiIL-6-VEGFR, AntiIL6-VEGFR or other similar term or inverse terms (e.g. “VEGF Trap-IL-6 Ab,” “VEGF Trap-IL-6 antibody fusion,” etc.) denotes the arrangement of the Ab fused to the relevant domains of a VEGF binding protein so as to provide a VEGF trap. As noted above, this section of the VEGF binding protein is one that prevents VEGF from binding to VEGF receptors. As described herein, the arrangement (ordering) of the Trap and antibody sections can be varied. Thus, unless denoted otherwise explicitly or by context, the phrases used herein regarding Ab-Trap (or I1-6 / VEGF Trap, etc.) fusions, denote all disclosed embodiments for the positioning of the antibody and the Trap. Thus, unless explained otherwise, the phrase Ab-Trap (or I1-6 / VEGF Trap, etc.), denotes the left embodiment in FIG. 6, and the right embodiment in FIG. 6, and both embodiments in FIG. 6. Thus, the general language is denoted as disclosing all three options for convenience. If the orientation is specifically denoted, it can be denoted, for example, by stating that the “arrangement” can be one of: Trap-Ab, Trap IL-6 Ab, VEGF Trap Ab IL-6, VEGF Trap Ab IL6. Similarly, it will be appreciated that the context of some of the present Examples specific orientations or arrangements of the molecules, which are denoted by the context of the Example. Both arrangements (in the alternative and combined) are explicitly contemplated for all discussions of fusion proteins provided herein. In addition, due to the ordering, it is appreciated that the phrase IL-6 Ab, when used in the context of the fusion protein, includes both the option where the antibody is contiguous, FIG. 6, left-hand side, and where the TRAP is positioned “within” the Ab (FIG. 6 right-hand side). Again, the term “Ab” or “antibody”, when used in the fusion protein context (or other similar term), encompasses all three options (left-hand side of FIG. 6, right-hand side of FIG. 6, and both options), unless otherwise noted. In some embodiments, the VEGF Trap is fused to IL-6 in one of the following manners: at an N-terminal end of a heavy chain comprising IL-6 VH; or between a hinge region and after a CHI domain of a heavy chain comprising IL-6 VH. There is no difference between the designations of Ab, antibody, “anti”’ or other similar term when used in a name to designate and antibody or fragment thereof. There is no difference between the designations of “11-6” or “IL6” or “IL-6”. As used herein, when referencing a fusion construct with IL-6, the terms “VEGF”, “VEGFR”, “VEGF Trap”, “VEGFR Trap” arc used interchangeably. The terms can have differentmeanings when used separately from the IL-6 fusion arrangement, which will depend upon the context of the term in question.

[0215] As used herein, the term “biopolymer” denotes that a polymer has been linked to the protein of interest. The term can also be described as the “conjugated” form of the protein. This can be done for all of the proteins described herein. Thus, IL-6 Ab biopolymers and IL-6 antibody-VEGF Trap biopolymers are contemplated for all such IL-6 Ab and IL-6 antibody-VEGF Trap provided herein. In addition, VEGF Trap biopolymers are also provided.

[0216] As used herein, “Antagonistic antibody” denotes an antibody that blocks one or more function or activity of the molecule that the antibody binds to.

[0217] As used herein, an “effective dosage” or “effective amount” of drug, compound, or pharmaceutical composition is an amount sufficient to effect any one or more beneficial or desired results. In more specific aspects, an effective amount prevents, alleviates or ameliorates symptoms of disease, and / or prolongs the survival of the subject being treated. For prophylactic use, beneficial or desired results include eliminating or reducing the risk, lessening the severity, or delaying the outset of the disease, including biochemical, histological and / or behavioral symptoms of the disease, its complications and intermediate pathological phenotypes presenting during development of the disease. For therapeutic use, beneficial or desired results include clinical results such as reducing one or more symptoms of a disease such as, for example, AMD including, for example without limitation, dry AMD and wet AMD, decreasing the dose of other medications required to treat the disease, enhancing the effect of another medication, and / or delaying the progression of AMD in patients. An effective dosage can be administered in one or more administrations. For purposes of this invention, an effective dosage of drug, compound, or pharmaceutical composition is an amount sufficient to accomplish prophylactic or therapeutic treatment either directly or indirectly. As is understood in the clinical context, an effective dosage of a drug, compound, or pharmaceutical composition may or may not be achieved in conjunction with another drug, compound, or pharmaceutical composition. Thus, an “effective dosage” may be considered in the context of administering one or more therapeutic agents, and a single agent may be considered to be given in an effective amount if, in conjunction with one or more other agents, a desirable result may be or is achieved.

[0218] Anti-IL-6 antibodies are administered in an effective regimen meaning a dosage, route of administration and frequency of administration that delays the onset, reduces theseverity, inhibits further deterioration, and / or ameliorates at least one sign or symptom of a disorder. If a patient is already suffering from a disorder, the regimen can be referred to as a therapeutically effective regimen. If the patient is at elevated risk of the disorder relative to the general population but is not yet experiencing symptoms, the regimen can be referred to as a prophylactically effective regimen. In some instances, therapeutic or prophylactic efficacy can be observed in an individual patient relative to historical controls or past experience in the same patient. In other instances, therapeutic or prophylactic efficacy can be demonstrated in a preclinical or clinical trial in a population of treated patients relative to a control population of untreated patients.

[0219] Anti-IL-6-VEGF Traps are administered in an effective regimen meaning a dosage, route of administration and frequency of administration that delays the onset, reduces the severity, inhibits further deterioration, and / or ameliorates at least one sign or symptom of a disorder. If a patient is already suffering from a disorder, the regimen can be referred to as a therapeutically effective regimen. If the patient is at elevated risk of the disorder relative to the general population but is not yet experiencing symptoms, the regimen can be referred to as a prophylactically effective regimen. In some instances, therapeutic or prophylactic efficacy can be observed in an individual patient relative to historical controls or past experience in the same patient. In other instances, therapeutic or prophylactic efficacy can be demonstrated in a preclinical or clinical trial in a population of treated patients relative to a control population of untreated patients.

[0220] The “biological half-life” of a substance is a pharmacokinetic parameter which specifies the time required for one half of the substance to be removed from an organism following introduction of the substance into the organism.

[0221] The term “preventing” or “prevent” refers to (a) keeping a disorder from occurring (b) delaying the onset of a disorder or onset of symptoms of a disorder, or (c) slowing the progression of an existing condition. Unless denoted otherwise, “preventing” does not require the absolute prohibition of the event from occurring.

[0222] An “individual” or a “subject” is a mammal or bird, more preferably, a human. Mammals also include, but are not limited to, farm animals (e.g., cows, pigs, horses, chickens, etc.), sport animals, pets, primates, horses, dogs, cats, mice and rats.

[0223] As used herein, “vector” means a construct, which is capable of delivering, and, preferably, expressing, one or more gcnc(s) or scqucncc(s) of interest in a host cell. Examples of vectors include, but are not limited to, viral vectors, naked DNA or RNA expression vectors, plasmid, cosmid or phage vectors, DNA or RNA expression vectors associated with cationic condensing agents, DNA or RNA expression vectors encapsulated in liposomes, and certain eukaryotic cells, such as producer cells.

[0224] As used herein, “expression control sequence” means a nucleic acid sequence that directs transcription of a nucleic acid. An expression control sequence can be a promoter, such as a constitutive or an inducible promoter, or an enhancer. The expression control sequence is operably linked to the nucleic acid sequence to be transcribed.

[0225] As used herein, “pharmaceutically acceptable carrier” or “pharmaceutical acceptable excipient” includes any material which, when combined with an active ingredient, allows the ingredient to retain biological activity and is non-reactive with the subject’s immune system. Examples include, but are not limited to, any of the standard pharmaceutical carriers such as a phosphate buffered saline solution, water, emulsions such as oil / water emulsion, various types of wetting agents, detergents such as polysorbate 20 to prevent aggregation, and sugars such as sucrose as cryoprotectant. Preferred diluents for aerosol or parenteral administration are phosphate buffered saline (PBS) or normal (0.9%) saline. Compositions comprising such carriers are formulated by well-known conventional methods (see, for example, Remington’s Pharmaceutical Sciences, 18th edition, A. Gennaro, ed., Mack Publishing Co., Easton, PA, 1990; and Remington, The Science and Practice of Pharmacy 20th Ed. Mack Publishing, 2000).

[0226] The term “kOn”, as used herein, refers to the rate constant for association of an antibody (or bioconjugate) to an antigen. Specifically, the rate constants (kouand koff) and equilibrium dissociation constants are measured using full-length antibodies and / or Fab antibody fragments (i.e. univalent) and IL-6.

[0227] The term “koff “, as used herein, refers to the rate constant for dissociation of an antibody (or bioconjugate) from the antibody / antigen complex.

[0228] The term “KD”, as used herein, refers to the equilibrium dissociation constant of an antibody-antigen (or bioconjugate-antigen) interaction.

[0229] Reference to “about” a value or parameter herein includes (and describes) embodiments that are directed to that value or parameter per se. For example, description referringto “about X” includes description of “X.” Numeric ranges are inclusive of the numbers defining the range. “About” denotes a range 10% above or below the specified value. For example, “about 15 mM” denotes a range of 13.5 to 16.5 mM, and “about 0.025%” denotes a range of 0.0225% to 0.0275%.

[0230] The term “patient” includes human and other subjects (including mammals) that receive either prophylactic or therapeutic treatment.

[0231] For purposes of classifying amino acids substitutions as conservative or nonconservative, amino acids are grouped as follows: Group I (hydrophobic side chains): met, ala, val, leu, ile; Group II (neutral hydrophilic side chains): cys, ser, thr; Group III (acidic side chains): asp, glu; Group IV (basic side chains): asn, gin, his, lys, arg; Group V (residues influencing chain orientation): gly, pro; and Group VI (aromatic side chains): trp, tyr, phe. Conservative substitutions involve substitutions between amino acids in the same class. Non-conservative substitutions constitute exchanging a member of one of these classes for a member of another.

[0232] Percentage sequence identities are determined with antibody sequences maximally aligned by the Kabat numbering convention for a variable region or EU numbering for a constant region. After alignment, if a subject antibody region (e.g., the entire mature variable region of a heavy or light chain) is being compared with the same region of a reference antibody, the percentage sequence identity between the subject and reference antibody regions is the number of positions occupied by the same amino acid in both the subject and reference antibody region divided by the total number of aligned positions of the two regions, with gaps not counted, multiplied by 100 to convert to percentage. Sequence identities of other sequences can be determined by aligning sequences using algorithms, such as BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package Release 7.0, Genetics Computer Group, 575 Science Dr., Madison, WI, using default gap parameters, or by inspection, and the best alignment (i.e., resulting in the highest percentage of sequence similarity over a comparison window). Percentage of sequence identity is calculated by comparing two optimally aligned sequences over a window of comparison, determining the number of positions at which the identical residues occur in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison (i.e., the window size), and multiplying the result by 100 to yield the percentage of sequence identity.

[0233] The term “antibody-dependent cellular cytotoxicity”, or ADCC, is a mechanism for inducing cell death that depends upon the interaction of antibody-coated target cells (i.c., cells with bound antibody) with immune cells possessing lytic activity (also referred to as effector cells). Such effector cells include natural killer cells, monocytes / macrophages and neutrophils. ADCC is triggered by interactions between the Fc region of an antibody bound to a cell and Fey receptors, particularly FcyRI and FcyRIII, on immune effector cells such as neutrophils, macrophages and natural killer cells. The target cell is eliminated by phagocytosis or lysis, depending on the type of mediating effector cell. Death of the antibody-coated target cell occurs as a result of effector cell activity.

[0234] A humanized antibody is a genetically engineered antibody in which the CDRs from a non-human “donor” antibody are grafted into a human “acceptor” antibody sequence (see, e.g., Queen, US 5,530,101 and 5,585,089; Winter, US 5,225,539, Carter, US 6,407,213, Adair, US 5,859,2056,881,557, Foote, US 6,881,557). The acceptor antibody sequences can be, for example, a mature human antibody sequence, a composite of such sequences, a consensus sequence of human antibody sequences, or a germline region sequence. Thus, a humanized antibody is an antibody having some or all CDRs entirely or substantially from a donor antibody and variable region framework sequences and constant regions, if present, entirely or substantially from human antibody sequences. Similarly, a humanized heavy chain has at least one, two and usually all three CDRs entirely or substantially from a donor antibody heavy chain, and a heavy chain variable region framework sequence and heavy chain constant region, if present, substantially from human heavy chain variable region framework and constant region sequences. Similarly, a humanized light chain has at least one, two and usually all three CDRs entirely or substantially from a donor antibody light chain, and a light chain variable region framework sequence and light chain constant region, if present, substantially from human light chain variable region framework and constant region sequences. Other than nanobodies and dAbs, a humanized antibody comprises a humanized heavy chain and a humanized light chain. A CDR in a humanized antibody is substantially from a corresponding CDR in a non-human antibody when at least 85%, 90%, 95% or 100% of corresponding residues (as defined by Kabat) are identical between the respective CDRs. The variable region framework sequences of an antibody chain or the constant region of an antibody chain are substantially from a human variable region framework sequence or human constantregion respectively when at least 85, 90, 95 or 100% of corresponding residues defined by Kabat arc identical.

[0235] Although humanized antibodies often incorporate all six CDRs (preferably as defined by Kabat) from a mouse antibody, they can also be made with less than all CDRs (e.g., at least 3, 4, or 5 CDRs from a mouse antibody) (e.g., Pascalis et al, J. Immunol. 169:3076, 2002; Vajdos et al., Journal of Molecular Biology, 320: 415-428, 2002; Iwahashi et al., Mol. Immunol. 36: 1079-1091, 1999; Tamura et al, Journal of Immunology, 164: 1432-1441, 2000).

[0236] A chimeric antibody is an antibody in which the mature variable regions of light and heavy chains of a non-human antibody (e.g., a mouse) are combined with human light and heavy chain constant regions. Such antibodies substantially or entirely retain the binding specificity of the mouse antibody and are about two-thirds human sequence.

[0237] A veneered antibody is a type of humanized antibody that retains some and usually all of the CDRs and some of the non-human variable region framework residues of a non- human antibody but replaces other variable region framework residues that may contribute to B- or T-cell epitopes, for example exposed residues (Padlan, Mol. Immunol. 28:489, 1991) with residues from the corresponding positions of a human antibody sequence. The result is an antibody in which the CDRs are entirely or substantially from a non-human antibody and the variable region frameworks of the non-human antibody are made more human-like by the substitutions. A human antibody can be isolated from a human, or otherwise result from expression of human immunoglobulin genes (e.g., in a transgenic mouse, in vitro or by phage display). Methods for producing human antibodies include the trioma method of Oestberg et al., Hybridoma 2:361-367 (1983); Oestberg, U.S. Patent No. 4,634,664; and Engleman et al., US Patent 4,634,666, use of transgenic mice including human immunoglobulin genes (see, e.g., Lonberg et al., W093 / 12227 (1993); US 5,877,397, US 5,874,299, US 5,814,318, US 5,789,650, US 5,770,429, US 5,661,016, US 5,633,425, US 5,625,126, US 5,569,825, US 5,545,806, Nature 148, 1547-1553 (1994), Nature Biotechnology 14, 826 (1996), Kucherlapati, WO 91 / 10741 (1991) and phage display methods (see, e.g., Dower et al., WO 91 / 17271 and McCafferty et al., WO 92 / 01047, US 5,877,218, US 5,871,907, US 5,858,657, US 5,837,242, US 5,733,743 and US 5,565,332.

[0238] A “polymer” is a molecule composed of many repeating subunits. The subunits also sometimes referred to as “monomers” can be the same or different. There are both natural and synthetic polymers. DNA, protein and complex carbohydrates are examples of naturalpolymers. Poly-styrene and poly-acrylamide are examples of synthetic polymers. A polymer composed of repeating units of a single monomer is called a homopolymcr. A polymer composed of two or more monomers is called a copolymer or sometimes a heteropolymer. A copolymer in which certain monomer types are clustered together are sometimes called block copolymers. Polymers can be linear or branched. When the polymer is branched, polymer chains having a common origin are sometimes referred to as a polymer arm(s).

[0239] An “initiator” is a compound capable of serving as a substrate on which one or more polymerizations can take place using monomers or comonomers as described herein. The polymerization can be a conventional free radical polymerization or preferably a controlled / ”living” radical polymerization, such as Atom Transfer Radical Polymerization (ATRP), Reversible Addition-Fragmentation-Termination (RAFT) polymerization or nitroxide mediated polymerization (NMP). The polymerization can be a “pseudo” controlled polymerization, such as degenerative transfer. Initiators suitable for ATRP contain one or more labile bonds which can be homolytically cleaved to form an initiator fragment, I, being a radical capable of initiating a radical polymerization, and a radical scavenger, I’, which reacts with the radical of the growing polymer chain to reversibly terminate the polymerization. The radical scavenger I’ is typically a halogen, but can also be an organic moiety, such as a nitrile. Also provided herein, the initiator can contain one or more 2-bromoisobutyrate groups as sites for polymerization via ATRP.

[0240] A “chemical linker” refers to a chemical moiety that links two groups together, such as a half-life extending moiety and a protein. The linker can be cleavable or non-cleavable. Cleavable linkers can be hydrolysable, enzymatically cleavable, pH sensitive, photolabile, or disulfide linkers, among others. Other linkers include homobifunctional and heterobifunctional linkers. A “linking group” is a functional group capable of forming a covalent linkage consisting of one or more bonds to a bioactive agent. Non-limiting examples include those illustrated in Table 1 of WO2013059137 (incorporated by reference).

[0241] The term “reactive group” refers to a group that is capable of reacting with another chemical group to form a covalent bond, i.e. is covalently reactive under suitable reaction conditions, and generally represents a point of attachment for another substance. The reactive group is a moiety, such as maleimide or succinimidyl ester, is capable of chemically reacting witha functional group on a different moiety to form a covalent linkage. Reactive groups generally include nucleophiles, electrophiles and photoactivatablc groups.

[0242] As used herein, “phosphorylcholine,” also denoted as “PC,” refers to the ofollowing: o' where * denotes the point of attachment. The phosphorylcholine is a zwitterionic group and includes salts (such as inner salts), and protonated and deprotonated forms thereof.

[0243] As used herein, “phosphorylcholine-based polymer” is a polymer that contains phosphorylcholine. “Zwitterion containing polymer” refers to a polymer that contains a zwitterion.

[0244] Poly(acryloyloxyethyl phosphorylcholine) containing polymer refers to a polymer containing 2-(acryloyloxy)ethyl-2-(trimethylammonium)ethyl phosphate as monomer.

[0245] Poly(methacryloyloxyethyl phosphorylcholine) containing polymer refers to a polymer containing 2-(methacryloyloxy)ethyl-2-(trimethylammonium)ethyl phosphate as monomer.

[0246] As used herein, “molecular weight” in the context of the polymer can be expressed as either a number average molecular weight, or a weight average molecular weight or a peak molecular weight. Unless otherwise indicated, all references to molecular weight herein refer to the peak molecular weight. These molecular weight determinations, number average (Mn), weight average (Mw) and peak (Mp), can be measured using size exclusion chromatography or other liquid chromatography techniques. Other methods for measuring molecular weight values can also be used, such as the use of end-group analysis or the measurement of colligative properties (e.g., freezing-point depression, boiling-point elevation, or osmotic pressure) to determine number average molecular weight, or the use of light scattering techniques, ultracentrifugation or viscometry to determine weight average molecular weight. Also provided herein, the molecular weight can be measured by SEC-MALS (size exclusion chromatography - multi angle light scattering). The polymeric reagents provided herein are typically polydisperse (z.e., number average molecular weight and weight average molecular weight of the polymers are not equal). The Poly Dispersity Index (PDI) provides a measure for the dispersity of polymers in a mixture. PDI is given by the formula Mw / Mn. In this regard a homogenous protein will have a PDI of 1.0 (Mn is the same as Mw). Typically, the PDI for polymers will be above 1.0. Polymers providedherein preferably have relatively low polydispersity (PDI) values of, for example, less than about 1.5, as judged, for example, by SEC-MALS. In other embodiments, the polydispcrsitics (PDI) arc more preferably in the range of about 1.4 to about 1.2, still more preferably less than about 1.15, and still more preferably less than about 1.10, yet still more preferably less than about 1.05, and most preferably less than about 1.03.

[0247] As used herein, “protected,” “protected form,” “protecting group” and “protective group” refer to the presence of a group (z'.e., the protecting group) that prevents or blocks reaction of a particular’ chemically reactive functional group in a molecule under certain reaction conditions. Protecting groups vary depending upon the type of chemically reactive group being protected as well as the reaction conditions to be employed and the presence of additional reactive or protecting groups in the molecule, if any. Suitable protecting groups include those such as found in the treatise by Greene et al., “Protective Groups In Organic Synthesis,” 3rdEdition, John Wiley and Sons, Inc., New York, 1999.

[0248] As used herein, “alkyl” refers to a straight or branched, saturated, aliphatic radical having the number of carbon atoms indicated. For example, Ci-Ce alkyl includes, but is not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, hexyl, etc. Other alkyl groups include, but are not limited to heptyl, octyl, nonyl, decyl, etc. Alkyl can include any number of carbons, such as 1-2, 1-3, 1-4, 1-5, 1-6, 1-7, 1-8, 1-9, 1-10, 2-3, 2-4, 2-5, 2-6, 3-4, 3-5, 3-6, 4-5, 4-6 and 5-6 carbons.

[0249] The term “lower” referred to above and hereinafter in connection with organic radicals or compounds respectively defines a compound or radical which can be branched or unbranched with up to and including 7, preferably up to and including 4 and (as unbranched) one or two carbon atoms.

[0250] As used herein, “alkylene” refers to an alkyl group, as defined above, linking at least two other groups, z.e., a divalent hydrocarbon radical. The two moieties linked to the alkylene can be linked to the same atom or different atoms of the alkylene. For instance, a straight chain alkylene can be the bivalent radical of -(CHijn, where n is 1, 2, 3, 4, 5 or 6. Alkylene groups include, but are not limited to, methylene, ethylene, propylene, isopropylene, butylene, isobutylene, sec -butylene, pentylene and hexylene.

[0251] Substituents for the alkyl, alkenyl, alkylene, heteroalkyl, heteroalkylene, heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, and heterocycloalkenylradicals can be one or more of a variety of groups selected from, but not limited to: -OR’, =0, =NR’,=N-0R’, -NR’R”, -SR’, -halogen, -SiR’R”R”‘, -0C(0)R’, -C(0)R’, -C02R’, -CONR’R”, -0C( 0)NR’R”, -NR”C(0)R’, -NR’-C(0)NR”R”‘, -NR”C(0)2R’, -NR-C(NR’R”R’”)=NR”“, -NR-C( NR’R”)=NR”’, -S(O)R’, -S(O)2R’, -S(O)2NR’R”, -NRSO2R’, -CN and -N02in a number ranging from 1 to (2m’+l), where m’ is the total number of carbon atoms in such radical. Each of R’, R”, R”‘ and R”“ independently refers to hydrogen, substituted or unsubstituted heteroalkyl, substituted or unsubstituted aryl, e.g., aryl substituted with 1-3 halogens, substituted or unsubstituted alkyl, alkoxy or thioalkoxy groups, or arylalkyl groups. When R’ and R” are attached to the same nitrogen atom, they can be combined with the nitrogen atom to form a 5-, 6-, or 7-membered ring. For example, -NR’R” is meant to include, but not be limited to, 1-pyrrolidinyl and 4-morpholinyl.

[0252] As used herein, “alkoxy” refers to alkyl group attached to an oxygen atom and forms radical -O-R, wherein R is alkyl. Alkoxy groups include, for example, methoxy, ethoxy, propoxy, iso-propoxy, butoxy, 2-butoxy, iso-butoxy, sec-butoxy, tert-butoxy, pentoxy, hexoxy, etc. The alkoxy groups can be further substituted with a variety of substituents described herein. For example, the alkoxy groups can be substituted with halogens to form a “halo-alkoxy” group.

[0253] As used herein, “carboxyalkyl” means an alkyl group (as defined herein) substituted with a carboxy group. The term “carboxycycloalkyl” means an cycloalkyl group (as defined herein) substituted with a carboxy group. The term alkoxyalkyl means an alkyl group (as defined herein) substituted with an alkoxy group. The term “carboxy” employed herein refers to carboxylic acids and their esters.

[0254] As used herein, “haloalkyl” refers to alkyl as defined above where some or all of the hydrogen atoms are substituted with halogen atoms. Halogen (halo) preferably represents chloro or fluoro, but may also be bromo or iodo. For example, haloalkyl includes trifluoromethyl, fluoromethyl, 1,2,3,4,5-pentafluoro-phenyl, etc. The term “perfluoro” defines a compound or radical which has all available hydrogens that are replaced with fluorine. For example, perfluorophenyl refers to 1,2,3,4,5-pentafluorophenyl, perfluoromethyl refers to 1,1,1 -trifluoromethyl, and perfluoromethoxy refers to 1,1,1 -trifluoromethoxy. Haloalkyl can also be referred to as halo- substitute alkyl, such as fluoro-substituted alkyl.

[0255] As used herein, “cytokine” in the context provided herein is a member of a group of protein signaling molecules that may participate in cell-cell communication in immuneand inflammatory responses. Cytokines are typically small, water-soluble glycoproteins that have a mass of about 8-35 kDa.

[0256] As used herein, “cycloalkyl” refers to a saturated mono- or multi- cyclic aliphatic ring system that contains from about 3 to 12, from 3 to 10, from 3 to 7, or from 3 to 6 carbon atoms. When cycloalkyl group is composed of two or more rings, the rings may be joined together with a fused ring or a spiro ring structure. When cycloalkyl group is composed of three or more rings, the rings may also join together forming a bridged ring structure. Monocyclic rings include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cyclooctyl. Bicyclic and polycyclic rings include, for example, bicyclo [l.l.l]pentane, bicyclco[2.1.1]heptane, norbornane, decahydronaphthalene and adamantane. For example, C3-8 cycloalkyl includes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclooctyl, and norbornane.

[0257] As used herein, “endocyclic” refers to an atom or group of atoms which comprise part of a cyclic ring structure.

[0258] As used herein, “exocyclic” refers to an atom or group of atoms which are attached but do not define the cyclic ring structure.

[0259] As used herein, “cyclic alkyl ether” refers to a 4 or 5 member cyclic alkyl group having 3 or 4 endocyclic carbon atoms and 1 endocyclic oxygen or sulfur atom (e.g., oxetane, thietane, tetrahydro furan, tetrahydrothiophene); or a 6 to 7 member cyclic alkyl group having 1 or 2 endocyclic oxygen or sulfur atoms (e.g., tetrahydropyran, 1,3-dioxane, 1,4-dioxane, tetrahydro thiopyran, 1,3-dithiane, 1,4-dithiane, 1,4-oxathiane).

[0260] As used herein, “alkenyl” refers to either a straight chain or branched hydrocarbon of 2 to 6 carbon atoms, having at least one double bond. Examples of alkenyl groups include, but are not limited to, vinyl, propenyl, isopropenyl, 1-butenyl, 2-butenyl, isobutenyl, butadienyl, 1-pentenyl, 2-pentenyl, isopentenyl, 1,3-pentadienyl, 1 ,4-pentadienyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 1,3-hexadienyl, 1,4-hexadienyl, 1,5-hexadienyl, 2,4-hexadienyl, or 1,3,5-hexatrienyl. Alkenyl groups can also have from 2 to 3, 2 to 4, 2 to 5, 3 to 4, 3 to 5, 3 to 6, 4 to 5, 4 to 6 and 5 to 6 carbons.

[0261] As used herein, “alkenylene” refers to an alkenyl group, as defined above, linking at least two other groups, i.e., a divalent hydrocarbon radical. The two moieties linked to the alkenylene can be linked to the same atom or different atoms of the alkenylene. Alkenylenegroups include, but are not limited to, ethenylene, propenylene, isopropenylene, butenylene, isobutcnylcnc, sec -butenylene, pcntcnylcnc and hcxcnylcnc.

[0262] As used herein, “alkynyl” refers to either a straight chain or branched hydrocarbon of 2 to 6 carbon atoms, having at least one triple bond. Examples of alkynyl groups include, but are not limited to, acetylenyl, propynyl, 1-butynyl, 2-butynyl, isobutynyl, sec-butynyl, butadiynyl, 1 -pentynyl, 2-pentynyl, isopentynyl, 1,3-pentadiynyl, 1,4-pentadiynyl, 1 -hexynyl, 2-hexynyl, 3-hexynyl, 1,3-hexadiynyl, 1 ,4-hexadiynyl, 1,5 -hexadiynyl, 2,4-hexadiynyl, or 1,3,5-hexatriynyl. Alkynyl groups can also have from 2 to 3, 2 to 4, 2 to 5, 3 to 4, 3 to 5, 3 to 6, 4 to 5, 4 to 6 and 5 to 6 carbons.

[0263] As used herein, “alkynylene” refers to an alkynyl group, as defined above, linking at least two other groups, i.e., a divalent hydrocarbon radical. The two moieties linked to the alkynylene can be linked to the same atom or different atoms of the alkynylene. Alkynylene groups include, but are not limited to, ethynylene, propynylene, butynylene, sec-butynylene, pentynylene and hexynylene.

[0264] As used herein, “cycloalkylene” refers to a cycloalkyl group, as defined above, linking at least two other groups, i.e., a divalent hydrocarbon radical. The two moieties linked to the cycloalkylene can be linked to the same atom or different atoms of the cycloalkylene. Cycloalkylene groups include, but are not limited to, cyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene, and cyclooctylene.

[0265] As used herein, “heterocycloalkyl” refers to a ring system having from 3 ring members to about 20 ring members and from 1 to about 5 heteroatoms such as N, O and S. Additional heteroatoms can also be useful, including, but not limited to, B, Al, Si and P. The heteroatoms can also be oxidized, such as, but not limited to, -S(O)- and -S(O)2-. For example, heterocycle includes, but is not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, morpholino, pyrrolidinyl, pyrrolinyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl, piperazinyl, piperidinyl, indolinyl, quinuclidinyl and l,4-dioxa-8-aza-spiro[4.5]dec-8-yl.

[0266] As used herein, “heterocycloalkylene” refers to a heterocyclalkyl group, as defined above, linking at least two other groups. The two moieties linked to the heterocycloalkylene can be linked to the same atom or different atoms of the heterocyclo alkylene.

[0267] As used herein, “aryl” refers to a monocyclic or multicyclic (e.g., fused bicyclic, tricyclic or greater) aromatic ring assembly containing 6 to 16 carbon atoms. For example, arylmay be phenyl, benzyl or naphthyl, preferably phenyl. Aryl groups can be mono-, di- or tri-substitutcd by one, two or three radicals selected from alkyl, alkoxy, aryl, hydroxy, halogen, cyano, amino, amino-alkyl, trifluoromethyl, alkylenedioxy and oxy-Ci-Ca-alkylcnc; all of which are optionally further substituted, for instance as hereinbefore defined; or 1- or 2-naphthyl; or 1- or 2-phenanthrenyl. Alkylenedioxy is a divalent substitute attached to two adjacent carbon atoms of phenyl, e.g. methylenedioxy or ethylenedioxy. Oxy-Ci-Cs-alkylcnc is also a divalent substituent attached to two adjacent carbon atoms of phenyl, e.g. oxyethylene or oxypropylene. An example for oxy- Ci-Ca-alkylene-phenyl is 2,3-dihydrobenzofuran-5-yl.

[0268] Preferred as aryl is naphthyl, phenyl or phenyl mono- or disubstituted by alkoxy, phenyl, halogen, alkyl or trifluoromethyl, especially phenyl or phenyl-mono- or disubstituted by alkoxy, halogen or trifluoromethyl, and in particular phenyl.

[0269] Examples of substituted phenyl groups as R are, e.g. 4-chlorophen-l-yl, 3,4-dichlorophen-l-yl, 4-methoxyphen-l-yl, 4-methylphen-l-yl, 4-aminomethylphen-l-yl, 4-methoxyethylaminomethylphen- 1 -yl, 4-hydroxyethylaminomethylphen- 1 -yl,4-hydroxyethyl-(methyl)-aminomethylphen-l-yl, 3-aminomethylphen-l-yl,4-N-acetylaminomethylphen-l-yl, 4-aminophen-l-yl, 3-aminophen-l-yl, 2-aminophen-l-yl, 4-phenyl-phen- 1 -yl, 4-(imidazol- 1 -yl)-phenyl, 4-(imidazol- 1 -ylmethyl)-phen- 1 -yl,4-(morpholin- 1 -yl)-phen- 1 -yl, 4-(morpholin- 1 -ylmethylj-phen- 1 -yl,4-(2-methoxyethylaminomethyl)-phen- 1-yl and 4-(pyrrolidin- 1 -ylmethyl)-phen- 1 -yl,4-(thiophenyl)-phen- 1 -yl, 4-(3-thiophenyl)-phen- 1 -yl, 4-(4-methylpiperazin- 1 -yl)-phen- 1 -yl, and 4-(piperidinyl)-phenyl and 4-(pyridinyl)-phenyl optionally substituted in the heterocyclic ring.

[0270] As used herein, “arylene” refers to an aryl group, as defined above, linking at least two other groups. The two moieties linked to the arylene are linked to different atoms of the arylene. Arylene groups include, but are not limited to, phenylene.

[0271] As used herein, “arylene-oxy” refers to an arylene group, as defined above, where one of the moieties linked to the arylene is linked through an oxygen atom. Arylene-oxy groups include, but are not limited to, phenylene-oxy.

[0272] Similarly, substituents for the aryl and heteroaryl groups are varied and are selected from: -halogen, -OR’, -OC(O)R’, -NR’R”, -SR’, -R’, -CN, -NO2, -CO2R’, -CONR’R”, -C(O)R’, -OC(O)NR’R”, -NR”C(O)R’, -NR”C(O)2R’, -NR’-C(O)NR”R”‘, -NH-C(NH2)=NH, -NR’C(NH2)=NH, -NH-C(NH2)=NR’, -S(O)R’, -S(O)2R’, -S(O)2NR’R”, -N3, -CH(Ph)2, pcrfluoro(Ci-C4)alkoxy, and pcrfluoro(Ci-C4)alkyl, in a number ranging from zero to the total number of open valences on the aromatic ring system; and where R’ , R” and R” ‘ are independently selected from hydrogen, (Ci-Cs)alkyl and heteroalkyl, unsubstituted aryl and heteroaryl, (unsubstituted aryl)-(Ci-C4)alkyl, and (unsubstituted aryl)oxy-(Ci-C4)alkyl.

[0273] Two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced with a substituent of the formula -T-C(O)-(CH2)q-U-, wherein T and U are independently -NH-, -O-, -CH2- or a single bond, and q is an integer of from 0 to 2. Alternatively, two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced with a substituent of the formula -A-(CH2)r-B-, wherein A and B are independently -CH2-, -O-, -NH-, -S-, -S(O)-, -S(O)2-, -S(O)2NR’- or a single bond, and r is an integer of from 1 to 3. One of the single bonds of the new ring so formed may optionally be replaced with a double bond. Alternatively, two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced with a substituent of the formula -(CH2)s-X-(CH2)t-, where s and t are independently integers of from 0 to 3, and X is -O-, -NR’-, -S-, -S(O)-, -S(O)2-, or -S(O)2NR’-. The substituent R’ in -NR’- and -S(O)2NR’- is selected from hydrogen or unsubstituted (Ci-Cfi)alkyl.

[0274] As used herein, “heteroaryl” refers to a monocyclic or fused bicyclic or tricyclic aromatic ring assembly containing 5 to 16 ring atoms, where from 1 to 4 of the ring atoms are a heteroatom each N, O or S. For example, heteroaryl includes pyridyl, indolyl, indazolyl, quinoxalinyl, quinolinyl, isoquinolinyl, benzothienyl, benzofuranyl, furanyl, pyrrolyl, thiazolyl, benzothiazolyl, oxazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazolyl, imidazolyl, thienyl, or any other radicals substituted, especially mono- or di-substituted, by e.g. alkyl, nitro or halogen. Pyridyl represents 2-, 3- or 4-pyridyl, advantageously 2- or 3-pyridyl. Thienyl represents 2- or 3-thienyl. Quinolinyl represents preferably 2-, 3- or 4-quinolinyl. Isoquinolinyl represents preferably 1-, 3- or 4-isoquinolinyl. Benzopyranyl, benzothiopyranyl represents preferably3-benzopyranyl or 3-benzothiopyranyl, respectively. Thiazolyl represents preferably 2- or4-thiazolyl, and most preferred, 4-thiazolyl. Triazolyl is preferably 1-, 2- or 5-(l,2,4-triazolyl). Tetrazolyl is preferably 5-tetrazolyl.

[0275] Preferably, heteroaryl is pyridyl, indolyl, quinolinyl, pyrrolyl, thiazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazolyl, imidazolyl, thienyl, furanyl, benzo thiazolyl,benzofuranyl, isoquinolinyl, benzothienyl, oxazolyl, indazolyl, or any of the radicals substituted, especially mono- or di-substituted.

[0276] The term “heteroalkyl” refers to an alkyl group having from 1 to 3 heteroatoms such as N, O and S. Additional heteroatoms can also be useful, including, but not limited to, B, Al, Si and P. The heteroatoms can also be oxidized, such as, but not limited to, -S(O)- and -S(O)2-. For example, heteroalkyl can include ethers, thioethers, alkyl-amines and alkyl-thiols.

[0277] The term “heteroalkylene” refers to a heteroalkyl group, as defined above, linking at least two other groups. The two moieties linked to the heteroalkylene can be linked to the same atom or different atoms of the heteroalkylene.

[0278] As used herein, “electrophile” refers to an ion or atom or collection of atoms, which may be ionic, having an electrophilic center, z.e., a center that is electron seeking, capable of reacting with a nucleophile. An electrophile (or electrophilic reagent) is a reagent that forms a bond to its reaction partner (the nucleophile) by accepting both bonding electrons from that reaction partner.

[0279] As used herein, “nucleophile” refers to an ion or atom or collection of atoms, which may be ionic, having a nucleophilic center, i.e., a center that is seeking an electrophilic center or capable of reacting with an electrophile. A nucleophile (or nucleophilic reagent) is a reagent that forms a bond to its reaction partner (the electrophile) by donating both bonding electrons. A “nucleophilic group” refers to a nucleophile after it has reacted with a reactive group. Non limiting examples include amino, hydroxyl, alkoxy, haloalkoxy and the like.

[0280] As used herein, “maleimido” refers to a pyrrole-2, 5-dione- 1-yl group having the structure:which upon reaction with a sulfhydryl (e.g., a thio alkyl) forms an -S-maleimido group having the structurewhereindicates the point of attachment for the maleimido group and“indicates the point of attachment of the sulfur atom the thiol to the remainder of the original sulfhydryl bearing group.

[0281] For the purpose of this disclosure, “naturally occurring amino acids” found in proteins and polypeptides are L-alanine, L-arginine, L-asparagine, L-aspartic acid, L-cysteine, L- glutamine, L-glutamic acid, L-glycine, L-histidine, L-isoleucine, L-leucine, L-lysine, L- methionine, L-phenylalanine, L-proline, L-serine, L-threonine, L-tryptophan, L-tyrosine, and or L-valine. “Nonnaturally occurring amino acids” found in proteins are any amino acid other than those recited as naturally occurring amino acids. Nonnaturally occurring amino acids include, without limitation, the D isomers of the naturally occurring amino acids, and mixtures of D and L isomers of the naturally occurring amino acids. Other amino acids, such as 4hydroxyproline, desmosine, isodesmosine, 5hydroxylysine, epsilonNmethyllysine, 3methylhistidine, although found in naturally occurring proteins, are considered to be nonnaturally occurring amino acids found in proteins for the purpose of this disclosure as they are generally introduced by means other than ribosomal translation of mRNA.

[0282] As used herein, “linear” in reference to the geometry, architecture or overall structure of a polymer, refers to polymer having a single polymer arm.

[0283] As used herein, “branched,” in reference to the geometry, architecture or overall structure of a polymer, refers to a polymer having 2 or more polymer “arms” extending from a core structure contained within an initiator. The initiator may be employed in an atom transfer radical polymerization (ATRP) reaction. A branched polymer may possess 2 polymer chains (arms), 3 polymer arms, 4 polymer arms, 5 polymer arms, 6 polymer arms, 7 polymer arms, 8 polymer arms, 9 polymer arms or more. Each polymer arm extends from a polymer initiation site. Each polymer initiation site is capable of being a site for the growth of a polymer chain by the addition of monomers. For example and not by way of limitation, using ATRP, the site of polymer initiation on an initiator is typically an organic halide undergoing a reversible redox process catalyzed by a transition metal compound such as cuprous halide. Preferably, the halide is a bromine.

[0284] As used herein, “pharmaceutically acceptable excipient” refer to an excipient that can be included in the compositions provided herein and that causes no significant adverse toxicological effect on the patient and is approved or approvable by the FDA for therapeutic use,particularly in humans. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, normal saline solutions, lactated Ringer’s, normal sucrose, normal glucose and the like.

[0285] As used herein, “OG1786” is a 9-arm initiator used for polymer synthesis with the structure shown in FIG. 2D, which depicts that salt form of OG1786 with trifluororacetic acid. OG1786 may also be used as provided herein as other salts or as the free base.

[0286] As used herein, “OG1801” is an approximately (+ / - 25%) 800 kDa polymer (either by Mn or Mp) made using OG1786 as an initiator for ATRP synthesis using the monomer HEMA-PC. The structure of OG1801 is shown in FIG. 2J.

[0287] As used herein, “OG1802” is OG1801 with a maleimide functionality added, and it has the structure shown in FIG. 2K, wherein each of m, , na, ru, 115, no. n?, ng and ng is an integer (positive) (from 0 up to about 3000) such that the total molecular weight of the polymer is (Mw) 800,000 ± 20% Daltons. When the term OG1802 is used to modify a protein term (such as VEGF trap or anti-IL-6 antibody), it designates that the protein is the conjugate protein.

[0288] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art as provided herein. In case of conflict, the present specification, including definitions, will control. Throughout this specification and claims, the word “comprise,” or variations such as “comprises” or “comprising” will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers. Unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. Any example(s) following the term “e.g.” or “for example” is not meant to be exhaustive or limiting.

[0289] The phrase “a” or “an” entity refers to one or more of that entity; for example, a compound refers to one or more compounds or at least one compound. As such, the terms “a” (or “an”), “one or more”, and “at least one” can be used interchangeably herein.

[0290] As used herein, “about” means variation one might see in measurements taken among different instruments, samples, and sample preparations.

[0291] As used herein “unconjugated protein” and “free protein” are used interchangeably to denote the protein that is not conjugated to a polymer (e.g., not conjugated to a phosphorylcholine-containing polymer).

[0292] As used herein, “molar amount” denotes a measure of the molar quantity of a molecule. In some embodiments, molar amount is a molar concentration (c.g., M, mM, pM, nM, etc.). In some embodiments, molar amount is expressed in units of moles (e.g., moles, millimoles, micromoles, etc.).

[0293] As used herein, CDR positions follow their order of appearance in variable domain when described. For example, heavy chain positions can be described as S35H or G66D. As used herein, Fc positions follow EU numbering when described or when noted to be in EU numbering. For example, mutations of antibodies can be described as L234A or L235A, which would be according to EU numbering. Positions may also be defined according to specified positions within a specific SEQ ID or sequence provided herein.

[0294] Multi-angle light scattering (MALS) is a technique of analyzing macromolecules where the laser light impinges on the molecule, the oscillating electric field of the light induces an oscillating dipole within it. This oscillating dipole will re-radiate light and can be measured using a MALS detector such as Wyatt miniDawn TREOS. The intensity of the radiated light depends on the magnitude of the dipole induced in the macromolecule which in turn is proportional to the polarizability of the macromolecule, the larger the induced dipole, and hence, the greater the intensity of the scattered light. Therefore, in order to analyze the scattering from a solution of such macromolecules, one should know their polarizability relative to the surrounding medium (e.g., the solvent). This may be determined from a measurement of the change, A / z, of the solution’s refractive index n with the molecular concentration change, Ac, by measuring the dn / dc (=An / Ac) value using a Wyatt Optilab T-rEX differential refractometer. Two molar weight parameters that MALS determination employ are number average molecular weight (Mn) and weight average molecular weight (Mw) where the poly dispersity index (PDI) equals Mw divided by Mn. SEC also allows another average molecular weight determination of the peak molecular weight Mp which is defined as the molecular weight of the highest peak at the SEC.

[0295] The PDI is used as a measure of the broadness of a molecular weight distribution of a polymer and bioconjugate which is derived from conjugation of a discrete protein to a polydisperse biopolymer (e.g., OG1802). For a protein sample, its polydispersity is close to 1.0 due to the fact that it is a product of translation where every protein molecule in a solution is expected to have almost the same length and molar mass. In contrast, due to the polydisperse nature of the biopolymer where the various length of polymer chains are synthesized during thepolymerization process, it is very important to determine the PDI of the sample as one of its quality attribute for narrow distribution of molecular weight.

[0296] Size exclusion chromatography (SEC) is a chromatography technique in which molecules in solution are separated by their size. Typically an aqueous solution is applied to transport the sample through the column which is packed with resins of various pore sizes. The resin is expected to be inert to the analyte when passing through the column and the analytes separate from each other based on their unique size and the pore size characteristics of the selected column.

[0297] Coupling the SEC with MALS or SEC / MALS provides accurate distribution of molar mass and size (root mean square radius) as opposed to relying on a set of SEC calibration standards. This type of arrangement has many advantages over traditional column calibration methods. Since the light scattering and concentration are measured for each eluting fraction, the molar mass and size can be determined independently of the elution position. This is particularly relevant for species with non-globular shaped macromolecules such as the biopolymers (OG1802) or bioconjugates; such species typically do not elute in a manner that might be described by a set of column calibration standards.

[0298] In some embodiments, a SEC / MALS analysis includes a Waters HPLC system with Alliance 2695 solvent delivery module and Waters 2996 Photodiole Array Detector equipped with a Shodex SEC-HPLC column (7.8x300mm). This is connected online with a Wyatt miniDawn TREOS and Wyatt Optilab T-rEX differential refractometer. The Empower software from Waters can be used to control the Waters HPLC system and the ASTRA V 6.1.7.16 software from Wyatt can be used to acquire the MALS data from the Wyatt miniDawn TREOS, dn / dc data from the T-rEX detector and the mass recovery data using the A280 absorbance signal from the Waters 2996 Photodiole Array detector. SEC can be carried out at Iml / min in IxPBS pH 7.4, upon sample injection, the MALS and RI signals can be analyzed by the ASTRA software for determination of absolute molar mass (Mp, Mw, Mn) and polydisperse index (PDI). In addition, the calculation also involves the input dn / dc values for polymer and protein as 0.142 and 0.183, respectively. For bioconjugates dn / dc value, the dn / dc is calculated based on the weighted MW of the polymer and the protein to be about 0.148 using the formula below:Conjugate dn / dc = 0.142 x [ MWpolymer / (M'Wpolymer+M'Wprotein)]+ 0.183 x [MWprotein / (MW polymer+MW protein)]where M\\rpolymer for OG1802 measured by SEC-MALS is about 800 kDa and the M'Wprotein for anti-IL-6 measured by SEC-MALS is about 145 kDa, the expected total molecular weight of the bioconjugate measured by SEC-MALS is about 1000 kDa. The M prolein for the antiIL-6 VEGF Trap or VEGF Trap-antiIL-6 is about 192 kDa, and the expected total molecular weight of the bioconjugate is 1000-1100 kDa.

[0299] In some embodiments, pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein, a buffer; a surfactant, and optionally a tonicity agent. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114. In some embodiments, the fusion protein is conjugated to a polymer. In some embodiments, the fusion protein comprises an antagonist antibody or a fragment thereof, that specifically binds to IL-6 that is conjugated to a polymer. In some embodiments, the fusion protein comprises an antagonistic IL-6 antibody or fragment thereof comprising: a) a heavy chain amino acid variable region that comprises a heavy chain that has a sequence of at least one of SEQ ID NOs: 7-13,19-27, 89, 90, 256-262; and b) a light chain amino acid variable region that comprises the light chain that has a sequences of at least one of SEQ ID NOs: 91-93, 28-30. In some embodiments, the fusion protein comprises an antagonist IL-6 antibody or fragment thereof comprising: a heavy chain variable region (VH) comprising 3 complementarity determining regions: VH (CDR1), VH CDR2, and VH CDR3 having an amino acid sequence from the CDRs listed in SEQ ID NO: 256; and a light chain variable region (VL) comprising a VL CDR1, VL CDR2, and VL CDR3 having an amino acid sequence selected from the group of CDRs listed in SEQ ID NO: 91-93. In some embodiments, the fusion protein further comprises an antagonistic antibody or fragment thereof that binds to IL-6, the antibody comprising: a CDRH1 that is a CDRH1 in SEQ ID NO: 172; a CDRH2 that is a CDRH2 in SEQ ID NO: 173; a CDRL1 that is a CDRL1 in SEQ ID NO: 199; a CDRL2 that is a CDRL2 in SEQ ID NO: 200; a CDRL3 that is a CDRL3 in SEQ ID NO: 201; at least one of the following mutations (EU numbering): L234A, L235A, and G237A; and at least one of the following mutations (EU numbering): Q347C or L443C. In some embodiments, the VEGF Trap is positioned either: a) at an N-terminal end of a heavy chain comprising IL-6 VH; or b) between a hinge region and after a CHI domain of a heavy chain comprising IL-6 VH. In some embodiments, the buffer comprises sodium acetate. In someembodiments, the surfactant comprises polysorbate 20. Tn some embodiments, the polysorbate 20 is between about 0.01% (w / w) and about 0.05% (w / w) of the formulation. In some embodiments, the surfactant comprises polysorbate 80. In some embodiments, the polysorbate 80 is between about 0.01% (w / w) and about 0.05% (w / w) of the formulation. In some embodiments, the polysorbate 20 is about 0.025% (w / w) of the formulation. In some embodiments, the polysorbate 80 is about 0.025% (w / w) of the formulation. In some embodiments, the sodium acetate concentration is between about 0.1 mM to about 75 mM. In some embodiments, the sodium acetate concentration is about 50 mM. In some embodiments, the formulation is between about pH 4.5 and about pH 6.5. In some embodiments, the formulation is about pH 5.0. In some embodiments, the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer. In some embodiments, the polydispersity index (PDI) is between about 0.5 and about 2. In some embodiments, the poly dispersity index (PDI) is 1. In some embodiments, the tonicity agent is between about 2%-10% (weight / volume). In some embodiments, the tonicity agent is any of sucrose, trehalose, glucose, fructose, maltose, lactose, mannitol, and sorbitol. In some embodiments, the tonicity agent is sucrose or trehalose. In some embodiments, the tonicity agent comprises about 4% (weight / volume) sucrose. In some embodiments, the tonicity agent comprises about 4% (weight / volume) trehalose. In some embodiments, the tonicity agent comprises about 5.2% (weight / volume) sucrose. In some embodiments, the tonicity agent comprises about 5.2% (weight / volume) trehalose. In some embodiments, the tonicity agent comprises about 6% (weight / volume) sucrose. In some embodiments, the tonicity agent comprises about 6% (weight / volume) trehalose. In some embodiments, the concentration of fusion protein is between about 20-75 mg / mL. In some embodiments, the concentration of fusion protein is about 50 mg / mL. In some embodiments, the formulation is not the formulation of any one of arrangements 1-98.

[0300] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of a fusion protein; a buffer solution, the buffer solution comprising sodium acetate, a surfactant, and optionally a tonicity agent. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114. In some embodiments, the fusion protein is conjugated to a polymer. In some embodiments, the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer. In someembodiments, the fusion protein comprises an antagonist antibody or a fragment thereof, that specifically binds to IL-6 that is conjugated to a polymer. In some embodiments, the fusion protein comprises an antagonistic IL-6 antibody or fragment thereof comprising: a) a heavy chain amino acid variable region that comprises a heavy chain that has a sequence of at least one of SEQ ID NOs: 7-13,19-27, 89, 90, 256-262; and b) a light chain amino acid variable region that comprises the light chain that has a sequences of at least one of SEQ ID NOs: 91-93, 28-30. In some embodiments, the fusion protein comprises an antagonist IL-6 antibody or fragment thereof comprising: a heavy chain variable region (VH) comprising 3 complementarity determining regions: VH (CDR1), VH CDR2, and VH CDR3 having an amino acid sequence from the CDRs listed in SEQ ID NO: 256; and a light chain variable region (VL) comprising a VL CDR1, VL CDR2, and VL CDR3 having an amino acid sequence selected from the group of CDRs listed in SEQ ID NO: 91-93. In some embodiments, the fusion protein further comprises an antagonistic antibody or fragment thereof that binds to IL-6, the antibody comprising: a CDRH1 that is a CDRH1 in SEQ ID NO: 172; a CDRH2 that is a CDRH2 in SEQ ID NO: 173; a CDRL1 that is a CDRL1 in SEQ ID NO: 199; a CDRL2 that is a CDRL2 in SEQ ID NO: 200; a CDRL3 that is a CDRL3 in SEQ ID NO: 201; at least one of the following mutations (EU numbering): L234A, L235A, and G237A; and at least one of the following mutations (EU numbering): Q347C or L443C. In some embodiments, the VEGF Trap is positioned either: a) at an N-terminal end of a heavy chain comprising IL-6 VH; or b) between a hinge region and after a CHI domain of a heavy chain comprising IL-6 VH. In some embodiments, the VEGF Trap consists of the sequence of SEQ ID NO: 114. In some embodiments, the surfactant comprises between about 0.01% to about 0.05% (weight / weight) polysorbate 20. In some embodiments, the surfactant comprises about 0.025% (weight / weight) polysorbate 20. In some embodiments, the surfactant comprises between about 0.01% to about 0.05% (weight / weight) polysorbate 80. In some embodiments, the surfactant comprises about 0.025% (weight / weight) polysorbate 80. In some embodiments, the formulation is between about pH 4.5 to about pH 6.5. In some embodiments, the formulation is about pH 5.0. In some embodiments, the formulation is stable for at least 52 weeks. In some embodiments, the formulation is stable for at least 52 weeks at 4 centigrade. In some embodiments, the formulation is stable for at least 8 weeks at 37 centigrade. In some embodiments, the formulation is stable in acidic conditions. In some embodiments, the formulation configured for intravitreal injection. In some embodiments, the sodium acetate concentration in the buffer solution is about 0.1 mM toabout 75 mM. In some embodiments, the concentration of fusion protein is between about 20-75 mg / mL. In some embodiments, the concentration of fusion protein is about 50 mg / mL. In some embodiments, the tonicity agent comprises about 4% (weight / volume) sucrose or trehalose. In some embodiments, the formulation is not the formulation of any one of arrangements 1-98.

[0301] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein, wherein the concentration of the fusion protein is between about 30 to about 85 mg / mL; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising sodium acetate mixed with acetic acid, wherein the buffer solution is between about 0.1 mM to about 75 mM, wherein the buffer solute on pH is about 5.0; a tonicity agent, wherein the tonicity agent comprises about 4% (weight / volume) sucrose or trehalose; and a surfactant, wherein the surfactant comprises between about 0.01% to about 0.05% (weight / weight) polysorbate 20 or polysorbate 80, wherein the formulation comprises unconjugated fusion protein and fusion protein conjugate; and wherein the fusion protein comprises: an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap, wherein the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114, wherein the fusion protein comprises the following structure:Formula (17)

[0302] In some embodiments, each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, 118 and n9 is about 3500 plus or minus about 10% to about 20%, wherein if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-terminal end of the heavy chain; orbetween a hinge region and a Fab region (after the CHI domain) of the heavy chain. In some embodiments, the fusion protein comprises an antagonist antibody or a fragment thereof, that specifically binds to IL-6 that is conjugated to the polymer. In some embodiments, the fusion protein comprises an antagonistic IL-6 antibody or fragment thereof comprising: a) a heavy chain amino acid variable region that comprises a heavy chain that has a sequence of at least one of SEQ ID NOs: 7-13,19-27, 89, 90, 256-262; and b) a light chain amino acid variable region that comprises the light chain that has a sequence of at least one of SEQ ID NOs: 91-93, 28-30. In some embodiments, the fusion protein comprises an antagonist IL-6 antibody or fragment thereof comprising: a heavy chain variable region (VH) comprising 3 complementarity determining regions: VH (CDR1), VH CDR2, and VH CDR3 having an amino acid sequence from the CDRs listed in SEQ ID NO: 256; and a light chain variable region (VL) comprising a VL CDR1, VL CDR2, and VL CDR3 having an amino acid sequence selected from the group of CDRs listed in SEQ ID NO: 91-93. In some embodiments, the fusion protein further comprises an antagonistic antibody or fragment thereof that binds to IL-6, the antibody comprising: a CDRH1 that is a CDRH1 in SEQ ID NO: 172; a CDRH2 that is a CDRH2 in SEQ ID NO: 173; a CDRL1 that is a CDRL1 in SEQ ID NO: 199; a CDRL2 that is a CDRL2 in SEQ ID NO: 200; a CDRL3 that is a CDRL3 in SEQ ID NO: 201; at least one of the following mutations (EU numbering): L234A, L235A, and G237A; and at least one of the following mutations (EU numbering): Q347C or L443C. In some embodiments, the VEGF Trap is positioned either: a) at an N-terminal end of a heavy chain comprising IL-6 VH; or b) between a hinge region and after a CHI domain of a heavy chain comprising IL-6 VH. In some embodiments, comprising a mutation at position 94 or 95 in the VEGF Trap sequence, wherein if the mutation occurs at position 94, the mutation is T94I and wherein if the mutation occurs at position 95, the mutation is H95I. In some embodiments, the fusion protein comprises a VEGFR- Anti-IL-6 dual inhibitor, wherein the VEGFR-Anti-IL-6 dual inhibitor comprises a trap antibody fusion of an anti-IL 6 antibody or a fragment thereof and an anti- VEGF trap (VEGFR 1 / 2), wherein the dual inhibitor includes at least one point mutation within a VEGFR sequence to reduce cleavage of the VEGFR protein, wherein the VEGFR-Anti- IL-6 dual inhibitor comprises a constant heavy, a constant light, a fragment antigen binding, a fragment crystallizable (Fc), a vascular endothelial growth factor receptor (VEGFR), a variable heavy, and a variable light regions. In some embodiments, the Anti-IL-6 heavy chain variable region sequences is selected from options SEQ ID NO: 7-13, 89, 90, and / or 256-262, wherein theVEGF trap sequences is selected from at least one of SEQ ID NOs: 145, 15, 16, or 17, wherein the linker sequence is SEQ ID NO: 18, or wherein the light chain sequence for Anti-IL-6 molecules comprises at least 1, 2, or 3 light chain CDRs from at least one of SEQ ID NOs 76-84. In some embodiments, comprising a VEGFR-Fc sequence from at least one of SEQ ID NOs 85-88. In some embodiments, the unconjugated fusion protein is present in the formulation in a range between 10- 60% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the unconjugated fusion protein is present in the formulation at about 30% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the formulation is not the formulation of any one of arrangements 1-98. In some embodiments, X is Br.

[0303] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein, wherein the concentration of the fusion protein is about 50.0 mg / mL; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising sodium acetate, wherein the sodium acetate is about 50 mM; and a surfactant, wherein the surfactant comprises about 0.025% (weight / weight) polysorbate 20 or polysorbate 80, a tonicity agent, wherein the tonicity agent comprises about 4% (weight / volume) sucrose or trehalose, wherein the formulation comprises unconjugated fusion protein and fusion protein conjugate, and, wherein the unconjugated fusion protein is present in the formulation at about 10-60% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein, and wherein the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap, wherein the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114. In some embodiments, the fusion protein comprises the following structure:Formula (17)

[0304] In some embodiments, each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments,if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-terminal end of the heavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain. In some embodiments, the fusion protein comprises an antagonist antibody or a fragment thereof, that specifically binds to IL-6 that is conjugated to the polymer. In some embodiments, the fusion protein comprises an antagonistic IL-6 antibody or fragment thereof comprising: a) a heavy chain amino acid variable region that comprises a heavy chain that has a sequence of at least one of SEQ ID NOs: 7-13,19-27, 89, 90, 256-262; and b) a light chain amino acid variable region that comprises the light chain that has a sequence of at least one of SEQ ID NOs: 91-93, 28-30. In some embodiments, the fusion protein comprises an antagonist IL-6 antibody or fragment thereof comprising: a heavy chain variable region (VH) comprising 3 complementarity determining regions: VH (CDR1), VH CDR2, and VH CDR3 having an amino acid sequence from the CDRs listed in SEQ ID NO: 256; and a light chain variable region (VL) comprising a VL CDR1, VL CDR2, and VL CDR3 having an amino acid sequence selected from the group of CDRs listed in SEQ ID NO: 91-93. In some embodiments, the fusion protein further comprises an antagonistic antibody or fragment thereof that binds to IL-6, the antibody comprising: a CDRH1 that is a CDRH1 in SEQ ID NO: 172; a CDRH2 that is a CDRH2 in SEQ ID NO: 173; a CDRL1 that is a CDRL1 in SEQ ID NO: 199; a CDRL2 that is a CDRL2 in SEQ ID NO: 200; a CDRL3 that is a CDRL3 in SEQ ID NO: 201; at least one of the following mutations (EU numbering): L234A, L235A, and G237A; and at least one of the following mutations (EU numbering): Q347C or L443C. In some embodiments, the VEGF Trap is positioned either: a) at an N-terminal end of a heavy chain comprising IL-6 VH; or b) between a hinge region and after a CHI domain of a heavy chain comprising IL-6 VH. In some embodiments, comprising a mutation at position 94 or 95 in the VEGF Trap sequence, wherein if the mutation occurs at position 94, the mutation is T94I and wherein if the mutation occurs at position 95, the mutation is H95I. In some embodiments, the fusion protein comprises a VEGFR- Anti-IL-6 dual inhibitor, wherein the VEGFR-Anti-IL-6 dual inhibitor comprises a trap antibody fusion of an anti-IL 6 antibody or a fragment thereof and an anti- VEGF trap (VEGFR 1 / 2), wherein the dual inhibitor includes at least one point mutation within a VEGFR sequence to reduce cleavage of the VEGFR protein, wherein the VEGFR-Anti- IL-6 dual inhibitor comprises a constant heavy, a constant light, a fragment antigen binding, a fragment crystallizable (Fc), a vascular endothelial growth factor receptor (VEGFR), a variable heavy, and a variable light regions. In some embodiments, the Anti-IL-6 heavy chain variableregion sequences is selected from options SEQ ID NO: 7-13, 89, 90, and / or 256-262, wherein the VEGF trap sequences is selected from at least one of SEQ ID NOs: 145, 15, 16, or 17, wherein the linker sequence is SEQ ID NO: 18, or wherein the light chain sequence for Anti-IL-6 molecules comprises at least 1, 2, or 3 light chain CDRs from at least one of SEQ ID NOs 76-84. In some embodiments, comprising a VEGFR-Fc sequence from at least one of SEQ ID NOs 85-88. In some embodiments, the formulation is configured for intravitreal administration. In some embodiments, the formulation is not the formulation of any one of arrangements 1-98. In some embodiments, X is Br.

[0305] In some embodiments, a method for producing a formulation is provided, the method comprising: culturing a cell line that recombinantly produces a fusion protein under conditions wherein the fusion protein is produced; recovering the fusion protein; conjugating the fusion protein to a polymer to form a fusion protein conjugate, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; separating the fusion protein conjugate from the polymer and fusion proteins; and transferring a pharmaceutically effective amount of the fusion protein conjugate to a formulation comprising: a buffer solution, the buffer solution comprising sodium acetate, wherein the sodium acetate is about 50 mM; and a surfactant, wherein the surfactant comprises about 0.025% (weight / weight) polysorbate 20 or polysorbate 80, a tonicity agent, wherein the tonicity agent comprises about 4% (weight / volume) sucrose or trehalose, In some embodiments, the fusion protein conjugate comprises the following structure:Formula (17)

[0306] In some embodiments, each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-terminal end of theheavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain. In some embodiments, transferring a pharmaceutically effective amount of the fusion protein conjugate comprises ultrafiltration or diafiltration. In some embodiments, transferring a pharmaceutically effective amount of the fusion protein conjugate comprises dialysis. In some embodiments, the composition comprises unconjugated fusion protein and fusion protein conjugate. In some embodiments, the unconjugated fusion protein is present in the formulation at about 10-60% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar- amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the formulation is not the formulation of any one of arrangements 1-98. In some embodiments, X is Br.

[0307] In some embodiments, a method for producing a formulation is provided, the method comprising: culturing a cell line that recombinantly produces a fusion protein under conditions wherein the fusion protein is produced; recovering the fusion protein; preparing the fusion protein by: removing thiolates from a cysteine residue, wherein removing thiolates comprises a reduction with TCEP, wherein the resulting TCEP and unbound thiolates are filtered; and re-oxidizing the fusion protein with DHAA, wherein the excess DHAA is removed by filtration conjugating the fusion protein to a polymer to form a fusion protein conjugate, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; separating the fusion protein conjugate from polymer and fusion protein via chromatography; and dialyzing 20 mg / mL of the fusion protein conjugate with a buffer solution to yield a dialyzed buffer solution. In some embodiments, the buffer solution comprises: sodium acetate, wherein the sodium acetate is about 20 mM; a tonicity agent, wherein the tonicity agent comprises about 1.5% sucrose or trehalose; and a surfactant, wherein the surfactant comprises about 0.01% (weight / weight) polysorbate 20 or polysorbate 80. In some embodiments, the fusion protein conjugate comprises the following structure:Formula (17)

[0308] In some embodiments, each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-terminal end of theheavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain. In some embodiments, the reduction with TCEP comprises 30x molar excess of TCEP (tris(2-carboxyethyl)phosphine over the fusion protein concentration. In some embodiments, the reduction with re-oxidizing the fusion protein with DHAA (dehydroascorbic acid) comprises 15x molar excess of DHAA over the fusion protein concentration. In some embodiments, filtration of resulting TCEP and unbound thiolates, and removal of excess DHAA are accomplished by tangential flow filtration (TFF). In some embodiments, dialyzing is accomplished through buffer exchange using TFF. In some embodiments, the dialyzed buffer solution is concentrated into a concentrated dialyzed buffer solution, the concentrated dialyzed buffer solution comprising about 50.0 mg / mL of the fusion protein conjugate, 50 mM sodium acetate, about 0.025% ( weight / weight) polysorbate 20 or polysorbate 80, about 4.0% (weight / volume) tonicity agent, where the tonicity agent is sucrose or trehalose. In some embodiments, the buffer comprises about 50 mM sodium acetate, about 0.025% (weight / weight) polysorbate 20, at about pH 5. In some embodiments, the formulation is not the formulation of any one of arrangements 1-98. In some embodiments, X is Br.

[0309] In some embodiments, a method for producing a formulation is provided, the method comprising: culturing a cell line that recombinantly produces a fusion protein under conditions wherein the fusion protein is produced; recovering the fusion protein; preparing the fusion protein by: removing thiolates from a cysteine residue, wherein removing thiolates comprises a reduction with TCEP, wherein the resulting TCEP and unbound thiolates are filtered; re-oxidizing the fusion protein with DHAA, wherein the excess DHAA is removed by filtration; and, conjugating the fusion protein to a polymer to form a fusion protein conjugate, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; separating the fusion protein conjugate from polymer and fusion protein via chromatography; and dialyzing a pharmaceutically effective amount of the fusion protein conjugate with a buffer solution. In some embodiments, the buffer solution comprises: sodium acetate, wherein the sodium acetate is about 20 mM; a tonicity agent, wherein the tonicity agent comprises about 1.5% sucrose or trehalose and a surfactant, wherein the surfactant comprises about 0.01% (weight / weight) polysorbate 20 or polysorbate 80; and concentrating the resulting prepared fusion protein, wherein the sodium acetate is concentrated to about 50 mM, wherein the surfactant is concentrated to about 0.025% (weight / weight) polysorbate 20 or polysorbate 80, and wherein the tonicity agent is concentratedto about 4% (weight / volume) sucrose or trehalose. In some embodiments, the fusion protein conjugate comprises the following structure:Formula (17)

[0310] In some embodiments, each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4,n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-tcrminal end of the heavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain. In some embodiments, the sodium acetate concentration is between about 0.1 mM to about 75 mM. In some embodiments, the Sodium Acetate concentration is between about 0.1 mM to about 50 mM. In some embodiments, the Anti-IL-6 heavy chain variable region sequences is selected from options SEQ ID NO: 7-13, 89, 90, and / or 256-262, wherein the VEGF trap sequences is selected from at least one of SEQ ID NOs: 114, 145, 15, 16, or 17, or wherein the light chain sequence for Anti-IL-6 molecules comprises at least 1, 2, or 3 light chain CDRs from at least one of SEQ ID NOs 76-84. In some embodiments, a pharmaceutically effective amount is an amount sufficient to effect any one or more beneficial or desired results. In some embodiments, a pharmaceutically effective amount is a concentration greater than 10 mg / mL. In some embodiments, a pharmaceutically effective amount is a concentration greater than 30 mg / mL. In some embodiments, the formulation is not the formulation of any one of arrangements 1-98. In some embodiments, Xis Br.

[0311] In some embodiments, a pharmaceutical formulation, the formulation comprising: a pharmaceutically effective amount of fusion protein, wherein the concentration of the fusion protein is about 50 mg / mL; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising sodium acetate, wherein the sodium acetate is about 50 mM, and a tonicity agent, wherein the tonicity agent is about 4% (w / v) sucrose or trehalose; and a surfactant, wherein the surfactant comprises about 0.025% (weight / weight) polysorbate 20 or polysorbate 80, wherein the formulation comprises unconjugated fusion protein and fusion protein conjugate, and wherein the unconjugated fusion protein is present in the formulation at about 10-60% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114, wherein the fusion protein has a light chain with at least 80% identity to the sequence of SEQ ID NO: 169, whereinthe fusion protein has a heavy chain with at least 80% identity to the sequence of SEQ ID NO:170. In some embodiments, the fusion protein comprises the following structure:Formula (17)

[0312] In some embodiments, each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4,n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-tcrminal end of the heavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain. In some embodiments, the formulation is not the formulation of any one of arrangements 1- 98. In some embodiments, X is Br.

[0313] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of a fusion protein, wherein the concentration of the fusion protein is about 50 mg / mL; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising sodium acetate, wherein the sodium acetate is about 50 mM, and a tonicity agent, wherein the tonicity agent comprises about 4% (w / v) sucrose or trehalose; and a surfactant, wherein the surfactant comprises about 0.025% (weight / weight) polysorbate 20 or polysorbate 80, wherein the formulation comprises unconjugated fusion protein and fusion protein conjugate, and, wherein the unconjugated fusion protein is present in the formulation at about 10-60% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114, wherein the fusion protein has a light chain with at least 80% identity to the sequence of SEQ ID NO: 169, wherein the fusion protein has a heavy chain with at least 80% identity to the sequence of SEQ ID NO: 170. In some embodiments, the fusion protein comprises the following structure:Formula (17A)

[0314] In some embodiments, part of each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), whichbond is depicted on one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-terminal end of the heavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain. In some embodiments, the fusion protein comprises the amino acid sequences of SEQID NOs: 169 and 170, conjugated to a polymer, wherein the polymer is the polymer depicted in the structure of Formula 17, Formula 17A, or 17Br. In some embodiments, the formulation is not the formulation of any one of arrangements 1-98. In some embodiments, X is Br.

[0315] In some embodiments, the formulation comprises: a pharmaceutically effective amount of a fusion protein, wherein the concentration of the fusion protein is 50 mg / mL; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising sodium acetate, wherein the sodium acetate is 50 mM, and a tonicity agent, wherein the tonicity agent comprises 4% (weight / volume) sucrose or trehalose; and a surfactant, wherein the surfactant comprises 0.025% (weight / weight) polysorbate 20 or polysorbate 80. The formulation comprises unconjugated fusion protein and fusion protein conjugate. In some embodiments the unconjugated fusion protein is present in the formulation at about 30% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments the fusion protein comprises an anti-IL-6 antibody and a VEGF trap, wherein the fusion protein comprises SEQ ID NOs: 169 and 170. In some embodiments the fusion protein comprises the following structure:Formula (17A)

[0316] In some embodiments, part of each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), whichbond is depicted on one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, the formulation is not the formulation of any one of arrangements 1-98. In some embodiments X is Br.-I l l-

[0317] In some embodiments a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein, a buffer, wherein the buffer comprises histidine; a surfactant, and optionally a tonicity agent. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114. In some embodiments, the fusion protein is conjugated to a polymer. In some embodiments, the fusion protein comprises an antagonist antibody or a fragment thereof, that specifically binds to IL-6 that is conjugated to a polymer; In some embodiments, the fusion protein comprises an antagonistic IL-6 antibody or fragment thereof comprising: a heavy chain amino acid variable region that comprises a heavy chain that has a sequence of at least one of SEQ ID NOs: 7-13,19-27, 89, 90, 256-262; and a light chain amino acid variable region that comprises the light chain that has a sequence of at least one of SEQ ID NOs: 91-93, 28-30. In some embodiments, the fusion protein comprises an antagonist IL-6 antibody or fragment thereof comprising: a heavy chain variable region (VH) comprising 3 complementarity determining regions: VH (CDR1), VH CDR2, and VH CDR3 having an amino acid sequence from the CDRs listed in SEQ ID NO: 256; and a light chain variable region (VL) comprising a VL CDR1, VL CDR2, and VL CDR3 having an amino acid sequence selected from the group of CDRs listed in SEQ ID NO: 91-93. In some embodiments, the fusion protein further comprises an antagonistic antibody or fragment thereof that binds to IL-6, the antibody comprising: a CDRH1 that is a CDRH1 in SEQ ID NO: 172; a CDRH2 that is a CDRH2 in SEQ ID NO: 173; a CDRL1 that is a CDRL1 in SEQ ID NO: 199; a CDRL2 that is a CDRL2 in SEQ ID NO: 200; a CDRL3 that is a CDRL3 in SEQ ID NO: 201; at least one of the following mutations (EU numbering): L234A, L235A, and G237A; and at least one of the following mutations (EU numbering): Q347C or L443C. In some embodiments, the VEGF Trap is positioned either: at an N-terminal end of a heavy chain comprising IL-6 VH; or between a hinge region and after a CHI domain of a heavy chain comprising IL-6 VH. In some embodiments, the buffer comprises histidine acetate. In some embodiments, the surfactant comprises polysorbate 20. In some embodiments, the polysorbate 20 is between about 0.01% (w / w) and about 0.1% (w / w) of the formulation. In some embodiments, the surfactant comprises polysorbate 80. In some embodiments, the polysorbate 80 is between about 0.01% (w / w) and about 0.1% (w / w) of the formulation. In some embodiments, the polysorbate 20 is about 0.025% (w / w) of the formulation.In some embodiments, the polysorbate 80 is about 0.025% (w / w) of the formulation. In some embodiments, the histidine acetate concentration is between about 10 mM to about 50 mM. In some embodiments, the Histidine Acetate concentration is about 25 mM. In some embodiments, the formulation is between about pH 4.5 and about pH 6.5. In some embodiments, the formulation is about pH 5.5. In some embodiments, the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer. In some embodiments, the poly dispersity index (PDI) is between about 0.5 and about 2. In some embodiments, the polydispersity index (PDI) is 1. In some embodiments, the tonicity agent is about 2-10% (weight / volume) of the formulation. In some embodiments, the tonicity agent is sucrose or trehalose. In some embodiments, the sucrose is about 6% (weight / volume) of the formulation. In some embodiments, the trehalose is about 6% (weight / volume) of the formulation. In some embodiments, the concentration of fusion protein is between about 20-75 mg / mL. In some embodiments, the concentration of fusion protein is about 50 mg / mL. In some embodiments, the formulation is not the formulation of any one of arrangements 1-98.

[0318] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of a fusion protein; a buffer solution, the buffer solution comprising histidine acetate, a surfactant, and optionally a tonicity agent. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114. In some embodiments, the fusion protein is conjugated to a polymer. In some embodiments, the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer. In some embodiments, the fusion protein comprises an antagonist antibody or a fragment thereof, that specifically binds to IL-6 that is conjugated to a polymer. In some embodiments, the fusion protein comprises an antagonistic IL-6 antibody or fragment thereof comprising: a) a heavy chain amino acid variable region that comprises a heavy chain that has a sequence of at least one of SEQ ID NOs: 7-13,19-27, 89, 90, 256-262; and b) a light chain amino acid variable region that comprises the light chain that has a sequences of at least one of SEQ ID NOs: 91-93, 28-30. In some embodiments, the fusion protein comprises an antagonist IL-6 antibody or fragment thereof comprising: a heavy chain variable region (VH) comprising 3 complementarity determining regions: VH (CDR1), VH CDR2, and VH CDR3 having an amino acid sequence from the CDRslisted in SEQ ID NO: 256; and a light chain variable region (VL) comprising a VL CDR1 , VL CDR2, and VL CDR3 having an amino acid sequence selected from the group of CDRs listed in SEQ ID NO: 91-93. In some embodiments, the fusion protein further comprises an antagonistic antibody or fragment thereof that binds to IL-6, the antibody comprising: a CDRH1 that is a CDRH1 in SEQ ID NO: 172; a CDRH2 that is a CDRH2 in SEQ ID NO: 173; a CDRL1 that is a CDRL1 in SEQ ID NO: 199; a CDRL2 that is a CDRL2 in SEQ ID NO: 200; a CDRL3 that is a CDRL3 in SEQ ID NO: 201; at least one of the following mutations (EU numbering): L234A, L235A, and G237A; and at least one of the following mutations (EU numbering): Q347C or L443C. In some embodiments, the VEGF Trap is positioned either: at an N-terminal end of a heavy chain comprising IL-6 VH; or between a hinge region and after a CHI domain of a heavy chain comprising IL-6 VH. In some embodiments, the VEGF Trap consists of the sequence of SEQ ID NO: 114. In some embodiments, the surfactant comprises between about 0.01% to about 0.1% (weight / weight) polysorbate 20. In some embodiments, the surfactant comprises about 0.025% (weight / weight) polysorbate 20. In some embodiments, the surfactant comprises between about 0.01% to about 0.05% (weight / weight) polysorbate 80. In some embodiments, the surfactant comprises about 0.025% (weight / weight) polysorbate 80. In some embodiments, the formulation is between about pH 4.5 to about pH 6.5. In some embodiments, the formulation is about pH 5.5. In some embodiments, the formulation is stable for at least 52 weeks. In some embodiments, the formulation is stable for at least 52 weeks at 4 centigrade. In some embodiments, the formulation is stable for at least 8 weeks at 37 centigrade. In some embodiments, the formulation is stable in acidic conditions. In some embodiments, the formulation configured for intravitreal injection. In some embodiments, the histidine acetate concentration in the buffer solution is about 0.1 mM to about 50 mM. In some embodiments, the concentration of fusion protein is between about 20-100 mg / mL. In some embodiments, the concentration of fusion protein is about 50 mg / mL. In some embodiments, the tonicity agent comprises between about 2-10% (weight / volume) sucrose or trehalose. In some embodiments, the formulation is not the formulation of any one of arrangements 1-98.

[0319] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein, wherein the concentration of the fusion protein is between about 20 to about 100 mg / mL; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises aphosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising histidine mixed with acetic acid, wherein the buffer solution is between about 10 mM to about 50 mM, wherein the buffer solute on pH is about 5.5; a tonicity agent, wherein the tonicity agent comprises about 6% (weight / volume) sucrose or trehalose; and a surfactant, wherein the surfactant comprises between about 0.01% to about 0.1% (weight / weight) polysorbate 20 or polysorbate 80. In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate. In some embodiments, the fusion protein comprises: an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114. In some embodiments, the fusion protein comprises the following structure:Formula (17)

[0320] In some embodiments, each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, 116, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-tenninal end of the heavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain. In some embodiments, the fusion protein comprises an antagonist antibody or a fragment thereof, that specifically binds to IL-6 that is conjugated to the polymer. In some embodiments, the fusion protein comprises an antagonistic IL-6 antibody or fragment thereof comprising: a heavy chain amino acid variable region that comprises a heavy chain that has a sequence of at least one of SEQ ID NOs: 7-13,19-27, 89, 90, 256-262; and a light chain amino acid variable region that comprises the light chain that has a sequence of at least one of SEQ ID NOs: 91-93, 28-30. In some embodiments, the fusion protein comprises an antagonist IL-6 antibody or fragment thereof comprising: a heavy chain variable region (VH) comprising 3 complementarity determining regions: VH (CDR1), VH CDR2, and VH CDR3 having an amino acid sequence from the CDRs listed in SEQ ID NO: 256; and a light chain variable region (VL) comprising a VL CDR1, VL CDR2, and VL CDR3 having an amino acid sequence selected from the group of CDRs listed in SEQ ID NO: 91-93. In some embodiments, the fusion protein further comprises an antagonistic antibody or fragment thereof that binds to IL-6, the antibody comprising: a CDRH1 that is a CDRH1 in SEQ ID NO: 172; a CDRH2 that is a CDRH2 in SEQ ID NO: 173; a CDRL1 that is a CDRL1 in SEQ ID NO: 199; a CDRL2 that is a CDRL2 in SEQ ID NO: 200; a CDRL3 that is a CDRL3 in SEQ ID NO: 201; at least one of the following mutations (EU numbering): L234A, L235A, and G237A; and at least one of the following mutations (EU numbering): Q347C or L443C. In some embodiments, the VEGF Trap is positioned either: at an N-terminal end of a heavy chain comprising IL-6 VH; or between a hinge region and after a CHI domain of a heavy chaincomprising IL-6 VH. In some embodiments, comprising a mutation at position 94 or 95 in the VEGF Trap sequence, wherein if the mutation occurs at position 94, the mutation is T94I and wherein if the mutation occurs at position 95, the mutation is H95I. In some embodiments, the fusion protein comprises a VEGFR- Anti-IL-6 dual inhibitor, wherein the VEGFR-Anti-IL-6 dual inhibitor comprises a trap antibody fusion of an anti-IL 6 antibody or a fragment thereof and an anti- VEGF trap (VEGFR 1 / 2), wherein the dual inhibitor includes at least one point mutation within a VEGFR sequence to reduce cleavage of the VEGFR protein, wherein the VEGFR-Anti- IL-6 dual inhibitor comprises a constant heavy, a constant light, a fragment antigen binding, a fragment crystallizable (Fc), a vascular endothelial growth factor receptor (VEGFR), a variable heavy, and a variable light regions. In some embodiments, the Anti-IL-6 heavy chain variable region sequences is selected from options SEQ ID NO: 7-13, 89, 90, and / or 256-262, wherein the VEGF trap sequences is selected from at least one of SEQ ID NOs: 145, 15, 16, or 17, wherein the linker sequence is SEQ ID NO: 18, or wherein the light chain sequence for Anti-IL-6 molecules comprises at least 1, 2, or 3 light chain CDRs from at least one of SEQ ID NOs 76-84. In some embodiments, comprising a VEGFR-Fc sequence from at least one of SEQ ID NOs 85-88. In some embodiments, the unconjugated fusion protein is present in the formulation in a range between 10- 60% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the unconjugated fusion protein is present in the formulation at about 30% of a total molar amount of the antibody conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the antibody conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the formulation is not the formulation of any one of arrangements 1-98. In some embodiments, X is Br.

[0321] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein, wherein the concentration of the fusion protein is about 50.0 mg / mL; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising histidine acetate, wherein the histidine acetate is about 25 mM; a surfactant, wherein the surfactant comprises about 0.025% (weight / weight) polysorbate 20 or polysorbate 80, and optionally atonicity agent, wherein the tonicity agent comprises about 6% (w / v) sucrose or trehalose. In some embodiments, the formulation comprises unconjugatcd fusion protein and fusion protein conjugate, and wherein the unconjugated fusion protein is present in the formulation at about 10- 60% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114. In some embodiments, the fusion protein comprises the following structure:Formula (17)

[0322] In some embodiments, each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-tenninal end of the heavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain. In some embodiments, the fusion protein comprises an antagonist antibody or a fragment thereof, that specifically binds to IL-6 that is conjugated to the polymer. In some embodiments, the fusion protein comprises an antagonistic IL-6 antibody or fragment thereof comprising: a heavy chain amino acid variable region that comprises a heavy chain that has a sequence of at least one of SEQ ID NOs: 7-13,19-27, 89, 90, 256-262; and a light chain amino acid variable region that comprises the light chain that has a sequence of at least one of SEQ ID NOs: 91-93, 28-30. In some embodiments, the fusion protein comprises an antagonist IL-6 antibody or fragment thereof comprising: a heavy chain variable region (VH) comprising 3 complementarity determining regions: VH (CDR1), VH CDR2, and VH CDR3 having an amino acid sequence from the CDRs listed in SEQ ID NO: 256; and a light chain variable region (VL) comprising a VL CDR1, VL CDR2, and VL CDR3 having an amino acid sequence selected from the group of CDRs listed in SEQ ID NO: 91-93. In some embodiments, the fusion protein further comprises an antagonistic antibody or fragment thereof that binds to IL-6, the antibody comprising: a CDRH1 that is a CDRH1 in SEQ ID NO: 172;a CDRH2 that is a CDRH2 in SEQ ID NO: 173; a CDRL1 that is a CDRL1 in SEQ ID NO: 199; a CDRL2 that is a CDRL2 in SEQ ID NO: 200; a CDRL3 that is a CDRL3 in SEQ ID NO: 201; at least one of the following mutations (EU numbering): L234A, L235A, and G237A; and at least one of the following mutations (EU numbering): Q347C or L443C. In some embodiments, the VEGF Trap is positioned either: at an N-terminal end of a heavy chain comprising IL-6 VH; or between a hinge region and after a CHI domain of a heavy chaincomprising TL-6 VH. In some embodiments, comprising a mutation at position 94 or 95 in the VEGF Trap sequence, wherein if the mutation occurs at position 94, the mutation is T94I and wherein if the mutation occurs at position 95, the mutation is H95I. In some embodiments, the fusion protein comprises a VEGFR- Anti-IL-6 dual inhibitor, wherein the VEGFR-Anti-IL-6 dual inhibitor comprises a trap antibody fusion of an anti-IL 6 antibody or a fragment thereof and an anti- VEGF trap (VEGFR 1 / 2), wherein the dual inhibitor includes at least one point mutation within a VEGFR sequence to reduce cleavage of the VEGFR protein, wherein the VEGFR-Anti- IL-6 dual inhibitor comprises a constant heavy, a constant light, a fragment antigen binding, a fragment crystallizable (Fc), a vascular endothelial growth factor receptor (VEGFR), a variable heavy, and a variable light regions. In some embodiments, the Anti-IL-6 heavy chain variable region sequences is selected from options SEQ ID NO: 7-13, 89, 90, and / or 256-262, wherein the VEGF trap sequences is selected from at least one of SEQ ID NOs: 145, 15, 16, or 17, wherein the linker sequence is SEQ ID NO: 18, or wherein the light chain sequence for Anti-IL-6 molecules comprises at least 1, 2, or 3 light chain CDRs from at least one of SEQ ID NOs 76-84. In some embodiments, a VEGFR-Fc sequence from at least one of SEQ ID NOs 85-88. In some embodiments, the formulation is configured for intravitreal administration. In some embodiments, the formulation is configured for intravitreal administration. In some embodiments, the formulation is not the formulation of any one of arrangements 1-98. In some embodiments, X is Br.

[0323] In some embodiments, a method for producing a formulation is provided, the method comprising: culturing a cell line that recombinantly produces a fusion protein under conditions wherein the fusion protein is produced; recovering the fusion protein; conjugating the fusion protein to a polymer to form a fusion protein conjugate, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; separating the fusion protein conjugate from the polymer and fusion proteins; and transferring a pharmaceutically effective amount of the fusion protein conjugate to a formulation comprising: a buffer solution, the buffer solution comprising histidine acetate, wherein the histidine acetate is about 25 mM; a surfactant, wherein the surfactant comprises about 0.025% (weight / weight) polysorbate 20 or polysorbate 80, and a tonicity agent, wherein the tonicity agent comprises about 6% (weight / volume) sucrose or trehalose. In some embodiments, the fusion protein conjugate comprises the following structure:Formula (17)

[0324] In some embodiments, each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-terminal end of theheavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain. In some embodiments, transferring a pharmaceutically effective amount of the fusion protein conjugate comprises ultrafiltration or diafiltration. In some embodiments, transferring a pharmaceutically effective amount of the fusion protein conjugate comprises dialysis. In some embodiments, the composition comprises unconjugated fusion protein and fusion protein conjugate. In some embodiments, the unconjugated fusion protein is present in the formulation at about 10-60% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar- amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the formulation is not the formulation of any one of arrangements 1-98. In some embodiments, X is Br.

[0325] In some embodiments, a method for producing a formulation, the method comprising: culturing a cell line that recombinantly produces a fusion protein under conditions wherein the fusion protein is produced; recovering the fusion protein; preparing the fusion protein by: removing thiolates from a cysteine residue, wherein removing thiolates comprises a reduction with TCEP, wherein the resulting TCEP and unbound thiolates are filtered; and re-oxidizing the fusion protein with DHAA, wherein the excess DHAA is removed by filtration conjugating the fusion protein to a polymer to form a fusion protein conjugate, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; separating the fusion protein conjugate from polymer and fusion protein via chromatography; and dialyzing 20 mg / mL of the fusion protein conjugate with a buffer solution to yield a dialyzed buffer solution, wherein the buffer solution comprises: histidine acetate, wherein the histidine acetate is about 10 mM; a tonicity agent, wherein the tonicity agent comprises about 2.4% (weight / volume) sucrose or trehalose; and a surfactant, wherein the surfactant comprises about 0.01% (weight / weight) polysorbate 20 or polysorbate 80. In some embodiments, the fusion protein conjugate comprises the following structure:Formula (17)

[0326] In some embodiments, each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-terminal end of theheavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain. In some embodiments, the reduction with TCEP comprises 30x molar excess of TCEP (tris(2-carboxyethyl)phosphine) over the fusion protein concentration. In some embodiments, the reduction with re-oxidizing the fusion protein with DHAA (dehydroascorbic acid) comprises 15x molar excess of DHAA over the fusion protein concentration. In some embodiments, filtration of resulting TCEP and unbound thiolates, and removal of excess DHAA are accomplished by tangential flow filtration (TFF). In some embodiments, dialyzing is accomplished through buffer exchange using TFF. In some embodiments, the dialyzed buffer solution is concentrated into a concentrated dialyzed buffer solution, the concentrated dialyzed buffer solution comprising about 50.0 mg / mL of the fusion protein conjugate, 25 mM histidine acetate, about 0.025% (weight / weight) polysorbate 20 or polysorbate 80, about 6.0% (weight / volume) of a tonicity agent, wherein the tonicity agent is sucrose or trehalose. In some embodiments, the buffer comprises about 25 mM histidine acetate, about 0.025% (weight / weight) PS20, at about pH 5.5. In some embodiments, the formulation is not the formulation of any one of arrangements 1-98. In some embodiments, X is Br.

[0327] A method for producing a formulation, the method comprising; culturing a cell line that recombinantly produces a fusion protein under conditions wherein the fusion protein is produced; recovering the fusion protein; preparing the fusion protein by: removing thiolates from a cysteine residue, wherein removing thiolates comprises a reduction with TCEP, wherein the resulting TCEP and unbound thiolates are filtered; re-oxidizing the fusion protein with DHAA, wherein the excess DHAA is removed by filtration; and, conjugating the fusion protein to a polymer to form a fusion protein conjugate, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; separating the fusion protein conjugate from polymer and fusion protein via chromatography; and dialyzing a pharmaceutically effective amount of the fusion protein conjugate with a buffer solution, wherein the buffer solution comprises: histidine acetate, wherein the histidine acetate is about 10 mM; a tonicity agent, wherein the tonicity agent comprises about 2.4% (weight / volume) sucrose or trehalose; and a surfactant, wherein the surfactant comprises about 0.01% (weight / weight) polysorbate 20 or polysorbate 80; and concentrating the resulting prepared fusion protein, wherein the histidine acetate is concentrated to about 25 mM, wherein the surfactant is concentrated to about 0.025% (weight / weight) polysorbate 20 or polysorbate 80, and wherein the tonicity agent concentrated toabout 6% (weight / volume) sucrose or trehalose. In some embodiments, the fusion protein conjugate comprises the following structure:Formula (17)

[0328] In some embodiments, each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; andnl , n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of n 1 , n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-terminal end of the heavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain. In some embodiments, the Anti-IL-6 heavy chain variable region sequences is selected from options SEQ ID NO: 7-13, 89, 90, and / or 256-262, wherein the VEGF trap sequences is selected from at least one of SEQ ID NOs: 114, 145, 15, 16, or 17, or wherein the light chain sequence for Anti-IL-6 molecules comprises at least 1, 2, or 3 light chain CDRs from at least one of SEQ ID NOs 76-84. In some embodiments, a pharmaceutically effective amount is an amount sufficient to effect any one or more beneficial or desired results. In some embodiments, a pharmaceutically effective amount is a concentration greater than 10 mg / mL. In some embodiments, a pharmaceutically effective amount is a concentration greater than 30 mg / mL. In some embodiments, the formulation is not the formulation of any one of arrangements 1-98. In some embodiments, X is Br.

[0329] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein, wherein the concentration of the fusion protein is about 50 mg / mL; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising histidine acetate, wherein the histidine acetate is about 25 mM, and a tonicity agent, wherein the tonicity agent is about 6% (weight / volume) sucrose or trehalose; and a surfactant, wherein the surfactant comprises about 0.025% (weight / weight) polysorbate 20 or polysorbate 80. In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate, and wherein the unconjugated fusion protein is present in the formulation at about 10-60% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114, wherein the fusion protein has a light chain with at least 80% identity to the sequence of SEQ ID NO: 169, wherein the fusion protein has a heavy chain with at least 80%identity to the sequence of SEQ ID NO: 170. In some embodiments, the fusion protein comprises the following structure:Formula (17)

[0330] In some embodiments, each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), which bond is depictedon one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4,n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-tcrminal end of the heavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain. In some embodiments, the formulation is not the formulation of any one of arrangements 1- 98. In some embodiments, X is Br.

[0331] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of a fusion protein, wherein the concentration of the fusion protein is about 50 mg / mL; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising histidine acetate, wherein the histidine acetate is about 25 mM, and a tonicity agent, wherein the tonicity agent is about 6% (w / v) sucrose or trehalose; and a surfactant, wherein the surfactant comprises about 0.025% (weight / weight) polysorbate 20 or polysorbate 80. In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate, and wherein the unconjugated fusion protein is present in the formulation at about 10-60% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114, wherein the fusion protein has a light chain with at least 80% identity to the sequence of SEQ ID NO: 169, wherein the fusion protein has a heavy chain with at least 80% identity to the sequence of SEQ ID NO: 170. In some embodiments, the fusion protein comprises the following structure:Formula (17A)

[0332] In some embodiments, part of each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), whichbond is depicted on one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, if the conjugate comprises a VEGF Trap, the VEGF Trap is fused: to the N-terminal end of the heavy chain; or between a hinge region and a Fab region (after the CHI domain) of the heavy chain. In some embodiments, the fusion protein comprises the amino acid sequences of SEQID NOs: 169 and 170, conjugated to a polymer, wherein the polymer is the polymer depicted in the structure of Formula 17, Formula 17 A, or Formula 17Br. In some embodiments, the formulation is not the formulation of any one of arrangements 1-98. In some embodiments, X is Br.

[0333] In some embodiments, the formulation comprises: a pharmaceutically effective amount of a fusion protein, wherein the concentration of the fusion protein is 50 mg / mL; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising histidine acetate, wherein the histidine acetate is 25 mM, a tonicity agent, wherein the tonicity agent is about 6% (weight / volume) sucrose or trehalose; and a surfactant, wherein the surfactant comprises 0.025% (weight / weight) polysorbate 20 or polysorbate 80. In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate, and wherein the unconjugated fusion protein is present in the formulation at about 30% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the fusion protein comprises an anti-IL-6 antibody and a VEGF trap, wherein the fusion protein comprises SEQ ID NOs: 169 and 170, wherein the fusion protein comprises the following structure:-BOFormula (17A)

[0334] In some embodiments, part of each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), whichbond is depicted on one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) H, or c) any halogen, including -Br, -Cl, or -I, d) -SCN, or e) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%. In some embodiments, the formulation is not the formulation of any one of arrangements 1-98. In some embodiments, X is Br.

[0335] In some embodiments, a pharmaceutical formulation is provided comprising: a pharmaceutically effective amount of a fusion protein, wherein the concentration of the fusion protein is 50 mg / mL; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; a buffer solution, the buffer solution comprising sodium acetate, wherein the sodium acetate is 50 mM, and a tonicity agent, wherein the tonicity agent is 4% (weight / volume) sucrose or trehalose; and a surfactant, wherein the surfactant comprises 0.025% (weight / weight) polysorbate 20 or polysorbate 80, wherein the pH is 5.0. In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate, and wherein the unconjugated fusion protein is present in the formulation at about 30% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the fusion protein comprises an anti-IL-6 antibody and a VEGF trap, wherein the fusion protein comprises SEQ ID NOs: 169 and 170. In some embodiments, the fusion protein comprises the following structure:Formula (17Br)

[0336] In some embodiments, part of each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L; the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), whichbond is depicted on one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is Br; and n 1 , n2, n3, n4, n5, n6, n7, n8 and n9 arc the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%.

[0337] In some embodiments, a pharmaceutical formulation is provided comprising: a pharmaceutically effective amount of a fusion protein, wherein the concentration of the fusion protein is 50 mg / mL; a polymer, wherein the fusion protein is conjugated to the polymer, wherein the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer; abuffer solution, the buffer solution comprising histidine acetate, wherein the histidine acetate is 25 mM, and a tonicity agent, wherein the tonicity agent is about 6% (weight / volume) sucrose or trehalose; and a surfactant, wherein the surfactant comprises 0.025% (weight / weight) polysorbate 20 or polysorbate 80, wherein the pH is 5.5. In some embodiments, the formulation comprises unconjugated fusion protein and fusion protein conjugate, and wherein the unconjugated fusion protein is present in the formulation at about 30% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein. In some embodiments, the fusion protein comprises an anti-IL-6 antibody and a VEGF trap, wherein the fusion protein comprises SEQ ID NOs: 169 and 170. In some embodiments, the fusion protein comprises the following structure:Formula (17Br)

[0338] In some embodiments, part of each heavy chain of the anti-IL-6 antibody is denoted by the letter H, and each light chain of the anti-IL-6 antibody is denoted by the letter L;the polymer is bonded to the antibody through the sulfhydryl of C443 (EU numbering), whichbond is depicted on one of the heavy chains; PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is Br; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6,n7, n8 and n9 is about 3500 plus or minus about 10% to about 20%.

[0339] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of fusion protein, a buffer, wherein the buffer comprises histidine; a surfactant, and optionally a tonicity agent. In some embodiments, the fusion protein comprises an anti-IL-6 antibody and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114. In some embodiments, the fusion protein is conjugated to a polymer. In some embodiments, the fusion protein comprises an antagonist antibody or a fragment thereof, that specifically binds to IL-6 that is conjugated to a polymer. In some embodiments, the fusion protein comprises an antagonistic IL-6 antibody or fragment thereof comprising: a heavy chain amino acid variable region that comprises a heavy chain that has a sequence of at least one of SEQ ID NOs: 7-13,19-27, 89, 90, 256-262; and a light chain amino acid variable region that comprises the light chain that has a sequence of at least one of SEQ ID NOs: 91-93, 28-30. In some embodiments, the fusion protein comprises an antagonist IL-6 antibody or fragment thereof comprising: a heavy chain variable region (VH) comprising 3 complementarity determining regions: VH (CDR1), VH CDR2, and VH CDR3 having an amino acid sequence from the CDRs listed in SEQ ID NO: 256; and a light chain variable region (VL) comprising a VL CDR1, VL CDR2, and VL CDR3 having an amino acid sequence selected from the group of CDRs listed in SEQ ID NO: 91-93. In some embodiments, the fusion protein further comprises an antagonistic antibody or fragment thereof that binds to IL-6, the antibody comprising: a CDRH1 that is a CDRH1 in SEQ ID NO: 172; a CDRH2 that is a CDRH2 in SEQ ID NO: 173; a CDRL1 that is a CDRL1 in SEQ ID NO: 199; a CDRL2 that is a CDRL2 in SEQ ID NO: 200; a CDRL3 that is a CDRL3 in SEQ ID NO: 201; at least one of the following mutations (EU numbering): L234A, L235A, and G237A; and at least one of the following mutations (EU numbering): Q347C or L443C. In some embodiments, the VEGF Trap is positioned either: at an N-terminal end of a heavy chain comprising IL-6 VH; or between a hinge region and after a CHIdomain of a heavy chain comprising IL-6 VH. In some embodiments, the buffer comprises histidine acetate. In some embodiments, the emulsifier comprises polysorbate 20. In some embodiments, the polysorbate 20 is between about 0.01% (w / w) and about 0.1% (w / w) of the formulation. In some embodiments, the emulsifier comprises polysorbate 80. In some embodiments, the polysorbate 80 is between about 0.01% (w / w) and about 0.1% (w / w) of the formulation. In some embodiments, the polysorbate 20 is about 0.025% (w / w) of the formulation. In some embodiments, the polysorbate 80 is about 0.025% (w / w) of the formulation. In some embodiments, the histidine acetate concentration is between about 10 mM to about 50 mM. In some embodiments, histidine acetate concentration is about 25 mM. In some embodiments, the formulation is between about pH 4.5 and about pH 6.5. In some embodiments, the formulation is about pH 5.5. In some embodiments, the polymer comprises a phosphorylcholine containing polymer or a zwitterionic monomer. In some embodiments, the polydispersity index (PDI) is between about 0.5 and about 2. In some embodiments, the polydispersity index (PDI) is 1. In some embodiments, the tonicity agent is about 2-10% (weight / volume) of the formulation. In some embodiments, the tonicity agent is sucrose or trehalose. In some embodiments, the sucrose is about 6.0% (weight / volume) of the formulation. In some embodiments, the trehalose is about 6.0% (weight / volume) of the formulation. In some embodiments, the formulation is not the formulation of any one of arrangements 1-98.

[0340] In some embodiments, the formulation comprising: a pharmaceutically effective amount of a fusion protein; a buffer solution, the buffer solution comprising sodium acetate, a surfactant, and optionally a tonicity agent. In some embodiments, the fusion protein comprises an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap. In some embodiments, the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114. In some embodiments, the fusion protein comprises an antagonist antibody or a fragment thereof, that specifically binds to IL-6 that is conjugated to a polymer. In some embodiments, the fusion protein comprises an antagonistic IL-6 antibody or fragment thereof comprising: a) a heavy chain amino acid variable region that comprises a heavy chain that has a sequence of at least one of SEQ ID NOs: 7-13,19-27, 89, 90, 256-262; and b) a light chain amino acid variable region that comprises the light chain that has a sequence of at least one of SEQ ID NOs: 91-93, 28-30. In some embodiments, the fusion protein comprises an antagonist IL-6 antibody or fragment thereof comprising: a heavy chain variable region (VH)comprising 3 complementarity determining regions: VH (CDR1), VH CDR2, and VH CDR3 having an amino acid sequence from the CDRs listed in SEQ ID NO: 256; and a light chain variable region (VL) comprising a VL CDR1, VL CDR2, and VL CDR3 having an amino acid sequence selected from the group of CDRs listed in SEQ ID NO: 91-93. In some embodiments, the fusion protein further comprises an antagonistic antibody or fragment thereof that binds to IL-6, the antibody comprising: a CDRH1 that is a CDRH1 in SEQ ID NO: 172; a CDRH2 that is a CDRH2 in SEQ ID NO: 173; a CDRL1 that is a CDRL1 in SEQ ID NO: 199; a CDRL2 that is a CDRL2 in SEQ ID NO: 200; a CDRL3 that is a CDRL3 in SEQ ID NO: 201; at least one of the following mutations (EU numbering): L234A, L235A, and G237A; and at least one of the following mutations (EU numbering): Q347C or L443C. In some embodiments, the VEGF Trap is positioned either: a) at an N-terminal end of a heavy chain comprising IL-6 VH; or b) between a hinge region and after a CHI domain of a heavy chain comprising IL-6 VH. In some embodiments, the VEGF Trap consists of the sequence of SEQ ID NO: 114. In some embodiments, the surfactant comprises between about 0.01% to about 0.05% (weight / weight) polysorbate 20. In some embodiments, the surfactant comprises about 0.025% (weight / weight) polysorbate 20. In some embodiments, the surfactant comprises between about 0.01% to about 0.05% (weight / weight) polysorbate 80. In some embodiments, the surfactant comprises about 0.025% (weight / weight) polysorbate 80. In some embodiments, the formulation is between about pH 4.5 to about pH 6.5. In some embodiments, the formulation is about pH 5.5. In some embodiments, the formulation is stable for at least 52 weeks. In some embodiments, the formulation is stable for at least 52 weeks at 4 centigrade. In some embodiments, the formulation is stable for at least 8 weeks at 37 centigrade. In some embodiments, the formulation is stable in acidic conditions. In some embodiments, the formulation configured for intravitreal injection. In some embodiments, the sodium acetate concentration in the buffer solution is about 0.1 mM to about 50 mM. In some embodiments, the sodium acetate concentration in the buffer solution is about 15 mM sodium acetate. In some embodiments, the concentration of fusion protein is between about 20-150 mg / mL. In some embodiments, the concentration of fusion protein is about 100 mg / mL. In some embodiments, the tonicity agent is about 2-10% (weight / volume) of the formulation. In some embodiments, the tonicity agent is sucrose or trehalose. In some embodiments, the sucrose is about 5.2% (weight / volume) of the formulation. In some embodiments, the trehalose is about 5.2%(weight / volume) of the formulation. In some embodiments, the formulation is not the formulation of any one of arrangements 1-98.

[0341] In some embodiments, a pharmaceutical formulation is provided, the formulation comprising: a pharmaceutically effective amount of a fusion protein, wherein the concentration of the fusion protein is 100 mg / mL a buffer solution, the buffer solution comprising sodium acetate, wherein the concentration of sodium acetate is 15 mM, and a tonicity agent, wherein the tonicity agent is 5.2% (weight / volume) sucrose or trehalose; a surfactant, and wherein the surfactant comprises 0.025% (weight / weight) polysorbate 20 or polysorbate 80, wherein the pH is 5.5; and wherein the fusion protein comprises an anti-IL-6 antibody and a VEGF trap, wherein the fusion protein comprises SEQ ID NOs: 169 and 170 (with or without the C-terminal lysine in SEQ ID NO: 170).

[0342] In some embodiments, the formulation excludes the formulations recited in any one of or all of arrangements 1-98.

[0343] In some embodiments, the formulation is a formulation disclosed herein but that excludes the specific formulations disclosed in any one of Examples 1-33.

[0344] In some embodiments, the formulation can include histidine. In some embodiments, the formulation can include histidine acetate. In some embodiments, the formulation can include the features as seen in FIG. 52 In some embodiments, the formulation can replace sodium acetate with histidine acetate. In some embodiments, the formulation can include a histidine acetate range from 10-50 nM. In some embodiments, the formulation can include a sucrose or trehalose range from 2-10% (w / v). In some embodiments, the formulation can include a polysorbate 20 range from 0.01-0.1% (w / w). In some embodiments, the formulation can include a protein concentration range from 20-100 mg / mL. In some embodiments, the formulation can include a pH range from 5.0-6.5. In some embodiments, the formulation can include a percent of unconjugated fusion protein range from 10-50%. In some embodiments, the formulation can include 25 mM histidine acetate. In some embodiments, the formulation can include 6% (w / v) of sucrose or trehalose. In some embodiments, the formulation can include 0.025 % (w / w) of polysorbate 20. In some embodiments, the formulation can include a protein concentration of 50 mg / mL. In some embodiments, the formulation can include a pH of 5.5. In some embodiments, the formulation can include an unconjugated fusion protein concentration of 30%. In someembodiments, the polymer can be any polymer disclosed herein. In some embodiments, the fusion protein can be any fusion protein disclosed herein.

[0345] In some embodiments, the formulation can comprise a pH range of 4.5-5.5. In some embodiments, the formulation can comprise a tonicity agent percent concentration range from 0-6%. In some embodiments, the formulation can comprise a sodium chloride concentration range from 0-40 mM. In some embodiments, the formulation can comprise a sodium acetate concentration range from 10-50 mM. In some embodiments, the formulation can comprise a protein concentration range from 50-100 g / L. In some embodiments, the formulation can comprise a buffer further supplemented with 0.025% polysorbate 20.

[0346] In some embodiment, the formulation comprises 15 mM sodium acetate, pH 5.5; 5.2% tonicity agent; 100 g / L OG2072 mAb. In some embodiments, parameters for some exemplary formulations of OG2072 and combination of OG2072 and OG2074 are shown in FIG. 52. In some embodiments, preferred formulations can be the formulations on the right side of FIG. 52. OG2072 denotes a fusion construct that includes an anti-IL-6 antibody fused to a VEGF trap, and that includes a heavy chain of SEQ ID NO: 170 (with or without the C-terminal lysine) and a light chain of SEQ ID NO: 169.

[0347] Exemplary methods and materials are described herein, although methods and materials similar or equivalent to those described herein can also be used. The materials, methods, and examples are illustrative only and not intended to be limiting.IL-6 ANTAGONIST ANTIBODIES, IL-6 Ab-VEGF traps and / or CONJUGATES THEREOF

[0348] Provided herein are anti-IL-6 antibodies that block, suppress or reduce (including significantly reduces) IL-6 biological activity, including downstream events mediated by IL-6. In some embodiments, the IL-6 antagonist antibody will have one or more of the CDR sequences provided herein.

[0349] In some embodiments, the isolated antagonist antibody specifically binds to IL- 6.

[0350] In some embodiments, the antibody preferably reacts with IL-6 in a manner that inhibits IL-6 signaling function. In some embodiments, the IL-6 antagonist antibody specifically binds primate IL-6.

[0351] Also provided are antibodies that encompass monoclonal antibodies, polyclonal antibodies, antibody fragments (c.g., Fab, Fab’, F(ab’)i, Fv, Fc, etc.), chimeric antibodies, bispecific antibodies, heteroconjugate antibodies, single chain (ScFv), mutants thereof, fusion proteins comprising an antibody portion (e.g., a domain antibody), humanized antibodies, and any other modified configuration of the immunoglobulin molecule that comprises an antigen recognition site of the required specificity, including glycosylation variants of antibodies, amino acid sequence variants of antibodies, and covalently modified antibodies. The antibodies may be murine, rat, human, or any other origin (including chimeric or humanized antibodies). In some embodiments, the IL-6 antagonist antibody is a monoclonal antibody. In some embodiments, the antibody is a human or humanized antibody.

[0352] In some embodiments, the antibody comprises a heavy chain amino acid variable region as shown in Tables 1, 2, 6, 7, 8 and / or 9 or FIG. 5. In some embodiments, an isolated antagonist antibody comprises a heavy chain variable region (VH) comprising a VH complementarity determining region one (CDR1), VH CDR2, and VH CDR3 of the VH having an amino acid sequence of that shown in Tables 1, 2, 6, 7, 8 and / or 9, and a light chain variable region (VL) comprising a VL CDR1, VL CDR2, and VL CDR3 of the VL having an amino acid sequence of that shown in the table.

[0353] In some embodiments, an isolated antagonist anti-IL-6 antibody is provided. The antibody comprises a heavy chain constant domain comprising one or more mutations to reduce effector function. In some embodiments, the one or more mutations reduce effector functions of the antibody related to the complement cascade, for example, a reduced activation of the complement cascade. In some embodiments, the reduction in effector function is at least about 50%.

[0354] In some embodiments, an isolated antagonist antibody that specifically binds to IL-6 comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the antibody comprises the mutations L234A, L235A, and G237A (based on EU numbering) is provided. In some embodiments, the isolated antagonist antibody comprises the mutations L234A, L235A, and G237A. In some embodiments, the isolated antagonist antibody with mutations has minimized binding to FC gamma receptors or Clq. In some embodiments, the isolated antagonist antibody with mutations L234A, L235A, and G237A has minimized binding to FC gamma receptors or Clq. In some embodiments, an isolated antagonist anti-IL-6 antibodyis provided, wherein the mutation(s) is located at one or more of the following amino acid positions (EU numbering): E233, L234, L235, G236, G237, A327, A330, and P33E In some embodiments, an isolated antagonist anti-IL-6 antibody is provided, wherein the mutation (s) is selected from the group consisting of E233P, L234V, L234A, L235A, G237A, A327G, A330S, and P331S.

[0355] In some embodiments, an isolated antagonist anti-IL-6 antibody is provided. The heavy chain constant domain further comprises a cysteine residue introduced by recombinant DNA technology. In some embodiments, the cysteine residue is selected from the group consisting of Q347C and L443C (EU numbering). In some embodiments, the cysteine residue is L443C (EU numbering).

[0356] In some embodiments, the antibody comprises all three of the following mutations (EU numbering) L234A, L235A, and G237A, and the antibody comprises L443C (EU numbering). In some embodiments, the antibody is a human IgGl, and a heavy chain constant domain of the antibody comprises one or more mutations that reduce an immune-mediated effector function.

[0357] In some embodiments, an isolated antagonist antibody is provided that binds an epitope on human IL-6 that is the same as or overlaps with the epitope recognized by an antibody comprising the amino acid sequences in any one or more of Tables: 1 and 2, and / or 6-9. In some embodiments, an IL-6 antibody with a cys and that is linked through that cysteine to a polymer is provided (as shown in Formula 17, 17 A, or 17Br herein).

[0358] In some embodiments, an isolated antagonist antibody that binds to IL-6 is provided. In some embodiments, the isolated antagonist antibody that binds to IL-6 comprises a heavy chain comprising the amino acid sequence shown in Tables 1, 2, 6, 7, 8 and / or 9, with or without the C-terminal lysine and a light chain comprising the amino acid sequence shown in Tables 1, 2, 6, 7, 8 and / or 9.

[0359] In some embodiments, an isolated antagonist antibody that binds to IL-6 is provided. The antibody comprises a VH comprising the amino acid sequence shown in Tables 1, 2, 6, 7, 8 and / or 9, or a sequence that is at least 90% identical thereto, having amino acid substitutions in residues that are not within a CDR. In some embodiments, the antibody comprises one or more of: HCDR1: in FIG. 5, HCDR2: in FIG. 5, HCDR3: in FIG. 5; LCDR1: in FIG. 5, LCDR2: in FIG. 5, LCDR3: in FIG. 5, for example, 1, 2, 3, 4, 5, or all 6 CDRs.

[0360] In some embodiments, an antibody that binds to IL-6 is provided, wherein the antibody comprises a CDRH1 that is the CDRH1 in Tables 1, 2, 6, 7, 8 and / or 9, a CDRH2 that is the CDRH2 in Tables 1, 2, 6, 7, 8 and / or 9, a CDRH3 that is the CDRH3 in Tables 1, 2, 6, 7, 8 and / or 9, a CDRL1 that is the CDRL1 in Tables 1, 2, 6, 7, 8 and / or 9, a CDRL2 that is the CDRL2 in Tables 1, 2, 6, 7, 8 and / or 9, a CDRL3 that is the CDRL3 in Tables 1, 2, 6, 7, 8 and / or 9, at least one of the following mutations: L234A, L235A, and G237A based on EU numbering, and at least one of the following mutations:Q347C or L443C based on EU numbering.

[0361] In some embodiments, an isolated antagonist anti-IL-6 antibody is provided. The heavy chain variable region of the antibody comprises three complementarity determining regions (CDRs) comprising the amino acid sequences shown in Table 1. In some embodiments, an isolated antagonist anti-IL-6 antibody is provided, wherein the light chain variable region of the antibody comprises three complementarity determining regions (CDRs) comprising the amino acid sequences shown in Table 2. In some embodiments, the antibody is one that contains one or more of the identified sequences in FIG. 5, e.g., one or more of the CDRS (including 2, 3, 4, 5 or 6 of the boxed CDRs) and / or the entire heavy and light chain variable regions.

[0362] In some embodiments, an isolated antagonist anti-IL-6 antibody comprises a heavy chain variable region (VH) that comprises three CDRs comprising the amino acid sequences shown in Table 1, and the light chain variable region (VL) of the antibody comprises three CDRs comprising the amino acid sequences shown in Table 2.

[0363] In some embodiments, an isolated antagonist anti-IL-6 antibody is provided. The VH comprises the amino acid sequences shown in Table 1 and the light chain variable region of the antibody comprises three CDRs comprising the amino acid sequences shown in Table 2.

[0364] In some embodiments, an isolated antagonist anti-IL-6 antibody is provided, wherein the antibody comprises a VL comprising the amino acid sequence shown in Table 2, or a variant thereof with one amino acid substitution in amino acids that are not within a CDR. In some embodiments, an isolated antagonist anti-IL-6 antibody is provided, wherein the antibody comprises a VH comprising the amino acid sequence shown in Table 1, or a variant thereof with several amino acid substitutions in amino acids that are not within a CDR.

[0365] In some embodiments, an isolated antagonist antibody is provided, wherein the antibody comprises a heavy chain comprising the amino acid sequence shown in Table 1, with a C-terminal lysine, and a light chain comprising the amino acid sequence shown in Table 2. Insome embodiments, an isolated antagonist antibody is provided, wherein the antibody comprises a heavy chain comprising the amino acid sequence shown in Tabic 1, without a C-tcrminal lysine, and a light chain comprising the amino acid sequence shown in Table 2.

[0366] The IL-6 antagonist antibodies may be made by any method known in the art. General techniques for production of human and mouse antibodies are known in the art and / or are described herein.

[0367] IL-6 antagonist antibodies can be identified or characterized using methods known in the ail, whereby reduction, amelioration, or neutralization of IL-6 biological activity is detected and / or measured. In some embodiments, an IL-6 antagonist antibody is identified by incubating a candidate antibody with IL-6 and monitoring binding to IL-6R or IL-6 / IL-6R binding to gpl30 and / or attendant reduction or neutralization of a biological activity of IL-6. The binding assay may be performed with, e.g., purified IL-6 polypeptide(s), or with cells naturally expressing (e.g., various strains), or transfected to express, IL-6 polypeptide(s). In one embodiment, the binding assay is a competitive binding assay, where the ability of a candidate antibody to compete with a known IL-6 antagonist antibody for IL-6 binding is evaluated. The assay may be performed in various formats, including the ELISA format.

[0368] Following initial identification, the activity of a candidate IL-6 antagonist antibody can be further confirmed and refined by bioassays, known to test the targeted biological activities. In some embodiments, an in vitro cell assay is used to further characterize a candidate IL-6 antagonist antibody.

[0369] IL-6 antagonist antibodies may be characterized using methods well known in the art. For example, one method is to identify the epitope to which it binds, or “epitope mapping.” There are many methods known in the art for mapping and characterizing the location of epitopes on proteins, including solving the crystal structure of an antibody-antigen complex, competition assays, gene fragment expression assays, and synthetic peptide-based assays, as described, for example, in Chapter 11 of Harlow and Lane, Using Antibodies, a Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1999. In an additional example, epitope mapping can be used to determine the sequence to which an IL-6 antagonist antibody binds. IL-6 antagonist antibody Epitope mapping is commercially available from various sources, for example, Pepscan Systems (Edelhertweg 15, 8219 PH Lelystad, The Netherlands). The epitope can be a linear epitope, i.e., contained in a single stretch of amino acids, or a conformational epitope formedby a three-dimensional interaction of amino acids that may not necessarily be contained in a single stretch. Peptides of varying lengths (c.g., at least 4-6 amino acids long) can be isolated or synthesized (e.g., recombinantly) and used for binding assays with an IL-6 antagonist antibody. In another example, the epitope to which the IL-6 antagonist antibody binds can be determined in a systematic screening by using overlapping peptides derived from the IL-6 sequence and determining binding by the IL-6 antagonist antibody. According to the gene fragment expression assays, the open reading frame encoding IL-6 is fragmented either randomly or by specific genetic constructions and the reactivity of the expressed fragments of IL-6 with the antibody to be tested is determined. The gene fragments may, for example, be produced by PCR and then transcribed and translated into protein in vitro, in the presence of radioactive amino acids. The binding of the antibody to the radioactively labeled IL-6 fragments is then determined by immunoprecipitation and gel electrophoresis. Certain epitopes can also be identified by using large libraries of random peptide sequences displayed on the surface of phage particles (phage libraries) or yeast (yeast display). Alternatively, a defined library of overlapping peptide fragments can be tested for binding to the test antibody in simple binding assays. In an additional example, mutagenesis of an antigen, domain swapping experiments and alanine scanning mutagenesis can be performed to identify residues required, sufficient, and / or necessary for epitope binding. For example, alanine scanning mutagenesis experiments can be performed using a mutant IL-6 in which various residues of the IL-6 polypeptide have been replaced with alanine. By assessing binding of the antibody to the mutant IL-6, the importance of the particular IL-6 residues to antibody binding can be assessed.

[0370] In some embodiments, an isolated antagonist anti-IL-6 antibody is provided, wherein the antibody binds human IL-6 with an affinity of between about 0.01 pM to about 10 nM. In some embodiments, an isolated antagonist anti-IL-6 antibody is provided, wherein the antibody binds human IL-6 with an affinity of between about 0.1 pM to about 2 nM. In some embodiments, an isolated antagonist anti-IL-6 antibody is provided, wherein the antibody binds human IL-6 with an affinity of about 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000 pM.

[0371] The binding affinity (KD) of an IL-6 antagonist antibody to IL-6 can be about 0.001 to about 200 nM. In some embodiments, the binding affinity is any of about 200 nM, about 100 nM, about 50 nM, about 10 nM, about 1 nM, about 500 pM, about 100 pM, about 60 pM, about 50 pM, about 20 pM, about 15 pM, about 10 pM, about 5 pM, about 2 pM, or about 1 pM.In some embodiments, the binding affinity is less than any of about 250 nM, about 200 nM, about 100 nM, about 50 nM, about 10 nM, about 1 nM, about 500 pM, about 100 pM, about 50 pM, about 20 pM, about 10 pM, about 5 pM, about 2 pM, about 1 pM, about 0.5 pM, about 0.1 pM, about 0.05 pM, about 0.01 pM, about 0.005 pM, or about 0.001 pM.

[0372] In some embodiments, an isolated antagonist anti-IL-6 antibody is provided, wherein the antibody binds human IL-6 with a koff that is at least 5.0E-03 at 37 degrees. In some embodiments the koff is 5E-04. In some embodiments, an isolated antagonist anti-IL-6 antibody is provided, wherein the antibody binds human IL-6 with a koff that is better than 5.0E-04 at 37 degrees.

[0373] In some embodiments, binding affinity can be defined in terms of one or more of association constant (ka), dissociation constant (ka), and analyte concentration that achieves half-maximum binding capacity (KD). In some embodiments, kacan range from about 0.50E+05 to about 5.OOE+O8. In some embodiments, kd can range from about 0.50E-06 to about 5.00E-03. In some embodiments, KD can range from about 0.50E-12 to about 0.50E-07.

[0374] In some embodiments, a pharmaceutical composition comprising any of the antibodies disclosed herein is provided. In some embodiments, a pharmaceutical composition comprising any of the conjugates disclosed herein is provided. In some embodiments, a pharmaceutical composition comprising any of the antibodies disclosed herein and any of the conjugates disclosed herein is provided. In some embodiments, the pharmaceutical composition comprises one or more pharmaceutically acceptable carriers. In some embodiments, the pharmaceutical composition is a liquid. In some embodiments, the pharmaceutical composition has an endotoxin level less than about 0.2 EU / ml. In some embodiments, the pharmaceutical composition is a liquid and has an endotoxin level less than about 0.2 EU / ml. In some embodiments, the pharmaceutical composition is a liquid and has an endotoxin level less than about 2.0, 1, 0.5, 0.2 EU / ml. In some embodiments, for example in intravitreal injection, the endotoxin limit is 0.01-0.02 EU / injection / eye.

[0375] Some embodiments provide any of the following, or compositions (including pharmaceutical compositions) comprising an antibody having a partial light chain sequence and a partial heavy chain sequence as found in Tables 1 and 2, or variants thereof. In Tables 1 and 2, the underlined sequences are some embodiments of CDR sequences as provided herein.TABLE 1Tabic 1. Anti-IL-6 heavy chain variable region sequences. CDRs arc underlined.Table 2. Anti-IL-6 light chain variable region sequences. CDRs are underlined.

[0376] In some embodiments, the antibody does not have one or more (or any) of the following CDRs, Tables 3, 4, and / or 5.TABLE 3TABLE 4TABLE 5

[0377] In some embodiments, a composition as disclosed herein comprises an antibody having a partial or complete light chain sequence and a partial or complete heavy chain sequence from any of the options provided in Tables 1, 2, 6, 7, 8 and / or 9, or variants thereof. In some embodiments, the antibody (or binding fragment thereof) can include any one or more of the CDRs provided in Tables 1, 2, 6, 7, 8 and / or 9. In some embodiments, the antibody (or binding fragment thereof) can include any three or more of the CDRs provided in Tables 1, 2, 6, 7, 8 and / or 9. In some embodiments, the antibody (or binding fragment thereof) can include any all six of the CDRs provided in Tables 1, 2, 6, 7, 8 and / or 9. In some embodiments, the heavy and / or light chain can be any one or more of the other antibody constructs provided herein, including, for example, those provided in FIGs. 5, 18, and / or 21-24 and Tables 1, 3, 4, 5, 6, 7, 8 and / or 9.

[0378] In some embodiments, a composition as disclosed herein comprises an antibody having a partial or complete light chain CDR sequence and a partial or complete heavy chain CDR sequence from any of the options provided in Tables 1, 2, 6, 7, 8 and / or 9.

[0379] In some embodiments, CDR portions of IL-6 antagonist antibodies are also provided. Determination of CDR regions is well within the skill of the art. It is understood that in some embodiments, CDRs can be a combination of the IMGT and Paratome CDRs (also termed “combined CDRs” or “extended CDRs”). Determination of CDRs is well within the skill of the art. In some embodiments, the CDRs are the IMGT CDRs. In other embodiments, the CDRs are the Paratome CDRs. In other embodiments, the CDRs are the extended, AbM, conformational, Kabat, or Chothia CDRs. In embodiments with more than one CDR, the CDRs may be any of IMGT, Paratome, extended, Kabat, Chothia, AbM, conformational CDRs, or combinations thereof. In some embodiments, other CDR definitions may also be used. In some embodiments, only residues that are in common between 2, 3, 4, 5, 6, or 7 of the above definitions are used (resulting in a shorter sequence). In some embodiments, any residue in any of 2, 3, 4, 5, 6, or 7 of the above definitions can be used (resulting in a longer sequence).

[0380] In some embodiments, an IL-6 antagonist antibody comprises three CDRs of any one of the heavy chain variable regions shown in Tables 1, 2, 6, 7, 8 and / or 9. In some embodiments, the antibody comprises three CDRs of any one of the light chain variable regions shown in Tables 1, 2, 6, 7, 8 and / or 9. In some embodiments, the antibody comprises three CDRs of any one of the heavy chain variable regions shown in Table 1, and three CDRs of any one of the light chain variable regions shown in Table 2. In some embodiments, the CDRs are one or more of those designated in Tables 6 and / or 7, or 8 and / or 9 below:Table 6 ANTI IL-6 HEAVY CHAIN CDR SEQUENCES.Table 7. ANTI IL-6 HEAVY CHAIN CDR SEQUENCES. KabatTable 8. ANTI IL-6 LIGHT CHAIN CDR SEQUENCES.Table 9 ANTI IL-6 LIGHT CHAIN CDR SEQUENCES.

[0381] In some embodiments, the antibody used for binding to IL-6 can be one that includes one or more of the sequences in Tables 1, 2, 6, 7, 8 and / or 9. In some embodiments, the antibody used for binding to IL-6 can be one that includes three or more of the sequences in Tables 1, 2, 6, 7, 8 and / or 9. In some embodiments, the antibody used for binding to IL-6 can be one that includes six of the sequences in any one of Tables 1, 2, 6, 7, 8 and / or 9. In some embodiments, the antibody that binds to IL-6 can be one that competes for binding with an antibody that includes 6 of the specified CDRs in any one of Tables 1, 2, 6, 7, 8 and / or 9.

[0382] In some embodiments, the antibody can be linked or fused to a VEGF Trap sequence. In some embodiments, this Trap sequence can be as shown in Table 10. In some embodiments, the sequence is at least 80% identical to that shown in Table 10, e.g., at least 80, 85, 90, 95, 96, 97, 98, 99% identical to that shown in Table 10. In some embodiments, any of the VEGF Trap molecules in U.S. Pub. No. 20150376271 can be employed herein. In some embodiments, the VEGF Trap sequence is fused to IL-6 in one of the following manners: at an N- terminal end of a heavy chain comprising IL-6 VH (FIG. 6 left), or between a hinge region and after a CHI domain of a heavy chain comprising IL-6 VH (FIG. 6 right). Unless designated otherwise, both options in the alternative and together are contemplated for the embodiments provided herein wherein any Ab-Trap fusion is discussed. In some embodiments, the term “Trap” refers to a full length extracellular region or any portion thereof, or combination of portions from different VEGF receptors that can antagonize signaling between at least one VEGF and VEGFR. Preferably, the extracellular trap segment includes at least one domain from one of VEGFR- 1, -2 or -3, and more preferably at least two contiguous domains, such as D2 and D3. Optionally, an extracellular’ domain includes at least one domain from at least two different VEGFRs. A preferred extracellular domain comprises or consists essentially of D2 of VEGFR- 1 and D3 of VEGFR-2.TABLE 10. VEGFR1 , Domain 2 and VEGFR2, Domain 3 Fusion sequence

[0383] In some embodiments, the IL-6 Ab VEGF Trap construct can have any of the sequences provided in TABLE 11. In some embodiments, the construct can be at least identical to the sequences in Table 11, e.g., 80, 85, 90, 95, 96, 97, 98, 99 or higher. In some embodiments, the fusion protein can be in line with those percentages, with the exception that the antibody IL-6 domain does not contain one or more of the CDRs in Tables 3, 4, and / or 5. In some embodiments, the fusion protein is one that contains one or more of the identified sequences in FIG. 5, e.g., one of more of the CDRS (including 2, 3, 4, 5 or 6 of the boxed CDRs) and / or the entire heavy and light chain variable regions, along with a VEGF Trap sequence (e.g., Table 10). In some embodiments, the sequences can be directly fused to one another. In some embodiments, one or more flexible linking sequences or sections can be used. The linking sequence can be positioned between the Ab sequence and the VEGF Trap sequence. These sequences can be 5 to 30 amino acids in length. In some embodiments, the linking sequence can include G and S in a ratio of about 4:1. In some embodiments, the linker includes the following sequence: GGGGSGGGGS (SEQ ID NO: 115). In some embodiments, any flexible linker can be employed. In some embodiments, the Fc portion of the 11-6 Ab is IgGl.TABLE 11.Heavy and light chain sequences for dual inhibitor molecules. CDRs are underlined in the heavy and light chains, VEGF trap sequence is bolded in black, Gly-Ser linker is italicized.

[0384] To express the anti-IL-6 antibodies and / or IL-6 VEGF Traps provided herein, DNA fragments encoding VH and VL regions described can first be obtained. Variousmodifications, e.g. mutations, deletions, and / or additions can also be introduced into the DNA sequences using standard methods known to those of skill in the art. For example, mutagenesis can be carried out using standard methods, such as PCR-mediated mutagenesis, in which the mutated nucleotides are incorporated into the PCR primers such that the PCR product contains the desired mutations or site-directed mutagenesis.

[0385] Also provided are modifications to the variable regions shown herein. For example, also provided are antibodies comprising functionally equivalent variable regions and CDRs which do not significantly affect their properties as well as variants which have enhanced or decreased activity and / or affinity. For example, the amino acid sequence may be mutated to obtain an antibody with the desired binding affinity to IL-6. Modification of polypeptides is routine practice in the art and need not be described in detail herein. Examples of modified polypeptides include polypeptides with conservative substitutions of amino acid residues, one or more deletions or additions of amino acids which do not significantly deleteriously change the functional activity, or which mature (enhance) the affinity of the polypeptide for its ligand, or use of chemical analogs.

[0386] Amino acid sequence insertions include amino- and / or carboxyl-terminal fusions ranging in length from one residue to polypeptides containing a hundred or more residues, as well as intra- sequence insertions of single or multiple amino acid residues. Examples of terminal insertions include an antibody with an N-terminal methionyl residue or the antibody fused to an epitope tag. Other insertional variants of the antibody molecule include the fusion to the N- or C- terminus of the antibody of an enzyme or a polypeptide which increases the half-life of the antibody in the blood circulation.

[0387] Substitution variants have at least one amino acid residue in the antibody molecule removed and a different residue inserted in its place. The sites of greatest interest for substitutional mutagenesis include the hypervariable regions, but framework alterations are also contemplated. Conservative substitutions are shown in Table 12 under the heading of “conservative substitutions.” If such substitutions result in a change in biological activity, then more substantial changes, denominated “exemplary substitutions” in Table 12, or as further described below in reference to amino acid classes, may be introduced and the products screened.Table 12 - Amino Acid. Substitutions

[0388] Substantial modifications in the biological properties of the antibody are accomplished by selecting substitutions that differ significantly in their effect on maintaining (a) the structure of the polypeptide backbone in the area of the substitution, for example, as a P-sheet or helical conformation, (b) the charge or hydrophobicity of the molecule at the target site, or (c) the bulk of the side chain. Naturally occurring residues are divided into groups based on common side-chain properties:(1) Non-polar: Norleucine, Met, Ala, Vai, Leu, He;(2) Polar without charge: Cys, Ser, Thr, Asn, Gin;(3) Acidic (negatively charged): Asp, Glu;(4) Basic (positively charged): Lys, Arg;(5) Residues that influence chain orientation: Gly, Pro; and(6) Aromatic: Tip, Tyr, Phe, His.

[0389] Non-conservative substitutions are made by exchanging a member of one of these classes for another class.

[0390] One type of substitution, for example, that may be made is to change one or more cysteines in the antibody, which may be chemically reactive, to another residue, such as, without limitation, alanine or serine. For example, there can be a substitution of a non-canonical cysteine. The substitution can be made in a CDR or framework region of a variable domain or in the constant region of an antibody. In some embodiments, the cysteine is canonical. Any cysteine residue not involved in maintaining the proper conformation of the antibody also may be substituted, generally with serine, to improve the oxidative stability of the molecule and prevent aberrant cross-linking. Conversely, cysteine bond(s) may be added to the antibody to improve its stability, particularly where the antibody is an antibody fragment such as an Fv fragment.

[0391] The antibodies may also be modified, e.g. in the variable domains of the heavy and / or light chains, e.g., to alter a binding property of the antibody. Changes in the variable region can alter binding affinity and / or specificity. In some embodiments, no more than one to five conservative amino acid substitutions are made within a CDR domain. In other embodiments, no more than one to three conservative amino acid substitutions are made within a CDR domain. For example, a mutation may be made in one or more of the CDR regions to increase or decrease the KD of the antibody for IL-6, to increase or decrease koff, or to alter the binding specificity of the antibody. Techniques in site-directed mutagenesis are well-known in the art. See, e.g., Sambrook et al. and Ausubel et al., supra.

[0392] According to an aspect, the IgG domain of an IL-6 antagonist antibody can be IgGl, IgG2, IgG3 or IgG4. According to another aspect, the IgG domain can be a composite in which a constant regions is formed from more than one of the above isotypes (e.g., CHi region from IgG2 or IgG4, hinge, CH2 and CH3 regions from IgGl). In choosing an isotype, it is known in the ail that human isotopes IgGl and IgG3 have complement-mediated cytotoxicity whereas human isotypes IgG2 and IgG4 have poor or no complement-mediated cytotoxicity. In some embodiments the IL-6 antagonist antibody isotype is IgGl.

[0393] The light chain constant region can be either human lambda or kappa. In some embodiments, the IL-6 antagonist antibody has a human kappa light chain constant region.

[0394] Human constant regions show allotypic variation and isoallotypic variation between different individuals, that is, the constant regions can differ in different individuals at one or more polymorphic positions. Isoallotypes differ from allotypes in that sera recognizing an isoallotype binds to a non-polymorphic region of one or more other isotypes. Reference to a human constant region includes a constant region with any natural allotype or any permutation of residues occupying polymorphic positions in natural allotypes or up to 3, 5 or 10 substitutions for reducing or increasing effector function as described below.

[0395] One or several amino acids at the amino or carboxy terminus of the light and / or heavy chains such as the C-terminal lysine of the heavy chain, may be missing or derivatized in a proportion or all of the molecules.

[0396] Substitutions can be made in the constant regions to reduce or increase effector function such as complement-mediated cytotoxicity (CDC), antibody-dependent cell-mediated cytotoxicity (ADCC) (see, e.g., Winter et al., US Patent No.5, 624, 821; Tso et al., US Patent No. 5,834,597; and Lazar et al., Proc. Natl. Acad. Sci. USA 103:4005, 2006), or to prolong half-life in humans (see, e.g., Hinton et al., J. Biol. Chem. 279:6213, 2004).

[0397] In some embodiments, the IL-6 antagonist antibodies provided herein include one more substitutions that reduce complement mediated cytotoxicity. Reduction in complement mediated cytotoxicity can be accomplished with or without reduction in Fc receptor binding depending on the nature of the mutation(s). Antibodies with reduced complement mediated cytotoxicity but little or no reduction in Fc receptor allow a desired effect of Fc-mediated phagocytosis of iC3b without activating complement, which may contribute to side effects. Exemplary mutations known to reduce complement-mediated cytotoxicity in human constant regions include mutations at positions 241, 264, 265, 270, 296, 297, 322, 329 and 331 by EU numbering. Mutations in positions 318, 320, and 322 have been reported to reduce complement activation in mouse antibodies. Alanine is a preferred residue to occupy these positions in a mutated constant region. Some exemplary human mutations that have been used include F241A, V264A, D265A, V296A, N297A, K322A, and P331S in human IgG3 and D270A or E, N297Q, K322A, P329A, and P331S in human IgGl (EU numbering).

[0398] Here, as elsewhere, the EU numbering scheme is used for numbering amino acids in the constant region of an antibody. When a residue in a variable region is referenced herein (unless designated otherwise) the residue numbering is according to the variable domain(or, if designated, the SEQ ID NO). Substitution at any or all of positions 234, 235, 236 and / or 237 reduce affinity for Fey receptors, particularly FcyRI receptor and also reduces complement binding and activation (see, e.g., US 6,624,821 WO / 2009 / 052439). An alanine substitution at positions 234, 235 and 237 reduces effector functions, particularly in the context of human IgGl. Optionally, positions 234, 236 and / or 237 in human IgG2 are substituted with alanine and position 235 with glutamine. (See, e.g., US 5,624,821) to reduce Fc receptor binding. Exemplary substitutions for increasing half-life include a Gin at position 250 and / or a Feu at position 428. Also provided are aspects where the anti-IE-6 antibody presented has a human IgGl isotype, it is preferred that the antibody has at least one mutation in the constant region. Preferably, the mutation reduces complement fixation or activation by the constant region. Also provided are particularly preferred aspects, where the antibody has one or more mutations at positions E233, E234, E235, G236, G237, A327, A330 and P331 by EU numbering. Still more preferably, the mutations constitute one or more of the following E233P, E234V, E234A, E235A, G237A, A327G, A330S and P331S by EU numbering. In the most preferred embodiments the human IgGl has the following mutations L234A, L235A and G237A by EU numbering.Conjugates

[0399] The half-life of IL-6 antagonist antibodies and / or IL-6 Ab VEGF Traps can be extended by attachment of a “half-life extending moieties” or “half-life extending groups,” which terms are herein used interchangeably to refer to one or more chemical groups attached to one or more amino acid site chain functionalities such as -SH, -OH, -COOH, -CONH2, -NH2, or one or more N- and / or O-glycan structures and that can increase in vivo circulatory half-life of proteins / peptides when conjugated to these proteins / peptides. Examples of half-life extending moieties include polymers described herein, particularly those of zwitterionic monomers, such as HEMA-phosphorylcholine, PEG, biocompatible fatty acids and derivatives thereof, Hydroxy Alkyl Starch (HAS) e.g. Hydroxy Ethyl Starch (HES), Poly Ethylene Glycol (PEG), Poly (Glyx- Sery) (HAP), Hyaluronic acid (HA), Heparosan polymers (HEP), Fleximers, Dextran, Poly-sialic acids (PSA), Fc domains, Transferrin, 25 Albumin, Elastin like (ELP) peptides, XTEN polymers, PAS polymers, PA polymers, Albumin binding peptides, CTP peptides, FcRn binding peptides and any combination thereof.

[0400] In some embodiments, the antibody is conjugated with a phosphorylcholine containing polymer. In some embodiments, the antibody is conjugated with apoly(acryloyloxyethyl phosphorylcholine) containing polymer, such as a polymer of acrylic acid containing at least one acryloyloxycthyl phosphorylcholinc monomer such as 2- methacryloyloxyethyl phosphorylcholine (i.e., 2-methacryloyl-2’ -trimethylammonium ethyl phosphate).

[0401] In some embodiments, the antibody and / or antibody VEGF Trap fusion is conjugated with a water-soluble polymer, which refers to a polymer that is soluble in water. A solution of a water-soluble polymer may transmit at least about 75%, more preferably at least about 95% of light, transmitted by the same solution after filtering. On a weight basis, a water-soluble polymer or segment thereof may be at least about 35%, at least about 50%, about 70%, about 85%, about 95% or 100% (by weight of dry polymer) soluble in water.

[0402] In one embodiment a half-life extending moiety can be conjugated to an IL-6 antagonist antibodies and / or IL-6 Ab VEGF Trap via free amino groups of the protein using N- hydroxysuccinimide (NHS) esters. Reagents targeting conjugation to amine groups can randomly react to e-amine group of lysines, a-amine group of N-terminal amino acids, and 5-amine group of histidines.

[0403] However, also provided are IL-6 antagonist antibodies which have many amine groups available for polymer conjugation. Conjugation of polymers to free amino groups, thus, might negatively impact the ability of the antibody to bind to the epitope.

[0404] In another embodiment, a half-life extending moiety is coupled to one or more free SH groups using any appropriate thiol-reactive chemistry including, without limitation, maleimide chemistry, or the coupling of polymer hydrazides or polymer amines to carbohydrate moieties of the IL-6 antagonist antibodies and / or IL-6 Ab VEGF Traps after prior oxidation. The use of maleimide coupling is a particularly preferred embodiment. Coupling preferably occurs at cysteines naturally present or introduced via genetic engineering.

[0405] In some embodiments, polymers are covalently attached to cysteine residues introduced into IL-6 antagonist antibodies and / or IL-6 Ab VEGF Traps by site directed mutagenesis. In some embodiments, the cysteine residues in the Fc portion of the IL-6 antagonist antibody and / or IL-6 Ab VEGF Trap can be used. In some embodiments, sites to introduce cysteine residues into an Fc region are provided in WO 2013 / 093809, US 7,521,541, WO 2008 / 020827, US 8,008,453, US 8,455,622 and US2012 / 0213705, incorporated herein by reference for all purposes. In some embodiments, cysteine mutations are Q347C and L443Creferring to the human IgG heavy chain by the EU index of Kabat. In some embodiments, the cysteine added by directed mut...

Claims

What Is Claimed Is:

1. A pharmaceutical formulation comprising: a fusion protein conjugate comprising a first fusion protein conjugated to a phosphorylcholine-containing polymer; an unconjugated fusion protein comprising a second fusion protein that is not conjugated to a phosphorylcholine-containing polymer; a buffer, wherein the buffer comprises histidine; and a surfactant, wherein the first fusion protein and the second fusion protein each comprises: an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap, wherein the unconjugated fusion protein is present in the formulation at 10-60% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein.

2. The formulation of claim 1, wherein the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114.

3. The formulation of claim 1, wherein the first fusion protein and the second fusion protein each comprises: a light chain with at least 80% identity to the sequence of SEQ ID NO: 169; and a heavy chain with at least 80% identity to the sequence of SEQ ID NO: 170.

4. The formulation of claim 3, wherein the fusion protein conjugate comprises the following structure:Formula (17 A) wherein: part of each heavy chain of the first fusion protein is denoted by the letter H, and each light chain of the first fusion protein is denoted by the letter L; the polymer is bonded to the heavy chain through the sulfhydryl of C443 (EU numbering), which bond is depicted on one of the heavy chains;PC is , where the curvy line indicates the point of attachment to the rest of the polymer, where X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) -H, c) any halogen, including -Br, -Cl, or -I, d) -SCN, or c) - NCS; andnl , n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 2500 plus or minus 15%.

5. The formulation of claim 1, wherein the first fusion protein and the second fusion protein each comprises an antagonistic IL-6 antibody or an antigen-binding fragment thereof comprising: a heavy chain amino acid variable region that comprises a heavy chain comprising a sequence of at least one of SEQ ID NOs: 7-13,19-27, 89, 90, 256-262; and a light chain amino acid variable region that comprises a light chain comprising a sequence of at least one of SEQ ID NOs: 91-93, 28-30.

6. The formulation of claim 1, wherein the first fusion protein and the second fusion protein each comprises an antagonist IL-6 antibody or fragment thereof comprising: a heavy chain variable region (VH) comprising 3 complementarity determining regions: VH (CDR1), VH CDR2, and VH CDR3 having an amino acid sequence from the CDRs listed in SEQ ID NO: 256; and a light chain variable region (VL) comprising a VL CDR1, VL CDR2, and VL CDR3 having an amino acid sequence selected from the group of CDRs listed in SEQ ID NOs: 91-93.

7. The formulation of claim 1, wherein the first fusion protein and the second fusion protein each comprises an antagonistic antibody or fragment thereof that binds to IL-6, the antibody comprising: a CDRH1 that is a CDRH1 in SEQ ID NO: 172; a CDRH2 that is a CDRH2 in SEQ ID NO: 173; a CDRL1 that is a CDRL1 in SEQ ID NO: 199; a CDRL2 that is a CDRL2 in SEQ ID NO: 200; a CDRL3 that is a CDRL3 in SEQ ID NO: 201; and at least one of the following mutations (EU numbering): L234A, L235A, and G237A, wherein the first fusion protein comprises at least one of the following mutations (EU numbering): Q347C or L443C, and optionally wherein the second fusion protein comprises at least one of the following mutations (EU numbering): Q347C or L443C.

8. The formulation of claim 1, wherein the first fusion protein comprises an antagonistic IL-6 antibody or an antigen-binding fragment thereof, wherein the VEGF Trap is positioned either: at an N-terminal end of a heavy chain comprising IL-6 VH; orbetween a hinge region and after a CH 1 domain of a heavy chain comprising TL-6 VH.

9. The formulation of claim 1, wherein the surfactant comprises polysorbate 20, polysorbate 80, or poloxamer 188.

10. The formulation of claim 1 , wherein the surfactant is present in the formulation at 0.01% (w / w) to 0.1% (w / w).

11. The formulation of claim 1, wherein the buffer comprises histidine acetate.

12. The formulation of claim 11, wherein the histidine acetate concentration is 1 mM to 60 mM.

13. The formulation of claim 12, wherein the histidine acetate concentration is about 15 mM.

14. The formulation of claim 1, wherein the formulation has a pH from 4.9 to 6.2.

15. The formulation of claim 1, wherein the formulation has a pH from 5.6 to 5.9.

16. The formulation of claim 1, wherein the formulation has a pH of about 5.6.

17. The formulation of claim 1, wherein the concentration of the first fusion protein and second fusion protein combined is 40-60 mg / mL.

18. The formulation of claim 1, wherein the concentration of the first fusion protein and second fusion protein combined is about 50 mg / mL.

19. A pharmaceutical formulation comprising: a fusion protein conjugate comprising: a first fusion protein comprising: a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:170, with or without the C-terminal lysine; and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 169; and a phosphorylcholine-containing polymer conjugated to the heavy chain,wherein the fusion protein conjugate comprises the following structure:Formula (17A) wherein: part of each heavy chain of the first fusion protein is denoted by the letter H, and each light chain of the first fusion protein is denoted by the letter L; the polymer is bonded to the heavy chain of the first fusion protein through the sulfhydryl of C443 (EU numbering), which bond is depicted on one of the heavy chains;PC is , wherein the curvy line indicates the point of attachment to the rest of the polymer, wherein X is a) -OR where R is H, methyl,ethyl , propyl, or isopropyl, b) -H, or c) any halogen, including -Br, -Cl, or -I, d) - SCN, or c) -NCS; and nl, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, n8 and n9 is about 2500 plus or minus 15%; an unconjugated fusion protein comprising a second fusion protein that is not conjugated to a phosphorylcholine-containing polymer, the unconjugated fusion protein comprising: a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 170, with or without the C-terminal lysine, or a variant thereof having L449 as numbered according to SEQ ID NO: 170; and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 169, wherein the concentration of the first fusion protein and second fusion protein combined is 40-60 mg / mL, wherein the unconjugated fusion protein is present in the formulation at about 30% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, and wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein; histidine acetate at 1-60 mM; and a surfactant selected from polysorbate 20, polysorbate 80, and poloxamer 188, wherein the surfactant is present in the formulation at 0.01% (w / w) to 0.1% (w / w), wherein the formulation has a pH from 4.9 to 6.2.

20. The formulation of claim 19, consisting of or consisting essentially of: the fusion protein conjugate; the unconjugated fusion protein, wherein the concentration of the first fusion protein and second fusion protein combined is 40-60 mg / mL, wherein the unconjugated fusion protein is present in the formulation at about 30% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, and wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein; histidine acetate at 1-60 mM; and the surfactant present in the formulation at 0.01% (w / w) to 0.1% (w / w).

21. A pharmaceutical formulation, the formulation comprising: a pharmaceutically effective amount of a fusion protein, wherein the concentration of the fusion protein is about 50 mg / mL; a phosphorylcholine-containing polymer, wherein the fusion protein either is conjugated to the polymer or is unconjugated; histidine acetate at about 15 mM; and a surfactant at about 0.025% (weight / weight), wherein the surfactant comprises polysorbate 20, polysorbate 80, and / or poloxamer 188, wherein the formulation has a pH of about 5.6, wherein the unconjugated fusion protein is present in the formulation at about 30% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein, and wherein the fusion protein comprises: a light chain comprising an amino acid sequence at least 80% identical toSEQ ID NO: 169; and a heavy chain comprising an amino acid sequence at least 80% identical toSEQ ID NO: 170, wherein the fusion protein conjugate comprises the following structure:Formula (17 A) wherein: part of each heavy chain of the fusion protein is denoted by the letter H, and each light chain of the fusion protein is denoted by the letter L; the polymer is bonded to the heavy chain through the sulfhydryl of C443 (EU numbering), which bond is depicted on one of the heavy chains;PC is , wherein the curvy line indicates the point of attachment to the rest of the polymer, wherein X is a) -OR where R is H, methyl, ethyl, propyl, or isopropyl, b) -H, or c) any halogen, including -Br, -Cl, or -I, d) - SCN, or e) -NCS; andnl , n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of nl, n2, n3, n4, n5, n6, n7, 118 and n9 is about 2500 plus or minus 15%.

22. The formulation of claim 21, consisting of or consisting essentially of: the pharmaceutically effective amount of the fusion protein, wherein the concentration of the fusion protein is about 50 mg / mL; the phosphorylcholine-containing polymer, wherein the fusion protein either is conjugated to the polymer or is unconjugated; histidine acetate at about 15 mM; and the surfactant at about 0.025% ( weigh t / weight), wherein the unconjugated fusion protein is present in the formulation at about 30% of a total molar amount of the fusion protein conjugate and the unconjugated fusion protein, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated fusion protein.

23. The formulation of any one of the preceding claims, wherein the formulation is storage stable for at least 4 weeks at about 37 °C.

24. The formulation of any one of the preceding claims, wherein the formulation is storage stable for about 2 months at about 37 °C.

25. The formulation of any one of the preceding claims, wherein the formulation is substantially free of turbidity.

26. The formulation of any one of the preceding claims, wherein the formulation is substantially free of turbidity after storage at about 37 °C for at least 4 weeks.

27. The formulation of any one of the preceding claims, wherein the formulation is substantially free of turbidity after storage at about 37 °C for about 2 months.

28. The formulation of any one of the preceding claims that is a formulation for intravitreal injection.

29. A pharmaceutical formulation comprising: a fusion protein comprising: an CDRH3 having at least 80% identity with the amino acid sequence of SEQ IDNO: 174 (QAWGYYALDI); and a VEGF trap; and a buffer,wherein the pharmaceutical formulation has a pH in a range of 4.5-6.8, optionally, wherein the formulation comprises a surfactant.

30. The formulation of claim 29, wherein the VEGF Trap has at least 80% identity to the sequence of SEQ ID NO: 114.

31. The formulation of claim 29, wherein the fusion protein comprises an anti-IL-6 antibody or an antigen-binding fragment thereof comprising: a) a heavy chain amino acid variable region that comprises a heavy chain that has a sequence of at least one of SEQ ID NOs: 7-13,19-27, 89, 90, 256-262; and b) a light chain amino acid variable region that comprises the light chain that has a sequences of at least one of SEQ ID NOs: 91-93, 28-30.

32. The formulation of claim 29, wherein the fusion protein comprises an anti-IL-6 antibody or an antigen-binding fragment thereof comprising: a heavy chain variable region (VH) comprising 3 complementarity determining regions: VH (CDR1), VH CDR2, and VH CDR3 having an amino acid sequence from the CDRs listed in SEQ ID NO: 256; and a light chain variable region (VL) comprising a VL CDR1, VL CDR2, and VL CDR3 having an amino acid sequence selected from the group of CDRs listed in SEQ ID NO: 91-93.

33. The formulation of claim 29, wherein the fusion protein comprises an anti-IL-6 antibody or an antigen-binding fragment thereof comprising: a CDRH1 that is a CDRH1 in SEQ ID NO: 172; a CDRH2 that is a CDRH2 in SEQ ID NO: 173; a CDRL1 that is a CDRL1 in SEQ ID NO: 199; a CDRL2 that is a CDRL2 in SEQ ID NO: 200; a CDRL3 that is a CDRL3 in SEQ ID NO: 201; at least one of the following mutations (EU numbering): L234A, L235A, and G237A, optionally wherein the anti-IL-6 antibody or an antigen-binding fragment thereof comprises at least one of the following mutations (EU numbering): Q347C or L443C.

34. The formulation of claim 29, wherein the VEGF trap is positioned either: a) at an N-terminal end of a heavy chain comprising IL-6 VH; or b) between a hinge region and after a CHI domain of a heavy chain comprising IL-6VH.

35. The formulation of claim 29, wherein the fusion protein comprises: a light chain comprising SEQ ID NO: 169, or a sequence at least 80% identical thereto; and a heavy chain comprising SEQ ID NO: 170 (with or without a C-terminal lysine), or a sequence at least 80% identical thereto.

36. The formulation of claim 29, wherein the buffer is selected from sodium acetate, histidine acetate, and histidine hydrochloride.

37. The formulation of claim 36, wherein the buffer is present at 10-50 mM.

38. The formulation of claim 29, comprising the surfactant.

39. The formulation of claim 38, wherein the surfactant is selected from polysorbate 20, polysorbate 80, and poloxamer 188.

40. The formulation of claim 38, wherein the surfactant is present at 0.01-0.1% (w / w).

41. The formulation of claim 29, wherein the pH of the formulation is about 5.7.

42. The formulation of claim 29, wherein the pH of the formulation is about 6.3.

43. The formulation of claim 29, comprising a carbohydrate.

44. The formulation of claim 43, wherein the carbohydrate is sucrose or trehalose.

45. The formulation of claim 44, wherein the carbohydrate is present at a concentration of up to 20% (w / v).

46. The formulation of claim 29, wherein the fusion protein is present at a concentration in a range of 20-200 mg / mL.

47. A pharmaceutical formulation comprising: a fusion protein comprising: an CDRH3 having at least 80% identity with the amino acid sequence of SEQ ID NO: 174 (QAWGYYALDI); and a VEGF trap; sodium acetate or histidine acetate at 10-50 mM; polysorbate 20 at 0-0.1% (w / w); and sucrose or trehalose at 0-8% (w / v), wherein the pharmaceutical formulation has a pH in a range of 4.5-6.8, and the fusion protein is present at 20-200 mg / mL.

48. A pharmaceutical formulation comprising:a fusion protein comprising: a light chain comprising SEQ ID NO: 169, or a sequence at least 80% identical thereto; and a heavy chain comprising SEQ ID NO: 170 (with or without a C-terminal lysine), or a sequence at least 80% identical thereto; sodium acetate at about 15 mM; polysorbate 20 at about 0.025% (w / w); and sucrose about 5.2% (w / v), wherein the pharmaceutical formulation has a pH of about 5.7, and the fusion protein is present at about 100 mg / mL.

49. A pharmaceutical formulation comprising: a fusion protein comprising: a light chain comprising SEQ ID NO: 169, or a sequence at least 80% identical thereto; and a heavy chain comprising SEQ ID NO: 170 (with or without a C-terminal lysine), or a sequence at least 80% identical thereto; histidine acetate at about 15 mM; polysorbate 20 at about 0.025% (w / w); and sucrose about 5.2% (w / v), wherein the pharmaceutical formulation has a pH of about 6.3, and the fusion protein is present at about 100 mg / mL.

50. The formulation of any one of claims 29-49, wherein the fusion protein comprises a heavy chain comprising L443 (EU numbering) in an Fc domain.

51. The formulation of any one of claims 29-49, wherein the fusion protein comprises a heavy chain comprising a variant of SEQ ID NO: 170 (with or without the C-terminal lysine) having L449 as numbered according to SEQ ID NO: 170.

52. The formulation of any one of claims 29-51, wherein the formulation is storage stable for at least 4 weeks at about 37 °C.

53. The formulation of any one of claims 29-52, wherein the formulation is storage stable for about 2 months at about 37 °C.

54. The formulation of any one of claims 29-53, that is a formulation for intravitreal injection.