Compositions of conjugated and unconjugated proteins

Abstract

Provided herein are pharmaceutical compositions and formulations of a conjugate comprising a first protein, such as an anti-VEGF antibody, conjugated to a phosphorylcholine-containing polymer, and a second protein that is not conjugated to a phosphorylcholine-containing polymer. The second protein can be the same as the first protein, or can be different. In some embodiments, the pH of the formulation is different from the isoelectric point (pI) of the second protein so as to prevent or reduce turbidity of the formulation. Also provided are methods of preparing the formulations, methods of use thereof, and kits with pre-filled syringes containing the formulations.

Description

KDIAK.210WO PATENT APPLICATION COMPOSITIONS OF CONJUGATED AND UNCONJUGATED PROTEINS REFERENCE TO RELATED APPLICATIONS

[0001] The present application is claims priority to U.S. Provisional Application No. 63 / 506781, filed June 7, 2023. The content of each of the aforementioned related application(s) is incorporated herein by reference in its entirety. REFRENCE TO 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 KDIAK210WO_SEQLIST.xml, created May 29, 2024, which is 196,681 bytes in size. The information in the electronic format of the Sequence Listing is incorporated herein by reference in its entirety. FIELD

[0003] The present invention relates to a composition comprising a mixture of unconjugated and conjugated proteins (e.g., antibodies and conjugates thereof) and methods of using and manufacturing said composition. BACKGROUND

[0004] Diabetic retinopathy is a leading cause of blindness in people between the ages of about 20 to 64 years of age. Engelgau M, Geiss L, Saaddine J, Boyle J, et al. 2004. The Evolving Diabetes Burden in the United States. Ann of Int Med. 140 (11): 945-951. In the United States, diabetic retinopathy accounts for some 12% of new cases of blindness. Typically, in cases of diabetic retinopathy, retinal blood vessels will swell and leak fluid into the rear of the eye. Hyperglycemia induces intramural and thickening of the basement membrane, resulting in leaky or permeable blood vessels.

[0005] In diabetic retinopathy, changes in blood glucose level cause changes to retinal blood vessels. All people with diabetes mellitus are at risk. The longer a person has diabetes, the higher their risk of developing some ocular problem. Between 40 to 45 percent of Americans diagnosed with diabetes have some stage of diabetic retinopathy. Causes and Risk Factors. Diabetic Retinopathy. United States National Library of Medicine.15 September

[0006] Diabetic retinopathy is first exhibited in the development of microaneurysms in the retina. Microaneurysms occur when there is a swelling of capillaries (very small blood vessels) that feed the retina. The presence of relatively small numbers of microaneurysms will not usually cause problems with vision. However, if the retinopathy develops to later stages, there are significant chances of vision loss. Such early-stage retinopathy are referred to as background diabetic retinopathy or non-proliferative diabetic retinopathy (NPDR). While NPDR patients are generally asymptomatic, early detection of retinopathy is crucial because if the disease proceeds to later stages, significant vision loss is very likely.

[0007] In the next stage of diabetic retinopathy, neovascularization occurs in the back of the eye (proliferative diabetic retinopathy). The neovasculature is leaky and the vessels can burst, followed by bleeding and resulting in blurred or obscured vision. Due to lack of oxygen in the eye, still further neovascularization occurs. Blood vessels grow along the retina and in the vitreous humor. As these vessels burst, there is further bleeding and the retina can be badly damaged or destroyed. The accumulation of fluid in the macula due to leaking blood vessels is called diabetic macular edema. Many patients with diabetic retinopathy will develop diabetic macular edema.

[0008] There are generally three treatment pathways for patients with diabetic retinopathy: laser surgery, injection of corticosteroids and injection of biologics (e.g. AVASTIN®(bevacizumab), LUCENTIS®(ranibizumab), Eylea®(aflibercept), Beovu® (brocizumab), and Vabysmo™ (faricimab)). While laser surgery is generally effective in treating diabetic retinopathy, retinal damage induced by the laser is a frequent side effect. Steroid preparations such as triamcinolone acetonide have been administered via intravitreal injection to treat diabetic retinopathy. However, to treat diabetic retinopathy, the steroid solutions must be frequently administered. Moreover, intravitreal treatment with steroids has been associated with cataracts, steroid-induced glaucoma and endophthalmitis.

[0009] Another way to treat diabetic retinopathy is the intravitreal injection of anti- VEGF agents. In this regard, anti-VEGF therapies such as LUCENTIS®(ranibizumab) and EYLEA®(aflibercept) have been approved for treatment of diabetic retinopathy in patients with diabetic macular edema. VEGF-directed therapies are effective not just for diabeticretinopathy, but also for retinal vascular diseases such as Age-Related Macular Degeneration (AMD), neovascular (wet) AMD and Retinal Vein Occlusion (RVO).

[0010] In order to treat diseases such as retinal diseases, it can be useful to use biologics such as antibodies or antibody fragments. These proteins should be formulated in clear solutions. Such formulations can have a defined buffer system and some excipients added for further enhancement of protein stability. Proteins can be conjugated to other moieties to bring an enhanced property or set of properties to the protein. For example, the conjugation of the protein to a potent toxin to create an antibody drug conjugate that targets the toxin to particular receptor-containing cell type for enhanced potency or enhanced safety. The present disclosure is not limited to diabetic retinopathy, and can be applied to various indications as will be appreciated by those in the art. SUMMARY

[0011] Provided herein is a formulation comprising: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second molar amount of a second protein that is not conjugated to a phosphorylcholine- containing polymer; and a pharmaceutically acceptable carrier, wherein the formulation comprises the second protein at about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the formulation has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein. Also provided is a therapeutically acceptable composition comprising: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second molar amount of a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a therapeutically acceptable carrier, wherein the composition comprises the second protein at about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the composition has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein. Provided herein is a therapeutically acceptable composition comprising: a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein thepercent composition of the second protein relative to the total protein mass weight concentration of the first protein and the second protein in the composition is about 1% or more, wherein the composition has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein. Also provided is a formulation comprising: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine- containing polymer; a second molar amount of a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the formulation comprises the second protein at about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the formulation has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein, wherein the formulation has a reduced viscosity and / or an enhanced injectability compared to a reference formulation comprising the conjugate at the total molar amount.

[0012] Provided herein is a therapeutically acceptable composition comprising: a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the percent composition of the second protein is about 1% or more, with the remainder comprising the first protein, wherein the composition has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein, wherein the composition has a reduced viscosity and / or an enhanced injectability compared to a reference composition comprising the conjugate, wherein the first protein of the conjugate is present in the reference composition at the total mass weight concentration of the first and second proteins in the composition. Also provided is a low-viscosity formulation of a protein conjugate, comprising: a first molar amount of a conjugate comprising a protein conjugated to a phosphorylcholine-containing polymer; a second molar amount of the protein that is not conjugated to the phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the formulation has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the protein, wherein the formulation has reduced viscosity and / or an enhanced injectability compared to a reference formulation comprising the conjugate at a total molar amount that is the sum of the first and second molar amounts. Provided herein is a low-viscosity therapeutically acceptable composition of a protein conjugate, comprising afirst molar amount of a conjugate comprising a protein conjugated to a phosphorylcholine- containing polymer; a second molar amount of the protein that is not conjugated to the phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the composition has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the protein, wherein the composition has reduced viscosity and / or an enhanced injectability compared to a reference composition comprising the conjugate at a total molar amount that is the sum of the first and second molar amounts.

[0013] Also provided is a formulation comprising: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second molar amount of a second protein that is not conjugated to a phosphorylcholine- containing polymer; and a pharmaceutically acceptable carrier, wherein the second protein is present in the formulation at about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the formulation has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein, wherein the formulation has a reduced turbidity compared to a reference formulation comprising the first molar amount (e.g., the same first molar amount) of the conjugate and the second molar amount (e.g., the same second molar amount) of the second protein at a pH within 0.5 pH units of the pI of the second protein. In some embodiments, the formulation comprises: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second molar amount of a second protein that is not conjugated to a phosphorylcholine- containing polymer; and a pharmaceutically acceptable carrier, wherein the second protein is present in the formulation at about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the formulation has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein, wherein the formulation has a reduced turbidity compared to a reference formulation comprising the same first molar amount of the conjugate and the same second molar amount of the second protein at a pH about the same as (e.g., within 0.05, 0.1, 0.15, 0.2, 0.2, 0.3, 0.4, or 0.5 pH units of) the pI of the second protein. Provided herein is a therapeutically acceptable composition comprising: a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; asecond protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the percent composition of the second protein is about 1% or more, with the remainder comprising the first protein, wherein the composition has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein, wherein the composition has a reduced turbidity compared to a reference composition comprising the second protein at the percent composition (e.g., the same percent composition), with the remainder comprising the first protein, at a pH within 0.5 pH units of the pI of the second protein. In some embodiments, the therapeutically acceptable composition comprises: a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the percent composition of the second protein is about 1% or more, with the remainder comprising the first protein, wherein the composition has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein, wherein the composition has a reduced turbidity compared to a reference composition comprising the second protein at the same percent composition, with the remainder comprising the first protein, at a pH about the same as (e.g., within 0.05, 0.1, 0.15, 0.2, 0.3, 0.4, or 0.5 pH units of) the pI of the second protein. Provided herein is a pharmaceutical formulation comprising: a first molar amount of a conjugate comprising a protein conjugated to a phosphorylcholine-containing polymer; a second molar amount the protein that is not conjugated to the phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the formulation comprises the protein that is not conjugated to the phosphorylcholine-containing polymer at about 1% or more of a total molar amount of the conjugate and unconjugated proteins, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the formulation has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the protein, wherein the formulation is substantially free of turbidity. Also provided is a formulation comprising: a phosphorylcholine-containing polymer present in the formulation at 100 mg / mL or higher; and a protein that is not conjugated to a phosphorylcholine-containing polymer, wherein the protein is present in the formulation at a second molar amount, wherein the protein is present in the formulation at about 1% or more of a total molar amount of the polymer and the protein, wherein the total molar amount comprises a sum of the first molar amount and the secondmolar amount, wherein the formulation has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the protein.

[0014] Provided herein is a formulation comprising: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second molar amount of a second protein that is not conjugated to a phosphorylcholine- containing polymer; and a pharmaceutically acceptable carrier, wherein the difference between the pI of the second protein and the pH of the formulation in the acidic or basic direction is selected to be greater than the minimum difference in the corresponding acidic or basic direction between the pI of the second protein and the pH for a reference formulation comprising: a third molar amount of the conjugate comprising the first protein conjugated to the phosphorylcholine-containing polymer; a fourth molar amount of the second protein that is not conjugated to the phosphorylcholine-containing polymer; and the pharmaceutically acceptable carrier, wherein a first total molar amount comprising a sum of the first molar amount and the second molar amount, and a second total molar amount comprising a sum of the third molar amount and the fourth molar amount are substantially the same, wherein the second molar amount is greater than the fourth molar amount, wherein the reference formulation is substantially free of turbidity. Also provided is a therapeutically acceptable composition comprising: a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the difference between the pI of the second protein and the pH of the formulation is selected to be greater than the minimum difference between the pI of the second protein and the pH for a reference formulation comprising: the conjugate comprising the first protein conjugated to the phosphorylcholine-containing polymer; the second protein that is not conjugated to the phosphorylcholine-containing polymer; and the pharmaceutically acceptable carrier, wherein the percent composition of the second protein in the composition is higher than the percent composition of the second protein in the reference composition, wherein the reference composition is substantially free of turbidity.

[0015] Also provided is a formulation comprising: a first molar amount of a conjugate comprising a first protein conjugated to a polymer; and a second molar amount of a second protein that is not conjugated to a polymer, wherein the formulation comprises thesecond protein at about 1% or more of a total molar amount of the first protein and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount. Provided herein is a therapeutically acceptable composition comprising: a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the percent composition of the second protein is about 1% or more, with the remainder comprising the first protein. Also provided is a therapeutically acceptable composition comprising: a first molar amount of a conjugate comprising a first protein conjugated to a polymer; and a second molar amount of a second protein that is not conjugated to a polymer, wherein the composition comprises the second protein at about 1% or more of a total molar amount of the first protein and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount. Further provided herein is a formulation comprising: a first molar amount of a first protein that is conjugated to a polymer; and a second molar amount of a second protein that is not conjugated to a polymer, the further improvement comprising: the formulation comprises the second protein at about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount. Provided herein is a therapeutically acceptable composition comprising: a first protein that is conjugated to a polymer; and a second protein that is not conjugated to a polymer, the further improvement comprising: the percent composition of the second protein is about 1% or more, with the remainder comprising the first protein.

[0016] Also provided is a formulation comprising: a conjugate comprising a first protein conjugated to a polymer; and a second protein that is not conjugated to a polymer, wherein a first molar amount of the conjugate and a second molar amount of the second protein has been combined in the formulation such that the second molar amount is about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount. Also provided is a therapeutically acceptable composition comprising: a conjugate comprising a first protein conjugated to a polymer; and a second protein that is not conjugated to a polymer, wherein a first molar amount of the conjugate and a second molar amount of the second protein has been combined in the composition such that the second molar amount is about 1% or more of a totalmolar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount. Further provided is a therapeutically acceptable composition comprising: a conjugate comprising a first protein conjugated to a polymer; and a second protein that is not conjugated to a polymer, wherein the second protein at a percent composition relative to the total protein mass weight concentration of the first protein and the second protein in the composition of about 1% or more (e.g., about 5-90%, 15-25%, 25-35%, etc.) has been combined with the conjugate, wherein the remainder of the total protein mass weight concentration comprises the first protein.

[0017] Provided herein is a formulation comprising: a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer, wherein the polymer has 9 arms and a molecular weight of between 600,000 and 1,000,000 Da, wherein the polymer is present in the formulation at about 100 mg / mL or more; and a second protein that is not conjugated to a polymer, wherein the second protein is present in the formulation at 5-15 mg / mL. Also provided is an intraocular therapeutic composition comprising an anti-VEGF-A antibody at about 50 mg / mL of protein, the anti-VEGF-A antibody comprising: a heavy chain comprising a complementarity determining region 1 (CDRH1): GYDFTHYGMN (SEQ ID NO: 9), CDRH2: WINTYTGEPTYAADFKR (SEQ ID NO: 10), and CDRH3: YPYYYGTSHWYFDV (SEQ ID NO: 11); and a light chain comprising CDRL1: SASQDISNYLN (SEQ ID NO: 12), CDRL2: FTSSLHS (SEQ ID NO: 13), and CDRL3: QQYSTVPWT (SEQ ID NO: 14), wherein the anti-VEGF-A antibody is present in the composition as either an antibody conjugate or unconjugated antibody, wherein the unconjugated antibody is present in the formulation at between about 10% to about 30% of a total molar amount of the antibody conjugate and the unconjugated antibody, wherein the total molar amount is the sum of the molar amount of the antibody conjugate and the molar amount of the unconjugated antibody, wherein the antibody conjugate comprises the anti-VEGF-A antibody conjugated to a phosphorylcholine-containing polymer at a non-native cysteine outside a variable region of the antibody, wherein the phosphorylcholine-containing polymer is present in the composition at about 100 mg / mL or more, wherein the phosphorylcholine- containing polymer has 9 arms and a molecular weight of between 600,000 and 1,000,000 Da, wherein the pH of the composition is about 5.5 or lower.

[0018] Also provided is an intraocular therapeutic composition comprising an anti- VEGF-A antibody at about 50 mg / mL of protein, the anti-VEGF-A antibody comprising: a heavy chain comprising an amino acid sequence of SEQ ID NO: 1 (with or without the C- terminal lysine); and a light chain comprising an amino acid sequence of SEQ ID NO: 2, wherein the anti-VEGF-A antibody is present in the composition as either an antibody conjugate or unconjugated antibody, wherein the unconjugated antibody is present in the formulation at between about 10% to about 30% of a total molar amount of the antibody conjugate and the unconjugated antibody, wherein the total molar amount is the sum of the molar amount of the antibody conjugate and the molar amount of the unconjugated antibody, wherein the antibody conjugate comprises the following structure:wherein: each heavy chain of the conjugate is denoted by the letter H, and each light chain of the conjugate is denoted by the letter L; the polymer is bonded to the heavy chain of theconjugate through the sulfhydryl of C443 (EU numbering), which bond is depicted on one ofthe 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, isopropyl, b) –H, c) any halogen, including –Br, –Cl, or –I, d) –SCN, or e) –NCS; and n1, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of n1, n2, n3, n4, n5, n6, n7, n8 and n9 is 2500 plus or minus 15%, wherein the phosphorylcholine-containing polymer is present in the composition at about 100 mg / mL or more, wherein the pH of the composition is about 5.5 or lower.

[0019] Also provided is an intraocular therapeutic composition comprising a fusion construct at about 53 mg / mL of protein, the fusion construct comprising a VEGF trap fused to an anti-IL-6 antibody, wherein the fusion construct comprises: a heavy chain comprising an amino acid sequence of SEQ ID NO:105 (with or without the C-terminal lysine); and a light chain comprising an amino acid sequence of SEQ ID NO:106, wherein the fusion construct is present in the composition as either a conjugate or an unconjugated fusion construct, wherein the unconjugated fusion construct is present in the formulation at between about 20% to about 40% of a total molar amount of the conjugate and the unconjugated fusion construct, wherein the total molar amount is the sum of the molar amount of the conjugate and the molar amount of the unconjugated fusion construct, wherein the conjugate comprises the fusion construct conjugated to a phosphorylcholine-containing polymer, wherein the phosphorylcholine- containing polymer is present in the composition at about 100 mg / mL or more, wherein the pH of the composition is about 5 or lower.

[0020] Provided herein is an intraocular therapeutic composition comprising a fusion construct at about 50 mg / mL of protein, the fusion construct comprising a VEGF trap fused to an anti-IL-6 antibody, wherein the fusion construct comprises: a heavy chain comprising a complementarity determining region 1 (CDRH1): PFAMH (SEQ ID NO: 134), CDRH2: KISPGGSWTYYSDTVTD (SEQ ID NO: 135), and CDRH3: QAWGYYALDI (SEQ ID NO: 136); and a light chain comprising CDRL1: SASISVSYLY (SEQ ID NO: 137), CDRL2: DDSSLAS (SEQ ID NO: 138), and CDRL3: QQWSGYPYT (SEQ ID NO: 139), wherein the fusion construct is present in the composition as either a conjugate or an unconjugated fusionconstruct, wherein the unconjugated fusion construct is present in the formulation at between about 20% to about 40% of a total molar amount of the conjugate and the unconjugated fusion construct, wherein the total molar amount is the sum of the molar amount of the conjugate and the molar amount of the unconjugated fusion construct, wherein the conjugate comprises the following structure:wherein: each heavy chain of the conjugate is denoted by the letter H, and each light chain of the conjugate is denoted by the letter L; the polymer is bonded to the heavy chain of the conjugate through the sulfhydryl of C443 (EU numbering), which bond is depicted on one ofthe 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,isopropyl, b) –H, c) any halogen, including –Br, –Cl, or –I, d) –SCN, or e) –NCS; and n1, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of n1, n2, n3, n4, n5, n6, n7, n8 and n9 is 2500 plus or minus 15%, wherein the phosphorylcholine-containing polymer is present in the composition at about 100 mg / mL or more, wherein the pH of the composition is about 5 or lower. In some embodiments, the sum of n1, n2, n3, n4, n5, n6, n7, n8 and n9 is about 1500 to about 3500 plus or minus about 10% to about 20%. Also provided is an intraocular therapeutic composition comprising a fusion construct at about 50 mg / mL of protein, the fusion construct comprising a VEGF trap fused to an anti-IL-6 antibody, wherein the fusion construct comprises: a heavy chain comprising a complementarity determining region 1 (CDRH1): PFAMH (SEQ ID NO: 134), CDRH2: KISPGGSWTYYSDTVTD (SEQ ID NO: 135), and CDRH3: QAWGYYALDI (SEQ ID NO: 136); and a light chain comprising CDRL1: SASISVSYLY (SEQ ID NO: 137), CDRL2: DDSSLAS (SEQ ID NO: 138), and CDRL3: QQWSGYPYT (SEQ ID NO: 139), wherein the fusion construct is present in the composition as either a conjugate or an unconjugated fusion construct, wherein the unconjugated fusion construct is present in the formulation at between about 20% to about 40% of a total molar amount of the conjugate and the unconjugated fusion construct, wherein the total molar amount is the sum of the molar amount of the conjugate and the molar amount of the unconjugated fusion construct, wherein the conjugate comprises the following structure:O ) wherein: each heavy chain of the conjugate is denoted by the letter H, and each light chain of the conjugate is denoted by the letter L; the polymer is bonded to the heavy chain of the conjugate through the sulfhydryl of C443 (EU numbering), which bond is depicted on one ofthe 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, isopropyl, b) –H, c) any halogen, including –Br, –Cl, or –I, d) –SCN, or e) –NCS; and n1, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of n1, n2, n3, n4, n5, n6, n7, n8 and n9 is 2500 plus or minus 15%, wherein the phosphorylcholine-containing polymer is present in the composition at about 100 mg / mL or more, wherein the pH of the composition is about 5 or lower.

[0021] Provided herein is a method of preparing a formulation, comprising combining in a formulation: a first molar amount of a conjugate comprising a first proteinconjugated to a phosphorylcholine-containing polymer; and a second molar amount of a second protein that is not conjugated to a phosphorylcholine-containing polymer, wherein the formulation comprises the second protein at about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the formulation has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein. Also provided is a method of preparing a therapeutically acceptable composition, comprising combining in a therapeutically acceptable composition: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; and a second molar amount of a second protein that is not conjugated to a phosphorylcholine-containing polymer, wherein the composition comprises the second protein at about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the composition has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein. Further provided herein is a method of preparing a therapeutically acceptable composition, comprising combining in a therapeutically acceptable composition: a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; and a second protein that is not conjugated to a phosphorylcholine-containing polymer, wherein the percent composition of the second protein is about 1% or more, with the remainder comprising the first protein, wherein the composition has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein. Also provided is a method of preparing a formulation, comprising adjusting the pH of a formulation to be about 0.5 pH units away or more from the isoelectric point (pI) of an unconjugated protein comprised in the formulation, wherein the formulation comprises: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; and a second molar amount of the unconjugated protein, wherein the unconjugated protein is not conjugated to a phosphorylcholine-containing polymer, wherein the formulation comprises the unconjugated protein at about 1% or more of a total molar amount of the conjugate and the unconjugated protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount.

[0022] Provided herein is a method of preparing a therapeutically acceptable composition, comprising adjusting the pH of a composition to be about 0.5 pH units away or more from the isoelectric point (pI) of an unconjugated protein comprised in the composition, wherein the composition comprises: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; and a second molar amount of the unconjugated protein, wherein the unconjugated protein is not conjugated to a phosphorylcholine-containing polymer, wherein the composition comprises the unconjugated protein at about 0.1% or more of a total molar amount of the conjugate and the unconjugated protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount. Also provided is a method of preparing a therapeutically acceptable composition, comprising adjusting the pH of a therapeutically acceptable composition to be about 0.5 pH units away or more from the isoelectric point (pI) of an unconjugated protein comprised in the composition, wherein the composition comprises: a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; and a second protein that is not conjugated to a phosphorylcholine-containing polymer, wherein the percent composition of the second protein is about 1% or more, with the remainder comprising the first protein.

[0023] Provided herein is a method of preparing a low-viscosity formulation of a protein conjugated to a phosphorylcholine-containing polymer, comprising combining in a formulation: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; and a second molar amount of the protein that is not conjugated to a phosphorylcholine-containing polymer, wherein the formulation comprises the protein that is not conjugated to the phosphorylcholine-containing polymer at about 1% or more of a total molar amount of the conjugate unconjugated proteins, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the formulation has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein, wherein the formulation has reduced viscosity and / or an enhanced injectability compared to a reference formulation comprising the conjugate at the total molar amount. Also provided is a method of preparing a low-viscosity therapeutically acceptable composition of a protein conjugated to a phosphorylcholine-containing polymer, comprising combining in a therapeutically acceptable composition: a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; and a second protein that is notconjugated to a phosphorylcholine-containing polymer, wherein the percent composition of the second protein is about 1% or more, with the remainder comprising the first protein, wherein the composition has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein, wherein the composition has reduced viscosity and / or an enhanced injectability compared to a reference composition comprising the conjugate, wherein the first protein of the conjugate is present in the reference composition at the total mass weight concentration of the first and second proteins in the composition.

[0024] Also provided is a method of preparing a formulation, comprising combining in a formulation: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; and a second molar amount of a second protein that is not conjugated to a phosphorylcholine-containing polymer, wherein the formulation comprises the second protein at about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount. Provided herein is a method of preparing a therapeutically acceptable composition, comprising combining in a therapeutically acceptable composition: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; and a second molar amount of a second protein that is not conjugated to a phosphorylcholine-containing polymer, wherein the composition comprises the second protein at about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount. Further provided herein is a method of preparing a therapeutically acceptable composition, comprising combining in a therapeutically acceptable composition: a conjugate comprising a first protein conjugated to a phosphorylcholine- containing polymer; and a second protein that is not conjugated to a phosphorylcholine- containing polymer, wherein the percent composition of the second protein is about 1% or more, with the remainder comprising the first protein.

[0025] Provided herein is a formulation or composition made by any one of the methods described herein.

[0026] Also provided is a method of treating a subject, comprising: intraocularly administering a therapeutically effective amount of any one of the formulation or composition described herein to a subject in need thereof. Also provided is a method of treating a subject,comprising: intraocularly administering a therapeutically effective amount of a low-viscosity formulation to a subject in need thereof, wherein the formulation comprises: a first concentration of a conjugate comprising a first anti-VEGF antibody conjugated to a phosphorylcholine-containing polymer; a second concentration of an anti-VEGF agent that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the formulation has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the anti-VEGF agent.

[0027] Provided herein is a kit comprising: a pre-filled syringe comprising a low- viscosity formulation comprising: a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; and a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a syringe needle for injection of the low-viscosity formulation, wherein the gauge of the needle is 27 or higher.

[0028] Also provided 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. Provided herein is a 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.

[0029] Provided herein is a method of storing a protein, comprising maintaining a protein in a formulation for at least 2 months and up to 2 years, the formulation comprising: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine- containing polymer; a second molar amount of a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the formulation comprises the second protein at about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the formulation has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein, wherein the protein comprises an antibody or a fusion construct.BRIEF DESCRIPTION OF THE DRAWINGS

[0030] FIG. 1 shows Compound L.

[0031] FIG. 2 shows Compound K.

[0032] FIG. 3 shows the synthesis of OG1802 from R3707.

[0033] FIG. 4 shows OG1786.

[0034] FIG. 5 shows the synthesis of OG1546 from OG1550.

[0035] FIG. 6 shows the synthesis of OG1784 from OG1546 and OG1563.

[0036] FIG. 7 shows the synthesis of OG1405 from OG1784.

[0037] FIG. 8 shows the synthesis of OG 1785 from OG1405.

[0038] FIG. 9 shows the synthesis of OG1786 from OG1785.

[0039] FIG. 10 shows OG1802.

[0040] FIG. 11 shows Compound E.

[0041] FIG. 12 depicts some embodiments of anti-VEGF-A heavy chain with certain effector function mutations and L443C (EU numbering, which is position 449 in SEQ ID NO.1).

[0042] FIG. 13 depicts some embodiments of an anti-VEGF-A light chain (SEQ ID NO.2).

[0043] FIG.14 depicts some embodiments of a bevacizumab heavy chain (SEQ ID NO.3).

[0044] FIG. 15 depicts some embodiments of a bevacizumab light chain (SEQ ID NO.4).

[0045] FIG.16 depicts some embodiments of a ranibizumab heavy chain (SEQ ID NO.5).

[0046] FIG. 17 depicts some embodiments of a ranibizumab light chain (SEQ ID NO.6).

[0047] FIG. 18 depicts some embodiments of a method for preparing an antibody conjugate.

[0048] FIG. 19 depicts Ion Exchanger analysis (A280 absorbance) of reactions A through G.

[0049] FIG. 20 depicts the effect of various anti-VEGF molecules on binding of biotin-VEGF to plate bound VEGFR ECD-Fc protein, and their IC50 values.

[0050] FIG. 21 depicts the OG1950 binding affinity to VEGF measured by BIAcore single cycle kinetics.

[0051] FIG.22 depicts binding of the OG1950 to Fc gamma receptor I.

[0052] FIG. 23 depicts binding of the OG1950 to Fc gamma receptor IIIa.

[0053] FIG.24 depicts binding of QG 1950 to human complement protein C1q.

[0054] FIG.25 depicts the results of a proliferation assay (including IC50 values).

[0055] FIG.26 depicts the results of single cycle kinetics of VEGF binding to anti- VEGF agents.

[0056] FIG. 27 depicts some embodiments of nucleic acid sequences encoding heavy and light chain variable regions.

[0057] FIG.28 is a collection of images and a table depicting results of mixing the free antibody with either polymer or OG1953 composition.

[0058] FIG.29 is a collection of images showing visual appearance of 30 different formulation conditions in two design of experiment (DOE) screenings.

[0059] FIG. 30 is a graph depicting turbidity measurements of formulations antibody with OG1801 polymer (samples #1-#30) in the presence of different excipients.

[0060] FIG.31 is a plot produced using JMP SAS software depicting the turbidity changes over time for one embodiment of a DOE experiment with the OG1950 antibody mixed with OG1801 polymer in the presence of different excipients.

[0061] FIG. 32 is an image and a table showing the effect of Histidine and pH on turbidity of the formulation.

[0062] FIG. 33 is an image showing some embodiments of formulations at differing pHs and corresponding turbidity results.

[0063] FIG.34 is a graph showing some embodiments of formulations at differing pHs and corresponding turbidity results.

[0064] FIG.35A is a collection of images showing the appearances of four different preparations at 50mg / ml of conjugated antibody supplemented with unconjugated antibody at different pH setpoints. From left to right: A, preparation at pH 6.5 results in a cloudy mixture; B-D, preparations at 5.0 - 5.2 result in a clear solution irrespective of the formulation buffer and percent unconjugated antibody. The mixtures are first prepared by concentrating the purified conjugated antibody to approx. 6 mg / ml, adding a desired amount of unconjugatedantibody, buffer exchange the antibody solution into the formulation buffer followed by a concentration to 20 mg / ml. The protein solution is supplemented with polysorbate 20 and then concentrated to the final target combined concentration of 50 mg / ml.

[0065] FIG. 35B and 35C are a collection of chromatograms and tables showing cation exchanger chromatography (CEX) analysis and analytical size exclusion chromatography (SEC) analysis of several embodiments of formulations of OG1950 / OG1953.

[0066] FIG.36 is a table showing the turbidity result for an embodiment of mixing the OG2072 fusion protein with its conjugated form OG2074.

[0067] FIGS.37A and 37B are a collection of a table, images, and a graph showing the screening result of mixing various proteins other than OG1950 antibody with a fixed concentration of OG1801 polymer. This is a control in order to evaluate if the turbidity is a more general phenomenon related to the presence of polymer.

[0068] FIGs.38A and 38B are a collection of images and a plot showing the effect of pH adjustment on the turbidity of formulations. The result demonstrated that the turbidity is reversible or reproducible dependent on the pI of the protein.

[0069] FIG.39 is a table showing the sample setup for formulations with different percentages of free protein relative to conjugated protein.

[0070] FIG.40A is a collection of images showing the appearance of formulations having different percentages of free protein relative to conjugated protein.

[0071] FIG. 40B is a collection of traces showing analysis of different partitions during the tandem method analysis of a formulation.

[0072] FIG. 40C is a collection of plots showing analysis of different partitions during the tandem method analysis of a formulation.

[0073] FIG.40D is a table showing peak area comparison of CEX bound (free protein) to unbound fraction (conjugate).

[0074] FIG.40E is a collection of traces showing analysis of different partitions during the tandem method analysis of a formulation.

[0075] FIG. 40F is a collection of plots showing analysis of different partitions during the tandem method analysis of a formulation.

[0076] FIG. 40G is a table showing peak area comparison of CEX bound (free protein) to unbound fraction (conjugate).

[0077] FIG. 41 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.

[0078] FIGs. 42 and 43 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.

[0079] FIG.44 is a collection of a table and a graph showing protein concentration of formulations measured by SoloVPE OD280nm method.

[0080] FIG. 45 is a data table showing a summary of the results of the stability testing of formulations.

[0081] Figs. 46A 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. 46B provides a table and graphs showing size exclusion chromatography analysis of formulations for the aggregation and degradation level of the OG1953 conjugate.

[0082] Figs.47A shows a schematic diagram showing an overview of the Tandem HPLC method, which combines the CEX-HPLC in tandem with a SEC-HPLC column.

[0083] FIG. 47B is a collection of traces, a table, and graphs showing tandem method analysis of the OG1950 free protein and its aggregated forms (P1 and P2).

[0084] FIGs. 47C and 47D are a collection of traces and a table showing tandem method analysis of formulations.

[0085] Figs. 48A-48C are a collection of a table, image and graph showing the design matrix and visual result of the formulations.

[0086] Figs.49A and 49B are a collection of plots showing analysis of the turbidity results against the concentration of polymer, free protein and pH.

[0087] Figs. 49C and 49D are a collection of plots and schematic representations showing turbidity of formulations at various pH for the different levels of OG1801 polymer and free protein.

[0088] Fig. 49E is a plot showing turbidity measurements results against the concentration of polymer, free protein and pH. Figs.49F-49I are a collection of plots showingan overlay of the pH boundary curve plot with other OG1801 polymer with free protein only experimental data.

[0089] Figs.49J-49M are a collection of plots showing an overlay of the pH boundary curve plot with other OG1953 conjugate solution and free protein only experimental data. Fig. 49N is a plot showing analysis of turbidity measurements in OG1801 polymer solution with free protein against the concentration of polymer, free protein and pH.

[0090] Fig. 49O is a plot showing analysis of turbidity measurements in OG1953 conjugate solution with free protein against the concentration of polymer, free protein and pH.

[0091] Fig. 50A is a collection of graphs and schematic diagram showing formulation viscosity at 25ºC.

[0092] Figs.50B and 50C are a collection of graphs and a table showing impact of increasing free protein percent composition on viscosity of formulations at 25ºC.

[0093] Fig. 50D is a collection of plots and tables showing the viscosity of formulations with various concentrations of OG1801 polymer.

[0094] Fig. 51 is a collection of images showing air movement in syringes filled with different formulations.

[0095] Figs. 52A-52C are a collection of plots and tables showing potency comparison of various OG1953 formulations using ELISA or cell-based assay.

[0096] Figs. 53A-53D are a collection of graphs, charts, and tables showing improved vision in wet AMD patients administered with OG1953, aflibercept, or other anti- VEGF agents.

[0097] FIG. 54 is a collection of sequences showing some non-limiting embodiments of a light chain and heavy chain of a fusion construct.

[0098] FIG.55A and 55B are a collection of sequences showing some non-limiting embodiments of a heavy chain and light chain, respectively, of a fusion construct.

[0099] FIGs. 56A-56C are a collection of plots showing the level of impurities (e.g., aggregation and / or degradation level) in OG1953 conjugate formulations over time.

[0100] FIGs. 57A and 57B are a collection of plots and tables showing potency comparison of various OG1953 formulations using ELISA or cell-based assay.

[0101] FIGs.58A-58C are a collection of schematic diagrams showing components of non-limiting examples of formulations of the present disclosure.

[0102] FIGS. 59A-59D depict continuous 80min tandem method separation with PhotoDiol Array (PDA) detection set at 200-350nm. FIG. 59A depicts the 2D contour view of elution time versus wavelength; FIG.59B 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. 59C as a non-reducing gel and FIG.59D as a reducing gel.

[0103] FIGS.60A-60C depict KSI-301 stability data up to 9 months under different temperature conditions; -20 ± 5°C (FIG.60A), 5 ± 3°C (FIG.60B), and 25 ± 2°C / 60±5% RH (relative humidity) (FIG.60C).

[0104] FIG.61 depicts KSI-501DS Batches 1-3 Lot Release Data.

[0105] FIGS. 62 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.

[0106] FIG.63 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 KSI-501 mix formulation. DETAILED DESCRIPTION

[0107] Provided herein are formulations and compositions comprising a mixture of an unconjugated protein (e.g., an unconjugated anti-VEGF-A antibody) and conjugates thereof. In some embodiments, the conjugates can include a protein (which may or may not be the same as the unconjugated protein) conjugated to a phosphorylcholine-containing polymer. In some embodiments, the pH of the formulation can be different from the isoelectric point (pI) of the unconjugated protein in the formulation such that the formulation is not or is less turbid. In some embodiments, the formulation has reduced viscosity compared to a reference formulation of the conjugated protein (without the unconjugated protein). The reduced viscosity can improve injectability of the formulation (e.g. by a syringe) and / or handling of the formulation during manufacture. In some embodiments, the composition can be used for the treatment of certain conditions, such as eye disorders, including retinal vascular disorders. The formulations, compositions, and methods of the present disclosure can beprovided for treating any disease or disorder as described herein, and is not necessarily limited to diabetic retinopathy or diabetic macular edema.

[0108] Provided herein in some embodiments is a drug composition (a formulation) which is a mixture of a protein and a conjugate of that protein and which is a stable solution. Without being limited by theory, lowering the end concentration of biopolymer while keeping the amount of the antibody bioactive the same can result in (i) a decrease in solution viscosity which has many advantages for pharmaceutical manufacturing of drug substance and drug product, and (ii) better handling of the drug for dose preparation and handling in the physician office, and (iii) easier injectability into the patient for example in retina, where a needle is inserted into the vitreous of the eye and the syringe plunger is pushed with a maximum force of the hand, including shorter injection time, reduced force required to drive the plunger down to express out the drug, and a narrower needle such as 30G or 29G, a higher dose level with smaller volume. These properties (i), (ii), and (iii) in turn may lower the chances of unwanted side effects of intravitreal injection, such as but not limited to contamination of bacteria into the eye. In some cases, the side effect includes a cataract. Without being limited by theory, a mixture of unconjugated and conjugated protein can benefit from the direct activity of the protein and a modified activity of the protein as modified by conjugation to a polymer. For example, an immediacy of effect of the unconjugated protein at a defined ratio (for example, 20% of the administered antibody) as well as a modified durability of effect of the conjugated protein at a defined ratio (for example, the remaining 80% of the antibody is in a conjugated form) which modified effect can in combination provide a basal effect driven by the conjugated protein and a bolus effect driven by the unconjugated antibody. In some embodiments, the formulation is a clear solution containing the free (unconjugated) antibody, the antibody conjugated to the phosphorylcholine polymer, the buffer system and at a particular pH, where the formulation is stable and suitable for drug development, manufacturing and / or storage.

[0109] In some embodiments, an anti-VEGF antibody (OG1950) conjugated to a phosphorylcholine containing biopolymer (OG1802) which conjugate is called OG1953 can be formulated at 50 mg / mL by weight of antibody and formulated in sodium phosphate pH 6.5 that is clear and stable, and adding a desired amount of the free unconjugated anti-VEGF antibody (OG1950) to create a solution of 40 mg / mL of conjugate (OG1953) and 10 mg / mL of OG1950 without adjusting the formulation system as provided herein can result in turbidity.In some embodiments, formulations and methods provided herein can provide a clear solution of the unconjugated and conjugated antibody coformulation.

[0110] Provided herein in some embodiments is a clear and stable ‘formulation system’ of a protein with a phosphorylcholine-biopolymer-conjugated version of that protein, in which the protein can be soluble and clear and stable and avoids turbidity formation. Also provided, in some embodiments, is a method to reduce the turbidity.

[0111] In some embodiments, a solubility switch can be applied to OG1953 to create OG195380% + OG195020% at a pH 5.0, thereby preventing turbidity that forms at a pH of 7.5, which is at or close to the pI of the protein. In some embodiments, the solubility switch involves the application of pH to control this switch. In some embodiments, this solubility switch is applied to OG2074 to create OG207470% + OG207230% at pH 5.0, for ophthalmology (intravitreal) injection or for systemic diseases.

[0112] In some embodiments, in ophthalmology where dose volume is about 100 microliters, the concentration of the dose formulation, for OG1953 is for example 50 mg / mL (as measured by the antibody portion), and the bioconjugate has durability due to the conjugated biopolymer, but to improve manufacturability the same amount of protein bioactive (5.0 mg in 100 microliter dose, i.e.50 mg / mL in the formulation by weight of antibody) is kept while increasing the relative amount of unconjugated protein. For example, in some embodiments, OG1953 is 80% and OG1950 is 20%; or OG1953 is 75% and OG1950 is 25%; or OG1953 is 70% and OG1950 is 30%; or OG1953 is 65% and OG1950 is 35%; or OG1953 is 60% and OG1950 is 40%; or OG1953 is 50% and OG1950 is 50%. In some embodiments, the pH is at 5.0 (without the addition of histidine or sucrose or trehalose) by shifting the pH to 5.0 and adjusting the formulation buffering constituents to acetate from phosphate. In some embodiments, a high dose formulation with improved manufacturability is achieved because of decreased viscosity for drug substance and drug product (vials, prefilled syringes); improved usability and dose administration (because of decreased viscosity); improved clinical immediacy (so improved balance of clinical immediacy of the for example 20% mAb while retaining clinical durability of the for example 80% mAb-conjugate). In some embodiments, the composition can be tuned to achieve an optimal viscosity to enable large scale manufacturing (of drug substance, of drug product, of pre-filled syringes) and to enable safer dose handling and preparation (for example by physicians and patients) and to enable saferdose administration for example injectability requiring lower injection force by the doctor. In some embodiments, the injection force is less than 10 Newtons, or less than 5 Newtons, injection time is 10 seconds or less, or 5 seconds or less, and the injection needle has a bore size of between 30 gauge and 27 gauge, including 28 gauge or 29 gauge. In some embodiments, the composition can be tuned to achieve an optimal balance of durability of clinical effect (a basal activity) and immediacy of clinical effect (a bolus activity). In some embodiments, the therapeutics are formulated into a clear solution with long-term stability.

[0113] Further provided herein are methods for preparing conjugate compositions of antibodies (of any type of antibody and / or protein). In some embodiments, these methods allow for lower aggregate formation or higher efficiency of formation of the desired composition of antibody and antibody conjugate.

[0114] These and additional embodiments are provided below, following the definition section. Terms

[0115] All terms can have their customary and ordinary meaning to one of ordinary skill int the art, in view of the present disclosure. A “neovascular disorder” is a disorder or disease state characterized by altered, dysregulated or unregulated angiogenesis. Examples of neovascular disorders include neoplastic transformation (e.g., cancer) and ocular neovascular disorders including diabetic retinopathy and age-related macular degeneration.

[0116] An “ocular neovascular” disorder is a disorder characterized by altered, dysregulated or unregulated angiogenesis in the eye of a patient. Such disorders include optic disc neovascularization, iris neovascularization, retinal neovascularization, choroidal neovascularization, corneal neovascularization, vitreal neovascularization, glaucoma, pannus, pterygium, macular edema, diabetic retinopathy, diabetic macular edema, vascular retinopathy, retinal degeneration, uveitis, inflammatory diseases of the retina, and proliferative vitreoretinopathy.

[0117] The term “percent composition” refers to the percent amount (in mass or concentration units) of a component present in a composition. Percent composition is calculated by determining the amount of a component in mass units (e.g., μg) or in concentration units (e.g., mg / mL), dividing that amount by the total amount of all componentsin the composition in the corresponding unit, and multiplying by 100. For compositions and formulations of a conjugate and an unconjugated protein described herein, the amount of the unconjugated protein can be divided by the total amount of the protein component in the solution (excluding the contribution from the polymer component of the conjugate to the mass of the conjugate) to obtain a percent composition.

[0118] As used herein, “% total molar amount” denotes the proportion (in percent) of the amount (in moles or a molar concentration) of one component of a composition relative to the amount(s) (in moles or a molar concentration) of one or more other component of the composition, that together make up the whole (100%). It is understood that percent composition and % total molar amount can be converted between each other where the molecular weight of all of the relevant components is known.

[0119] The term antibody includes intact antibodies and binding fragments thereof. A binding fragment refers to a molecule other than an intact antibody that comprises a portion of an intact antibody that binds the antigen to which the intact antibody binds. Examples of binding fragments include Fv, Fab', Fab'-SH, F(ab')2; diabodies; linear antibodies; single-chain antibody molecules (e.g., scFv); and multispecific antibodies formed from antibody fragments. scFv antibodies are described in Houston JS. 1991. Methods in Enzymol. 203:46-96. In addition, antibody fragments comprise single chain polypeptides having the characteristics of a VH domain, namely being able to assemble together with a VL domain, or of a VL domain, namely being able to assemble together with a VH domain to a functional antigen binding site and thereby providing the antigen binding property of full-length antibodies.

[0120] Specific binding of an antibody to its target antigen(s) means an affinity of at least 106, 107, 108, 109, or 1010M-1. Specific binding is detectably higher in magnitude and distinguishable from non-specific binding occurring to at least one unrelated target. Specific binding can be the result of formation of bonds between particular functional groups or particular spatial fit (e.g., lock and key type) whereas nonspecific binding is usually the result of van der Waals forces. Specific binding does not however necessarily imply that an antibody or fusion protein binds one and only one target.

[0121] A basic antibody structural unit is a tetramer of subunits. Each tetramer includes two identical pairs of polypeptide chains, each pair having one "light" (about 25 kDa) and one "heavy" chain (about 50-70 kDa). The amino-terminal portion of each chain includesa variable region of about 100 to 110 or more amino acids primarily responsible for antigen recognition. This variable region is initially expressed linked to a cleavable signal peptide. The variable region without the signal peptide is sometimes referred to as a mature variable region. Thus, for example, a light chain mature variable region means a light chain variable region without the light chain signal peptide. However, reference to a variable region does not mean that a signal sequence is necessarily present; and in fact signal sequences are cleaved once the antibodies or fusion proteins have been expressed and secreted. A pair of heavy and light chain variable regions defines a binding region of an antibody. The carboxy-terminal portion of the light and heavy chains respectively defines light and heavy chain constant regions. The heavy chain constant region is primarily responsible for effector function. In IgG antibodies, the heavy chain constant region is divided into CH1, hinge, CH2, and CH3 regions. The CH1 region binds to the light chain constant region by disulfide and noncovalent bonding. The hinge region provides flexibility between the binding and effector regions of an antibody and also provides sites for intermolecular disulfide bonding between the two heavy chain constant regions in a tetramer subunit. The CH2 and CH3 regions are the primary site of effector functions and FcR binding.

[0122] Light chains are classified as either kappa or lambda. Heavy chains are classified as gamma, mu, alpha, delta, or epsilon, and define the antibody's isotype as IgG, IgM, IgA, IgD and IgE, respectively. Within light and heavy chains, the variable and constant regions are joined by a "J" segment of about 12 or more amino acids, with the heavy chain also including a "D" segment of about 10 or more amino acids. (See generally, Fundamental Immunology (Paul, W., ed., 2nd ed. Raven Press, N.Y., 1989), Ch. 7) (incorporated by reference in its entirety for all purposes).

[0123] The mature variable regions of each light / heavy chain pair form the antibody binding site. Thus, an intact antibody has two binding sites, i.e., is divalent. In natural antibodies, the binding sites are the same. However, bispecific antibodies can be made in which the two binding sites are different (see, e.g., Songsivilai S, Lachmann PC. 1990. Bispecific antibody: a tool for diagnosis and treatment of disease. Clin Exp Immunol.79:315- 321; Kostelny SA, Cole MS, Tso JY. 1992. Formation of bispecific antibody by the use of leucine zippers. J Immunol.148: 1547-1553). The variable regions all exhibit the same general structure of relatively conserved framework regions (FR) joined by three hypervariableregions, also called complementarity determining regions or CDRs. The CDRs from the two chains of each pair are aligned by the framework regions, enabling binding to a specific epitope. From N-terminal to C-terminal, both light and heavy chains comprise the domains FRl, CDRl, FR2, CDR2, FR3, CDR3 and FR4. For convenience, the variable heavy CDRs can be referred to as CDRH1, CDRH2 and CDRH3; the variable light chain CDRs can be referred to as CDRL1, CDRL2 and CDRL3. The assignment of amino acids to each domain is in accordance with the definitions of Kabat EA, et al. 1987 and 1991. Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, MD) or Chothia C, Lesk AM. 1987. Canonical Structures for the Hypervariable Regions of Immunoglobulins. J Mol Biol 196:901-917; Chothia C, et al. 1989. Conformations of Immunoglobulin Hypervariable Regions. Nature 342:877-883. Kabat also provides a widely used numbering convention (Kabat numbering) in which corresponding residues between different heavy chain variable regions or between different light chain variable regions are assigned the same number. Although Kabat numbering can be used for antibody constant regions, EU numbering is more commonly used, as is the case in this application. Although specific sequences are provided for exemplary antibodies disclosed herein, it will be appreciated that after expression of protein chains one to several amino acids at the amino or carboxy terminus of the light and / or heavy chain, particularly a heavy chain C-terminal lysine residue, may be missing or derivatized in a proportion or all of the molecules.

[0124] The term "epitope" refers to a site on an antigen to which an antibody or extracellular trap segment binds. An epitope on a protein can be formed from contiguous amino acids or noncontiguous amino acids juxtaposed by tertiary folding of one or more proteins. Epitopes formed from contiguous amino acids (also known as linear epitopes) are typically retained on exposure to denaturing solvents whereas epitopes formed by tertiary folding (also known as conformational epitopes) are typically lost on treatment with denaturing solvents. An epitope typically includes at least 3, and more usually, at least 5 or 8-10 amino acids in a unique spatial conformation. Methods of determining spatial conformation of epitopes include, for example, x-ray crystallography and 2-dimensional nuclear magnetic resonance. See, e.g., Epitope Mapping Protocols, in Methods in Molecular Biology, Vol. 66, Glenn E. Morris, Ed. (1996).

[0125] Antibodies that recognize the same or overlapping epitopes can be identified in a simple immunoassay showing the ability of one antibody to compete with the binding of another antibody to a target antigen. The epitope of an antibody can also be defined by X-ray crystallography of the antibody (or Fab fragment) bound to its antigen to identify contact residues.

[0126] Alternatively, two antibodies have the same epitope if all amino acid mutations in the antigen that reduce or eliminate binding of one antibody reduce or eliminate binding of the other. Two antibodies have overlapping epitopes if some amino acid mutations that reduce or eliminate binding of one antibody reduce or eliminate binding of the other.

[0127] Competition between antibodies is determined by an assay in which an antibody under test inhibits specific binding of a reference antibody to a common antigen (see, e.g., Junghans et al., Cancer Res. 50: 1495, 1990). A test antibody competes with a reference antibody if an excess of a test antibody (e.g., at least 2x, 5x, 10x, 20x or l00x) inhibits binding of the reference antibody by at least 50%. In some embodiments the test antibody inhibits binding of the reference antibody by 75%, 90%, or 99% as measured in a competitive binding assay. Antibodies identified by competition assay (competing antibodies) include antibodies binding to the same epitope as the reference antibody and antibodies binding to an adjacent epitope sufficiently proximal to the epitope bound by the reference antibody for steric hindrance to occur.

[0128] The term “conjugate” refers to a protein covalently linked to a polymer. In some embodiments, the protein is an antibody.

[0129] As used herein “unconjugated” and “free” with reference to a protein or antibody are used interchangeably to denote the protein or antibody that is not conjugated to a polymer (e.g., not conjugated to a phosphorylcholine-containing polymer).

[0130] The term “isotype” refers to a distinct class of antibody identifiable by the structure of its heavy chain, with each class differing in the (1) structure of the antibody’s hinge, (2) sequence (and thus domains), and (3) valency.

[0131] As used herein, “VEGF Trap” or similar term denotes the VEGF binding domains (e.g., 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 0.5. In some embodiments, theVEGF 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.

[0132] The term "patient" includes human and other mammalian subjects that receive either prophylactic or therapeutic treatment. In some embodiments, the patient is a human patient.

[0133] 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.

[0134] 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 bycomparing two optimally aligned sequences over a window of comparison, determining the number of positions at which the identical residues occurs 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.

[0135] Compositions or methods "comprising" one or more recited elements may include other elements not specifically recited. For example, a composition that comprises an antibody may contain the antibody alone or in combination with other ingredients, such as an antibody conjugate. Compositions can comprise a conjugated antibody and an unconjugated antibody.

[0136] 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.e., 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 Fcy receptors, particularly FcȖRI and FcȖRIII, 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.

[0137] The term opsonization also known as "antibody-dependent cellular phagocytosis", or ADCP, refers to the process by which antibody-coated cells are internalized, either in whole or in part, by phagocytic immune cells (e.g., macrophages, neutrophils and dendritic cells) that bind to an immunoglobulin Fc region.

[0138] The term "complement-dependent cytotoxicity" or CDC refers to a mechanism for inducing cell death in which an Fc effector domain(s) of a target-bound antibody activates a series of enzymatic reactions culminating in the formation of holes in the target cell membrane. Typically, antigen-antibody complexes such as those on antibody- coated target cells bind and activate complement component Clq which in turn activates the complement cascade leading to target cell death. Activation of complement may also result in deposition of complement components on the target cell surface that facilitate ADCC by binding complement receptors (e.g., CR3) on leukocytes.

[0139] A humanized antibody is a genetically engineered antibody in which the CDRs from a non-human "donor" antibody are grafted into human "acceptor" antibody sequences (see, e.g., Queen, US 5,530,101 and 5,585,089; Winter, US 5,225,539, Carter, US 6,407,213, Adair, US 5,859,205 6,881,557, 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 constant region respectively when at least 85, 90, 95 or 100% of corresponding residues defined by Kabat are identical.

[0140] Although humanized antibodies often incorporate all six CDRs (which can be 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., De Pascalis R, Iwahashi M, Tamura M, et al. 2002. Grafting “Abbreviated” Complementary-Determining Regions Containing Specificity-Determining Residues Essential for Ligand Contact to Engineer a Less Immunogenic Humanized Monoclonal Antibody. J Immunol. 169:3076-3084; Vajdos FF, Adams CW, Breece TN, Presta LG, de Vos AM, Sidhu, SS. 2002. Comprehensive functionalmaps of the antigen-binding site of an anti-ErbB2 antibody obtained with shotgun scanning mutagenesis. J Mol Biol. 320: 415–428; Iwahashi M, Milenic DE, Padlan EA, et al. 1999. CDR substitutions of a humanized monoclonal antibody (CC49): Contributions of individual CDRs to antigen binding and immunogenicity. Mol Immunol. 36:1079-1091; Tamura M, Milenic DE, Iwahashi M, et al. 2000. Structural correlates of an anticarcinoma antibody: Identification of specificity-determining regions (SDRs) and development of a minimally immunogenic antibody variant by retention of SDRs only. J Immunol.164:1432-1441).

[0141] 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.

[0142] 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 EA.1991. A possible procedure for reducing the immunogenicity of antibody variable domains while preserving their ligand- binding properties. Mol Immunol. 28:489-98) 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 Östberg L, Pursch E.1983. Human x (mouse x human) hybridomas stably producing human antibodies. Hybridoma 2:361-367; Östberg, 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.

[0143] “Polymer” refers to a series of monomer groups linked together. A polymer is composed of multiple units of a single monomer (a homopolymer) or different monomers (a heteropolymer). High MW polymers are prepared from monomers that include, but are not limited to, acrylates, methacrylates, acrylamides, methacrylamides, styrenes, vinylpyridine, vinylpyrrolidone and vinyl esters such as vinyl acetate. Additional monomers are useful in high MW polymers . When two different monomers are used, the two monomers are called “comonomers,” meaning that the different monomers are copolymerized to form a single polymer. In some embodiments, one monomer is a phosphorylcholine-containing monomer, and a second comonomer is a different comonomer with a different pendant group chemistry (for example a click chemistry to be a reactive group / recipient of a chemical reaction to be conjugated, for example, to a small molecule bioactive or to a chemical linker). The polymer can be linear or branched. When the polymer is branched, each polymer chain is referred to as a “polymer arm.” The end of the polymer arm linked to the initiator moiety is the proximal end, and the growing-chain end of the polymer arm is the distal end. On the growing chain- end of the polymer arm, the polymer arm end group can be the radical scavenger, or another group.

[0144] “Initiator” refers to a compound capable of initiating a polymerization using monomers or comonomers. The polymerization can be a conventional free radical polymerization or 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. When the initiator is suitable for ATRP, it contains a labile bond 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. In some embodiments , the initiator contains one of more 2-bromoisobutyrate groups as sites for polymerization via ATRP.

[0145] 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 hydrolyzable, 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).

[0146] 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 with a functional group on a different moiety to form a covalent linkage. Reactive groups generally include nucleophiles, electrophiles and photoactivatable groups.

[0147] “Phosphorylcholine,” also denoted as “PC,” refers to the following: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.

[0148] “Phosphorylcholine containing polymer” is a polymer that contains phosphorylcholine. “Zwitterion containing polymer” refers to a polymer that contains a zwitterion.

[0149] Poly(acryloyloxyethyl phosphorylcholine) containing polymer refers to a polymer containing 2-(acryloyloxy)ethyl-2-(trimethylammonium)ethyl phosphate (HEA-PC shown below in Example 6) as monomer.

[0150] Poly(methacryloyloxyethyl phosphorylcholine) containing polymer refers to a polymer containing 2-(methacryloyloxy)ethyl-2-(trimethylammonium)ethyl phosphate (HEMA-PC or MPC) as monomer (see below):.

[0151] As used herein, “MPC” and “HEMA-PC” are interchangeable.

[0152] “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 endgroup 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. In some embodiments, the molecular weight is measured by SEC-MALS (size exclusion chromatography – multi angle light scattering). In some embodiments, the multi-angle light scattering includes 18-angle MALS. In some embodiments, the multi-angle light scattering includes 3-angle and 18-angle MALS. In some embodiments, the polymeric reagents are typically polydisperse (i.e., number average molecular weight and weight average molecular weight of the polymers are not equal), and can possess low polydispersity values of, for example, less than about 1.5, as judged, for example, by the PDI value derived from the SEC-MALS measurement. In some embodiments, the polydispersities (PDI) are in the range of about 1.4 to about 1.2. In some embodiments the PDI is less than about 1.15, 1.10, 1.05, or 1.03.

[0153] 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.

[0154] “About” means variation one might see in measurements taken among different instruments, samples, and sample preparations.

[0155] “Protected,” “protected form,” “protecting group” and “protective group” refer to the presence of a group (i.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.

[0156] “Alkyl” refers to a straight or branched, saturated, aliphatic radical having the number of carbon atoms indicated. For example, C1-C6alkyl 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. The alkyl group is typically monovalent, but can be divalent, such as when the alkyl group links two moieties together.

[0157] 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 or up to and including 4 and (as unbranched) one or two carbon atoms.

[0158] “Alkylene” refers to an alkyl group, as defined above, linking at least two other groups, i.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 -(CH2)n, 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.

[0159] Substituents for the alkyl and heteroalkyl radicals (including those groups often referred to as alkylene, alkenyl, heteroalkylene, heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, and heterocycloalkenyl) can be a variety of groups selected from: -OR’, =O, =NR’, =N-OR’, -NR’R”, -SR’, -halogen, -SiR’R”R”’, -OC(O)R’, -C(O)R’, -CO2R’, -CONR’R”, - OC(O)NR’R”, -NR”C(O)R’, -NR’-C(O)NR”R”’, -NR”C(O)2R’, -NH-C(NH2)=NH, -NR’C( NH2)=NH, -NH-C(NH2)=NR’, -S(O)R’, -S(O)2R’, -S(O)2NR’R”, -CN and -NO2in a number ranging from zero to (2m’+1), where m’ is the total number of carbon atoms in such radical. R’, R” and R”’ each independently refer to hydrogen, unsubstituted (C1-C8)alkyl and heteroalkyl, unsubstituted aryl, aryl substituted with 1-3 halogens, unsubstituted alkyl, alkoxy or thioalkoxy groups, or aryl-(C1-C4)alkyl 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 1-pyrrolidinyl and 4-morpholinyl. The term “alkyl” includes groups such as haloalkyl (e.g., -CF3and -CH2CF3) and acyl (e.g., -C(O)CH3, -C(O)CF3, -C(O)CH2OCH3, and the like). In some embodiments, the substituted alkyl and heteroalkyl groups have from 1 to 4 substituents. In some embodiments, the substituted alkyl and heteroalkyl groups have 1, 2 or 3 substituents. Exceptions are those perhalo alkyl groups (e.g., pentafluoroethyl and the like) .

[0160] Substituents for the alkyl and heteroalkyl radicals (including those groups often referred to as alkylene, alkenyl, heteroalkylene, heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, and heterocycloalkenyl) can be one or more of a variety of groups selected from, but not limited to: -OR’, =O, =NR’, =N-OR’, -NR’R”, -SR’, -halogen, -SiR’R”R”’, -OC(O)R’, -C(O)R’, -CO2R’, -CONR’R”, -O C(O)NR’R”, -NR”C(O)R’, -NR’-C(O)NR”R”’, -NR”C(O)2R’, -NR-C(NR’R”R’”)=NR””, -N R-C(NR’R”)=NR’”, -S(O)R’, -S(O)2R’, -S(O)2NR’R”, -NRSO2R’, -CN and –NO2in a number ranging from zero to (2m’+1), where m’ is the total number of carbon atoms in such radical. R’, R”, R”’ and R”” each independently refer 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 a compound includes more than one R group, for example, each of the R groups is independently selected as are each R’, R”, R’” and R”” groups when more than one of these groups is present.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. From the above discussion of substituents, one of skill in the art will understand that the term “alkyl” is meant to include groups including carbon atoms bound to groups other than hydrogen groups, such as haloalkyl (e.g., -CF3and –CH2CF3) and acyl (e.g., -C(O)CH3, -C(O)CF3, -C(O)CH2OCH3, and the like).

[0161] “Alkoxy” refers to an alkyl group having an oxygen atom that either connects the alkoxy group to the point of attachment or is linked to two carbons of the alkoxy group. 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 within. For example, the alkoxy groups can be substituted with halogens to form a “halo-alkoxy” group.

[0162] “Carboxyalkyl” means an alkyl group (as defined herein) substituted with a carboxy group. The term “carboxycycloalkyl” means a 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.

[0163] “Haloalkyl” refers to alkyl as defined above where some or all of the hydrogen atoms are substituted with halogen atoms. Halogen (halo) 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.

[0164] “Fluoro-substituted alkyl” refers to an alkyl group where one, some, or all hydrogen atoms have been replaced by fluorine.

[0165] “Cytokine” is a member of a group of protein signaling molecules that may participate in cell-cell communication in immune and inflammatory responses. Cytokines are typically small, water-soluble glycoproteins that have a mass of about 8-35 kDa.

[0166] “Cycloalkyl” refers to a cyclic hydrocarbon group that contains from about 3 to 12, from 3 to 10, or from 3 to 7 endocyclic carbon atoms. Cycloalkyl groups include fused, bridged and spiro ring structures.

[0167] “Endocyclic” refers to an atom or group of atoms which comprise part of a cyclic ring structure.

[0168] “Exocyclic” refers to an atom or group of atoms which are attached but do not define the cyclic ring structure.

[0169] “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, tetrahydrofuran, 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, tetrahydrothiopyran, 1,3-dithiane, 1,4-dithiane, 1,4-oxathiane).

[0170] “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. The alkenyl group is typically monovalent, but can be divalent, such as when the alkenyl group links two moieties together.

[0171] “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. Alkenylene groups include, but are not limited to, ethenylene, propenylene, isopropenylene, butenylene, isobutenylene, sec-butenylene, pentenylene and hexenylene.

[0172] “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 to6, 4 to 5, 4 to 6 and 5 to 6 carbons. The alkynyl group is typically monovalent, but can be divalent, such as when the alkynyl group links two moieties together.

[0173] “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.

[0174] “Cycloalkyl” refers to a saturated or partially unsaturated, monocyclic, fused bicyclic or bridged polycyclic ring assembly containing from 3 to 12 ring atoms, or the number of atoms indicated. Monocyclic rings include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cyclooctyl. Bicyclic and polycyclic rings include, for example, norbornane, decahydronaphthalene and adamantane. For example, C3-8cycloalkyl includes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclooctyl, and norbornane.

[0175] “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.

[0176] “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 1,4-dioxa-8-aza-spiro[4.5]dec-8-yl.

[0177] “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 heterocycloalkylene.

[0178] “Aryl” refers to a monocyclic or fused bicyclic, tricyclic or greater, aromatic ring assembly containing 6 to 16 ring carbon atoms. For example, aryl may be phenyl, benzyl or naphthyl. “Arylene” means a divalent radical derived from an aryl group. Aryl groups canbe mono-, di- or tri-substituted by one, two or three radicals selected from alkyl, alkoxy, aryl, hydroxy, halogen, cyano, amino, amino-alkyl, trifluoromethyl, alkylenedioxy and oxy-C2-C3-alkylene; 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-C2-C3-alkylene is also a divalent substituent attached to two adjacent carbon atoms of phenyl, e.g. oxyethylene or oxypropylene. An example for oxy- C2-C3-alkylene-phenyl is 2,3-dihydrobenzofuran-5-yl.

[0179] In some embodiments the 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.

[0180] Examples of substituted phenyl groups as R are, e.g. 4-chlorophen-1-yl, 3,4-dichlorophen-1-yl, 4-methoxyphen-1-yl, 4-methylphen-1-yl, 4-aminomethylphen-1-yl, 4-methoxyethylaminomethylphen-1-yl, 4-hydroxyethylaminomethylphen-1-yl, 4-hydroxyethyl-(methyl)-aminomethylphen-1-yl, 3-aminomethylphen-1-yl, 4-N-acetylaminomethylphen-1-yl, 4-aminophen-1-yl, 3-aminophen-1-yl, 2-aminophen-1-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-ylmethyl)-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.

[0181] “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.

[0182] “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.

[0183] 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, perfluoro(C1-C4)alkoxy, and perfluoro(C1-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, (C1-C8)alkyl and heteroalkyl, unsubstituted aryl and heteroaryl, (unsubstituted aryl)-(C1-C4)alkyl, and (unsubstituted aryl)oxy-(C1-C4)alkyl.

[0184] 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 (C1-C6)alkyl.

[0185] “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. In some embodiments, quinolinyl represents 2-, 3- or 4-quinolinyl. In some embodiments, isoquinolinyl represents 1-, 3- or 4-isoquinolinyl. In some embodiments, benzopyranyl, benzothiopyranyl can represent 3-benzopyranyl or 3-benzothiopyranyl, respectively. In some embodiments, thiazolyl can represent 2- or4-thiazolyl. In some embodiments, triazolyl can be 1-, 2- or 5-(1,2,4-triazolyl). In some embodiments, tetrazolyl can be 5-tetrazolyl.

[0186] In some embodiments, heteroaryl is pyridyl, indolyl, quinolinyl, pyrrolyl, thiazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazolyl, imidazolyl, thienyl, furanyl, benzothiazolyl, benzofuranyl, isoquinolinyl, benzothienyl, oxazolyl, indazolyl, or any of the radicals substituted, especially mono- or di-substituted.

[0187] 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.

[0188] 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.

[0189] “Electrophile” refers to an ion or atom or collection of atoms, which may be ionic, having an electrophilic center, i.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.

[0190] “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.

[0191] “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 structureindicates 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.

[0192] 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. “Non-naturally occurring amino acids” found in proteins are any amino acid other than those recited as naturally occurring amino acids. Non-naturally 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 N-alpha- methyl amino acids (e.g. sarcosine), 4-hydroxyproline, desmosine, isodesmosine, 5-hydroxylysine, epsilon-N-methyllysine, 3-methylhistidine, although found in naturally occurring proteins, are considered to be non-naturally 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.

[0193] “Linear” in reference to the geometry, architecture, or overall structure of a polymer, refers to polymer having a single polymer arm.

[0194] “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. In some embodiments, the halide is a bromine.

[0195] “Pharmaceutically acceptable excipient” or “pharmaceutically acceptable carrier” refers to an excipient that can be included in compositions 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. Nonlimiting examples of pharmaceutically acceptable excipients or carriers include water, NaCl, normal saline solutions, lactated Ringer’s, normal sucrose, normal glucose and the like. In any embodiment, a pharmaceutically acceptable carrier can be acceptable for administering directly into the eye of a patient (e.g., acceptable for intravitreal administration).

[0196] Therapeutic proteins are administered in an effective regime 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 regime can be referred to as a therapeutically effective regime. If the patient is at elevated risk of the disorder relative to the general population but is not yet experiencing symptoms, the regime can be referred to as a prophylactically effective regime. 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 ina preclinical or clinical trial in a population of treated patients relative to a control population of untreated patients.

[0197] 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 a tissue or an organism following introduction of the substance.

[0198] “OG1786” is a 9-arm initiator used for polymer synthesis with the structure shown in FIG. 4, which depicts that salt form of OG1786 with trifluororacetic acid. OG1786 may be used as other salts are used or as the free base.

[0199] “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.

[0200] “OG1802” is OG1801 with a maleimide functionality added and is shown in FIG.10 wherein each of n1, n2, n3, n4, n5, n6, n7, n8 and n9 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.

[0201] 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, ǻn, of the solution's refractive index n with the molecular concentration change, ǻc, by measuring the dn / dc (=ǻn / ǻc) 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 polydispersity 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.

[0202] 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 (e.g. OG1950) 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 the polymerization process, it is very important to determine the PDI of the sample as one of its quality attribute for narrow distribution of molecular weight.

[0203] 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.

[0204] 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 (OG1953); such species typically do not elute in a manner that might be described by a set of column calibration standards.

[0205] 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 1ml / min in 1xPBS pH 7.4, upon sample injection, the MALS and RI signals can beanalyzed 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 OG1953 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 / (MWpolymer+MWprotein)]+ 0.183 x [MWprotein / (MWpolymer+MWprotein)] where MWpolymer for OG1802 is 800 kDa and the MWprotein for OG1950 is 150 kDa. GENERAL Protein and Antibody Compositions, and Methods of Preparing Same

[0206] In some embodiments, a formulation or composition (e.g., therapeutically acceptable composition) that includes a first protein that is conjugated to a polymer (e.g., a phosphorylcholine-containing polymer) and a second protein that is unconjugated is provided. In some embodiments, the first protein is an antibody and the second protein is an antibody. Both antibodies can be therapeutic antibodies. In some embodiments, the composition is for treating an eye disorder in a subject. In some embodiments, a formulation includes at least a polymer (e.g., a phosphorylcholine-containing polymer) and an unconjugated protein (e.g., an unconjugated antibody). As used herein, formulation and composition (e.g., therapeutically acceptable composition, pharmaceutical composition, or therapeutic composition) can be used interchangeably. In some embodiments, a formulation or therapeutically acceptable composition is safe for human use (e.g., administering to a human). In some embodiments, a formulation or therapeutically acceptable composition is not an intermediate product generated during manufacture of a final product (e.g., that may be suitable for use in a human).

[0207] In some embodiments, provided herein is a composition (e.g., therapeutically acceptable composition) comprising any two proteins that can be the same or different in function, wherein one is conjugated to a polymer and the other is not conjugated to the polymer (or conjugated to any polymer or conjugated to any effective amount of a polymer). The composition can be for the treatment of an eye disorder. In some embodiments, both of the proteins are therapeutics proteins for the treatment of an eye disorder. In some embodiments, one or both of the proteins are therapeutics, antibodies and / or therapeutic antibodies. In some embodiments, one antibody is conjugated to a polymer and the otherantibody is not conjugated to a polymer. In some embodiments, the antibody may be synthesized. In some embodiments, the antibody may be a native sequence antibody. In some embodiments, the antibody may be a Fab fragment. In some embodiments, the antibody may be a Trap fragment. In some embodiments, the antibody may be a fusion protein such as a Trap-antibody fusion protein. In some embodiments, the antibody may be a peptide fragment. In some embodiments, a non-antibody scaffold protein can be used instead of an antibody.

[0208] Provided herein is a formulation (or therapeutically acceptable composition) comprising: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer (the “conjugated protein”); a second molar amount of a second protein that is not conjugated to a phosphorylcholine-containing polymer (the “unconjugated protein”); and a pharmaceutically acceptable carrier, wherein the formulation comprises the second protein at about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the formulation has a pH that is about 0.5 pH units away (e.g., above or below) or more from the isoelectric point (pI) of the second protein. As used herein, “molar amount” denotes a measure of the molar quantity of a molecule. In some embodiments, molar amount is a molar concentration (e.g., M, mM, μM, nM, etc.). In some embodiments, molar amount is expressed in units of moles (e.g., moles, millimoles, micromoles, etc.). As used herein, the isoelectric point (pI) of a protein has its customary and ordinary meaning to one of ordinary skill int the art, in view of the present disclosure. The pI denotes the pH at which the protein carries no net charge. The pI can be a previously known value for the same or similar protein, or be determined based on a model, or empirically. In some embodiments, the pI is a theoretically determined pI. In some embodiments, the pI is an empirically determined pI.

[0209] Low-viscosity formulations of a protein conjugate (e.g., a protein conjugated to a phosphorylcholine-containing polymer) are also provided. A high concentration of the protein conjugate in the formulation can raise the viscosity of the formulation. In some embodiments, lowering the viscosity of the formulation (while maintaining the total amount of active protein) improves one or more of manufacturability, handling, storage, and injectability, for example when delivering the formulation with a syringe to a site of treatment (e.g., intraocular administration).

[0210] Formulations (or therapeutically acceptable compositions) of the present disclosure (e.g., having a polymer or polymer-conjugated protein and unconjugated protein) can include the unconjugated protein (e.g., protein that is not conjugated to the phosphorylcholine-containing polymer) at any suitable % molar amount of the total molar amount of the polymer / polymer conjugate and the unconjugated protein. In some embodiments, the formulation (or therapeutically acceptable composition) comprises the second protein (the unconjugated protein) at about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount of the conjugate and the second molar amount of the second protein. For example, where the combined concentration of the conjugate and the unconjugated protein is 100 μM, the unconjugated protein at 1% of the total molar amount is at 1 μM, and the conjugate is at 99 μM. In some embodiments, the formulation (or therapeutically acceptable compositions) comprises the second protein (the unconjugated protein) at about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount of the conjugate and the second molar amount of the second protein. In some embodiments, the formulation comprises the second protein (the unconjugated protein) at, or at about 1% or more, e.g., about 2% or more, about 5% or more, about 10% or more, about 15% or more, about 20% or more, about 25% or more, about 30% or more, about 35% or more, about 40% or more, about 45% or more, about 50% or more, about 55% or more, about 60% or more, about 65% or more, about 70% or more, about 75% or more, about 80% or more, about 85% or more, or about 95% or less, about 90% or less, about 85% or less, about 80% or less, about 75% or less, about 70% or less, about 65% or less, about 60% or less, about 55% or less, about 50% or less, about 45% or less, about 40% or less, about 35% or less, about 30% or less of a total molar amount of the conjugate and the second protein, or optionally, the formulation includes the second protein at a percentage in a range defined by any two of the preceding values (e.g., about 1-95%, 5-90%, 10-80%, 5-50%, 10- 40%, 15-35%, 15-25%, 25-35%, 25-40%, 40-95%, 50-80%, etc.) of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount of the conjugate and the second molar amount of the second protein. In some embodiments, the formulation comprises the second protein (the unconjugated protein) at between about 5% and about 50%, or between about 15% and about 30% of a total molaramount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount of the conjugate and the second molar amount of the second protein. In some embodiments, the formulation comprises the second protein (the unconjugated protein) at between about 15% and about 25% of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount of the conjugate and the second molar amount of the second protein. In some embodiments, the formulation comprises the second protein (the unconjugated protein) at between about 25% and about 35% of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount of the conjugate and the second molar amount of the second protein. In some embodiments, the formulation comprises the second protein (the unconjugated protein) at about 20% of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount of the conjugate and the second molar amount of the second protein. In some embodiments, the formulation comprises the second protein (the unconjugated protein) at about 30% of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount of the conjugate and the second molar amount of the second protein. In some embodiments, any formulation or composition provided herein comprises the second protein (the unconjugated protein) at more than 5% of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount of the conjugate and the second molar amount of the second protein. In some embodiments, any formulation or composition provided herein comprises the second protein (the unconjugated protein) at more than 10% of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount of the conjugate and the second molar amount of the second protein. In some embodiments, any composition or formulation herein includes two or more (e.g., 2, 3, 4, 5 or more) different second proteins (or unconjugated proteins), where the second molar amount is the sum of the molar amounts of the two or more different second proteins.

[0211] Provided herein is a formulation (or therapeutically acceptable compositions) comprising: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second molar amount of a second protein that is not conjugated to a phosphorylcholine-containing polymer; and apharmaceutically acceptable carrier, wherein the formulation (or therapeutically acceptable compositions) comprises the second protein at about 1% or more (e.g., about 5-90%, 15-25%, 25-35%, 25-40%, etc.) of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the formulation (or therapeutically acceptable compositions) has a pH that is about 0.5 pH units (e.g., above or below) or more from the isoelectric point (pI) of the second protein, wherein the formulation (or therapeutically acceptable compositions) has a reduced viscosity and / or an enhanced injectability compared to a reference formulation (or reference composition) comprising the conjugate at the total molar amount. In some embodiments, the reference formulation or composition is one that includes the conjugate at the total molar amount and effectively does not include the second protein (or includes the second protein at less than 1% of the total molar amount), but is otherwise the same as the formulation or composition for which it serves as a reference. In some embodiments, the reference formulation or composition is a therapeutically acceptable reference composition.

[0212] Also provided is a low-viscosity formulation (or therapeutically acceptable compositions) of a protein conjugate, comprising: a first molar amount of a conjugate comprising a protein conjugated to a phosphorylcholine-containing polymer; a second molar amount of the protein that is not conjugated to the phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the formulation (or therapeutically acceptable compositions) has a pH that is about 0.5 pH units away (e.g., above or below) or more from the isoelectric point (pI) of the protein, wherein the formulation (or therapeutically acceptable compositions) has reduced viscosity and / or an enhanced injectability compared to a reference formulation (or reference composition) comprising the conjugate at a total molar amount that is the sum of the first and second molar amounts. In some embodiments, the reference formulation or composition is one that includes the conjugate at the total molar amount and effectively does not include the second protein (or includes the second protein at less than 1% of the total molar amount), but is otherwise the same as the formulation or composition for which it serves as a reference. In some embodiments, the reference formulation or composition is a therapeutically acceptable reference composition.

[0213] The formulation (or therapeutically acceptable compositions) of the present disclosure can have any suitable viscosity. In some embodiments, the formulation has aviscosity of, or of about 1000 mPas•s or less, e.g., about 900 mPas•s or less, about 800 mPas•s or less, about 700 mPas•s or less, about 600 mPas•s or less, about 500 mPas•s or less, about 400 mPas•s or less, about 300 mPas•s or less, about 200 mPas•s or less, about 100 mPas•s or less, or about 200 mPas•s or more, 300 mPas•s or more, about 400 mPas•s or more, about 500 mPas•s or more, about 600 mPas•s or more, about 700 mPas•s or more, about 800 mPas•s or more, about 900 mPas•s or more, or a viscosity in a range defined by any two of the preceding values (e.g., 100-1000 mPas•s, 200-1000 mPas•s, 200-500 mPas•s, 300-800 mPas•s, 300-600 mPas•s, 200-300 mPas•s, etc.). In some embodiments, the formulation has a viscosity of about 100 to about 300 mPas•s or about 200 to about 500 mPas•s. In some embodiments, the formulation has a viscosity of about 300 to about 400 mPas•s. The viscosity can be measured using any suitable option, e.g., a rotational rheometer such as a TA Instruments DHR-20.

[0214] In some embodiments, the formulation (or therapeutically acceptable composition) has a viscosity that is reduced compared to a reference formulation having a molar amount of the conjugate that is the same as the total amount of the conjugate and unconjugated protein. In some embodiments, the viscosity is reduced by about 10% or more, e.g., about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or by about 90% or less, about 80% or more, about 70% or less, about 60% or less, about 50% or less, about 40% or less, about 30 % or less, or by a percentage in a range defined by any two of the preceding values (e.g., by about 10-90%, by about 20-80%, by about 50-80%, by about 30- 70%, by about 40-90%, etc.). In some embodiments, the viscosity is reduced by about 50- 80%. In some embodiments, the viscosity is reduced by about 70-80%.

[0215] In some embodiments, the reference formulation or composition (having the conjugate form only at the total molar amount) may include the conjugate (e.g., the polymer portion of the conjugate) at a sufficiently high molar amount to result in a formulation with high viscosity. In some embodiments, the reference formulation has a viscosity of, or of about 700 mPas•s or more, e.g., about 800 mPas•s or more, about 900 mPas•s or more, about 1000 mPas•s or more, about 1000 mPas•s or more, or about 1500 mPas•s or less, about 1400 mPas•s or less, about 1300 mPas•s or less, about 1200 mPas•s or less, about 1100 mPas•s or less, about 1000 mPas•s or less, or a viscosity in a range defined by any two of the preceding values (e.g., about 700-1500 mPas•s, about 800-1300 mPas•s, about 900-1200, mPas•s, about 1000-1300mPas•s, etc.). In some embodiments, where the first protein and the second protein are the same or substantially the same protein (having substantially the same activity), replacing a portion of the conjugated protein with the unconjugated protein, while keeping the total molar amount the same, provides a formulation having lower viscosity and the same total amount of the protein as the reference formulation.

[0216] Formulations (or therapeutically acceptable compositions) of the present disclosure can have low turbidity (e.g., a visually clear solution). In some embodiments, adding an unconjugated protein to a formulation of a conjugate (e.g., a protein conjugated to a phosphorylcholine-containing polymer as provided herein) results in a mixed formulation that is turbid (e.g., cloudy visual appearance). In some embodiments, the turbidity of a formulation with a mixture of unconjugated and conjugated proteins is reduced when the pH of the formulation is not at or is not around (e.g., at least 0.5 pH units away from) the isoelectric point (pI) of the unconjugated protein. Where it is desirable for the formulation to be clear (e.g., when the formulation is for intraocular administration), a low-turbidity formulation of a mixture of unconjugated and conjugated proteins is obtained when the pH of the formulation is different from (e.g., at least 0.5 pH units above or below) the pI of the unconjugated protein.

[0217] Provided is a formulation (or therapeutically acceptable composition) comprising: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second molar amount of a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the second protein is present in the formulation at about 1% or more (e.g., about 5-90%, 15-25%, 25-35%, 25-40%, etc.) of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the formulation has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein, wherein the formulation has a reduced turbidity compared to a reference formulation comprising the first molar amount of the conjugate and the second molar amount of the second protein at a pH about the same as (e.g., within 0.05, 0.1, 0.15, 0.2, 0.3, 0.4, or 0.5 pH units of) the pI of the second protein. In some embodiments, the formulation has a reduced turbidity compared to a reference formulation comprising the first molar amount of the conjugate and the second molar amount of the second protein at a pH within 0.5 pH units of the pI of the second protein. The turbiditycan be measured using any suitable option. For example, turbidity can be measured at OD 600 nm using a plate reader and calibrating the OD values against a suitable standard (e.g., a 4000 NTU Formazin calibration standard). In some embodiments, the turbidity is reduced by about 10% or more, e.g., about 20% or more, about 30% or more, about 40% or more about 50% or more about 60% or more about 70% or more, about 80% or more, about 90% or more, or about 100%, or by a percentage in a range defined by any two of the preceding values (e.g., 10-100%, 10-50%, 30-70%, 50-100%, 80-100%, etc.).

[0218] In some embodiments, the formulation (or therapeutically acceptable composition) is substantially free of turbidity. In some embodiments, the formulation is free of turbidity based on visual inspection. In some embodiments, a formulation or composition that is substantially free of turbidity is free of turbidity based on visual inspection. In some embodiments, the prepared formulation has a turbidity that, when expressed as NTU values as measured at OD 600 nm and calibrated against a 4000 NTU Formazin calibration standard, is (or corresponds to) about 500 Nephelometric Turbidity Units (NTU) or less, e.g., about 400 NTU or less, about 300 NTU or less, about 200 NTU or less, about 100 NTU or less, or about –50 NTU or more, about 0 NTU or more, about 100 NTU or more, about 200 NTU or more, about 300 NTU or more, about 400 NTU or more, or a turbidity measure in a range defined by any two of the preceding values (e.g., about -50-500 NTU, about 0-400 NTU, about 100-300 NTU, or about 50-400 NTU). In some embodiments, the prepared formulation has a turbidity that, when expressed as NTU values as measured at OD 600 nm and calibrated against a 4000 NTU Formazin calibration standard, is (or corresponds to) about 300 NTU or less. In some embodiments, the prepared formulation has a turbidity that, when expressed as NTU values as measured at OD 600 nm and calibrated against a 4000 NTU Formazin calibration standard, is (or corresponds to) about 200 NTU or less. In some embodiments, the prepared formulation has a turbidity that, when expressed as NTU values as measured at OD 600 nm and calibrated against a 4000 NTU Formazin calibration standard, is (or corresponds to) about 100 NTU or less. In some embodiments, the formulation or composition is free of turbidity based on visual inspection when the turbidity expressed as NTU values as measured at OD 600 nm and calibrated against a 4000 NTU Formazin calibration standard is, is about, or is at most 500, 450, 400, 350, 300, 250, 200, 150, 100, or 50 NTU.

[0219] Also provided herein is a pharmaceutical formulation that includes: a first molar amount of a conjugate comprising a protein conjugated to a phosphorylcholine- containing polymer; a second molar amount the protein that is not conjugated to the phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the formulation comprises the protein that is not conjugated to the phosphorylcholine-containing polymer at about 1% or more (e.g., about 5-90%, 15-25%, 25-35%, 25-40%, etc.) of a total molar amount of the conjugate and unconjugated proteins, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the formulation has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the protein, wherein the formulation is substantially free of turbidity.

[0220] Provided herein is a formulation that includes: a phosphorylcholine- containing polymer present in the formulation at about 100 mg / mL or higher; and a protein that is not conjugated to a phosphorylcholine-containing polymer, wherein the protein is present in the formulation at a second molar amount, wherein the protein is present in the formulation at about 1% or more (e.g., about 5-90%, 15-25%, 25-35%, 25-40%, etc.) of a total molar amount of the polymer and the protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the formulation has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the protein. In some embodiments, the polymer is conjugated to a protein (e.g., an antibody, fusion construct, etc.).

[0221] Also provided is a formulation (or therapeutically acceptable composition) comprising: a first molar amount of a conjugate comprising a first protein conjugated to a polymer; and a second molar amount of a second protein that is not conjugated to a polymer, wherein the formulation comprises the second protein at about 1% or more (e.g., about 5-90%, 15-25%, 25-35%, 25-40%, etc.) of a total molar amount of the first protein and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount. In some embodiments, the formulation comprises the second protein at about 1-90%, about 5-80%, about 10-95%, about 15-30%, about 5-50%, or about 10-40%, of the total molar amount of the conjugate and the second protein. In some embodiments, the polymer is a phosphorylcholine-containing polymer. In some embodiments, the formulation comprises the second protein at about 5-50%, or about 15-30% of the total molar amount ofthe conjugate and the second protein. In some embodiments, the polymer is a phosphorylcholine-containing polymer.

[0222] Also provided is a formulation (or therapeutically acceptable composition) comprising: a conjugate comprising a first protein conjugated to a polymer; and a second protein that is not conjugated to a polymer, wherein a first molar amount of the conjugate and a second molar amount of the second protein has been combined in the formulation such that the second molar amount is about 1% or more (e.g., about 5-90%, 15-25%, 25-35%, 25-40%, etc.) of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount. In some embodiments, the formulation is prepared by combining the first molar amount of the conjugate with the second molar amount of the second protein that is not conjugated to a polymer, such that the second molar amount is at the specified percentage of the sum of the first molar amount and the second molar amount (e.g., specified percentage of the total molar amount). In some embodiments, the second molar amount is about 1-90%, about 5-90%, about 5-80%, about 10-95%, about 15-30%, about 5-50%, or about 10-40%, of the total molar amount of the conjugate and the second protein. In some embodiments, the second molar amount is about 5-50% of the total molar amount of the conjugate and the second protein. In some embodiments, the second molar amount is about 15-30% of the total molar amount of the conjugate and the second protein. In some embodiments, the polymer is a phosphorylcholine-containing polymer.

[0223] In some embodiments, the formulation or composition has been prepared by combining the conjugate at a percent composition of about 1% or more (e.g., about 5-93%, 15-25%, 25-35%, 25-40%, etc.) of the second protein relative to the total protein mass weight concentration of the first protein and the second protein, where the remainder of the total protein mass weight concentration includes the first protein. For example, for a total mass weight concentration of 50 mg / mL, the therapeutically acceptable composition can be prepared by combining an amount of the second protein that corresponds to 10 mg / mL in the final composition (at percent composition of 20%) with an amount of the conjugate that corresponds to 40 mg / mL of the first protein (as the conjugate, excluding any contribution of the polymer to the mass weight concentration calculation) in the final composition.

[0224] In some embodiments, the conjugate includes a first protein conjugated to a polymer, wherein the polymer includes one or more of: polyethylene glycol (PEG), branched PEG, PolyPEG® (Warwick Effect Polymers; Coventry, UK), polysialic acid (PSA), starch, hydroxylethyl starch (HES), hydroxyalkyl starch (HAS), carbohydrate, polysaccharides, pullulane, chitosan, hyaluronic acid, chondroitin sulfate, dermatan sulfate, dextran, carboxymethyl-dextran, polyalkylene oxide (PAO), polyalkylene glycol (PAG), polypropylene glycol (PPG), polyoxazoline, polyacryloylmorpholine, polyvinyl alcohol (PVA), polycarboxylate, polyvinylpyrrolidone, polyphosphazene, polyoxazoline, polyethylene-co-maleic acid anyhydride, polystyrene-co-maleic acid anhydride, poly(1- hydroxymethyethylene hydroxymethylformal) (PHF), a zwitterionic polymer, a phosphorylcholine containing polymer and a polymer comprising MPC, Poly (Glyx-Sery), Hyaluronic acid (HA), Heparosan polymers (HEP), Fleximers, Dextran, and Poly-sialic acids (PSA).

[0225] Also provided is a formulation (or therapeutically acceptable composition) comprising: a conjugate comprising a first protein conjugated to a phosphorylcholine- containing polymer, wherein the polymer has 9 arms and a molecular weight of between 600,000 and 1,000,000 Da, wherein the polymer is present in the formulation at about 100 mg / mL or more; and a second protein that is not conjugated to a polymer, wherein the second protein is present in the formulation at about 5-15 mg / mL. In some embodiments, the first and second proteins are a therapeutic protein. In some embodiments, the first protein and second protein are the same (e.g., are at least, or at least about 85%, 90%, 95%, 97%, 98%, 99%, or are about 100% identical in amino acid sequence). In some embodiments, the first protein and second protein are different proteins.

[0226] In some embodiments, the phosphorylcholine-containing polymer is present in the formulation at about 100 mg / mL or more, e.g., about 150 mg / mL or more, about 200 mg / mL or more, about 250 mg / mL or more, about 300 mg / mL or more, about 350 mg / mL or more, about 400 mg / mL or more, about 450 mg / mL or more, or a concentration in range defined by any two of the preceding values (e.g., 100-450 mg / mL, 150-400 mg / mL, 200-400 mg / mL, 250-450 mg / mL, 300-450 mg / mL, etc.). In some embodiments, the phosphorylcholine-containing polymer is present in the formulation at between about 150-400mg / mL. In some embodiments, the phosphorylcholine-containing polymer is present in the formulation at between about 200-300 mg / mL.

[0227] In some embodiments, the polymer has a molecular weight of about 100,000 Da or more, e.g., about 150,000 Da or more, about 200,000 Da or more, about 350,000 Da or more, about 400,000 Da or more, about 450,000 Da or more, about 500,000 Da or more, about 550,000 Da or more, about 600,000 Da or more, about 650,000 Da or more, about 700,000 or more, about 750,000 Da or more, about 800,000 Da or more, about 850,000 Da or more, about 900,000 Da or more, about 950,000 Da or more, about 1,000,000 Da or more, or a molecular weight in a range defined by any two of the preceding values (e.g., 100,000-1,000,000 Da, 300,000-950,000 Da, 400,000-800,000 Da, 500,000-750.000 Da, 600,000-700,000 Da, 600,000-1,000,000 Da, etc.). In some embodiments, the polymer has a molecular weight in the range of about 700,000 to about 800,000 Da. In some embodiments, the polymer is any of the polymers disclosed herein. In some embodiments, the polymer is OG1801 or OG1802.

[0228] In some embodiments, formulations (or therapeutically acceptable compositions) of the present disclosure (e.g., having a polymer or polymer-conjugated protein and unconjugated protein) have a pH that is different (e.g., higher or lower) from the pI of the unconjugated protein (e.g., protein that is not conjugated to a phosphorylcholine-containing polymer). In some embodiments, the pH of the formulation is about 0.5 pH units away or more from the pI of the unconjugated protein (e.g., protein that is not conjugated to the phosphorylcholine-containing polymer). In some embodiments, the pH of the formulation is about 0.5 pH units away or more, e.g., about 0.6 pH units away or more, about 0.7 pH units away or more, about 0.8 pH units away or more, about 0.9 pH units away or more, about 1.0 pH units away or more, about 1.1 pH units away or more, about 1.2 pH units away or more, about 1.3 pH units away or more, about 1.4 pH units away or more, about 1.5 pH units away or more, about 1.6 pH units away or more, about 1.7 pH units away or more, about 1.8 pH units away or more, about 1.9 pH units away or more, about 2.0 pH units away or more, about 2.1 pH units away or more, about 2.2 pH units away or more, about 2.3 pH units away or more, about 2.4 pH units away or more, about 2.5 pH units away or more, about 2.6 pH units away or more, about 2.7 pH units away or more, about 2.8 pH units away or more, about 2.9 pH units away or more, about 3.0 pH units away or more, about 3.2 pH units away or more, about 3.4 pH units away or more, about 3.6 pH units away or more, about 3.8 pH units away or more,about 4.0 pH units away or more, about 4.5 pH units away or more, about 5.0 pH unit away or more, from the pI of the unconjugated protein (e.g., protein that is not conjugated to a phosphorylcholine-containing polymer), or away from the pI of the unconjugated protein by a pH unit in a range defined by any two of the preceding values (e.g., 0.5-5.0 pH units away, 1.0-4.0 pH units away, 1.5-3.0 pH units away, 2.0-3.0 pH units away, 1.5-3.6 pH units away, etc.). In some embodiments, the pH of the formulation is between about 2.0-3.0 pH units away from the pI of the unconjugated protein (e.g., protein that is not conjugated to the phosphorylcholine-containing polymer).

[0229] In some embodiments, formulations (or therapeutically acceptable compositions) of the present disclosure (e.g., having a polymer or polymer-conjugated protein and unconjugated protein) have a pH that is more acidic than the pI of the unconjugated protein (e.g., protein that is not conjugated to a phosphorylcholine-containing polymer). In some embodiments, the pH of the formulation is about 0.5 pH units less or lower, e.g., about 0.6 pH units less or lower, about 0.7 pH units less or lower, about 0.8 pH units less or lower, about 0.9 pH units less or lower, about 1.0 pH units less or lower, about 1.1 pH units less or lower, about 1.2 pH units less or lower, about 1.3 pH units less or lower, about 1.4 pH units less or lower, about 1.5 pH units less or lower, about 1.6 pH units less or lower, about 1.7 pH units less or lower, about 1.8 pH units less or lower, about 1.9 pH units less or lower, about 2.0 pH units less or lower, about 2.1 pH units less or lower, about 2.2 pH units less or lower, about 2.3 pH units less or lower, about 2.4 pH units less or lower, about 2.5 pH units less or lower, about 2.6 pH units less or lower, about 2.7 pH units less or lower, about 2.8 pH units less or lower, about 2.9 pH units less or lower, about 3.0 pH units less or lower, about 3.2 pH units less or lower, about 3.4 pH units less or lower, about 3.6 pH units less or lower, about 3.8 pH units less or lower, about 4.0 pH units less or lower, about 4.5 pH units less or lower, about 5.0 pH units less or lower from the pI of the unconjugated protein (e.g., protein that is not conjugated to the phosphorylcholine-containing polymer), or lower than the pI of the unconjugated protein by a pH unit in a range defined by any two of the preceding values (e.g., 0.5-5.0 pH units lower, 1.0-4.0 pH units lower, 1.5-3.0 pH units lower, 2.0-3.0 pH units lower, 1.5-3.6 pH units lower, etc.). In some embodiments, the pH of the formulation is between about 2.0-3.0 pH units lower than the pI of the unconjugated protein (e.g., protein that is not conjugated to the phosphorylcholine-containing polymer).

[0230] In some embodiments, formulations (or therapeutically acceptable compositions) of the present disclosure (e.g., having a polymer or polymer-conjugated protein and unconjugated protein) have a pH that is more basic than the pI of the unconjugated protein (e.g., protein that is not conjugated to the phosphorylcholine-containing polymer). In some embodiments, the pH of the formulation is about 0.5 pH units more or higher, e.g., about 0.6 pH units more or higher, about 0.7 pH units more or higher, about 0.8 pH units more or higher, about 0.9 pH units more or higher, about 1.0 pH units more or higher, about 1.1 pH units more or higher, about 1.2 pH units more or higher, about 1.3 pH units more or higher, about 1.4 pH units more or higher, about 1.5 pH units more or higher, about 1.6 pH units more or higher, about 1.7 pH units more or higher, about 1.8 pH units more or higher, about 1.9 pH units more or higher, about 2.0 pH units more or higher, about 2.1 pH units more or higher, about 2.2 pH units more or higher, about 2.3 pH units more or higher, about 2.4 pH units more or higher, about 2.5 pH units more or higher, about 2.6 pH units more or higher, about 2.7 pH units more or higher, about 2.8 pH units more or higher, about 2.9 pH units more or higher, about 3.0 pH units more or higher, about 3.2 pH units more or higher, about 3.4 pH units more or higher, about 3.6 pH units more or higher, about 3.8 pH units more or higher, about 4.0 pH units more or higher, about 4.5 pH units more or higher, about 5.0 pH units more or higher, than the pI of the unconjugated protein (e.g., protein that is not conjugated to the phosphorylcholine- containing polymer), or greater than the pI of the unconjugated protein by a pH unit in a range defined by any two of the preceding values (e.g., 0.5-4.0 pH units greater, 1.0-3.0 pH units greater, 1.5-3.0 pH units greater, 2.0-3.0 pH units greater, 1.5-3.6 pH units greater, etc.). In some embodiments, the pH of the formulation is between about 2.0-3.0 pH units greater than the pI of the unconjugated protein (e.g., protein that is not conjugated to the phosphorylcholine- containing polymer).

[0231] In some embodiments, the difference between the pH of a formulation (or therapeutically acceptable composition) containing a mixture of a polymer or polymer- conjugated protein and an unconjugated protein (e.g., protein that is not conjugated to a phosphorylcholine-containing polymer), as provided herein, and the pI of the unconjugated protein depends on the relative amounts (e.g., molar amounts) of the polymer / polymer- conjugate and the unconjugated protein in the formulation. Without being bound by theory, in general, the higher the relative amount (e.g., molar amount) of the unconjugated proteincompared to the polymer / polymer-conjugate, the further away the pH will be relative to the pI of the unconjugated protein (for a given concentration (e.g., molar amount) of the polymer or conjugate above a threshold in the formulation). Without being bound by theory, when the pH is too close to the pI of the unconjugated protein, the formulation can turn turbid. In some embodiments, a formulation having a pH at about 4.0 or lower is substantially free of turbidity regardless of the concentration of polymer / polymer conjugate, or the concentration of the unconjugated protein.

[0232] Also provided herein is a formulation (or therapeutically acceptable composition) that includes: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second molar amount of a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the difference between the pI of the second protein and the pH of the formulation in the acidic or basic direction is selected to be greater than the minimum difference in the corresponding acidic or basic direction between the pI of the second protein and the pH for a reference formulation comprising: a third molar amount of the conjugate comprising the first protein conjugated to the phosphorylcholine-containing polymer; a fourth molar amount of the second protein that is not conjugated to the phosphorylcholine-containing polymer; and the pharmaceutically acceptable carrier, wherein a first total molar amount comprising a sum of the first molar amount and the second molar amount, and a second total molar amount comprising a sum of the third molar amount and the fourth molar amount are substantially the same, wherein the second molar amount is greater than the fourth molar amount, wherein the reference formulation is substantially free of turbidity. Without being bound by theory, between two formulations each having an unconjugated protein (e.g., antibody or fusion construct) and a polymer conjugate, and the same total molar amount of the protein (unconjugated and conjugated), and when the concentration of polymer is sufficiently high, the difference between the pI of the unconjugated protein and the pH of the formulation having a higher percentage of the total molar amount of the unconjugated protein (in either the acidic or basic direction) will generally be greater than the minimum difference between the pI of the unconjugated protein and the pH of the formulation having the lower percentage of the total molar amount of the unconjugated protein (in the corresponding acidic or basic direction) to maintain a clear formulation that issubstantially free of turbidity. For example and without limitation, for two formulations of the same unconjugated / conjugated protein pair (where the protein component of the conjugate may or may not be the same as the unconjugated protein), the same total molar amount of unconjugated and conjugated protein and a sufficiently high concentration of polymer from the conjugate, the pH of the formulation may be lower in the formulation having a higher proportion of unconjugated to conjugated protein if the pH is below the pI of the unconjugated protein, to maintain a clear solution. For example and without limitation, a formulation containing the unconjugated protein (having a pI of around 7.4) at about 5% of the total molar amount of unconjugated protein and conjugate, and at least about 200 mg / mL polymer, may be clear up to around pH 6, and another formulation having the unconjugated protein at 10% or 15% of the total molar amount, and at least about 200 mg / mL polymer, may be clear up to around pH 5, but turbid at around pH 6. In some embodiments, the pH of the formulation and the reference formulation is each at least about 4.5 for this trend. In some embodiments, the pH of the formulation is selected to be lower than the maximum pH for the reference formulation (e.g., for maintaining a clear formulation), wherein the pH of the formulation and a maximum pH for the reference formulation are lower than the pI of the second protein. In some embodiments, the pH of the formulation is selected to be higher than a minimum pH for the reference formulation (e.g., for maintaining a clear formulation), wherein the pH of the formulation and the minimum pH for the reference formulation are higher than the pI of the second protein. In some embodiments, the pH of the formulation and the reference formulation is each at most about 8.5 for this trend. In some embodiments, the pH of the formulation and the reference formulation is between about 4.5 and 8.5 for this trend. In some embodiments, the concentration of the polymer component of the conjugate in the formulation and the reference formulation is each at least about 100 mg / mL (e.g., about 150 mg / mL, about 200 mg / mL, about 250 mg / mL, or about 300 mg / mL, or in a range of 150-300 mg / mL, e.g., 200- 300 mg / mL, or 250-300 mg / mL).

[0233] The formulation (or therapeutically acceptable composition) can have any suitable pH. In some embodiments, the pH of the formulation is about 3.0 or higher, about 3.5 or higher, about 4.0 or higher, about 4.5 or higher, about 5.0 or higher, about 5.5 or higher, about 6.0 or higher, about 6.5 or higher, about 7.0 or higher, about 7.5 or higher, about 8.0 or higher, about 8.5 or higher, about 9.0 or higher, about 9.5 or higher, about 10.0 or higher, orabout 10.5 or higher, or about 12.0 or lower, about 11.5 or lower, about 11.0 or lower, about 10.5 or lower, about 10.0 or lower, about 9.5 or lower, about 9.0 or lower, about 8.5 or lower, about 8.0 or lower, about 7.5 or lower, about 7.0 or lower, about 6.5 or lower, about 6.0 or lower, about 5.5 or lower, about 5.0 or lower, about 4.5 or lower, about 4.0 or lower, or a pH in a range defined by any two of the preceding values (e.g., 3.0-12.0, 3.5-10.0, 4.0-6.0, 8-12, 5.0-8.0, 9-10, etc.). In some embodiments, the pH of the formulation is in the range of about pH 4 to about pH 5.5. In some embodiments, the pH of the formulation is in the range of about pH 4.5 to about pH 5.3. In some embodiments, the pH of the formulation is about 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, or about 5.5. In some embodiments, the pH of the formulation is about 5.0. In some embodiments, the pH of the formulation is about 4.5.

[0234] In some embodiments, the unconjugated protein (e.g., the protein that is not conjugated to a phosphorylcholine-containing polymer) in the formulation has a pH of, or of about 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, or about 9.5 or higher. In some embodiments, the pI of the unconjugated protein (e.g., the protein that is not conjugated to a phosphorylcholine-containing polymer) in the formulation is between about 4.0 and about 9.5, between about 5.0 and about 9.5, between about 5.5 and about 8.5, between about 6 and about 8.5, between about 7.0 and about 9.5, or between about 5.0 and about 8.0. In some embodiments, the pI of the unconjugated protein (e.g., the protein that is not conjugated to a phosphorylcholine-containing polymer) in the formulation is between about 7.0 and about 8.5, the pH of the formulation is between about 4.0 and about 5.5, and the phosphorylcholine- containing polymer is present in the formulation (e.g., as a conjugate of the protein) at a concentration of between about 200 and about 300 mg / mL. In some embodiments, the pI of the unconjugated protein (e.g., the protein that is not conjugated to a phosphorylcholine- containing polymer) in the formulation is between about 7.0 and about 8.5, the pH of the formulation is between about 4.8 and about 5.2, and the phosphorylcholine-containing polymer is present in the formulation (e.g., as a conjugate of the protein) at a concentration of between about 200 and about 300 mg / mL.

[0235] In some embodiments, a formulation (or therapeutically acceptable composition, pharmaceutical composition, or therapeutic composition) provided herein is storage stable. In some embodiments, a formulation (or therapeutically acceptable composition, pharmaceutical composition, or therapeutic composition) provided herein shows long-term stability (e.g., when stored under standard storage conditions). In some embodiments, components of the formulation (the polymer / polymer conjugate and unconjugated protein) are soluble, and the formulation is a clear solution. In some embodiments, a storage stable formulation remains substantially free of turbidity. In some embodiments, the formulation is stable (e.g., clarity of the solution, structurally in terms of the protein and polymer components, and / or functionally with respect to the protein activity) after extended periods of time after being formulated at standard storage temperatures. The formulation (or therapeutic composition, pharmaceutical composition, or therapeutically acceptable composition) can be stored at any suitable temperature. In some embodiments, the formulation (or therapeutic composition, pharmaceutical composition, or therapeutically acceptable composition) is stored at, at about, or at most at 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40°C, or optionally it is stored at a temperature in a range defined by any two of the preceding values (e.g., 0-40°C, 0-10°C, 10- 20°C, 20-30°C, 30-40°C, etc.). In some embodiments, the formulation (or therapeutic composition, pharmaceutical composition, or therapeutically acceptable composition) is stored at a temperature in the range of 0-10°C. In some embodiments, the formulation (or therapeutic composition, pharmaceutical composition, or therapeutically acceptable composition) is stored at a temperature in the range of 10-20°C. In some embodiments, the formulation (or therapeutic composition or therapeutically acceptable composition) is stored at or at about 5°C. In some embodiments, the formulation (or therapeutic composition, pharmaceutical composition, or therapeutically acceptable composition) is stored at or at about 25°C. In some embodiments, the formulation (or therapeutic composition, pharmaceutical composition, or therapeutically acceptable composition) is stored at room temperature. In some embodiments, the formulation (or therapeutically acceptable composition) is stored under ambient atmospheric pressure. In some embodiments, the formulation (or therapeutic composition, pharmaceutical composition, or therapeutically acceptable composition) is stored at, at about, or at least at -5°C, -10°C, -15°C, -20°C, or -25°C.

[0236] In some embodiments, the formulation is stable when a measured feature of the formulation (e.g., turbidity, structural integrity and concentration of the protein and polymer components, activity of the protein) remains within at least about 20%, at least about 15%, at least about 10%, or at least about 5% of the originally measured level when the formulation was initially prepared. In some embodiments, the formulation is stable when a measured feature of the formulation (e.g., turbidity, structural integrity and concentration of the protein and polymer components, activity of the protein) remains within at least about 20%, at least about 15%, at least about 10%, or at least about 5% of the originally measured level at an initial time point. In some embodiments, the formulation (or therapeutically acceptable composition) is storage stable with respect to at least one of the following functional properties: turbidity, percent impurities, concentration of intact protein, concentration of intact polymer, activity of protein (e.g., inhibition of VEGF-A binding to VEGFR). In some embodiments, “impurities” refer to product related impurities such as degraded, aggregated, unconjugated (e.g., where the product of interest is conjugated), or modified proteins. In some embodiments, product unrelated impurities such as Host Cell Protein, endotoxin, Host cell DNA, are not considered “impurities” as defined herein. In some embodiments, the formulation is stable (e.g., less than about 5% or about 10% total impurities, and / or IC50for competitive binding to VEGF bound to a VEGFR that is no more than about 10% or about 15% lower than a control formulation of the conjugate only) for at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 9 months, at least about 12 months, at least about 16 months, at least about 20 months, at least about 24 months, at least about 36 months, at least about 48 months, or longer, or optionally, it is stable for a period of time in a range defined by any two of the preceding values (e.g., 1-48 months, 1-24 months, 1-12 months, 1-6 months, 3-9 months, 4-12 months, 12-24 months, 12-36 months, etc.), when stored at about 5°C. In some embodiments, the formulation is stable (e.g., less than about 5% or about 10% total impurities, and / or IC50 for competitive binding to VEGF bound to a VEGFR that is no more than about 10% or about 15% lower than a control formulation of the conjugate only) for at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 9 months, at least about 12 months, at least about 16 months, at least about 20 months, at least about 24 months, or longer, or optionally, it is stablefor a period of time in a range defined by any two of the preceding values (e.g., 1-24 months, 1-20 months, 1-12 months, 1-6 months, 3-9 months, 4-12 months, 12-24 months, etc.), when stored at about 25°C. In some embodiments, the formulation is stable (e.g., less than about 5% or about 10% total impurities, and / or IC50 for competitive binding to VEGF bound to a VEGFR that is no more than about 10% or about 15% lower than a control formulation of the conjugate only) for at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 9 months, at least about 12 months, at least about 16 months, at least about 20 months, at least about 24 months, at least about 36 months, or longer, or optionally, it is stable for a period of time in a range defined by any two of the preceding values (e.g., 1-12 months, 1-6 months, 3-9 months, 4-12 months, etc.), when stored at room temperature. In some embodiments, the formulation is stable for, for about, or for at least 3 months when stored at about 5°C. In some embodiments, the formulation is stable for, for about, or for at least 6 months when stored at about 5°C. In some embodiments, the formulation is stable for, for about, or for at least 9 months when stored at about 5°C. In some embodiments, the formulation is stable for, for about, or for at least 12 months when stored at about 5°C. In some embodiments, the formulation is stable for, for about, or for at least 24 months when stored at about 5°C. In some embodiments, the formulation is stable for, for about, or for at least 3 months when stored at about 25°C. In some embodiments, the formulation is stable for, for about, or for at least 6 months when stored at about 25°C. In some embodiments, the formulation is stable for, for about, or for at least 9 months when stored at about 25°C. In some embodiments, the formulation is stable for, for about, or for at least 12 months when stored at about 25°C.

[0237] In some embodiments, the formulation (or therapeutically acceptable composition) is storage stable with respect to at least one of the following measures: color and clarity by visual inspection; aggregation or degradation of the unconjugated protein and / or the conjugated protein at different temperature as measured by size exclusion chromatography (SEC-HPLC), ion-exchanger chromatography (IEX-HPLC), SEC with Multi-angle light scattering (MALS) detector, or a Tandem HPLC method that allows monitoring of the stability of either the conjugate or free protein population separately; potency; and maintenance of the percent unconjugated protein (e.g., percent composition or % of the total molar amount) in the formulation.

[0238] In some embodiments, the formulation or composition comprises a mixture, by mass weight concentration, of 5% to 10% unconjugated protein (e.g. unconjugated antibody or unconjugated fusion construct) with the remainder comprising the conjugated protein (e.g., conjugated antibody or conjugated fusion construct). In some embodiments, the amount of the unconjugated protein relative to the total amount of unconjugated and conjugated protein in the formulation is expressed as a percentage by mass weight concentration when the unconjugated protein and the protein portion of the conjugated protein has the same or similar (e.g., within about 10% of each other) molecular weight. In some embodiments, the composition comprises a mixture, by mass weight concentration of 5% to 15% unconjugated protein (e.g. unconjugated antibody) with the remainder comprising the conjugated protein. In some embodiments, the composition comprises a mixture of the two proteins (e.g., antibodies and / or fusion constructs) of the same molecular weight, of which 15% to 25% unconjugated protein (e.g. unconjugated antibody) by mass weight concentration with the remainder comprising the conjugated protein. In some embodiments, the composition comprises a mixture of two antibodies of the same molecular weight, of which 15-25% of unconjugated protein (e.g. unconjugated antibody) concentration in mass weight concentration (e.g. Gram / liter) of the total protein concentration in the mixture. In some embodiments, the composition comprises a mixture of the two proteins (e.g., antibodies and / or fusion constructs) by mass weight concentration of 25% to 35% unconjugated protein (e.g. unconjugated antibody) with the remainder comprising the conjugated protein. In some embodiments, the composition comprises a mixture of the two proteins (e.g., antibodies and / or fusion constructs) by mass weight concentration of 35% to 45% unconjugated protein (e.g. unconjugated antibody) with the remainder comprising the conjugated protein. In some embodiments, the composition comprises a mixture of the two proteins (e.g., two antibodies) by mass weight concentration of 5% to 20% unconjugated protein (e.g. unconjugated antibody) with the remainder comprising the conjugated protein. In some embodiments, the composition comprises a mixture of the two proteins (e.g., two antibodies) by mass weight concentration of 5% to 25% unconjugated protein (e.g. unconjugated antibody) with the remainder comprising the conjugated protein. In some embodiments, the composition comprises a mixture of the two proteins (e.g., antibodies and / or fusion constructs) by mass weight concentration of 5% to 30% unconjugated protein (e.g. unconjugated antibody) with the remainder comprising theconjugated protein. In some embodiments, the composition comprises a mixture of the two proteins (e.g., two antibodies) by mass weight concentration of 5% to 35% unconjugated protein (e.g., unconjugated antibody) with the remainder comprising the conjugated protein. In some embodiments, the composition comprises a mixture of the two proteins (e.g., antibodies and / or fusion constructs) by mass weight concentration of 5% to 40% unconjugated protein (e.g., unconjugated antibody) with the remainder comprising the conjugated protein. In some embodiments, the composition comprises a mixture of the two proteins (e.g., two antibodies) by mass weight concentration of 5% to 50% unconjugated protein (e.g., unconjugated antibody) with the remainder comprising the conjugated protein. In some embodiments, the composition comprises a mixture of the two proteins (e.g., antibodies and / or fusion constructs) by mass weight concentration of 5% to 55% unconjugated protein (e.g., unconjugated antibody) with the remainder comprising the conjugated protein.

[0239] In some embodiments, the composition or formulation comprises a mixture of the two proteins (e.g., antibodies and / or fusion constructs) by mass weight concentration of 5% to 55% unconjugated protein (e.g., unconjugated antibody or unconjugated fusion construct) with the remainder comprising the conjugated protein (e.g., the conjugated antibody or conjugated fusion construct). In some embodiments, the composition comprises a mixture of the two proteins (e.g., two antibodies) by mass weight concentration of 5% to 60% unconjugated protein (e.g., unconjugated antibody) with the remainder comprising the conjugated protein. In some embodiments, the composition comprises a mixture of the two proteins (e.g., antibodies and / or fusion constructs) by mass weight concentration of 5% to 65% unconjugated protein (e.g., unconjugated antibody) with the remainder comprising the conjugated protein. In some embodiments, the composition comprises a mixture of the two proteins (e.g., two antibodies) by mass weight concentration of 5% to 70% unconjugated protein (e.g., unconjugated antibody) with the remainder comprising the conjugated protein. In some embodiments, the composition comprises a mixture of the two proteins (e.g., antibodies and / or fusion constructs) by mass weight concentration of 5% to 75% unconjugated protein (e.g., unconjugated antibody) with the remainder comprising the conjugated protein. In some embodiments, the composition comprises a mixture of the two proteins (e.g., two antibodies) by mass weight concentration of 5% to 80% unconjugated protein (e.g., unconjugated antibody) with the remainder comprising the conjugated protein. In someembodiments, the composition comprises a mixture of the two proteins (e.g., antibodies and / or fusion constructs) by mass weight concentration of 5% to 85% unconjugated protein (e.g., unconjugated antibody) with the remainder comprising the conjugated protein. In some embodiments, the composition comprises a mixture of the two proteins (e.g., antibodies and / or fusion constructs) by mass weight concentration of 5% to 90% unconjugated protein (e.g., unconjugated antibody) with the remainder comprising the conjugated protein.

[0240] In some embodiments, the percent of the conjugated to unconjugated protein (e.g., % total molar amount) is calculated by (1) measuring the conjugated protein and unconjugated protein in mg / mL; (2) converting the mg / mL values of the conjugated protein and unconjugated protein into a molecular weight measured in kDa; and (3) dividing the molecular weight of each of the conjugated protein and unconjugated protein by the total molecular weight of the conjugated protein and unconjugated protein in the composition, and multiplied by 100 to achieve a percent of the total molar amount for each.

[0241] Any of the formulations and compositions provided herein, in some embodiments, can be defined as a percent composition (e.g., in mass weight concentration) of one component relative to the total mass weight concentration of the proteins (e.g., excluding any contribution of a polymer that may be conjugated thereto) in the composition. In some embodiments, a formulation or composition defined in % total molar amount of the second protein (e.g., the unconjugated protein) can be defined in percent composition (e.g., in mass weight concentration) of the second protein relative to the total mass weight concentration of the first and second proteins, given the relevant molecular weight of each protein. In some embodiments, percent composition is measured in mass weight concentration (in other words, gram per liter or milligram per milliliter) of the free protein relative to the total mass weight concentration of the proteins (e.g., excluding any contribution of a polymer that may be conjugated thereto) in the mixture.

[0242] Provided herein is a therapeutically acceptable composition comprising: a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the percent composition of the second protein relative to the total protein mass weight concentration of the first protein and the second protein in the composition is about 1% or more (e.g., about 5-93%, 15-25%, 25-35%, 25-40%, etc.),wherein the composition has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein.

[0243] Also provided is a therapeutically acceptable composition comprising: a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the percent composition of the second protein relative to the total protein mass weight concentration of the first protein and the second protein in the composition is about 1% or more (e.g., about 5-93%, 15-25%, 25-35%, 25-40%, etc.), wherein the composition has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein, wherein the composition has a reduced viscosity and / or an enhanced injectability compared to a reference composition comprising the conjugate, wherein the first protein of the conjugate is present in the reference composition at the total mass weight concentration of the first and second proteins in the composition. In some embodiments, the reference composition is one that includes the first protein of the conjugate at the total mass weight concentration and effectively does not include the second protein (or includes the second protein at a percent composition relative to the total mass weight concentration of the first protein and the second protein in the reference composition of less than 1%), but is otherwise the same as the composition for which it serves as a reference.

[0244] Also provided is a therapeutically acceptable composition comprising a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the percent composition of the second protein relative to the total protein mass weight concentration of the first protein and the second protein in the composition is about 1% or more (e.g., about 5-93%, 15-25%, 25-35%, 25-40%, etc.), wherein the composition has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein, wherein the composition has a reduced turbidity compared to a reference composition comprising the second protein at the percent composition relative to the total protein mass weight concentration of the first protein and the second protein in the composition, at a pH about the same as (e.g., within 0.1, 0.15, 0.2, 0.3, 0.4, or 0.5 pH units of) the pI of the second protein. In some embodiments, the composition has a reduced turbidity compared to a reference composition comprising the second protein at the percent compositionrelative to the total protein mass weight concentration of the first protein and the second protein in the composition. In some embodiments, the reference composition includes the first protein conjugated to a phosphorylcholine-containing polymer and the second protein that is not conjugated to a phosphorylcholine-containing polymer; and the pharmaceutically acceptable carrier, wherein the percent composition of the second protein relative to the total protein mass weight concentration of the first protein and the second protein in the reference composition is the same as the percent composition of the second protein in the therapeutically acceptable composition, and the pH is about the same as (e.g., within 0.05, 0.1, 0.15, 0.2, 0.3, 0.4, or 0.5 pH units of) the pI of the second protein. In some embodiments, the composition has a reduced turbidity compared to a reference composition comprising the second protein at the percent composition relative to the total protein mass weight concentration of the first protein and the second protein in the composition.

[0245] Provided herein is a therapeutically acceptable composition comprising: a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the difference between the pI of the second protein and the pH of the formulation is selected to be greater than the minimum difference between the pI of the second protein and the pH for a reference formulation comprising: the conjugate comprising the first protein conjugated to the phosphorylcholine-containing polymer; the second protein that is not conjugated to the phosphorylcholine-containing polymer; and the pharmaceutically acceptable carrier, wherein the percent composition of the second protein relative to the total protein mass weight concentration of the first protein and the second protein in the composition in the composition is higher than the percent composition of the second protein relative to the total protein mass weight concentration of the first protein and the second protein in the composition in the reference composition, wherein the reference composition is substantially free of turbidity.

[0246] Provided herein is a therapeutically acceptable composition comprising: a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the percent composition of the second proteinrelative to the total protein mass weight concentration of the first protein and the second protein in the composition is about 1% or more (e.g., about 5-90%, 15-25%, 25-35%, 25-40%, etc.).

[0247] Further provided is a therapeutically acceptable composition comprising: a conjugate comprising a first protein conjugated to a polymer; and a second protein that is not conjugated to a polymer, wherein the second protein at a percent composition relative to the total protein mass weight concentration of the first protein and the second protein in the composition of about 1% or more (e.g., about 5-93%, 15-25%, 25-35%, 25-40%, etc.) has been combined with the conjugate, wherein the remainder of the total protein mass weight concentration comprises the first protein. In some embodiments, the composition has been prepared by combining the second protein at a percent composition of about 1% or more (e.g., about 5-93%, 15-25%, 25-35%, 25-40%, etc.) relative to the total protein mass weight concentration of the first protein and the second protein, with the conjugate such that the first protein at a percent composition of at a remainder of the total protein mass weight concentration. For example, for a total mass weight concentration of 50 mg / mL, the therapeutically acceptable composition can be prepared by combining an amount of the second protein that corresponds to 10 mg / mL in the final composition (percent composition of 20%) with an amount of the conjugate that corresponds to 40 mg / mL of the first protein (as the conjugate, excluding any contribution of the polymer to the mass weight concentration calculation) in the final composition.

[0248] In any embodiment herein, the second protein that is not conjugated to a polymer (e.g., a phosphorylcholine-containing polymer) can be referred to as the unconjugated protein, and the first protein that is conjugated to a polymer (e.g., a phosphorylcholine- containing polymer) can be referred to as the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) relative to the total protein mass weight concentration of the first protein and the second protein in the composition is between 5% and 6%, with the remainder comprising the conjugated protein. As used herein, “the remainder” denotes the portion of the total protein mass weight concentration of the composition (excluding any contribution of a polymer conjugated to the first protein to the mass weight concentration) that is not the unconjugated protein (e.g., the second protein), where the percent composition of the unconjugated protein (e.g., the second protein) and the remainder adds up to 100% of the total protein mass weight concentration.For example, where the percent composition of the unconjugated protein is between 5% and 6%, with the remainder comprising the conjugated protein, between 5% and 6% of the total mass weight concentration of the total protein concentration in the mixture is the unconjugated protein, and between 94% and 95% of the total mass weight concentration of the proteins in the mixture is the conjugated protein, where the percentages add up to 100%. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 1% and 2%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 3% and 4%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 6% and 7%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 7% and 8%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 8% and 9%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 9% and 10%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 10% and 11%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 11% and 12%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 12% and 13%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 13% and 14%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 14% and 15%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 15% and 16%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 16% and 17%, with the remainder comprising the conjugated protein. Insome embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 17% and 18%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 18% and 19%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 19% and 20%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 20% and 21%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 21% and 22%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 22% and 23%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 23% and 24%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 24% and 25%.

[0249] In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody or unconjugated fusion construct) is between 25% and 26%, with the remainder comprising the conjugated protein (e.g., conjugated antibody or conjugated fusion construct). In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 26% and 27%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 27% and 28%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 28% and 29%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 29% and 30%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 30% and 31%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 31% and 32%, with the remainder comprising theconjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 32% and 33%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 33% and 34%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 34% and 35%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 35% and 36%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 36% and 37%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 37% and 38%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 38% and 39%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 39% and 40%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 40% and 41%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 41% and 42%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 42% and 43%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 43% and 44%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 44% and 45%.

[0250] In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody or fusion construct) is between 45% and 46%, with the remainder comprising the conjugated protein (e.g., conjugated antibody or unconjugated fusion construct). In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 46% and 47%, with the remainder comprising theconjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 47% and 48%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 48% and 49%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 49% and 50%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 50% and 51%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 51% and 52%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 52% and 53%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 53% and 54%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 54% and 55%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 55% and 56%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 56% and 57%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 57% and 58%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 58% and 59%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 59% and 60%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 60% and 61%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 61% and 62%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein(e.g., unconjugated antibody) is between 62% and 63%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 63% and 64%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 64% and 65%.

[0251] In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody or unconjugated fusion construct) is between 65% and 66%, with the remainder comprising the conjugated protein (e.g., conjugated antibody or conjugate fusion construct). In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 66% and 67%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 67% and 68%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 68% and 69%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 69% and 70%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 70% and 71%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 71% and 72%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 72% and 73%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 73% and 74%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 74% and 75%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 75% and 76%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 76% and 77%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein(e.g., unconjugated antibody) is between 77% and 78%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 78% and 79%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 79% and 80%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 80% and 81%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 81% and 82%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 82% and 83%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 83% and 84%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 84% and 85%.

[0252] In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody or unconjugated fusion construct) is between 85% and 86%, with the remainder comprising the conjugated protein (e.g., conjugated antibody or conjugated fusion construct). In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 86% and 87%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 87% and 88%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 88% and 89%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 89% and 90%, with the remainder comprising the conjugated protein.

[0253] In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody or unconjugated fusion construct) is between 5% and 25%, with the remainder comprising the conjugated protein (e.g., conjugated antibody or conjugated fusion construct). In some embodiments, the percent composition of the unconjugated protein(e.g., unconjugated antibody) is between 1% and 5%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 5% and 12.5%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 5% and 15%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 5% and 17.5%. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 5% and 20%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 5% and 25%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 5% and 30%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 5% and 35%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 5% and 45%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 5% and 50%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 5% and 55%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 5% and 60%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 5% and 65%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 5% and 70%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 5% and 75%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 5% and 80%, with the remainder comprising the conjugated protein. In some embodiments, the percentcomposition of the unconjugated protein (e.g., unconjugated antibody) is between 5% and 85%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 5% and 90%.

[0254] In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody or unconjugated fusion construct) is between 10% and 12.5%, with the remainder comprising the conjugated protein (e.g., conjugated antibody or conjugated fusion construct). In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 10% and 15%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 10% and 17.5%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 10% and 20%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 15% and 25%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 10% and 25%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 10% and 30%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 10% and 35%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 10% and 45%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 10% and 50%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 10% and 55%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 10% and 60%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 10% and 65%, with the remainder comprising theconjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 10% and 70%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 10% and 75%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 10% and 80%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 10% and 85%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 10% and 90%.

[0255] In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody or unconjugated fusion construct) is between 12.5% and 15%, with the remainder comprising the conjugated protein (e.g., the conjugated antibody or conjugated fusion construct). In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 12.5% and 17.5%. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 17.5% and 22.5%. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 12.5% and 20%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 12.5% and 25%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 12.5% and 30%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 12.5% and 35%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 12.5% and 45%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 12.5% and 50%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 12.5% and 55%, with the remainder comprising the conjugated protein. In some embodiments, the percent compositionof the unconjugated protein (e.g., unconjugated antibody) is between 12.5% and 60%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 12.5% and 65%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 12.5% and 70%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 12.5% and 75%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 12.5% and 80%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 12.5% and 85%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 12.5% and 90%.

[0256] In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody or unconjugated fusion construct) is between 15% and 17.5%, with the remainder comprising the conjugated protein (e.g., conjugated antibody or fusion construct). In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 15% and 20%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 15% and 25%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 15% and 30%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 15% and 35%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 15% and 45%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 15% and 50%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 15% and 55%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein(e.g., unconjugated antibody) is between 15% and 60%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 15% and 65%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 15% and 70%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 15% and 75%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 15% and 80%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 15% and 85%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 15% and 90%.

[0257] In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody or unconjugated fusion construct) is between 20% and 25%, with the remainder comprising the conjugated protein (e.g., conjugated antibody or fusion construct). In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 25% and 30%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 25% and 35%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 25% and 45%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 25% and 50%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 25% and 55%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 25% and 60%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 25% and 65%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein(e.g., unconjugated antibody) is between 25% and 70%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 25% and 75%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 25% and 80%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 25% and 85%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is between 25% and 90%, with the remainder comprising the conjugated protein.

[0258] In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody or unconjugated fusion construct) is 1%, with the remainder comprising the conjugated protein (e.g., conjugated antibody or conjugated fusion construct). In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody or unconjugated fusion construct) is 5%, with the remainder comprising the conjugated protein (e.g., conjugated antibody or conjugated fusion construct). In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 6%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 7%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 8%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 9%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 10%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 11%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 12%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 13%, with the remainder comprising the conjugated protein. In someembodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 14%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 15%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 16%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 17%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 18%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 19%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 20%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 21%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 22%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 23%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 24%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 25%.

[0259] In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody or unconjugated fusion construct) is 26%, with the remainder comprising the conjugated protein (e.g., conjugated antibody or conjugated fusion construct). In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 27%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 28%, with the remainder comprising the conjugated protein. In someembodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 29%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 30%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 31%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 32%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 33%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 34%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 35%.

[0260] In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody or unconjugated fusion construct) is 36%, with the remainder comprising the conjugated protein (e.g., conjugated antibody or fusion construct). In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 37%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 38%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 39%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 40%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 41%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 42%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 43%, with the remainder comprising the conjugated protein. In someembodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 44%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 45%.

[0261] In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody or unconjugated fusion construct) is 46%, with the remainder comprising the conjugated protein (e.g., conjugated antibody or conjugated fusion construct). In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 47%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 48%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 49%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 50%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 51%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 52%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 53%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 54%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 55%.

[0262] In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody or unconjugated fusion construct) is 56%, with the remainder comprising the conjugated protein (e.g., conjugated antibody or conjugated fusion construct). In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 57%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugatedantibody) is 58%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 59%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 60%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 51%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 62%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 63%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 64%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 65%.

[0263] In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody or unconjguated fusion construct) is 66%, with the remainder comprising the conjugated protein (e.g., conjugated antibody or conjugated fusion construct). In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 67%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 68%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 69%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 70%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 11%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 72%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugatedantibody) is 73%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 74%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 75%.

[0264] In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody or unconjugated) is 76%, with the remainder comprising the conjugated protein (e.g., conjugated antibody or conjugated fusion construct). In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 77%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 78%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 79%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 80%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 81%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 82%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 83%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 84%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 85%.

[0265] In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody or unconjugated fusion construct) is 86%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 87%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein(e.g., unconjugated antibody) is 88%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 89%, with the remainder comprising the conjugated protein. In some embodiments, the percent composition of the unconjugated protein (e.g., unconjugated antibody) is 90%.

[0266] In some embodiments, the ratio of the molecular weight of the unconjugated protein (e.g., second protein that is not conjugated to a phosphorylcholine-containing polymer) to the polymer in the formulation can be any suitable ratio. In some embodiments, the ratio is at most about 1:2, e.g., at most about 1:3, at most about 1:4, at most about 1:5, at most about 1:6, at most about 1:7, at most about 1:8, at most about 1:9 or at most about 1:10, or a ratio in a range defined by any two of the preceding values (e.g., 1:2-1:10, 1:3-1:8, 1:4-1:6). In some embodiments, the ratio is between about 1:4 and 1:6. In some embodiments, the ratio is about 1:5.33.

[0267] The protein conjugated to the phosphorylcholine-containing polymer and the unconjugated protein can each be any suitable protein. In some embodiments, the first protein (e.g., the protein conjugated to the phosphorylcholine-containing polymer) and the second protein (e.g., the protein that is not conjugated to a phosphorylcholine-containing polymer) have the same activity or function (e.g., bind the same epitope, inhibit the same target, catalyze the same reaction, etc.). In some embodiments, the first protein (e.g., the protein conjugated to the phosphorylcholine-containing polymer) and the second protein (e.g., the protein that is not conjugated to a phosphorylcholine-containing polymer) are the same protein. In some embodiments, the first protein (e.g., the protein conjugated to the phosphorylcholine- containing polymer) and the second protein (e.g., the protein that is not conjugated to a phosphorylcholine-containing polymer) are at least about 85%, at least about 90%, at least about 95%, at least about 97%, at least about 98%, at least about 99%, or about 100%, or by a percentage in a range defined by any two of the preceding values (e.g., 85-100%, 90-99%, 90- 95%, etc.) identical to each other in amino acid sequence. In some embodiments, where the first and second proteins each include two or more polypeptide chains, each of the corresponding polypeptide chains can independently have any of the noted sequence identity. In some embodiments, the second protein is or comprises a derivative of the first protein (e.g., the protein portion without the polymer). In some embodiments, the second protein isalkylated form of the first protein (e.g., the protein portion without the polymer). For example, the second protein is an iodoacetamide (IAM)- or N-ethylmaleimide (NEM)-treated form of the first protein (e.g., the protein portion without the polymer).

[0268] In some embodiments, the first protein (e.g., the protein conjugated to the phosphorylcholine-containing polymer) has a molecular weight (based on the protein portion) of about 5 kDa or more, about 10 kDa or more, about 15 kDa or more, about 25 kDa or more, about 50 kDa or more, about 75 kDa or more, about 100 kDa or more, about 125 kDa or more, about 150 kDa or more, about 175 kDa or more, about 200 kDa or more, about 250 kDa or more, about 300 kDa or more, or a molecular weight in a range defined by any two of the preceding values (e.g., 50-300 kDa, 100-300 kDa, 100-200 kDa, 150-250 kDa, etc.). In some embodiments, the first protein (e.g., the protein conjugated to the phosphorylcholine- containing polymer) has a molecular weight (based on the protein portion) of about 150 kDa. In some embodiments, the first protein (e.g., the protein conjugated to the phosphorylcholine- containing polymer) has a molecular weight of about 200 kDa. In some embodiments, the second protein (e.g., the unconjugated protein or protein that is not conjugated to a phosphorylcholine-containing polymer) has a molecular weight of about 50 kDa or more, e.g., about 75 kDa or more, about 100 kDa or more, about 125 kDa or more, about 150 kDa or more, about 175 kDa or more, about 200 kDa or more, about 250 kDa or more, about 300 kDa or more, or a molecular weight in a range defined by any two of the preceding values (e.g., 50- 300 kDa, 100-300 kDa, 100-200 kDa, 150-250 kDa, etc.). In some embodiments, the second protein (e.g., the unconjugated protein or protein that is not conjugated to a phosphorylcholine- containing polymer) has a molecular weight of about 150 kDa. In some embodiments, the second protein (e.g., the unconjugated protein or protein that is not conjugated to a phosphorylcholine-containing polymer) has a molecular weight of about 200 kDa.

[0269] In some embodiments, one or both of the first protein (e.g., the protein conjugated to the phosphorylcholine-containing polymer) and the second protein (e.g., the unconjugated protein or protein that is not conjugated to a phosphorylcholine-containing polymer) is a therapeutic protein. In some embodiments, at least one of the first protein (e.g., the protein conjugated to the phosphorylcholine-containing polymer) and the second protein (e.g., the unconjugated protein or protein that is not conjugated to a phosphorylcholine- containing polymer) is a therapeutic protein that is FDA approved as of May 2023. In someembodiments, one or both of the first protein (e.g., the protein conjugated to the phosphorylcholine-containing polymer) and the second protein (e.g., the unconjugated protein or protein that is not conjugated to a phosphorylcholine-containing polymer) is an antibody (e.g., therapeutic antibody). Any suitable antibody can be used in the formulations. In some embodiments, one or both of the first protein (e.g., the protein conjugated to the phosphorylcholine-containing polymer) and the second protein (e.g., the unconjugated protein or protein that is not conjugated to a phosphorylcholine-containing polymer) is a fusion construct. Any suitable fusion construct can be used in the formulations. The antibody or fusion construct of any of the formulation or composition herein may or may not include a C-terminal lysine. “Fusion protein” and “fusion construct” are used interchangeably herein.

[0270] In some embodiments, one or both of the first protein (e.g., the protein conjugated to the phosphorylcholine-containing polymer) and the second protein (e.g., the unconjugated protein or protein that is not conjugated to a phosphorylcholine-containing polymer) is an anti-VEGF-A antibody, a fusion construct comprising a VEGF Trap fused to the heavy chain of an anti-IL-6 antibody, an anti-IL-6 antibody, a fusion construct comprising a PDGFR extracellular trap fused to the heavy chain of an anti-VEGF-A antibody, a VEGF trap-Fc fusion protein, an anti-HTRA1 antibody, or an anti-complement factor D (CFD) antibody. In some embodiments, both of the first protein (e.g., the protein conjugated to the phosphorylcholine-containing polymer) and the second protein (e.g., the unconjugated protein or protein that is not conjugated to a phosphorylcholine-containing polymer) are an anti- VEGF-A antibody. In some embodiments, both of the first protein (e.g., the protein conjugated to the phosphorylcholine-containing polymer) and the second protein (e.g., the unconjugated protein or protein that is not conjugated to a phosphorylcholine-containing polymer) are a fusion construct comprising a VEGF Trap fused to the heavy chain of an anti-IL-6 antibody. In some embodiments, the anti-VEGF-A antibody is a full-length antibody. In some embodiments, the anti-VEGF-A antibody is or includes an anti-VEGF-A Fab fragment. In some embodiments, the anti-VEGF-A antibody is OG1950, e.g., as described herein and in US patent publication no.2017 / 0190766, the entirety of which is incorporated herein by reference. In some embodiments, the anti-VEGF-A antibody is selected from bevacizumab, ranibizumab, brolucizumab, or faricimab. In some embodiments, the VEGF trap-Fc fusion protein is aflibercept. In some embodiments, the fusion construct is OG2072, e.g., as described hereinand in US patent publication no. 2019 / 0270806, the entirety of which is incorporated herein by reference. In some embodiments, the first protein comprises any one of the amino sequences or a combination thereof set forth in Tables 0.1, 0.2, 0.3, 0.4, 0.5, 0.6 and 0.7, and Figs. 12-17, 54, 55A, and 55B. In some embodiments, the second protein comprises any one of the amino sequences or a combination thereof set forth in Tables 0.1, 0.2, 0.3, 0.4, 0.5, 0.6 and 0.7, and Figs.12-17, 54, 55A, and 55B. In some embodiments, the first protein comprises any one of the amino sequences or a combination thereof set forth in Tables 0.1, 0.2, 0.3, 0.4, 0.5, 0.6 and 0.7, and Figs.12-17, 54, 55A, and 55B, and the second protein comprises any one of the amino sequences or a combination thereof set forth in Tables 0.1, 0.2, 0.3, 0.4, 0.5, 0.6 and 0.7, and Figs.12-17, 54, 55A, and 55B. In some embodiments, the first protein comprises any one of the amino sequences or a combination thereof set forth in SEQ ID NOs: 1-6, 15- 133, and 156-161. In some embodiments, the first protein comprises any one of the amino sequences or a combination thereof set forth in SEQ ID NOs: 1-6, 15-133, and 156-161. In some embodiments, the first protein comprises any one of the amino sequences or a combination thereof set forth in SEQ ID NOs: 1-6, 15-133, and 156-161, and the second protein comprises any one of the amino sequences or a combination thereof set forth in SEQ ID NOs: 1-6, 15-133, and 156-161. In some embodiments, the amino acid sequences of the first protein and second protein are paired as they are arranged in Tables 0.1, 0.2, 0.3, 0.4, 0.5, 0.6 and 0.7, and Figs.12-17, 54, 55A, and 55B).

[0271] In some embodiments, the conjugated and unconjugated proteins are antibodies, and one or both antibodies are therapeutic. In some embodiments, one or both antibodies are the same or variants of each other, wherein one antibody is conjugated to a polymer at a cysteine outside a variable region of the antibody and the other protein is unconjugated. In some embodiments, both antibodies are therapeutic antibodies. In some embodiments, at least one of the antibodies is a therapeutic antibody that is FDA approved as of May 2023. In some embodiments, both antibodies are therapeutic for the treatment of an eye disorder. In some embodiments, both antibodies (conjugated and unconjugated) share at least the same CDR sequences in the heavy and light chains. In some embodiments, both antibodies (conjugated and unconjugated) have at least one (e.g., 1, 2, 3, 4, 5, or 6) CDR in the heavy or light chains that is different from each other. In some embodiments, the composition comprises (a) an antibody conjugate comprising an anti-VEGF-A antibody and aphosphorylcholine-containing polymer wherein the polymer is covalently bonded to the antibody at a cysteine outside a variable region of the antibody; and (b) an unconjugated anti- VEGF-A antibody.

[0272] In some embodiments, the anti-VEGF-A antibody of the antibody conjugate comprises a light chain and a heavy chain, said heavy chain comprising an Fc region. In some embodiments, a cysteine of the antibody conjugate is in the Fc region of the heavy chain. In some embodiments, the anti-VEGF-A antibody of the antibody conjugate is an immunoglobulin G (IgG).

[0273] In some embodiments, the anti-VEGF-A antibody heavy chain of the antibody conjugate or the unconjugated antibody comprises: CDRH1: GYDFTHYGMN (SEQ ID NO: 9), CDRH2: WINTYTGEPTYAADFKR (SEQ ID NO: 10), and CDRH3: YPYYYGTSHWYFDV (SEQ ID NO: 11), and the anti-VEGF-A light chain of the antibody conjugate or the unconjugated antibody comprises CDRL1: SASQDISNYLN (SEQ ID NO: 12), CDRL2: FTSSLHS (SEQ ID NO: 13), and CDRL3: QQYSTVPWT (SEQ ID NO: 14). In some embodiments, the anti-VEGFA heavy chain isotype of the antibody conjugate or the unconjugated antibody is human IgG1. In some embodiments, the heavy chain constant domain of the anti-VEGF-A antibody of the antibody conjugate or the unconjugated antibody has one or more mutations relative to the constant domain of human IgG1 to modulate effector function. T

[0274] In some embodiments, mutations of the antibody conjugate (and / or unconjugated antibody) may be to one or more of the following amino acid positions (EU numbering): E233X, L234X, L235X, G236X, G237X, A327X, A330X, and P331X wherein X is any natural or unnatural amino acid. In some embodiments, mutations of the antibody conjugate (and / or unconjugated antibody) are selected from the group consisting of (EU numbering): E233P, L234V, L234A, L235A, G237A, A327G, A330S, and P331S. In some embodiments, mutations of the antibody conjugate (and / or unconjugated antibody) comprise: L234A, L235A, and G237A (EU numbering). In some embodiments, the cysteine of the antibody conjugate (and / or unconjugated antibody) is in the anti-VEGF-A antibody heavy chain and is Q347C (EU numbering) or L443C (EU numbering).

[0275] In some embodiments, the sequence of the anti-VEGF-A antibody heavy chain of the antibody conjugate or the unconjugated antibody is SEQ ID NO: 1 (with or withoutthe C-terminal lysine) and the sequence of the anti-VEGF-A light chain of the antibody conjugate or the unconjugated antibody is SEQ ID NO: 2. In some embodiments, the sequence of the anti-VEGF-A antibody heavy chain of the antibody conjugate or the unconjugated antibody is at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90-97%, etc.) identical to SEQ ID NO: 1, and the sequence of the anti-VEGF-A light chain of the antibody conjugate or the unconjugated antibody is at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90-97%, etc.) identical to SEQ ID NO: 2. In some embodiments, the cysteine of the antibody conjugate or the unconjugated antibody is L443C (EU numbering).

[0276] In some embodiments, the solution pH of the composition depends on the isoelectric point of a desired protein, wherein the solution pH is at least 1-2 pH units away from the desired protein’s isoelectric point.

[0277] In some embodiments, a composition comprising a first antibody and a second antibody is provided. The first antibody is conjugated to a phosphorylcholine- containing polymer, wherein the polymer is covalently bonded to the first antibody at a cysteine outside a variable region of the first antibody; wherein the second antibody is not conjugated to the phosphorylcholine-containing polymer.

[0278] In some embodiments, the first antibody comprises CDRH1: GYDFTHYGMN (SEQ ID NO: 9), CDRH2: WINTYTGEPTYAADFKR (SEQ ID NO: 10), and CDRH3: YPYYYGTSHWYFDV (SEQ ID NO: 11), CDRL1: SASQDISNYLN (SEQ ID NO: 12), CDRL2: FTSSLHS (SEQ ID NO: 13), and CDRL3: QQYSTVPWT (SEQ ID NO: 14). In some embodiments, mutations of the first antibody are to one or more of the following amino acid positions (EU numbering): E233X, L234X, L235X, G236X, G237X, A327X, A330X, and P331X wherein X is any natural or unnatural amino acid. In some embodiments, mutations of the first antibody are selected from the group consisting of (EU numbering): E233P, L234V, L234A, L235A, G237A, A327G, A330S, and P331S. In some embodiments, the following mutations of the first antibody: L234A, L235A, and G237A (EU numbering). In some embodiments, the cysteine of the first antibody is in the anti-VEGF-A antibody heavy chain and is Q347C (EU numbering) or L443C (EU numbering). In some embodiments, thesequence of the anti-VEGF-A antibody heavy chain of the first antibody is SEQ ID NO: 1 (with or without the C-terminal lysine) and the sequence of the anti-VEGF-A light chain of the first antibody is SEQ ID NO: 2. Any or all the foregoing embodiments in this paragraph could apply to the second antibody. OG1950 denotes an anti-VEGF-A antibody having a heavy chain of SEQ ID NO:1 (with or without the C-terminal lysine) and a light chain of SEQ ID NO: 2.

[0279] In some embodiments, the cysteine of the first antibody is L443C (EU numbering).

[0280] In some embodiments, the first and second antibody have the same CDRs (1, 2, 3, 4, 5, or all 6), same VH, VL, same VH and same VL, and / or same HC and LC, with the only difference being one is conjugated to a polymer and one is not conjugated to the polymer.

[0281] In some embodiments, one or both of the first protein (e.g., the protein conjugated to the phosphorylcholine-containing polymer) and the second protein (e.g., the unconjugated protein or protein that is not conjugated to a phosphorylcholine-containing polymer) is an anti-complement factor D (CFD) antibody. In some embodiments, the anti-CFD antibody includes a heavy chain having any 3 of the heavy chain CDRs in any one of SEQ ID NOs: 15-47, and a light chain having any 3 of the light chain CDRs in any one of SEQ ID NOs: 48-80. In some embodiments, the anti-CFD antibody includes a heavy chain having any 3 of the heavy chain CDRs in any one of SEQ ID NOs: 15-47, and a light chain having any 3 of the light chain CDRs in any one of SEQ ID NOs: 48-80, as they are paired in Table 0.1. In some embodiments, the anti-CFD antibody includes a heavy chain variable region having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90-97%, etc.) identical to any one of SEQ ID NOs: 15-47, and a light chain variable region having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90-97%, etc.) identical to any one of SEQ ID NOs: 48-80. In some embodiments, the anti-CFD antibody includes a heavy chain variable region having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or apercentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90- 97%, etc.) identical to any one of SEQ ID NOs: 15-47, and a light chain variable region having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90-97%, etc.) identical to any one of SEQ ID NOs: 48-80, as they are paired in Table 0.1. Table 0.1

[0282] In some embodiments, the anti-CFD antibody includes a heavy chain variable region having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90-97%, etc.) identical to SEQ ID NO: 129 (or a heavy chain variable region thereof), and a light chain variable region having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90-97%, etc.) identical to SEQ ID NO: 130 (or a light chain variable region thereof). In some embodiments, the anti-CFD antibody heavy chain includes the sequence of SEQ ID NO: 129 (with or without the C-terminal lysine). In some embodiments, the CFD antibody light chain can comprise the sequence of SEQ ID NO: 130.

[0283] In some embodiments, one or both of the first protein (e.g., the protein conjugated to the phosphorylcholine-containing polymer) and the second protein (e.g., the unconjugated protein or protein that is not conjugated to a phosphorylcholine-containing polymer) is an anti-IL-6 antibody. In some embodiments, the anti-IL-6 antibody includes a heavy chain having any 3 of the heavy chain CDRs in any one of SEQ ID NOs: 81-89, and a light chain having any 3 of the light chain CDRs in any one of ID NOs: 90-92. In some embodiments, the anti-IL-6 antibody includes a heavy chain variable region having an aminoacid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90-97%, etc.) identical to any one of SEQ ID NOs: 81-89, and a light chain variable region having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90-97%, etc.) identical to any one of SEQ ID NOs: 90-92. Table 0.2Table 0.3

[0284] In some embodiments, one or both of the first protein (e.g., the protein conjugated to the phosphorylcholine-containing polymer) and the second protein (e.g., the unconjugated protein or protein that is not conjugated to a phosphorylcholine-containingpolymer) is a fusion construct comprising a VEGF Trap fused to the heavy chain of an anti- IL-6 antibody. In some embodiments, the fusion construct includes a heavy chain variable region having any 3 of the heavy chain CDRs (or all 3 heavy chain CDRs) in any one of SEQ ID NOs: 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, and 127, and a light chain variable region having any 3 of the light chain CDRs (or all 3 light chain CDRs) in any one of ID NOs: 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, and 128. In some embodiments, the fusion construct includes a heavy chain variable region having any 3 of the heavy chain CDRs (or all 3 heavy chain CDRs) in any one of SEQ ID NOs: 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, and 127, and a light chain variable region having any 3 of the light chain CDRs (or all 3 light chain CDRs) in any one of SEQ ID NOs: 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, and 128, as they are paired in Table 0.4. In some embodiments, the fusion construct includes a heavy chain variable region having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90-97%, etc.) identical to a heavy chain variable region in any one of SEQ ID NOs: 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, and 127, and a light chain variable region having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90-97%, etc.) identical to a light chain variable region in any one of SEQ ID NOs: 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, and 128. In some embodiments, the fusion construct includes a heavy chain variable region having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90-97%, etc.) identical to a heavy chain variable region in any one of SEQ ID NOs: 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, and 127, and a light chain variable region having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80- 100%, 85-95%, 90-97%, etc.) identical to a light chain variable region in any one of SEQ IDNOs: 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, and 128, as they are paired in Table 0.4. In some embodiments, the fusion construct includes a heavy chain having any 3 of the heavy chain CDRs in any one of SEQ ID NOs: 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, and 127, and a light chain having any 3 of the light chain CDRs in any one of ID NOs: 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, and 128. In some embodiments, the fusion construct includes a heavy chain having any 3 of the heavy chain CDRs in any one of SEQ ID NOs: 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, and 127, and a light chain having any 3 of the light chain CDRs in any one of SEQ ID NOs: 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, and 128, as they are paired in Table 0.4. In some embodiments, the fusion construct includes a heavy chain variable region having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90-97%, etc.) identical to any one of SEQ ID NOs: 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, and 127, and a light chain variable region having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90- 97%, etc.) identical to any one of SEQ ID NOs: 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, and 128. In some embodiments, the fusion construct includes a heavy chain variable region having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90- 97%, etc.) identical to any one of SEQ ID NOs: 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, and 127, and a light chain variable region having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90-97%, etc.) identical to any one of SEQ ID NOs: 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, and 128, as they are paired in Table 0.4. In some embodiments, the fusion construct includes: a heavy chain variable region having an amino acid sequence at least 80%, at least 85%, at least 90%, at least95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90-97%, etc.) identical to a heavy chain variable region in any one of SEQ ID NOs: 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, and 127, and heavy chain CDRs of the 3 heavy chain CDRs in the any one of SEQ ID NOs: 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, and 127; and a light chain variable region having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80- 100%, 85-95%, 90-97%, etc.) identical to a light chain variable region in any one of SEQ ID NOs: 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, and 128, and light chain CDRs of the 3 light chain CDRs in the any one of ID NOs: 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, and 128, where the SEQ ID NOs of the light chain and heavy chain are paired as they are in Table 0.4. In some embodiments, the fusion construct includes a heavy chain having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80- 100%, 85-95%, 90-97%, etc.) identical to any one of SEQ ID NOs: 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, and 127 (each with or without the C- terminal lysine), and a light chain having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90- 97%, etc.) identical to any one of SEQ ID NOs: 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, and 128, as they are paired in Table 0.4. In some embodiments, the fusion construct includes a heavy chain having an amino acid sequence of any one of SEQ ID NOs: 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, and 127 (each with or without the C-terminal lysine), and a light chain having an amino acid sequence of any one of SEQ ID NOs: 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, and 128, as they are paired in Table 0.4. Table 0.4

[0285] In some embodiments, the VEGF Trap comprises human VEGFR1 domain 2 and human VEGFR2 domain 3. In some embodiments, the VEGF Trap includes the amino acid sequence of SEQ ID NO:133. In some embodiments, the VEGF Trap comprises an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90-97%, etc.) identical to SEQ ID NO:133. Table 0.5

[0286] In some embodiments a fusion construct includes a heavy chain that includes a CDRH1 that is the CDRH1 in SEQ ID NO: 105; a CDRH2 that is the CDRH2 in SEQ ID NO: 105; a CDRH3 that is the CDRH3 in SEQ ID NO: 105; a CDRL1 that is the CDRL1 in SEQ ID NO: 106; a CDRL2 that is the CDRL2 in SEQ ID NO: 106; and a CDRL3 that is the CDRL3 in SEQ ID NO: 106. In some embodiments, the fusion construct includes a heavy chain comprising a complementarity determining region 1 (CDRH1): PFAMH (SEQ ID NO: 134), CDRH2: KISPGGSWTYYSDTVTD (SEQ ID NO: 135), and CDRH3: QAWGYYALDI (SEQ ID NO: 136); and a light chain comprising CDRL1: SASISVSYLY (SEQ ID NO: 137), CDRL2: DDSSLAS (SEQ ID NO: 138), and CDRL3: QQWSGYPYT (SEQ ID NO: 139). In some embodiments, the heavy chain comprises an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90- 97%, etc.) identical to SEQ ID NO:105, and the light chain comprises an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80- 100%, 85-95%, 90-97%, etc.) identical to SEQ ID NO:106. In some embodiments, the heavy chain comprises the amino acid sequence of SEQ ID NO:105 (with or without the C-terminal lysine), and the light chain comprises the amino acid sequence of SEQ ID NO:106. 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:105 (with or without the C-terminal lysine) and a light chain of SEQ ID NO:106.

[0287] In some embodiments, one or both of the first protein (e.g., the protein conjugated to the phosphorylcholine-containing polymer) and the second protein (e.g., the unconjugated protein or protein that is not conjugated to a phosphorylcholine-containing polymer) is a fusion construct comprising a PDGFR extracellular trap fused to the heavy chain of an anti-VEGF-A antibody. In some embodiments, the fusion construct includes a heavy chain variable region having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90-97%, etc.) identical to SEQ ID NO: 131, and a light chain variable region having an amino acid sequence at least80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90-97%, etc.) identical to SEQ ID NO: 132. In some embodiments, the heavy chain comprises the amino acid sequence of SEQ ID NO:131 (with or without the C-terminal lysine), and the light chain comprises the amino acid sequence of SEQ ID NO:132.

[0288] In some embodiments, one or both of the first protein (e.g., the protein conjugated to the phosphorylcholine-containing polymer) and the second protein (e.g., the unconjugated protein or protein that is not conjugated to a phosphorylcholine-containing polymer) is an anti-HTRA1 antibody. In some embodiments, the anti-HTRA1 antibody includes a heavy chain comprising a complementarity determining region 1 CDRH1: FYHVH (SEQ ID NO: SEQ ID NO:140), CDRH2: SIYTSGYTEYASALES (SEQ ID NO:141), and CDRH3: EGLQRVGVLDA (SEQ ID NO:142) or EGLQRVGVFDA (SEQ ID NO:143) or EGLQRVGVMDA (SEQ ID NO:144), and a light chain comprising a CDRL1: RSSQSLLDEAGETYLA (SEQ ID NO:145), CDRL2: EVSLLES (SEQ ID NO:146), and CDRL3: QQATYFPYT (SEQ ID NO:147). In some embodiments, the anti-HTRA1 antibody includes a heavy chain comprising a complementarity determining region 1 CDRH1: GFSLTFYH (SEQ ID NO: SEQ ID NO:148), CDRH2: IYTSGYT (SEQ ID NO:149), and CDRH3: AREGLQRVGVFDA (SEQ ID NO:150) or AREGLQRVGVMDA (SEQ ID NO:151) or AREGLQRVGVLDA (SEQ ID NO:152), and a light chain comprising a CDRL1: QSLLDEAGETY (SEQ ID NO:153), CDRL2: EV, and CDRL3: QQATYFPYT (SEQ ID NO:147). In some embodiments, the anti-HTRA1 antibody includes a heavy chain comprising a complementarity determining region 1 CDRH1: GFSLTFY (SEQ ID NO: SEQ ID NO:154), CDRH2: YTSGY (SEQ ID NO:155), and CDRH3: EGLQRVGVLDA (SEQ ID NO:142) or EGLQRVGVFDA (SEQ ID NO:143) or EGLQRVGVMDA (SEQ ID NO:144), and a light chain comprising a CDRL1: RSSQSLLDEAGETYLA (SEQ ID NO:145), CDRL2: EVSLLES (SEQ ID NO:146), and CDRL3: QQATYFPYT (SEQ ID NO:147). In some embodiments, the heavy chain comprises an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90-97%, etc.) identical to any one of the VH sequences set forth in Table 0.6, and the light chain comprises an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, atleast 99%, or about 100%, or a percentage in a range defined by any two of the preceding values (e.g., 80-100%, 85-95%, 90-97%, etc.) identical to the VL sequence set forth in Table 0.7. In some embodiments, the heavy chain comprises any one of the VH sequences set forth in Table 0.6, and the light chain comprises the VL sequence set forth in Table 0.7. Table 0.6Table 0.7

[0289] In some embodiments, the unconjugated protein (e.g., unconjugated antibody or unconjugated fusion construct) can have a conjugation cysteine or an engineered cysteine or a cysteine for conjugation that is blocked—thereby preventing polymer conjugation. In some embodiments, the unconjugated species within the composition is separate from the conjugated species by the fact that the conjugated species was subjected to a conjugation reaction and then unconjugated protein (e.g., unconjugated antibody or unconjugated fusion construct) was then added to the mix to create the composition. In some embodiments, the unconjugated protein does not have a non-native cysteine that can be conjugated to a polymer. In some embodiments, the conjugate includes a polymer conjugated to a cysteine, or free amino groups of the protein, e.g., using N-hydroxysuccinimide (NHS)esters. In some embodiments, the conjugate includes a polymer conjugated to ^-amine group of lysines, Į-amine group of N-terminal amino acids, and / or į-amine group of histidines.

[0290] In some embodiments, the formulation or therapeutically acceptable composition comprises KSI-301. In some embodiments, KSI-301 comprises OG1950 and its conjugated form with OG1802, OG1953. OG1950 denotes an anti-VEGF-A antibody having a heavy chain of SEQ ID NO:1 (with or without the C-terminal lysine) and a light chain of SEQ ID NO: 2. OG1802 may be bonded to the heavy chain of OG1950 at C443 (EU numbering) to form OG1953. In some embodiments, the first protein (e.g., the protein conjugated to the phosphorylcholine-containing polymer) is or comprises OG1950 conjugated with OG1802 (to form OG1953), and the second protein (e.g., the unconjugated protein or protein that is not conjugated to a phosphorylcholine-containing polymer) is or comprises OG1950. In some embodiments, the formulation or therapeutically acceptable composition comprises 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 (e.g., by molar amount or percent composition by total protein mass weight concentration), at pH about 4.5 to about 5.5. In some embodiments, the formulation or therapeutically acceptable composition comprises 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 (e.g., by molar amount or percent composition by total protein mass weight concentration), at about pH 5. In some embodiments, the formulation or therapeutically acceptable composition consists of, or consists 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 (e.g., by molar amount or percent composition by total protein mass weight concentration), at about pH 5. In some embodiments, the second protein is or comprises OG1950IAM (e.g., OG1950 treated with iodoacetamide). FIG. 58A depicts components of a non-limiting example of a formulation of the present disclosure, that can include, among other components, a mixture of an anti-VEGF-A antibody and an anti-VEGF-A antibody conjugate, as described herein.

[0291] In some embodiments, the formulation or therapeutically acceptable composition comprises KSI-501. In some embodiments, KSI-501 comprises fusion protein OG2072 and its conjugated form with OG1802, OG2074. 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:105 (with or without the C-terminal lysine) and a light chain of SEQ ID NO:106. OG1802 may be bonded to the heavy chain of OG2072 at C443 (EU numbering) to form OG2074. In some embodiments, the first protein (e.g., the protein conjugated to the phosphorylcholine-containing polymer) is or comprises OG2072 conjugated with OG1802 (to form OG2074), and the second protein (e.g., the unconjugated protein or protein that is not conjugated to a phosphorylcholine-containing polymer) is or comprises OG2072. In some embodiments, the formulation or therapeutically acceptable composition comprises about 50 to about 60 mM sodium acetate, polysorbate 20 (e.g., 0.025% polysorbate 20), sucrose (e.g., 4% sucrose), about 50 mg / mL (total protein concentration) of a mixture of OG2072 and OG2074, the mixture containing 20-40% OG2072 and 60-80% OG2074 (e.g., by molar amount or percent composition by total protein mass weight concentration), at pH about 4.5 to about 5.5. In some embodiments, the formulation or therapeutically acceptable composition comprises about 50 to about 60 mM sodium acetate, about 0.025% polysorbate 20, about 4% sucrose, about 50 mg / mL (total protein concentration) of a mixture of OG2072 and OG2074, the mixture containing 30% OG2072 and 70% OG2074 (e.g., by molar amount or percent composition by total protein mass weight concentration), at about pH 5 (e.g., about pH 4.8 to about pH 5). In some embodiments, the formulation or therapeutically acceptable composition consists of, or consists essentially of, about 50 to about 60 mM mM sodium acetate, about 0.025% polysorbate 20, about 4% sucrose, about 50 mg / mL (total protein concentration) of a mixture of OG2072 and OG2074, the mixture containing 30% OG2072 and 70% OG2074 (e.g., by molar amount or percent composition by total protein mass weight concentration), at about pH 5 (e.g., about pH 4.8 to about pH 5). In some embodiments, the formulation or therapeutically acceptable composition comprises about 50 mM sodium acetate, about 0.025% polysorbate 20, about 4% sucrose, about 50 mg / mL (total protein concentration) of a mixture of OG2072 and OG2074, the mixture containing 30% OG2072 and 70% OG2074 (e.g., by molar amount or percent composition by total protein mass weight concentration), at about pH 5. In some embodiments, the formulation or therapeutically acceptable composition consistsof, or consists essentially of, about 50 mM sodium acetate, about 0.025% polysorbate 20, about 4% sucrose, about 50 mg / mL (total protein concentration) of a mixture of OG2072 and OG2074, the mixture containing 30% OG2072 and 70% OG2074 (e.g., by molar amount or percent composition by total protein mass weight concentration), at about pH 5. In some embodiments, the formulation or therapeutically acceptable composition does not include sucrose. In some embodiments, the formulation or therapeutically acceptable composition does not include a sugar. In some embodiments, the formulation or therapeutically acceptable composition comprises about 50 to about 60 mM sodium acetate, about 0.025% polysorbate 20, about 50 mg / mL (total protein concentration) of a mixture of OG2072 and OG2074, the mixture containing 30% OG2072 and 70% OG2074 (e.g., by molar amount or percent composition by total protein mass weight concentration), at about pH 5 (e.g., about pH 4.8 to about pH 5), optionally where the formulation or therapeutically acceptable composition does not include sucrose. In some embodiments, the formulation or therapeutically acceptable composition consists of, or consists essentially of, 50-60 mM sodium acetate, about 0.025% polysorbate 20, about 50 mg / mL (total protein concentration) of a mixture of OG2072 and OG2074, the mixture containing 30% OG2072 and 70% OG2074 (e.g., by molar amount or percent composition by total protein mass weight concentration), at about pH 5 (e.g., about pH 4.8 to about pH 5), optionally where the formulation or therapeutically acceptable composition does not include sucrose. In some embodiments, the formulation or therapeutically acceptable composition includes a histidine acetate buffer, about 0.025% polysorbate 20, about 4% sucrose, about 50 mg / mL (total protein concentration) of a mixture of OG2072 and OG2074, the mixture containing 30% OG2072 and 70% OG2074 (e.g., by molar amount or percent composition by total protein mass weight concentration), at about pH 5.2 to about pH 6.2. FIGs. 58B and 58C depict components of a non-limiting examples of a formulation of the present disclosure, that can include, among other components, a mixture of a fusion construct and a fusion construct conjugate, as described herein.

[0292] In some embodiments, the polymer component of the conjugated protein has a molecular weight of about 100,000 Da or more, e.g., about 150,000 Da or more, about 200,000 Da or more, about 350,000 Da or more, about 400,000 Da or more, about 450,000 Da or more, about 500,000 Da or more, about 550,000 Da or more, about 600,000 Da or more, about 650,000 Da or more, about 700,000 or more, about 750,000 Da or more, about 800,000Da or more, about 850,000 Da or more, about 900,000 Da or more, about 950,000 Da or more, about 1,000,000 Da or more, or a molecular weight in a range defined by any two of the preceding values (e.g., 100,000-1,000,000 Da, 300,000-950,000 Da, 400,000-800,000 Da, 500,000-750.000 Da, 600,000-700,000 Da, 600,000-1,000,000 Da, etc.). In some embodiments, the polymer has a molecular weight of between about 700,000 to about 800,000 Da. In some embodiments, the polymer component of the conjugated protein is any of the polymers disclosed herein. In some embodiments, the polymer component of the conjugated protein is OG1801.

[0293] In some embodiments, the phosphorylcholine-containing polymer of the conjugatecomprises 2-(methacryloyloxyethyl)-2’-(trimethylammonium)ethyl phosphate (MPC) monomers as set forth below:

[0294] In some embodiments, the polymer of the conjugate has three or more arms or is synthesized with an initiator comprising 3 or more polymer initiation sites. In some embodiments, the polymer of the conjugate has 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 arms or is synthesized with an initiator comprising 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 polymer initiation sites. In some embodiments, the polymer of the conjugate has 9 arms or is synthesized with an initiator comprising 9 polymer initiation sites. In some embodiments, the polymer of the conjugate has a polydispersity value (PDI) of less than about 1.2.

[0295] In some embodiments, the polymer of the first protein that is conjugated (e.g., first antibody and / or conjugated antibody) has three or more arms or is synthesized withan initiator comprising 3 or more polymer initiation sites. In some embodiments, the polymer of the first protein that is conjugated (e.g., antibody) has 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 arms or is synthesized with an initiator comprising 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 polymer initiation sites. In some embodiments, the polymer of the first antibody has a polydispersity index (PDI) of less than about 1.2. In some embodiments, the polymer comprises MPC monomers, wherein first antibody comprises: an amino acid sequence of SEQ ID NO: 1 (with or without the C- terminal lysine); an amino acid sequence of SEQ ID NO: 2, and wherein the antibody is bonded at C449 in SEQ ID NO: 1 to the polymer. In some embodiments, the polymer has 9 arms, and wherein the polymer has a molecular weight of between about 600,000 to about 900,000 Da.

[0296] In some embodiments, the first protein or the conjugated protein (e.g., the antibody conjugated to the polymer) is an antibody or a fusion construct, and has the following structure:wherein each heavy chain of the antibody or fusion construct is denoted by the letter H, and each light chain of the antibody or fusion construct is denoted by the letter L; the polymer is bonded to the antibody or fusion construct 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, isopropyl, b) –H, c) any halogen, including –Br, –Cl, or –I, d) –SCN, or e) –NCS; and n1, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of n1, n2, n3, n4, n5, n6, n7, n8 and n9 is 2500 plus or minus 15%. In some embodiments, the sum of n1, n2, n3, n4, n5, n6, n7, n8 and n9 is about 1500 to about 3500 plus or minus about 10% to about 20%. In some embodiments, the first protein or the conjugated protein (e.g., the antibody conjugated to the polymer) is an antibody or a fusion construct, and has the following structure:wherein each heavy chain of the antibody or fusion construct is denoted by the letter H, and each light chain of the antibody or fusion construct is denoted by the letter L; the polymer is bonded to the antibody or fusion construct through the sulfhydryl of C443 (EUnumbering), 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, isopropyl, b) –H, c) any halogen, including –Br, –Cl, or –I, d) –SCN, or e) –NCS; and n1, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of n1, n2, n3, n4, n5, n6, n7, n8 and n9 is 2500 plus or minus 15%.

[0297] In some embodiments, the second (and / or the unconjugated) antibody comprises: a CDRH1 that is the CDRH1 in SEQ ID NO: 1; a CDRH2 that is the CDRH2 in SEQ ID NO: 1; a CDRH3 that is the CDRH3 in SEQ ID NO: 1; a CDRL1 that is the CDRL1 in SEQ ID NO: 2; a CDRL2 that is the CDRL2 in SEQ ID NO: 2; a CDRL3 that is the CDRL3 in SEQ ID NO: 2; 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.

[0298] Formulations and compositions provided herein can include a suitable pharmaceutically acceptable carrier. In some embodiments, the pharmaceutically acceptable carrier includes water or a buffer. In some embodiments, the pharmaceutically acceptable carrier comprises a buffer comprising a buffering agent having a pKa that is within, or within about 2, 1, or 0.5 pH units of the pH of the formulation. In some embodiment, the formulation includes as a pharmaceutically acceptable carrier a buffer selected from: acetate, phosphate, citrate, glycine, histidine, HEPES, and Tris buffers. In some embodiments, the formulation includes a sodium acetate buffer. In some embodiments, the formulation includes, includes about, or includes at least 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 90, 100 mM or more sodium acetate, or optionally, the formulation includes sodium acetate at a concentration in a range defined by any two of the preceding values (e.g., 10-100 mM, 30-80 mM, 40-60 mM, 40-70 mM, etc.). In some embodiments, the formulation includes sodium acetate at a concentration in a range of 40-60 mM. In some embodiments, the formulation includes sodium acetate at a concentration in a range of 30-80 mM. In some embodiments, the formulation includes a histidine acetate buffer. In some embodiments, the formulation includes, includes about, or includes at least 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 90, 100 mM or more histidine acetate, or optionally, the formulation includeshistidine acetate at a concentration in a range defined by any two of the preceding values (e.g., 5-100 mM, 20-80 mM, 10-60 mM, 15-70 mM, 10-30 mM, 15-25 mM, etc.). In some embodiments, the formulation includes histidine acetate at a concentration in a range of 10-60 mM. In some embodiments, the formulation includes histidine acetate at a concentration in a range of 10-30 mM. In some embodiments, the formulation includes histidine acetate at a concentration in a range of 15-25 mM.

[0299] In any of the composition or formulation described herein, in some embodiments, the composition or formulation includes a surfactant, such as but not limited to a polysorbate (e.g., polysorbate 20). The composition or formulation can include any suitable amount of the surfactant. In some embodiments, the composition or formulation includes, includes about, or includes at least 0.005, 0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.05, 0.0.6, 0.07, 0.08, 0.09, 0.1% (w / w) or more of the surfactant, or optionally, the composition or formulation includes a percentage (w / w) of the surfactant in a range defined by any two of the preceding values (e.g., 0.005-0.1%, 0.01-0.05%, 0.02-0.03%, 0.015-0.08%, etc.). In some embodiments, the composition or formulation includes the surfactant at about 0.01% to about 0.04% (w / w). In some embodiments, the composition or formulation includes the surfactant at about 0.02% to about 0.03% (w / w). In some embodiments, the composition or formulation includes about 0.025% (w / w) surfactant. In some embodiments, the surfactant is or comprises polysorbate 20 or polysorbate 80 or poloxamer 188. In some embodiments, the surfactant is polysorbate 20. In some embodiments, the composition or formulation includes about 0.01- 0.05% (w / w) polysorbate 20. In some embodiments, the composition or formulation includes about 0.02-0.03% (w / w) polysorbate 20. In some embodiments, the composition or formulation includes about 0.025% (w / w) polysorbate 20. In some embodiments, the surfactant is poloxamer 188.

[0300] In any of the composition or formulation described herein, in some embodiments, the composition or formulation includes a tonicity agent. In some embodiments, the tonicity agent is or includes a sugar. In some embodiments, the tonicity agent is or includes sucrose or trehalose. The composition or formulation can include any suitable amount of the tonicity agent. In some embodiments, the composition or formulation includes, includes about, or includes at least 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.5%, 8.0%, 8.5%, 9.0%, 10.0% (w / v) or more of the tonicity agent, or optionally, the compositionor formulation includes a percentage (w / v) of the tonicity agent in a range defined by any two of the preceding values (e.g., 2-10%, 2.5-6%, 3-5%, 3.5-8.5%, etc.). In some embodiments, the composition or formulation includes the tonicity agent at about 2% to about 10% (w / v).some embodiments, the composition or formulation includes the tonicity agent at about 3% to about 5% (w / v). In some embodiments, the composition or formulation includes about 4% (w / v) tonicity agent. In some embodiments, the composition or formulation includes about 2% to about 10% (w / v) sucrose. In some embodiments, the composition or formulation includes about 3% to about 5% (w / v) sucrose. In some embodiments, the composition or formulation includes about 4% (w / v) sucrose.

[0301] Also provided is an intraocular therapeutic composition comprising an anti- VEGF-A antibody at about 50 mg / mL of protein, the anti-VEGF-A antibody comprising: a heavy chain comprising a complementarity determining region 1 (CDRH1): GYDFTHYGMN (SEQ ID NO: 9), CDRH2: WINTYTGEPTYAADFKR (SEQ ID NO: 10), and CDRH3: YPYYYGTSHWYFDV (SEQ ID NO: 11); and a light chain comprising CDRL1: SASQDISNYLN (SEQ ID NO: 12), CDRL2: FTSSLHS (SEQ ID NO: 13), and CDRL3: QQYSTVPWT (SEQ ID NO: 14), wherein the anti-VEGF-A antibody is present in the composition as either an antibody conjugate or unconjugated antibody, wherein the unconjugated antibody is present in the formulation at between about 10% to about 30% of a total molar amount of the antibody conjugate and the unconjugated antibody, wherein the total molar amount is the sum of the molar amount of the antibody conjugate and the molar amount of the unconjugated antibody, wherein the antibody conjugate comprises the anti-VEGF-A antibody conjugated to a phosphorylcholine-containing polymer at a non-native cysteine outside a variable region of the antibody, wherein the phosphorylcholine-containing polymer is present in the composition at about 100 mg / mL or more, wherein the phosphorylcholine- containing polymer has a molecular weight of between 300,000 and 1,200,000 Da, wherein the pH of the composition is about 5.5 or lower.

[0302] Provided herein is an intraocular therapeutic composition comprising an anti-VEGF-A antibody at about 50 mg / mL of protein, the anti-VEGF-A antibody comprising: a heavy chain comprising an amino acid sequence of SEQ ID NO: 1 (with or without the C- terminal lysine); and a light chain comprising an amino acid sequence of SEQ ID NO: 2, wherein the anti-VEGF-A antibody is present in the composition as either an antibodyconjugate or unconjugated antibody, wherein the unconjugated antibody is present in the formulation at between about 10% to about 30% of a total molar amount of the antibody conjugate and the unconjugated antibody, wherein the total molar amount is the sum of the molar amount of the antibody conjugate and the molar amount of the unconjugated antibody, wherein the antibody conjugate comprises the following structure:wherein: each heavy chain of the conjugate is denoted by the letter H, and each light chain of the conjugate is denoted by the letter L; the polymer is bonded to the heavy chain of the conjugate through the sulfhydryl of C443 (EU numbering), which bond is depicted on oneof 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, isopropyl, b) –H, c) any halogen, including –Br, –Cl, or –I, d) –SCN, or e) –NCS; and n1, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of n1, n2, n3, n4, n5, n6, n7, n8 and n9 is 2500 plus or minus 15%, wherein the phosphorylcholine-containing polymer is present inthe composition at about 100 mg / mL or more, wherein the pH of the composition is about 5.5 or lower.

[0303] Also provided is an intraocular therapeutic composition comprising a fusion construct at about 53 mg / mL of protein, the fusion construct comprising a VEGF trap fused to an anti-IL-6 antibody, wherein the fusion construct comprises: a heavy chain comprising an amino acid sequence of SEQ ID NO:105 (with or without the C-terminal lysine), and a light chain comprising an amino acid sequence of SEQ ID NO:106, wherein the fusion construct is present in the composition as either a conjugate or an unconjugated fusion construct, wherein the unconjugated fusion construct is present in the formulation at between about 20% to about 40% of a total molar amount of the conjugate and the unconjugated fusion construct, wherein the total molar amount is the sum of the molar amount of the conjugate and the molar amount of the unconjugated fusion construct, wherein the conjugate comprises the fusion construct conjugated to a phosphorylcholine-containing polymer, wherein the phosphorylcholine- containing polymer is present in the composition at about 100 mg / mL or more, wherein the pH of the composition is about 5 or lower.

[0304] Provided herein is an intraocular therapeutic composition comprising a fusion construct at about 53 mg / mL of protein, the fusion construct comprising a VEGF trap fused to an anti-IL-6 antibody, wherein the fusion construct comprises: a heavy chain comprising a complementarity determining region 1 (CDRH1): PFAMH (SEQ ID NO: 134), CDRH2: KISPGGSWTYYSDTVTD (SEQ ID NO: 135), and CDRH3: QAWGYYALDI (SEQ ID NO: 136); and a light chain comprising CDRL1: SASISVSYLY (SEQ ID NO: 137), CDRL2: DDSSLAS (SEQ ID NO: 138), and CDRL3: QQWSGYPYT (SEQ ID NO: 139), wherein the fusion construct is present in the composition as either a conjugate or an unconjugated fusion construct, wherein the unconjugated fusion construct is present in the formulation at between about 20% to about 40% of a total molar amount of the conjugate and the unconjugated fusion construct, wherein the total molar amount is the sum of the molar amount of the conjugate and the molar amount of the unconjugated fusion construct, wherein the conjugate comprises the following structure:

[0305] wherein: each heavy chain of the conjugate is denoted by the letter H, and each light chain of the conjugate is denoted by the letter L; the polymer is bonded to the heavy chain of the conjugate 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, isopropyl, b) –H, c) any halogen, including –Br, –Cl, or –I, d) –SCN, or e) –NCS; and n1, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of n1, n2, n3, n4, n5, n6, n7, n8 and n9 is 2500 plus or minus 15%, wherein the phosphorylcholine-containing polymer is present in the composition at about 100 mg / mL or more, wherein the pH of the composition is about 5 or lower. Also provided is an intraocular therapeutic composition comprising a fusion construct at about 53 mg / mL of protein, the fusion construct comprising a VEGF trap fused to an anti-IL-6 antibody, wherein the fusion construct comprises: a heavy chain comprising acomplementarity determining region 1 (CDRH1): PFAMH (SEQ ID NO: 134), CDRH2: KISPGGSWTYYSDTVTD (SEQ ID NO: 135), and CDRH3: QAWGYYALDI (SEQ ID NO: 136); and a light chain comprising CDRL1: SASISVSYLY (SEQ ID NO: 137), CDRL2: DDSSLAS (SEQ ID NO: 138), and CDRL3: QQWSGYPYT (SEQ ID NO: 139), wherein the fusion construct is present in the composition as either a conjugate or an unconjugated fusion construct, wherein the unconjugated fusion construct is present in the formulation at between about 20% to about 40% of a total molar amount of the conjugate and the unconjugated fusion construct, wherein the total molar amount is the sum of the molar amount of the conjugate and the molar amount of the unconjugated fusion construct, wherein the conjugate comprises the following structure: )wherein: each heavy chain of the conjugate is denoted by the letter H, and each light chain of the conjugate is denoted by the letter L; the polymer is bonded to the heavy chain of the conjugate through the sulfhydryl of C443 (EU numbering), which bond is depicted on oneof 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, isopropyl, b) –H, c) any halogen, including –Br, –Cl, or –I, d) –SCN, or e) –NCS; and n1, n2, n3, n4, n5, n6, n7,n8 and n9 are the same or different such that the sum of n1, n2, n3, n4, n5, n6, n7, n8 and n9 is 2500 plus or minus 15%, wherein the phosphorylcholine-containing polymer is present in the composition at about 100 mg / mL or more, wherein the pH of the composition is about 5 or lower.

[0306] In some embodiments, a method of making a composition comprising an anti-VEGF-A antibody conjugated to a phosphorylcholine-containing polymer is provided. The method comprises: conjugating a first antibody to a phosphorylcholine-containing polymer, wherein the polymer is covalently bonded to the first antibody at a cysteine outside a variable region of the first antibody; combining a second antibody that is not conjugated to the phosphorylcholine-containing polymer with the first antibody. In some embodiments, the second antibody is any anti-VEGF Fab. In some embodiments, the second antibody is a Lucentis antibody (ranibizumab). In some embodiments, the second antibody is present in the composition at a percent composition of 5% to 93% relative to the total protein mass weight concentration of the first antibody and the second antibody in the composition. In some embodiments, the second antibody is present in the composition at a percent composition of 5% to 50% relative to the total protein mass weight concentration of the first antibody and the second antibody in the composition. In some embodiments, the second antibody is present in the composition at a percent composition of 5% to 25% relative to the total protein mass weight concentration of the first antibody and the second antibody in the composition. In some embodiments, the purified antibody conjugate has at least a 1.5-fold increase in half-life relative to an unconjugated anti-VEGF-A antibody.

[0307] In some embodiments, unconjugated biologics have rapid efficacy in wAMD. In some embodiments, unconjugated biologics have deep efficacy in wAMD. In some embodiments, for any of the compositions herein, clinical durability of the composition resulting in little to no disease recurrence is desirable.

[0308] In some embodiments, ranibizumab port delivery reservoir implant delivery efficacy in wAMD is desirable. In some embodiments, any of the compositions herein can result in longer durability without sacrificing immediate efficacy in wAMD by combining a conjugated version of the antibody in combination with an unconjugated version of the antibody. In some embodiments, the combination of the conjugated and unconjugated antibody provides a desired balance of immediacy (or bolus activity), and of durability (or basal activity). In some embodiments, any of the formulations and compositions herein can act to slow progress of retinal disease. In some embodiments, any of the formulations and compositions herein can act to stop progress of retinal disease. In some embodiments, any of the formulations and compositions herein are prepared with, or with about a 10% loading, or a 15% loading, or a 20% loading or a 25% loading, or a 30% loading, or a 35% loading of OG1950 mAb.

[0309] In some embodiments, the composition comprises a first antibody and a second antibody: wherein the first antibody is a conjugate; wherein the first antibody and second antibody have the same molecular weight, but the first antibody and second antibody can be the same or different proteins. In some embodiments, the composition comprises a first antibody and a second antibody: wherein the first antibody is a conjugate; wherein the first antibody and second antibody have a similar molecular weight, but the first antibody and second antibody can be the same or different proteins. In some embodiments, the composition comprises a first antibody and a second antibody: wherein the first antibody is a conjugate; wherein the first antibody and second antibody have different molecular weights, but the first antibody and second antibody can be the same or different proteins. In some embodiments, this composition comprises a first active moiety and a second active moiety that can be any protein. In some embodiments, this composition comprises a first active moiety and a second active moiety in which the first active moiety is a full antibody and a second active moiety is an antibody fragment. In some embodiments, this composition comprises a first active moiety and a second active moiety in which the first active moiety is a full antibody and a second active moiety is an antibody fragment of the fi...

Claims

WHAT IS CLAIMED IS:

1. A formulation comprising: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second molar amount of a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the formulation comprises the second protein at about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the formulation has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein.

2. A therapeutically acceptable composition comprising: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second molar amount of a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a therapeutically acceptable carrier, wherein the composition comprises the second protein at about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the composition has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein.

3. A therapeutically acceptable composition comprising: a conjugate comprising a first protein conjugated to a phosphorylcholine- containing polymer; a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier,wherein the percent composition of the second protein relative to the total protein mass weight concentration of the first protein and the second protein in the composition is about 1% or more, wherein the composition has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein.

4. A formulation comprising: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second molar amount of a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the formulation comprises the second protein at about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the formulation has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein, wherein the formulation has a reduced viscosity and / or an enhanced injectability compared to a reference formulation comprising the conjugate at the total molar amount.

5. A therapeutically acceptable composition comprising: a conjugate comprising a first protein conjugated to a phosphorylcholine- containing polymer; a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the percent composition of the second protein relative to the total protein mass weight concentration of the first protein and the second protein in the composition is about 1% or more, wherein the composition has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein,wherein the composition has a reduced viscosity and / or an enhanced injectability compared to a reference composition comprising the conjugate, wherein the first protein of the conjugate is present in the reference composition at the total mass weight concentration of the first and second proteins in the composition.

6. A low-viscosity formulation of a protein conjugate, comprising: a first molar amount of a conjugate comprising a protein conjugated to a phosphorylcholine-containing polymer; a second molar amount of the protein that is not conjugated to the phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the formulation has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the protein, wherein the formulation has reduced viscosity and / or an enhanced injectability compared to a reference formulation comprising the conjugate at a total molar amount that is the sum of the first and second molar amounts.

7. A formulation comprising: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second molar amount of a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the second protein is present in the formulation at about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the formulation has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein, wherein the formulation has a reduced turbidity compared to a reference formulation comprising the first molar amount of the conjugate and the second molar amount of the second protein at a pH about the same as (e.g., within 0.05, 0.1, 0.15, 0.2, 0.3, 0.4, or 0.5 pH units of) the pI of the second protein.

8. A pharmaceutical formulation comprising:a first molar amount of a conjugate comprising a protein conjugated to a phosphorylcholine-containing polymer; a second molar amount the protein that is not conjugated to the phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the formulation comprises the protein that is not conjugated to the phosphorylcholine-containing polymer at about 1% or more of a total molar amount of the conjugate and unconjugated proteins, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the formulation has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the protein, wherein the formulation is substantially free of turbidity.

9. A formulation comprising: a phosphorylcholine-containing polymer present in the formulation at 100 mg / mL or higher; and a protein that is not conjugated to a phosphorylcholine-containing polymer, wherein the protein is present in the formulation at a second molar amount, wherein the protein is present in the formulation at about 1% or more of a total molar amount of the polymer and the protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the formulation has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the protein.

10. A formulation comprising: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second molar amount of a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the difference between the pI of the second protein and the pH of the formulation in the acidic or basic direction is selected to be greater than the minimumdifference in the corresponding acidic or basic direction between the pI of the second protein and the pH for a reference formulation comprising: a third molar amount of the conjugate comprising the first protein conjugated to the phosphorylcholine-containing polymer; a fourth molar amount of the second protein that is not conjugated to the phosphorylcholine-containing polymer; and the pharmaceutically acceptable carrier, wherein a first total molar amount comprising a sum of the first molar amount and the second molar amount, and a second total molar amount comprising a sum of the third molar amount and the fourth molar amount are substantially the same, wherein the second molar amount is greater than the fourth molar amount, wherein the reference formulation is substantially free of turbidity.

11. A therapeutically acceptable composition comprising: a conjugate comprising a first protein conjugated to a phosphorylcholine- containing polymer; a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the difference between the pI of the second protein and the pH of the formulation is selected to be greater than the minimum difference between the pI of the second protein and the pH for a reference formulation comprising: the conjugate comprising the first protein conjugated to the phosphorylcholine- containing polymer; the second protein that is not conjugated to the phosphorylcholine-containing polymer; and the pharmaceutically acceptable carrier, wherein the percent composition of the second protein relative to the total protein mass weight concentration of the first protein and the second protein in the composition is higher than the percent composition of the second protein relative to the total protein mass weight concentration of the first protein and the second protein in the reference composition,wherein the reference composition is substantially free of turbidity.

12. A formulation comprising: a first molar amount of a conjugate comprising a first protein conjugated to a polymer; and a second molar amount of a second protein that is not conjugated to a polymer, wherein the formulation comprises the second protein at about 1% or more of a total molar amount of the first protein and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount.

13. A therapeutically acceptable composition comprising: a conjugate comprising a first protein conjugated to a phosphorylcholine- containing polymer; a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the percent composition of the second protein relative to the total protein mass weight concentration of the first protein and the second protein in the composition is about 1% or more.

14. A therapeutically acceptable composition comprising: a first molar amount of a conjugate comprising a first protein conjugated to a polymer; and a second molar amount of a second protein that is not conjugated to a polymer, wherein the composition comprises the second protein at about 1% or more of a total molar amount of the first protein and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount.

15. A formulation comprising: a first molar amount of a first protein that is conjugated to a polymer; and a second molar amount of a second protein that is not conjugated to a polymer, the further improvement comprising: the formulation comprises the second protein at about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount.

16. A formulation comprising: a conjugate comprising a first protein conjugated to a polymer; and a second protein that is not conjugated to a polymer, wherein a first molar amount of the conjugate and a second molar amount of the second protein has been combined in the formulation such that the second molar amount is about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount.

17. A therapeutically acceptable composition comprising: a conjugate comprising a first protein conjugated to a polymer; and a second protein that is not conjugated to a polymer, wherein the second protein at a percent composition relative to the total protein mass weight concentration of the first protein and the second protein in the composition of about 1% or more has been combined with the conjugate, wherein the remainder of the total protein mass weight concentration comprises the first protein.

18. A formulation comprising: a conjugate comprising a first protein conjugated to a phosphorylcholine- containing polymer, wherein the polymer has 9 arms and a molecular weight of between 600,000 and 1,000,000 Da, wherein the polymer is present in the formulation at about 100 mg / mL or more; and a second protein that is not conjugated to a polymer, wherein the second protein is present in the formulation at 5-15 mg / mL.

19. An intraocular therapeutic composition comprising an anti-VEGF-A antibody at about 50 mg / mL of protein, the anti-VEGF-A antibody comprising: a heavy chain comprising a complementarity determining region 1 (CDRH1): GYDFTHYGMN (SEQ ID NO: 9), CDRH2: WINTYTGEPTYAADFKR (SEQ ID NO: 10), and CDRH3: YPYYYGTSHWYFDV (SEQ ID NO: 11); and a light chain comprising CDRL1: SASQDISNYLN (SEQ ID NO: 12), CDRL2: FTSSLHS (SEQ ID NO: 13), and CDRL3: QQYSTVPWT (SEQ ID NO: 14), wherein the anti-VEGF-A antibody is present in the composition as either an antibody conjugate or unconjugated antibody, wherein the unconjugated antibody is present in theformulation at between about 10% to about 30% of a total molar amount of the antibody conjugate and the unconjugated antibody, wherein the total molar amount is the sum of the molar amount of the antibody conjugate and the molar amount of the unconjugated antibody, wherein the antibody conjugate comprises the anti-VEGF-A antibody conjugated to a phosphorylcholine-containing polymer at a non-native cysteine outside a variable region of the antibody, wherein the phosphorylcholine-containing polymer is present in the composition at about 100 mg / mL or more, wherein the phosphorylcholine-containing polymer has 9 arms and a molecular weight of between 600,000 and 1,000,000 Da, wherein the pH of the composition is about 5.5 or lower.

20. An intraocular therapeutic composition comprising an anti-VEGF-A antibody at about 50 mg / mL of protein, the anti-VEGF-A antibody comprising: a heavy chain comprising an amino acid sequence of SEQ ID NO: 1 (with or without the C-terminal lysine); and a light chain comprising an amino acid sequence of SEQ ID NO: 2, wherein the anti-VEGF-A antibody is present in the composition as either an antibody conjugate or unconjugated antibody, wherein the unconjugated antibody is present in the formulation at between about 10% to about 30% of a total molar amount of the antibody conjugate and the unconjugated antibody, wherein the total molar amount is the sum of the molar amount of the antibody conjugate and the molar amount of the unconjugated antibody, wherein the antibody conjugate comprises the following structure:wherein: each heavy chain of the conjugate is denoted by the letter H, and each light chain of the conjugate is denoted by the letter L; the polymer is bonded to the heavy chain of the conjugate 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, isopropyl, b) –H, c) any halogen, including –Br, –Cl, or –I, d) –SCN, or e) – NCS; andn1, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of n1, n2, n3, n4, n5, n6, n7, n8 and n9 is 2500 plus or minus 15%, wherein the phosphorylcholine-containing polymer is present in the composition at about 100 mg / mL or more, wherein the pH of the composition is about 5.5 or lower.

21. An intraocular therapeutic composition comprising a fusion construct at about 53 mg / mL of protein, the fusion construct comprising a VEGF trap fused to an anti-IL-6 antibody, wherein the fusion construct comprises: a heavy chain comprising an amino acid sequence of SEQ ID NO:105 (with or without the C-terminal lysine); and a light chain comprising an amino acid sequence of SEQ ID NO:106, wherein the fusion construct is present in the composition as either a conjugate or an unconjugated fusion construct, wherein the unconjugated fusion construct is present in the formulation at between about 20% to about 40% of a total molar amount of the conjugate and the unconjugated fusion construct, wherein the total molar amount is the sum of the molar amount of the conjugate and the molar amount of the unconjugated fusion construct, wherein the conjugate comprises the fusion construct conjugated to a phosphorylcholine-containing polymer, wherein the phosphorylcholine-containing polymer is present in the composition at about 100 mg / mL or more, wherein the pH of the composition is about 5 or lower.

22. An intraocular therapeutic composition comprising a fusion construct at about 50 mg / mL of protein, the fusion construct comprising a VEGF trap fused to an anti-IL-6 antibody, wherein the fusion construct comprises: a heavy chain comprising a complementarity determining region 1 (CDRH1): PFAMH (SEQ ID NO: 134), CDRH2: KISPGGSWTYYSDTVTD (SEQ ID NO: 135), and CDRH3: QAWGYYALDI (SEQ ID NO: 136); and a light chain comprising CDRL1: SASISVSYLY (SEQ ID NO: 137), CDRL2: DDSSLAS (SEQ ID NO: 138), and CDRL3: QQWSGYPYT (SEQ ID NO: 139), wherein the fusion construct is present in the composition as either a conjugate or an unconjugated fusion construct, wherein the unconjugated fusion construct is present in the formulation at between about 20% to about 40% of a total molar amount of the conjugate andthe unconjugated fusion construct, wherein the total molar amount is the sum of the molar amount of the conjugate and the molar amount of the unconjugated fusion construct, wherein the conjugate comprises the following structure: )wherein: each heavy chain of the conjugate is denoted by the letter H, and each light chain of the conjugate is denoted by the letter L; the polymer is bonded to the heavy chain of the conjugate 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, isopropyl, b) –H, c) any halogen, including –Br, –Cl, or –I, d) –SCN, or e) – NCS; and n1, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different such that the sum of n1, n2, n3, n4, n5, n6, n7, n8 and n9 is 2500 plus or minus 15%, wherein the phosphorylcholine-containing polymer is present in the composition at about 100 mg / mL or more, wherein the pH of the composition is about 5 or lower.

23. A method of preparing a formulation, comprising combining in a formulation: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; and a second molar amount of a second protein that is not conjugated to a phosphorylcholine-containing polymer, wherein the formulation comprises the second protein at about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the formulation has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein.

24. A method of preparing a therapeutically acceptable composition, comprising combining in a therapeutically acceptable composition: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; and a second molar amount of a second protein that is not conjugated to a phosphorylcholine-containing polymer, wherein the composition comprises the second protein at about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the composition has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein.

25. A method of preparing a therapeutically acceptable composition, comprising combining in a therapeutically acceptable composition: a conjugate comprising a first protein conjugated to a phosphorylcholine- containing polymer; anda second protein that is not conjugated to a phosphorylcholine-containing polymer, wherein the percent composition of the second protein relative to the total protein mass weight concentration of the first protein and the second protein in the composition is about 1% or more, wherein the composition has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein.

26. A method of preparing a formulation, comprising adjusting the pH of a formulation to be about 0.5 pH units away or more from the isoelectric point (pI) of an unconjugated protein comprised in the formulation, wherein the formulation comprises: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; and a second molar amount of the unconjugated protein, wherein the unconjugated protein is not conjugated to a phosphorylcholine-containing polymer, wherein the formulation comprises the unconjugated protein at about 1% or more of a total molar amount of the conjugate and the unconjugated protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount.

27. A method of preparing a therapeutically acceptable composition, comprising adjusting the pH of a composition to be about 0.5 pH units away or more from the isoelectric point (pI) of an unconjugated protein comprised in the composition, wherein the composition comprises: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; and a second molar amount of the unconjugated protein, wherein the unconjugated protein is not conjugated to a phosphorylcholine-containing polymer, wherein the composition comprises the unconjugated protein at about 0.1% or more of a total molar amount of the conjugate and the unconjugated protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount.

28. A method of preparing a therapeutically acceptable composition, comprising adjusting the pH of a therapeutically acceptable composition to be about 0.5 pH units away or more from the isoelectric point (pI) of an unconjugated protein comprised in the composition, wherein the composition comprises: a conjugate comprising a first protein conjugated to a phosphorylcholine- containing polymer; and a second protein that is not conjugated to a phosphorylcholine-containing polymer, wherein the percent composition of the second protein relative to the total protein mass weight concentration of the first protein and the second protein in the composition is about 1% or more, wherein the composition has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein.

29. A method of preparing a low-viscosity formulation of a protein conjugated to a phosphorylcholine-containing polymer, comprising combining in a formulation: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; and a second molar amount of the protein that is not conjugated to a phosphorylcholine-containing polymer, wherein the formulation comprises the protein that is not conjugated to the phosphorylcholine-containing polymer at about 1% or more of a total molar amount of the conjugate unconjugated proteins, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the formulation has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein, wherein the formulation has reduced viscosity and / or an enhanced injectability compared to a reference formulation comprising the conjugate at the total molar amount.

30. A method of preparing a formulation, comprising combining in a formulation: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; anda second molar amount of a second protein that is not conjugated to a phosphorylcholine-containing polymer, wherein the formulation comprises the second protein at about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount.

31. A method of preparing a therapeutically acceptable composition, comprising combining in a therapeutically acceptable composition: a conjugate comprising a first protein conjugated to a phosphorylcholine- containing polymer; and a second protein that is not conjugated to a phosphorylcholine-containing polymer, wherein the percent composition of the second protein relative to the total protein mass weight concentration of the first protein and the second protein in the composition is about 1% or more.

32. A method of treating a subject, comprising: intraocularly administering a therapeutically effective amount of the formulation or composition of any one of claims 1-22 to a subject in need thereof.

33. A kit comprising: a pre-filled syringe comprising a low-viscosity formulation comprising: a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; and a second protein that is not conjugated to a phosphorylcholine- containing polymer; and a syringe needle for injection of the low-viscosity formulation, wherein the gauge of the needle is 27 or higher.

34. 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.

35. A 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.

36. A method of storing a protein, comprising maintaining a protein in a formulation for at least 2 months and up to 2 years, the formulation comprising: a first molar amount of a conjugate comprising a first protein conjugated to a phosphorylcholine-containing polymer; a second molar amount of a second protein that is not conjugated to a phosphorylcholine-containing polymer; and a pharmaceutically acceptable carrier, wherein the formulation comprises the second protein at about 1% or more of a total molar amount of the conjugate and the second protein, wherein the total molar amount comprises a sum of the first molar amount and the second molar amount, wherein the formulation has a pH that is about 0.5 pH units away or more from the isoelectric point (pI) of the second protein, wherein the protein comprises an antibody or a fusion construct.

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