Methods of treating eye disorders

Abstract

Provided herein are methods of treating various eye disorders, including, for example, wet age-related macular degeneration (wAMD) or diabetic retinopathy (DR), using an anti-VEGF therapy, such as an anti-VEGF antibody conjugate.

Description

METHODS OF TREATING EYE DISORDERS REFERENCE TO RELATED APPLICATIONS

[0001] This Application claims priority to U.S. Provisional Application No. 63 / 697362, filed September 20, 2024, which is incorporated herein by reference in its entirety. REFERENCE 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 SEQLIST_KDIAK232WO.xml created on September 18, 2025, which is 15,942 bytes in size. The information in the electronic format of the Sequence Listing is incorporated herein by reference in its entirety. BACKGROUND

[0003] The present disclosure relates to the field of eye disorders (e.g., neovascular eye disease) and treatments thereof.

[0004] The present disclosure relates to anti-VEGF antibodies and conjugates thereof and methods of using the anti-VEGF antibodies and conjugates thereof to treat eye disorders, including wet Age-Related Macular Degeneration (wAMD) and diabetic retinopathy (DR). SUMMARY

[0005] Provided herein are methods of treating eye disorders, including wet age- related macular degeneration (wAMD), and diabetic retinopathy (DR), with anti-VEGF antibody conjugates.

[0006] Provided herein is a method of treating wet age-related macular degeneration (wAMD), the method comprising: identifying a subject with wAMD; administering a first dose of an anti-VEGF antibody conjugate to the subject; administering a second dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the first dose; administering a third dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the second dose; administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the third dose; and administering an individualized dose of the anti-VEGF antibody conjugate to the subject at least 4 weeks (+ / - 7 days) after the fourth dose if the subject has a decline in eye health, wherein the individualized dose is administeredno more frequently than about once every four weeks (or no more frequently than Q4W or QM), optionally wherein one or more subsequent doses of the anti-VEGF antibody conjugate are administered to the subject no less frequently than about once every 24 weeks (or no less frequently than Q24W or Q6M) after the fourth and / or the last dose.

[0007] Also provided is a method of treating wAMD, the method comprising: identifying a subject with wAMD; administering a first dose of an anti-VEGF antibody conjugate to the subject; administering a second dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the first dose; administering a third dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the second dose; administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the third dose; evaluating the subject’s eye health no more frequently than about once every four weeks (or no more frequently than Q4W or QM) after the fourth dose; and if the subject has a decline in eye health upon evaluating, administering an individualized dose of the anti-VEGF antibody conjugate to the subject, optionally wherein one or more subsequent doses of the anti-VEGF antibody conjugate are administered to the subject no less frequently than about once every 24 weeks (or no less frequently than Q24W or Q6M) after the fourth and / or the last dose.

[0008] Provided herein is a method of treating wAMD, the method comprising: identifying a subject with wAMD; administering a first dose of an anti-VEGF antibody conjugate to the subject; administering a second dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the first dose; administering a third dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the second dose; administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the third dose; evaluating the subject’s eye health no more frequently than about once every four weeks (or no more frequently than Q4W or QM) after the fourth dose; and administering an individualized dose to the subject based on the subject’s evaluated eye health, optionally wherein one or more subsequent doses of the anti-VEGF antibody conjugate are administered to the subject no less frequently than about once every 24 weeks (or no less frequently than Q24W or Q6M) after the fourth and / or the last dose.

[0009] Also provided is a method of treating wAMD, the method comprising: identifying a subject with wAMD; administering a first dose of an anti-VEGF antibody conjugate to the subject; administering a second dose of the anti-VEGF antibody conjugate tothe subject 4 weeks after the first dose; and administering a third dose of the anti-VEGF antibody conjugate to the subject 4 weeks after the second dose; administering a fourth dose of the anti-VEGF antibody conjugate to the subject 4 weeks after the third dose; and if the subject has a decline in eye health within about 24 weeks after the fourth dose, administering an individualized dose of the anti-VEGF antibody conjugate, optionally wherein one or more subsequent doses of the anti-VEGF antibody conjugate are administered to the subject no less frequently than about once every 24 weeks (or no less frequently than Q24W or Q6M) after the fourth and / or the last dose.

[0010] Further provided herein is a method of treating eye disease, the method comprising: identifying a subject with a neovascular eye disease; administering a first dose of an anti-VEGF antibody conjugate to the subject; administering a second dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the first dose; administering a third dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the second dose; administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the third dose; administering an individualized dose of the anti-VEGF antibody conjugate to the subject at least 4 weeks (+ / - 7 days) after the fourth dose if the subject has a decline in eye health, wherein the individualized dose is administered no more frequently than about once every four weeks (or no more frequently than Q4W or QM).

[0011] Also provided herein is a method of treating a subject with wAMD, the method comprising: administering to a subject in need thereof 4 loading doses of an anti-VEGF antibody conjugate at frequency of one loading dose every four weeks (or at Q4W or at QM); determining, at least 4 weeks (+ / - 7 days) after a last loading dose, a presence or absence of intraretinal fluid (IRF) and / or subretinal fluid (SRF) in an eye of the subject, and / or if macular hemorrhage due to wAMD activity has clinically worsened; and if there is IRF or SRF in the eye, or the subject has clinically worsened macular hemorrhage due to wAMD activity, administering a maintenance dose.

[0012] Further provided is a method of treating diabetic retinopathy (DR), the method comprising: identifying a subject with DR; administering a first dose of an anti-VEGF antibody conjugate to the subject; administering a second dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the first dose; administering a third dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the second dose;administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 12 weeks after the third dose; and administering a fifth dose of the anti-VEGF antibody conjugate to the subject about 24 weeks after the fourth dose.

[0013] Also provided is a method of treating diabetic retinopathy (DR), the method comprising: (a) identifying a treatment naive subject with moderately severe to severe NPDR; (b) administering a first dose of an anti-VEGF antibody conjugate to the subject; (c) administering a second dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the first dose; (d) administering a third dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the second dose; (e) administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 12 weeks after the third dose; and (f) administering a fifth dose of the anti-VEGF antibody conjugate to the subject about 24 weeks after the fourth dose, (g) performing (b)-(f) unless the subject develops DME, PDR, and / or ASNV, in which case: (h) administering a fifth dose of an anti-VEGF antibody conjugate to the subject when the subject is identified as having developed DME, PDR, and / or ASNV; (i) administering a sixth dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the fifth dose; (j) administering a seventh dose of the anti-VEGF antibody conjugate to the subject about 12 weeks after the sixth dose; (k) administering an eighth dose of the anti-VEGF antibody conjugate to the subject about 12 weeks after the seventh dose; and (l) administering a ninth dose of the anti-VEGF antibody conjugate to the subject about 12 weeks after the eighth dose.

[0014] Provided herein is a method of treating wet age-related macular degeneration (wAMD) comprising: identifying a subject with wAMD; administering a first dose of an anti-VEGF antibody conjugate (e.g., KSI-301) to the subject; administering a second dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the first dose; administering a third dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the second dose; administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the third dose; and administering an individualized dose of the anti-VEGF antibody conjugate to the subject at least 4 weeks (+ / - 7 days) after the fourth dose if the subject has a decline in eye health, wherein the individualized dose is administered no more frequently than, once every four weeks (or no more frequently than Q4W or QM), wherein the first, second, third, fourth doses and the individualized dose each comprises about 5 mg of protein of the anti-VEGF antibody conjugate, wherein the anti-VEGF antibodyconjugate comprises: (1) an anti-VEGF-A antibody that includes: a light chain that has the amino acid sequence set forth in SEQ ID NO: 2; and a heavy chain that has the amino acid sequence set forth in SEQ ID NO :1 (or any of the variants thereof in FIG. 10, e.g., SEQ ID NOs: 9, 10, 11, or 12); and (2) a phosphorylcholine containing polymer, wherein the polymer is covalently bonded to the heavy chain of the anti-VEGF-A antibody, where the antibody conjugate has the following structure:where: each heavy chain of the anti-VEGF-A antibody is denoted by the letter H, and each light chain of the anti-VEGF-A antibody is denoted by the letter L; the polymer is bonded to the heavy chain through a sulfhydryl at C443 according to EU numbering, which bond isdepicted on one of the heavy chains above; PC is, where the curvyline indicates the point of attachment to the rest of the polymer; 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%.

[0015] Also provided is a method of treating wet age-related macular degeneration (wAMD) comprising: identifying a subject with wAMD; administering a first dose of an anti- VEGF antibody conjugate (e.g., KSI-301) to the subject; administering a second dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the first dose; administering a third dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the second dose; administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the third dose; and if the subject has a decline in eye health within about 24 weeks after the fourth dose, administering an individualized dose of the anti-VEGF antibody conjugate, wherein the first, second, third, fourth doses and the individualized dose each comprises about 5 mg of protein of the anti-VEGF antibody conjugate, wherein the anti-VEGF antibody conjugate comprises: (1) an anti-VEGF-A antibody that includes: a light chain that has the amino acid sequence set forth in SEQ ID NO: 2; and a heavy chain that has the amino acid sequence set forth in SEQ ID NO :1 (or any of the variants thereof in FIG. 10, e.g., SEQ ID NOs: 9, 10, 11, or 12); and (2) a phosphorylcholine containing polymer, wherein the polymer is covalently bonded to the heavy chain of the anti-VEGF-A antibody, where the antibody conjugate has the following structure:where: each heavy chain of the anti-VEGF-A antibody is denoted by the letter H, and each light chain of the anti-VEGF-A antibody is denoted by the letter L; the polymer is bonded to the heavy chain through a sulfhydryl at C443 according to EU numbering, which bond isdepicted on one of the heavy chains above; PC is, where the curvy line indicates the point of attachment to the rest of the polymer; 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%.

[0016] Further provided herein is a method of treating a subject with wet age- related macular degeneration (wAMD) comprising: administering to a subject in need thereof 4 loading doses of an anti-VEGF antibody conjugate at frequency of one loading dose everyfour weeks (or at Q4W or at QM); determining, at least 4 weeks (+ / - 7 days) after a last loading dose, a presence or absence of intraretinal fluid (IRF) and / or subretinal fluid (SRF) in an eye of the subject, and / or if macular hemorrhage due to wAMD activity has clinically worsened; and if there is IRF or SRF in the eye, or the subject has clinically worsened macular hemorrhage due to wAMD activity, administering a maintenance dose, wherein the loading doses and the maintenance dose each comprises about 5 mg of protein of the anti-VEGF antibody conjugate, wherein the anti-VEGF antibody conjugate comprises: (1) an anti-VEGF-A antibody that includes: a light chain that has the amino acid sequence set forth in SEQ ID NO: 2; and a heavy chain that has the amino acid sequence set forth in SEQ ID NO :1 (or any of the variants thereof in FIG. 10, e.g., SEQ ID NOs: 9, 10, 11, or 12); and (2) a phosphorylcholine containing polymer, wherein the polymer is covalently bonded to the heavy chain of the anti-VEGF-A antibody, where the antibody conjugate has the following structure:where: each heavy chain of the anti-VEGF-A antibody is denoted by the letter H, and each light chain of the anti-VEGF-A antibody is denoted by the letter L; the polymer is bonded to the heavy chain through a sulfhydryl at C443 according to EU numbering, which bond isdepicted on one of the heavy chains above; PC is, where the curvy line indicates the point of attachment to the rest of the polymer; 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%.

[0017] Also provided is a method of treating eye disease, comprising: administering one or more doses of an anti-VEGF therapy no more frequently than Q4W (+ / - 7 days) to a subject with a neovascular retinal disease or condition; and evaluating the subject’s eye health after administering the one or more doses of the anti-VEGF therapy to determine if the subject has a decline in eye health, wherein the evaluating comprises determining a presence or absence of intraretinal fluid (IRF) and / or subretinal fluid (SRF) in an eye of the subject.

[0018] Provided herein is a method of treating eye disease, comprising administering an anti-VEGF therapy to a subject with a neovascular retinal disease or condition according to a treatment regimen, to thereby treat the eye disease, wherein the treatment regimen comprises administering one or more doses of the anti-VEGF therapy no more frequently than Q4W (+ / - 7 days) to the subject, and wherein an efficacy of the treatment regimen has been determined by evaluating an eye health of a cohort of patients with the neovascular retinal disease or condition that have been administered one or more doses of the anti-VEGF therapy no more frequently than Q4W (+ / - 7 days), by determining a change in an intraretinal fluid (IRF) and / or subretinal fluid (SRF) in an eye of each patient of the cohort after administration of the one or more doses of the anti-VEGF therapy. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 shows OG1786.

[0020] Figure 2 shows OG1801.

[0021] Figure 3 shows OG1802.

[0022] Figure 4 is a flow chart depicting a non-limiting method of treating a subject with wet age-related macular degeneration.

[0023] Figure 5 is a flow chart depicting a non-limiting method of treating a subject with wet age-related macular degeneration.

[0024] Figure 6 is a flow chart depicting a non-limiting method of treating a subject with wet age-related macular degeneration.

[0025] Figure 7 is a flow chart depicting a non-limiting method of treating a subject with wet age-related macular degeneration.

[0026] Figure 8 is a flow chart depicting a non-limiting method of treating a subject with eye disease.

[0027] Figure 9 is a flow chart depicting a non-limiting method of treating a subject with wet age-related macular degeneration.

[0028] Figure 10 is an amino acid sequence for some embodiments of OG1950.

[0029] Figure 11 depicts an amino acid sequence of the heavy and light chains of KSI-301, according to some non-limiting embodiments of the present disclosure.

[0030] Figure 12 is schematic diagram of a protocol of a dosing regimen for an anti-VEGF antibody conjugate treatment, according to some non-limiting embodiments of the present disclosure.

[0031] Figure 13 is a set of graphs showing the schedule of administration of tarcocimab to be received by individual patients treated for Non-proliferative Diabetic Retinopathy (NPDR), according to some embodiments of the present disclosure.

[0032] Figure 14 shows sight threatening complications of diabetic retinopathy (DR) that may occur during the clinical study, according to some embodiments of the present disclosure.

[0033] Figure 15 is a set of graphs showing the modified schedule of administration of tarcocimab to be received by individual patients treated for NPDR who have developed sight threatening complications, according to some embodiments of the present disclosure

[0034] Figure 16A is a flow chart depicting the schema for study participants not diagnosed with DME, PDR, and / or ASNV in their Study Eye during the study. Figure 16B isa flow chart depicting the schema for study participants diagnosed with DME, PDR, and / or ASNV in their Study Eye during the study.

[0035] Figure 17 is a flow chart depicting a non-limiting method of treating eye disease of the present disclosure. DETAILED DESCRIPTION

[0036] Age-related macular degeneration (AMD) is a progressive disease that affects the macula, which is the central portion of the retina and is the region of the eye responsible for sharp, central vision and color perception. Neovascular (wet) AMD (wAMD) is an advanced form of AMD that is characterized by choroidal neovascularization and fluid leakage within and under the retina.

[0037] Vascular endothelial growth factor (VEGF) stimulates vascular endothelial cell growth and induces vascular permeability. These biologic activities give it a central role in angiogenesis, both in normal and pathologic conditions. VEGF plays a critical role in the pathophysiology of choroidal and retinal neovascular diseases such as wAMD; DR, including DME; and ME due to RVO (Rubio 2016).

[0038] Inhibition of pathologic VEGF activity is both an “anti-angiogenic” and “anti-permeability” approach to treat choroidal and retinal neovascular diseases and has been shown to be effective in preserving and improving visual acuity (Campochiaro 2015b; Cheung 2010). Among the VEGF isoforms, VEGF-A is the most strongly linked to angiogenesis and vascular leakage (Campochiaro 2015a).

[0039] Provided herein are methods of treating eye disorders, including but not limited to wet age-related macular degeneration (wAMD) and diabetic macular edema (DME), with an anti-VEGF therapy, such as anti-VEGF antibody conjugates, the timing of dosing, and methods of evaluating eye health and vision. Terms

[0040] Unless defined otherwise or the context clearly dictates otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs when read in light of the current disclosure.

[0041] An “anti-VEGF therapy” or “VEGF antagonist” denotes a compound, molecule, agent, or composition that inhibits or interferes with VEGF signaling and thereby inhibits, abrogates, reduces or interferes with angiogenesis or an angiogenic process mediated by VEGF signaling. In the various embodiments disclosed herein, in some embodiments, an anti-VEGF therapy targets at least VEGF-A signaling. In the various embodiments disclosed herein, in some embodiments, an anti-VEGF antibody or conjugate thereof targets at least VEGF-A signaling.

[0042] As used herein, any time “anti-VEGF antibody conjugate” is referenced, a mixture of the anti-VEGF antibody conjugate and an unconjugated anti-VEGF antibody is also contemplated. In the various embodiments disclosed herein, any reference to an anti-VEGF antibody conjugate therapy, also contemplates a combination anti-VEGF antibody conjugate / unconjugated anti-VEGF antibody therapy.

[0043] The term "dose" as used herein has its plain and ordinary meaning as understood in light of the specification, and denotes an amount of a therapeutic, e.g., an anti- VEGF antibody conjugate, that is administered to a subject for a therapeutic effect. A single dose may be administered in a single administration session, e.g., in a day, in a single visit to a health care provider, etc. A single dose may be administered in a single volume, or may be divided into multiple volumes (e.g., that add up to the single dose). In some embodiments doses are administered in fixed time intervals, for example a set number of weeks or days. In some embodiments, a time interval may be modified to include extra days or to be a shorter time interval by being earlier than the original set time interval. In some embodiments, the modified time interval is shorter by seven or fewer days. In some embodiments, the modified time interval is longer by 7 or fewer days. In some embodiments, the dose is administered every Q4W. In some embodiments, Q4W means every four (4) weeks.

[0044] A “loading dose” has its ordinary and customary meaning as understood by a person of ordinary skill in the art, in view of the present disclosure. A loading dose may refer to an amount of a therapeutic agent administered to a subject, either before a therapeutic effect of the agent is observed in the subject, or before a desired level of therapeutic effect of the agent is achieved in the subject. In some embodiments, a loading dose is administered at the beginning of a course of treatment with the therapy. In some embodiments, the loading dose is administered more frequently or at shorter intervals compared to later doses that are formaintenance of a therapeutic result. The time period during which a subject receives one or more loading doses may be referred to as a loading phase. In some embodiments, a subject is not monitored for disease progression or status (e.g., not assessed for visual acuity, retinal thickness, etc.) during the loading phase. In some embodiments, a therapeutic result (as disclosed herein) of the anti-VEGF antibody conjugate therapy (e.g., KSI-301 therapy) is observed at least at the end of the loading phase. The loading dose may be one of a series of loading doses administered to the subject, e.g., during the loading phase. In some embodiments, a subject is administered 4 loading doses. In some embodiments a subject is administered more than 4 loading doses, for example, 5 or 6 loading doses. In some embodiments, a subject is administered fewer than 4 loading doses. In some embodiments, a subject is administered 3, 2, or 1 loading doses. A “final loading dose” may refer to the last loading dose in a series of loading doses administered to the subject, at and / or after which a desired level of therapeutic effect of the agent is achieved. Thus, where the subject is given only one loading dose, the final loading dose is the first loading dose. Where the subject is given only two loading doses, the final loading dose is the second loading dose. Where the subject is given only three loading doses, the final loading dose is the third loading dose. Likewise, where the subject is given only four loading doses, the final loading dose is the fourth loading dose, and so on. A dose of the therapeutic agent administered to a subject after the loading phase may be referred to as a maintenance dose or an individualized dose. In some embodiments provided herein, the loading doses can be adequate without as frequent need for, or any need for, subsequent maintenance or individualized doses. In some embodiments, a series of loading doses is administered to a subject at a higher frequency than a series of maintenance or individualized doses subsequently administered to the subject. In some embodiment, the loading dose(s) given may be sufficient to keep disease activity under control in the subject, without requiring a maintenance or individualized dose.

[0045] A “individualized dose” and “maintenance dose” are used interchangeably herein, and each has its ordinary and customary meaning as understood by a person of ordinary skill in the art, in view of the present disclosure. An individualized or maintenance dose may refer to one or more doses administered based on assessments of individual eye health and vision after the loading doses have been administered. In some embodiments, the timing of administering the individualized or maintenance doses for one subject is different from thetiming of administering the individualized or maintenance doses for another subject, e.g., when the two subjects differ in their response to the loading doses and / or the individualized or maintenance doses. In some embodiments, criteria for determining whether individualized or maintenance doses should be administered includes but is not limited to a change in retinal thickness, the presence of intraretinal fluid (IRF), the presence of subretinal fluid (SRF), and / or the presence of new or clinically worsening macular hemorrhage due to disease activity (e.g., wAMD activity), as determined by the investigator.

[0046] The term "administered" as used herein has its plain and ordinary meaning as understood by one of ordinary skill in the art in light of the specification, and refers to providing a dose of an anti-VEGF therapy, e.g., an anti-VEGF antibody conjugate, to a subject such that the dose comes into contact with the region of the eye in need of therapeutic treatment. In some embodiments, the dose is provided or administered intravitreally. In some embodiments, the dose is provided or administered by intravitreal injection.

[0047] A “dosing schedule” is a clinical regimen for administration of a therapeutic agent. Examples of dosing schedules include administration of a therapeutic agent every day, every week, every 4 weeks, or as needed due to monitoring of patient symptoms and symptomology. Dosing schedules can vary due to the severity of the underlying disorder or disease state.

[0048] A “dose frequency” is a clinical regimen for administration of a therapeutic agent. The dose frequency may be every day, every week, every 4 weeks, or as needed due to monitoring of patient symptoms and symptomology. Dose frequencies can vary due to the severity of the underlying disorder or disease state.

[0049] A “Ophthalmic Exam” is a comprehensive series of tests performed by a physician to assess vision and eye health of a patient. Commonly, Ophthalmic exams include patient histories, refraction tests to check vision, optical coherence tomography, and other assessments to aid a physician in rendering a professional opinion regarding a patient’s eye health. During an Ophthalmic Exam, a physician may use imaging techniques including but not limited to Fundus Photograph, SD-OCT, OCT-A, and Fluorescein Angiography (FA) to characterize anatomical features of the eye. A physician may further characterize whether anatomical structures within the eye are exudative or not, where structures that appear to “leak” bodily fluids would be “wet,” while non-exudative structures would be “dry”. On imaging aneye, the physician may also characterize finding excess intra and / or subretinal fluid and / or subretinal hyperreflective material (SHRM) affecting the central subfield of the eye as “wet”, while the lack of excess intra and / or subretinal fluid affecting the central subfield of the eye as “dry.”

[0050] The term "eye health" as used herein has its plain and ordinary meaning as understood by one of ordinary skill in the art in light of the specification and refers to the condition of a subject’s eye as determined using one or more criteria, e.g., measures of visual acuity and / or anatomical measures. In some embodiments, evaluating eye health includes evaluating a subject for ocular pain, discomfort or vision. In some embodiments, a change in eye health includes an increase or decrease in a subject’s ocular pain, discomfort or vision. In some embodiments, eye health is determined by evaluating a subject’s eye. A person trained in evaluating eye health and / or administering treatment may perform the evaluation. In some embodiments, a subject’s eye health may worsen without the subject experiencing and / or reporting any symptoms, including pain and / or a loss of vision. In some embodiments, eye health is determined by examining a subject’s eye. In some embodiments, the criteria used for determining whether a subject has a change in eye health are subjective. In some embodiments, one or more measurements are made to determine whether a subject has had a change in eye health, and the value is compared to a standard and / or to a measurement made of the subject’s eye during a previous evaluation. In some embodiments, intraretinal fluid (IRF) is measured. In some embodiments, subretinal fluid (SRF) is measured. In some embodiments, retinal thickness is measured. In some embodiments, optical coherence tomography (OCT) is used to evaluate a subject’s eye health. In some embodiments, optical coherence tomography central subfield thickness (OCT CST) is used to evaluate a subject’s eye health. In some embodiments, the subject’s eye health is not evaluated before each loading dose is administered.

[0051] In some embodiments, a subject’s eye health is determined by evaluating an image and / or a scan of the subject’s eye. In some embodiments, the image or scan is compared to a standard image and / or a previous image and / or scan of the subject’s eye. In some embodiments, eye images are captured using fundus photography. In some embodiments, artificial intelligence is used to evaluate the images and / or scans of the subject’s eye. In some embodiments, a subject’s eye health is determined through an indirect test, for example by testing a subject’s eye fluid including retinal fluid or tears. In some embodiments, a subject isdetermined to have a change in eye health if a change in macular hemorrhage due to wAMD activity is observed. In some embodiments, the criteria for determining whether a subject has had a change in eye health includes asking the subject questions about their vision or how their eye feels. In some embodiments, a vision test is administered to the subject to determine whether the subject has had a change in eye health. In some embodiments, the eye test used involves a response to changes in light.

[0052] The term “peptide,” “polypeptide,” or “protein,” as used interchangeably herein, generally refers to a polymer of at least two amino acid residues joined by peptide bond(s). This term does not connote a specific length of polymer, nor is it intended to imply or distinguish whether the peptide is produced using recombinant techniques, chemical or enzymatic synthesis, or is naturally occurring. The terms apply to naturally occurring amino acid polymers as well as amino acid polymers comprising at least one modified amino acid. In some cases, the polymer can be interrupted by non-amino acids. The terms include amino acid chains of any length, including full length proteins, and proteins with or without secondary and / or tertiary structure (e.g., domains). The terms also encompass an amino acid polymer that has been modified, for example, by disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, oxidation, and any other manipulation such as conjugation with a labeling component. The terms “amino acid” and “amino acids,” as used herein, generally refer to natural and non-natural amino acids, including, but not limited to, modified amino acids and amino acid analogues. Modified amino acids can include natural amino acids and non-natural amino acids, which have been chemically modified to include a group or a chemical moiety not naturally present on the amino acid. Amino acid analogues can refer to amino acid derivatives. The term “amino acid” includes both D-amino acids and L-amino acids.

[0053] 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 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 occurringamino 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.

[0054] Percentage sequence identities are determined with antibody sequences maximally aligned by the Kabat numbering convention for a variable region or EU numbering for a constant region. After alignment, if a subject antibody region (e.g., the entire mature variable region of a heavy or light chain) is being compared with the same region of a reference antibody, the percentage sequence identity between the subject and reference antibody regions is the number of positions occupied by the same amino acid in both the subject and reference antibody region divided by the total number of aligned positions of the two regions, with gaps not counted, multiplied by 100 to convert to percentage. Sequence identities of other sequences can be determined by aligning sequences using algorithms, such as BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package Release 7.0, Genetics Computer Group, 575 Science Dr., Madison, WI, using default gap parameters, or by inspection, and the best alignment (i.e., resulting in the highest percentage of sequence similarity over a comparison window). Percentage of sequence identity is calculated by comparing two optimally aligned sequences over a window of comparison, determining the number of positions at which the identical residues 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.

[0055] The term “polynucleotide,” “oligonucleotide,” or “nucleic acid,” as used interchangeably herein, generally refers to a polymeric form of nucleotides of any length, either deoxyribonucleotides or ribonucleotides, or analogs thereof, either in single-, double-, or multi- stranded form. A polynucleotide can be exogenous or endogenous to a cell. A polynucleotide can exist in a cell-free environment. A polynucleotide can be a gene or fragment thereof. A polynucleotide can be DNA. A polynucleotide can be RNA. A polynucleotide can have any three dimensional structure, and can perform any function, known or unknown. Apolynucleotide can comprise one or more analogs (e.g., altered backbone, sugar, or nucleobase).

[0056] A “neovascular disease” is a disease state characterized by altered, dysregulated or unregulated angiogenesis. Examples of neovascular diseases include neoplastic transformation (e.g. cancer) and ocular neovascular disorders including diabetic retinopathy (DR), age-related macular degeneration (AMD, e.g., wAMD), and retinal vein occlusion (RVO). Examples of neovascular disorders or disease include neoplastic transformation (e.g. cancer) and ocular neovascular disorders including diabetic retinopathy and age-related macular degeneration. An “ocular neovascular” disorder is 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.

[0057] 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.

[0058] 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 resultof van der Waals forces. Specific binding does not however necessarily imply that an antibody or fusion protein binds one and only one target.

[0059] 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 includes a 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.

[0060] 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).

[0061] 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 inwhich 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 hypervariable regions, 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.

[0062] 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 aminoacids 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).

[0063] 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.

[0064] 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.

[0065] 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.

[0066] The term "patient" includes human and other mammalian subjects that receive either prophylactic or therapeutic treatment.

[0067] 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.

[0068] Percentage sequence identities are determined with antibody sequences maximally aligned by the Kabat numbering convention for a variable region or EU numbering for a constant region. After alignment, if a subject antibody region (e.g., the entire mature variable region of a heavy or light chain) is being compared with the same region of a reference antibody, the percentage sequence identity between the subject and reference antibody regions is the number of positions occupied by the same amino acid in both the subject and reference antibody region divided by the total number of aligned positions of the two regions, with gaps not counted, multiplied by 100 to convert to percentage. Sequence identities of other sequences can be determined by aligning sequences using algorithms, such as BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package Release 7.0, Genetics Computer Group, 575 Science Dr., Madison, WI, using default gap parameters, or by inspection, and the best alignment (i.e., resulting in the highest percentage of sequence similarity over a comparison window). Percentage of sequence identity is calculated by comparing two optimally aligned sequences over a window of comparison, determining the number of positions at which the identical residues 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.

[0069] Compositions or methods "comprising" one or more recited elements may include other elements not specifically recited. For example, a composition that comprises antibody may contain the antibody alone or in combination with other ingredients.

[0070] 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 boundto a cell and Fcy receptors, particularly Fc RI and Fc RIII, on immune effector cells such asneutrophils, macrophages and natural killer cells. The target cell is eliminated by phagocytosisor lysis, depending on the type of mediating effector cell. Death of the antibody-coated target cell occurs as a result of effector cell activity.

[0071] 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.

[0072] 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.

[0073] 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 humanizedheavy 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.

[0074] 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 functional maps 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).

[0075] 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.

[0076] 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 positionsof 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.

[0077] “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, vinyl-pyridine, vinyl-pyrrolidone 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. 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.

[0078] “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.

[0079] 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).

[0080] 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.

[0081] “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.

[0082] “Phosphorylcholine containing polymer” is a polymer that contains phosphorylcholine. “Zwitterion containing polymer” refers to a polymer that contains a zwitterion.

[0083] 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.

[0084] 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):

[0085] As used herein, “MPC” and “HEMA-PC” are interchangeable.

[0086] “Molecular weight” in the context of the polymer can be expressed as either a number average molecular weight, or a weight average molecular weight or a peak molecular weight. Unless otherwise indicated, all references to molecular weight herein refer to the peak molecular weight. These molecular weight determinations, number average (Mn), weight average (Mw) and peak (Mp), can be measured using size exclusion chromatography or other liquid chromatography techniques. Other methods for measuring molecular weight values can also be used, such as the use of end-group analysis or the measurement of colligative properties , 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 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.

[0087] 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.

[0088] “About” means variation one might see in measurements taken among different instruments, samples, and sample preparations.

[0089] “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.

[0090] “Alkyl” refers to a straight or branched, saturated, aliphatic radical having the number of carbon atoms indicated. For example, C1-C6 alkyl includes, but is not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, hexyl, etc. Other alkyl groups include, but are not limited to heptyl, octyl, nonyl, decyl, etc. Alkyl can include any number of carbons, such as 1-2, 1-3, 1-4, 1-5, 1-6, 1-7, 1-8, 1-9, 1-10, 2-3, 2-4, 2-5, 2-6, 3-4, 3-5, 3-6, 4-5, 4-6 and 5-6. The alkyl group is typically monovalent, but can be divalent, such as when the alkyl group links two moieties together.

[0091] 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.

[0092] “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.

[0093] 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” is include 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 akyl and heteroalkyl groups have 1, 2 or 3 substituents. Exceptions are those perhalo alkyl groups (e.g., pentafluoroethyl and the like) .

[0094] 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 –NO2 in 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).

[0095] “Alkoxy” refers to 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.

[0096] “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.

[0097] “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.

[0098] “Fluoro-substituted alkyl” refers to an alkyl group where one, some, or all hydrogen atoms have been replaced by fluorine.

[0099] “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.

[0100] “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.

[0101] “Endocyclic” refers to an atom or group of atoms which comprise part of a cyclic ring structure.

[0102] “Exocyclic” refers to an atom or group of atoms which are attached but do not define the cyclic ring structure.

[0103] “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).

[0104] “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.

[0105] “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.

[0106] “Alkynyl” refers to either a straight chain or branched hydrocarbon of 2 to 6 carbon atoms, having at least one triple bond. Examples of alkynyl groups include, but are not limited to, acetylenyl, propynyl, 1-butynyl, 2-butynyl, isobutynyl, sec-butynyl, butadiynyl, 1-pentynyl, 2-pentynyl, isopentynyl, 1,3-pentadiynyl, 1,4-pentadiynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 1,3-hexadiynyl, 1,4-hexadiynyl, 1,5-hexadiynyl, 2,4-hexadiynyl, or 1,3,5-hexatriynyl. Alkynyl groups can also have from 2 to 3, 2 to 4, 2 to 5, 3 to 4, 3 to 5, 3 to 6, 4 to 5, 4 to 6 and 5 to 6 carbons. The alkynyl group is typically monovalent, but can be divalent, such as when the alkynyl group links two moieties together.

[0107] “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.

[0108] “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.

[0109] “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.

[0110] “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.

[0111] “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.

[0112] “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 can be 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.

[0113] 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.

[0114] 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.

[0115] “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.

[0116] “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.

[0117] 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.

[0118] 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 theformula -(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.

[0119] “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- or 4-thiazolyl. In some embodiments, triazolyl can be 1-, 2- or 5-(1,2,4-triazolyl). In some embodiments, tetrazolyl can be 5-tetrazolyl.

[0120] 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.

[0121] 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.

[0122] 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.

[0123] “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 formsa bond to its reaction partner (the nucleophile) by accepting both bonding electrons from that reaction partner.

[0124] “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.

[0125] “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 attachmentfor the maleimido group and “ “indicates the point of attachment of the sulfur atom the thiolto the remainder of the original sulfhydryl bearing group.

[0126] 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 occurringamino 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.

[0127] “Linear” in reference to the geometry, architecture or overall structure of a polymer, refers to polymer having a single polymer arm.

[0128] “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.

[0129] “Pharmaceutically acceptable excipient” 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. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, normal saline solutions, lactated Ringer’s, normal sucrose, normal glucose, a buffer, and the like.

[0130] As used herein, “OG1786” is a 9-arm initiator used for polymer synthesis with the structure shown in FIG.1, which depicts the salt form of OG1786 with trifluoroacetic acid. OG1786 may also be used as provided herein as other salts or as the free base.

[0131] As used herein, “OG1801” is an approximately (+ / - 25%) 800 kDa polymer (either by Mn or Mp) made using OG1786 as an initiator for ATRP synthesis using the monomer HEMA-PC. The structure of OG1801 is shown in FIG.2.

[0132] As used herein, “OG1802” is OG1801 with a maleimide functionality added, and it has the structure shown in FIG.3, 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.

[0133] 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, n, of the solution's refractive index n with the molecularconcentration change, c, by measuring the dn / dc () 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.

[0134] 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.

[0135] 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 poresizes. 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.

[0136] 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.

[0137] 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 be analyzed by the ASTRA software for determination of absolute molar mass (Mp, Mw, Mn) and polydisperse index (PDI). In addition, the calculation also involves the input dn / dc values for polymer and protein as 0.142 and 0.183, respectively. For 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.

[0138] Unless indicated otherwise, “about” denotes that deviation of ±10% from the recited value is encompassed. For example, “about 5 mg” encompasses a value in the range of 4.5-5.5 mg, unless indicated otherwise. The term “about” with reference to a frequency ofdosing can denote a deviation of up to 7 days (± 7 days) before or after the noted length of time.

[0139] As used herein, 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 components in 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.

[0140] 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. METHODS

[0141] Some embodiments disclosed herein (with reference to Figure 4) are a method of treating wet age-related macular degeneration (wAMD) (400), the method comprising identifying a subject with wAMD (401). The method can further include administering a first dose of an anti-VEGF antibody conjugate (e.g., KSI-301) to the subject (402). The method can also include administering a second dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the first dose (403). The method can include administering a third dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the second dose (404). The method can also include administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the third dose (405). Further, the method can include administering an individualized dose of the anti-VEGF antibody conjugate to the subject at least 4 weeks (+ / - 7 days) after the fourth dose. In some embodiments the individualized dose is administered to the subject if the subject has a declinein eye health, wherein the individualized dose is administered no more frequently than, or than about, once every four weeks (or no more frequently than Q4W or QM) (406). The first, second, third and fourth doses can each be a loading dose. In some embodiments, the subject’s eye health is not determined after administering the first dose and before administering the second, third, and / or fourth doses. In some embodiments, the method includes administering an individualized dose of the anti-VEGF antibody conjugate to the subject upon determining that the subject has a decline in eye health. In some embodiments, the method includes determining that the subject has a decline in eye health, and administering the individualized dose of the anti-VEGF antibody conjugate to the subject based on the determined decline in eye health. The decline in eye health can be based on any suitable anatomical and / or visual acuity-based measure of eye health, as described herein. In some embodiments, administering the dose (e.g., first, second, third, fourth dose) is performed no sooner than the designated point in time (e.g., no sooner than four weeks after the last dose). In some embodiments, administering the dose (e.g., first, second, third, fourth dose) is performed no sooner than four weeks after the last dose. In some embodiments, the dosing is ±1, 2, 3, 4, 5, 6, or 7 days from the designated point in time. In some embodiments, the first, second, third, and fourth doses are each a loading dose. In some embodiments, administering the first, second, third, and fourth doses does not depend on the eye health of the subject. In some embodiments, the subject’s eye health is not evaluated to determine whether to administer the first, second, third, and fourth doses. In some embodiments, the subject’s eye health is not evaluated until after the fourth dose. In some embodiments, the subject’s eye health is evaluated only after the fourth dose. In some embodiments, one or more subsequent doses of the anti-VEGF antibody conjugate are administered to the subject no less frequently than about once every 24 weeks after the fourth and / or the last dose (e.g., the last individualized dose). In some embodiments, the one or more subsequent doses are administered no less frequently than Q24W (+ / - 7 days) or Q6M (+ / - 7 days).

[0142] In some embodiments, any one or more of the dosing intervals between consecutive doses among the first, second, third and fourth doses (e.g., between the first and second doses, between the second and third doses, between the third and fourth doses) is 3, 4, or 5 weeks. In some embodiments, any one or more of the dosing intervals between consecutive doses (e.g., between the first and second doses, between the second and thirddoses, between the third and fourth doses) is in the range of 3-4 weeks, or 4-5 weeks. In some embodiments, any one or more of the dosing intervals between consecutive doses (e.g., between the first and second doses, between the second and third doses, between the third and fourth doses) is in the range of 3-5 weeks, e.g., 21-35 days, 22-34 days, 23-33 days, 24-32 days, 25-31 days, 26-30 days, or 27-29 days. In some embodiments, any one or more of the dosing intervals between consecutive doses (e.g., between the first and second doses, between the second and third doses, between the third and fourth doses) is 4 weeks. In some embodiments, any one or more of the dosing intervals between consecutive doses (e.g., between the first and second doses, between the second and third doses, between the third and fourth doses) is or is about one month. In some embodiments, the method includes administering the individualized dose of the anti-VEGF antibody conjugate to the subject at least one month (+ / - 7 days) after the fourth dose, wherein the individualized dose is administered no more frequently than about once every four weeks (or no more frequently than Q4W or QM). In some embodiments, the method includes administering the individualized dose of the anti-VEGF antibody conjugate to the subject at least one month (+ / - 7 days) after the fourth dose if the subject has a decline in eye health, wherein the individualized dose is administered no more frequently than about once every four weeks (or no more frequently than Q4W or QM).

[0143] In some embodiments, a subsequent dose of the anti-VEGF antibody conjugate is administered to the subject if the subject has not received any dose of the anti- VEGF antibody conjugate for, for about, or for at least 24 weeks after the last dose (e.g., the fourth dose, the last loading dose, the last individualized dose, the last maintenance dose, etc.). In some embodiments, the longest interval a subject can remain without being administered the anti-VEGF antibody conjugate is or is about 24 weeks. In some embodiments, the subsequent dose is administered to the subject regardless of the subject’s eye health (e.g., regardless of whether the subject’s eye health has or has not declined). In some embodiments, the subsequent dose is administered 23, 24 or 25 weeks after the last dose (e.g., the fourth dose, the last loading dose, the last individualized dose, the last maintenance dose, etc.). In some embodiments, the subsequent dose is administered to the subject at a frequency of, or of about, once every 24 weeks, if the subject does not have a decline in eye health after the fourth dose. In some embodiments, the subsequent dose is a maintenance dose. In some embodiments, thesubsequent dose is an individualized dose (e.g., a dose administered based on a decline in eye health).

[0144] Some non-limiting embodiments disclosed herein (with reference to Figure 5) are a method of treating wAMD (500), the method comprising identifying a subject with wAMD (501). The method can further include administering a first dose of an anti-VEGF antibody conjugate to the subject (502). The method can also include administering a second dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the first dose (503). The method can also include administering a third dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the second dose (504). The method can also include administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the third dose (505). The first, second, third and fourth doses can each be a loading dose. In some embodiments, the subject’s eye health is not determined after administering the first dose and before administering the second, third, and / or fourth doses. In some embodiments, the subject’s eye health is not determined until after the fourth dose. In some embodiments, the subject’s eye health is evaluated only after the fourth dose. Further, the method can include evaluating the subject’s eye health no more frequently than about once every four weeks (or no more frequently than Q4W or QM) after the fourth dose; and if the subject has a decline in eye health upon evaluating, administering an individualized dose of the anti-VEGF antibody conjugate to the subject (506). In some embodiments, the individualized dose is administered to the subject no more frequently than about once every four weeks (or no more frequently than Q4W or QM) after the fourth dose. In some embodiments, administering the dose is no sooner than the designated point in time. In some embodiments, the dosing is ±1, 2, 3, 4, 5, 6, or 7 days. The decline in eye health can be based on any suitable anatomical and / or visual acuity-based measure of eye health, as described herein. In some embodiments, one or more subsequent doses of the anti-VEGF antibody conjugate are administered to the subject no less frequently than about once every 24 weeks after the fourth and / or the last dose (e.g., the last individualized dose). In some embodiments, the one or more subsequent doses are administered no less frequently than Q24W (+ / - 7 days) or Q6M (+ / - 7 days). In some embodiments, a subsequent dose of the anti-VEGF antibody conjugate is administered to the subject if the subject has not received any dose of the anti- VEGF antibody conjugate for, for about, or for at least 24 weeks after the last dose (e.g., thefourth dose, the last loading dose, the last individualized dose, the last maintenance dose, etc.). In some embodiments, the subsequent dose is administered to the subject at a frequency of, or of about, once every 24 weeks, if the subject does not have a decline in eye health after the fourth dose.

[0145] Some non-limiting embodiments disclosed herein (with reference to Figure 6) are a method of treating wAMD (600), the method comprising identifying a subject with wAMD (601). The method can further include administering a first dose of an anti-VEGF antibody conjugate to the subject (602). The method can also include administering a second dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the first dose (603). The method can also include administering a third dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the second dose (604). The method can also include administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the third dose (605). The first, second, third and fourth doses can each be a loading dose. In some embodiments, the subject’s eye health is not determined after administering the first dose and before administering the second, third, and / or fourth doses. In some embodiments, the subject’s eye health is not determined until after the fourth dose. In some embodiments, the subject’s eye health is evaluated only after the fourth dose. The method can also include evaluating the subject’s eye health no more frequently than about once every four weeks (or no more frequently than Q4W or QM) after the fourth dose (606). Further, the method can include administering an individualized dose to the subject based on the subject’s evaluated eye health (607). In some embodiments, the dosing is no sooner than the designated point in time. In some embodiments, the dosing is ±1, 2, 3, 4, 5, 6, or 7 days. In some embodiments, one or more subsequent doses of the anti-VEGF antibody conjugate are administered to the subject no less frequently than about once every 24 weeks after the fourth and / or the last dose (e.g., the last individualized dose). In some embodiments, the one or more subsequent doses are administered no less frequently than Q24W (+ / - 7 days) or Q6M (+ / - 7 days). In some embodiments, a subsequent dose of the anti-VEGF antibody conjugate is administered to the subject if the subject has not received any dose of the anti-VEGF antibody conjugate for, for about, or for at least 24 weeks after the last dose (e.g., the fourth dose, the last loading dose, the last individualized dose, the last maintenance dose, etc.). In someembodiments, the subsequent dose is administered to the subject at a frequency of, or of about, once every 24 weeks, if the subject does not have a decline in eye health after the fourth dose.

[0146] Some non-limiting embodiments disclosed herein (with reference to Figure 7) are methods of treating wAMD (700), the method comprising identifying a subject with wAMD (701). The method can further include administering a first dose of an anti-VEGF antibody conjugate to the subject (702). The method can also include administering a second dose of the anti-VEGF antibody conjugate to the subject 4 weeks after the first dose (703). The method can also include administering a third dose of the anti-VEGF antibody conjugate to the subject 4 weeks after the second dose (704). The method can also include administering a fourth dose of the anti-VEGF antibody conjugate to the subject 4 weeks after the third dose (705). The first, second, third and fourth doses can each be a loading dose. In some embodiments, the subject’s eye health is not determined after administering the first dose and before administering the second, third, and / or fourth doses. In some embodiments, the subject’s eye health is not determined until after the fourth dose. In some embodiments, the subject’s eye health is evaluated only after the fourth dose. Further, if the subject has a decline in eye health within about 24 weeks after the fourth dose, administering an individualized dose of the anti-VEGF antibody conjugate (706). In some embodiments, the timing of administering the individualized dose of the anti-VEGF antibody conjugate to the subject is 4, 8, 12, 16, 20 or 24 weeks after the fourth dose. In some embodiments, the dosing is no sooner than the designated point in time. In some embodiments, the dosing is ±1, 2, 3, 4, 5, 6, or 7 days. The decline in eye health can be based on any suitable anatomical and / or visual acuity-based measure of eye health, as described herein. In some embodiments, one or more subsequent doses of the anti-VEGF antibody conjugate are administered to the subject no less frequently than about once every 24 weeks after the fourth and / or the last dose (e.g., the last individualized dose). In some embodiments, the one or more subsequent doses are administered no less frequently than Q24W (+ / - 7 days) or Q6M (+ / - 7 days). In some embodiments, a subsequent dose of the anti-VEGF antibody conjugate is administered to the subject if the subject has not received any dose of the anti-VEGF antibody conjugate for, for about, or for at least 24 weeks after the last dose (e.g., the fourth dose, the last loading dose, the last individualized dose, the last maintenance dose, etc.). In some embodiments, the subsequent dose is administered to thesubject at a frequency of, or of about, once every 24 weeks, if the subject does not have a decline in eye health after the fourth dose.

[0147] In some embodiments, the individualized dose is administered no more frequently than about once every 4 weeks (or no more frequently than Q4W or QM). In some embodiments, the dosing is no sooner than the designated point in time. In some embodiments, the dosing is ±1, 2, 3, 4, 5, 6, or 7 days from the designated time interval. In some embodiments, the individualized dose is administered 4 weeks after the last loading dose or last post-loading dose of the anti-VEGF conjugate was administered to the subject (e.g., after the fourth dose, after the last individualized dose). In some embodiments, the individualized dose is administered 8 weeks after the last loading dose or last post-loading dose of the anti-VEGF conjugate was administered to the subject (e.g., after the fourth dose, after the last individualized dose). In some embodiments, the individualized dose is administered 12 weeks after the last loading dose or last post-loading dose of the anti-VEGF conjugate was administered to the subject (e.g., after the fourth dose, after the last individualized dose). In some embodiments, the individualized dose is administered 16 weeks after the fourth dose of the anti-VEGF conjugate was administered to the subject. In some embodiments, the individualized dose is administered 20 weeks after the last loading dose or last post-loading dose of the anti-VEGF conjugate was administered to the subject (e.g., after the fourth dose, after the last individualized dose). In some embodiments, the individualized dose is administered 24 weeks after the last loading dose or last post-loading dose of the anti-VEGF conjugate was administered to the subject (e.g., after the fourth dose, after the last individualized dose). In some embodiments, the individualized dose is administered 28 weeks after the last loading dose or last post-loading dose of the anti-VEGF conjugate was administered to the subject (e.g., after the fourth dose, after the last individualized dose). In some embodiments, the individualized dose is administered 32 weeks after the last loading dose or last post-loading dose of the anti-VEGF conjugate was administered to the subject (e.g., after the fourth dose, after the last individualized dose). In some embodiments, the individualized dose is administered 4, 8, 12, 16, 20, 24, 28 and / or 32 weeks after the fourth dose or after the last individualized dose of the anti-VEGF conjugate was administered to the subject or a combination of these time points, for example 4, 12, and 28 weeks; 8, 16, 20 and 24 weeks; 16, 24 and 23 weeks; or 4 and 32 weeks.

[0148] Some non-limiting embodiments disclosed herein (with reference to Figure 8) are a method of treating eye disease (800), the method comprising identifying a subject with a neovascular eye disease (801). The method can further include administering a first dose of an anti-VEGF antibody conjugate to the subject (802). The method can also include administering a second dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the first dose (803). The method can also include administering a third dose of the anti- VEGF antibody conjugate to the subject about 4 weeks after the second dose (804). The method can also include administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the third dose (805). The first, second, third and fourth doses can each be a loading dose. In some embodiments, the subject’s eye health is not determined after administering the first dose and before administering the second, third, and / or fourth doses. In some embodiments, the subject’s eye health is not determined until after the fourth dose. In some embodiments, the subject’s eye health is evaluated only after the fourth dose. Further, the method can include administering an individualized dose of the anti-VEGF antibody conjugate to the subject at least 4 weeks (+ / - 7 days) after the fourth dose if the subject has a decline in eye health, wherein the individualized dose is administered no more frequently than about once every four weeks (or no more frequently than Q4W or QM) (806). The decline in eye health can be based on any suitable anatomical and / or visual acuity-based measure of eye health, as described herein. In some embodiments, a subsequent dose of the anti-VEGF antibody conjugate is administered to the subject if the subject has not received any dose of the anti-VEGF antibody conjugate for, for about, or for at least 24 weeks after the last dose (e.g., the fourth dose, the last loading dose, the last individualized dose, the last maintenance dose, etc.). In some embodiments, the subsequent dose is administered to the subject at a frequency of, or of about, once every 24 weeks, if the subject does not have a decline in eye health after the fourth dose.

[0149] The methods provided herein can treat any suitable neovascular eye disease. In some embodiments, the neovascular eye disease is diabetic retinopathy (DR), age-related macular degeneration (AMD, e.g., wAMD), diabetic macular edema (DME), or retinal vein occlusion (RVO). In some embodiments, the eye disease is wAMD. In some embodiments, the eye disease is DR (e.g., non-proliferative DR (NPDR) or proliferative DR (PDR)). In some embodiments, the eye disease is diabetic macular edema (DME).

[0150] In some embodiments, the individualized dose is administered 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 days after the fourth dose. In some embodiments, the dosing is no sooner than the designated point in time. In some embodiments, the dosing is ±1, 2, 3, 4, 5, 6, or 7 days. The individualized dose(s) can be administered at any suitable dosing interval, e.g., upon determining that the subject has a decline in eye health based on evaluating the subject’s eye health.

[0151] Any suitable number of individualized doses can be administered to the subject under the recited dosing regimen. In some embodiments, the method includes administering, or administering at least, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or more individualized doses to the subject, or optionally a number of individualized doses that is in a range defined by any two of the preceding values (e.g., 1- 24, 2-12, 3-21, 4-10, 2-6, etc.).

[0152] In some embodiments, the method achieves a therapeutic effect of the anti- VEGF therapy at least by the last loading dose. In some embodiments, the method achieves a therapeutic effect of the anti-VEGF therapy at least by Week 16, 20, 24, 28, 32, 36, 40, 44, or 48 from the first dose (e.g., first loading dose). In some embodiments, the therapeutic effect of the anti-VEGF therapy includes an improvement in visual acuity. In some embodiments, the therapeutic effect of the anti-VEGF therapy that is an improvement in visual acuity includes a BCVA increase by ETDRS letters of 3 letters or more, e.g., 4 letters or more, 5 letters or more, 6 letters or more, 7 letters or more, 8 letters or more, 9 letters or more, 10 letters or more, 12 letters or more, 15 letters or more, 18 letters or more, 20 letters or more, 22 letters, or more, including 25 letters or more compared to pre-treatment, or optionally an increase by a number within a range defined by any two of the preceding values (e.g., 3-25 letters, 4-20 letters, 5-15 letters, 3-18 letters, 4-10 letters, etc.) compared to pre-treatment. In some embodiments, the therapeutic effect of the anti-VEGF therapy includes a BCVA increase by ETDRS letters of, of about, or of at least 5, 10, or 15 letters. In some embodiments, the therapeutic effect of the anti-VEGF therapy includes an improvement in retinal anatomical features. In some embodiments, the therapeutic effect of the anti-VEGF therapy that is an improvement in retinal anatomical features includes a reduction, or at least a slowed increase, in retinal thickness (e.g., central subfield thickness). The retinal thickness may be measured using any suitable method, including, but not limited to, optical coherence tomography (OCT) or OCT-A. In someembodiments, the therapeutic result may include a reduction in retinal thickness of about 25 μm or more, e.g., about 30 μm or more, about 40 μm or more, about 50 μm or more, about 75 μm or more, about 100 μm or more, about 125 μm or more, about 150 μm or more, about 175 μm or more, about 200 μm or more, about 225 μm or more, about 250 μm or more, about 275 μm or more, about 300 μm or more, about 325 μm or more, about 350 μm or more, about 375 μm or more, about 400 μm or more compared to pre-treatment, or optionally a reduction within a range defined by any two of the preceding values (e.g., 25-400 μm, 30-300 μm, 40-250 μm, 30-200 μm, 40-375 μm, etc.) compared to pre-treatment.

[0153] In some embodiments, the method includes evaluating the subject’s eye health after the fourth dose (e.g., after the last loading dose). In some embodiments, the method includes evaluating the subject’s eye health after the fourth dose (or after the last loading dose) to determine if the subject has a decline in eye health. In some embodiments, the method includes evaluating the subject’s eye health no more frequently than about once every four weeks (or no more frequently than Q4W or QM). In some embodiments, the method includes evaluating the subject’s eye health no more frequently than once every 3-5 weeks. In some embodiments, the method includes evaluating the subject’s eye health about once every four weeks. In some embodiments, the method includes evaluating the subject’s eye health once every 4, 5, 6, 7, 8, 9, 10, 11, 12 weeks or less frequently, optionally evaluating the subject’s eye health at a frequency of once per a number of weeks in a range defined by any two of the preceding values (e.g., once per 4-12 weeks, once per 4-8 weeks, once per 4-6 weeks, etc.). In some embodiments, the method includes evaluating the subject’s eye health after administering an individualized dose. In some embodiments, the method includes evaluating the subject’s eye health after each administration of an individualized dose.

[0154] In some embodiments, the decline in eye health is based on the presence of intraretinal fluid (IRF), the presence of subretinal fluid (SRF), and / or new or worsening macular hemorrhage due to wAMD activity. In some embodiments, the subject does not have IRF or SRF before the decline in eye health (e.g., after administering the anti-VEGF therapy). As used herein, IRF or SRF is “present” when the amount of IRF or SRF in the subject’s eye (e.g., as determined by OCT) is above a threshold amount, which in some embodiments is a predetermined normal level. In some embodiments, the subject does not have IRF or SRF before the decline in eye health (e.g., after administering the anti-VEGF therapy). In someembodiments, before the decline in eye health (e.g., after administering the anti-VEGF therapy), the subject has an amount of IRF or SRF at or below the threshold amount, which in some embodiments is the predetermined normal level. In some embodiments, the subject is determined to have the presence of intraretinal fluid (IRF) and / or subretinal fluid (SRF) when the amount of fluid in the subject’s eye is above predetermined normal levels (for example, around 5-20 nanoliters for IRF at 3mm, or 10-50 nanoliters for SRF at 3mm, or IRF exceeding 5-10 nL at 1mm, or SRF exceeding 10-25 nL at 1mm), e.g., as determined by OCT. In some embodiments, the decline in eye health is based on the presence of intraretinal fluid (IRF) (e.g., presence of an amount of IRF above a threshold amount or above a predetermined normal level). In some embodiments, the subject is determined to have the presence of intraretinal fluid (IRF) when the amount of IRF in the subject’s eye is at or above 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nL at 3 mm. In some embodiments, the subject is determined to have the presence of intraretinal fluid (IRF) when the amount of IRF in the subject’s eye is at or above 5 nL at 3 mm. In some embodiments, the subject is determined to have the presence of intraretinal fluid (IRF) when the amount of IRF in the subject’s eye is at or above 20 nL at 3 mm. In some embodiments, the subject is determined to have the presence of intraretinal fluid (IRF) when the amount of IRF in the subject’s eye is at or above 5, 6, 7, 8, 9, or 10 nL at 1 mm. In some embodiments, the subject is determined to have the presence of intraretinal fluid (IRF) when the amount of IRF in the subject’s eye is at or above 5 nL at 1 mm. In some embodiments, the subject is determined to have the presence of intraretinal fluid (IRF) when the amount of IRF in the subject’s eye is at or above 10 nL at 1 mm. In some embodiments, the decline in eye health is based on the presence of subretinal fluid (SRF) (e.g., presence of an amount of SRF above a threshold amount or above a predetermined normal level). In some embodiments, the subject is determined to have the presence of SRF when the amount of SRF in the subject’s eye is at or above 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 nL at 3 mm. In some embodiments, the subject is determined to have the presence of SRF when the amount of SRF in the subject’s eye is at or above 10 nL at 3 mm. In some embodiments, the subject is determined to have the presence of SRF when the amount of SRF in the subject’s eye is at or above 50 nL at 3 mm. In some embodiments, the subject is determined to have the presence of SRF when the amount of SRF in the subject’s eye is at orabove 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nL at 1 mm. In some embodiments, the subject is determined to have the presence of SRF when the amount of SRF in the subject’s eye is at or above 10 nL at 1 mm. In some embodiments, the subject is determined to have the presence of SRF when the amount of SRF in the subject’s eye is at or above 25 nL at 1 mm. In some embodiments, the decline in eye health is based on new or worsening macular hemorrhage due to wAMD activity. In some embodiments, the decline in eye health is based on the presence of intraretinal fluid (IRF) and subretinal fluid (SRF).

[0155] In some embodiments, the decline in eye health comprises an increase in retinal thickness. In some embodiments, the retinal thickness is based on an optical coherence tomography central subfield thickness (OCT CST). In some embodiments, the decline in eye health comprises an increase in OCT CST compared to a previous measurement of OCT CST. In some embodiments, the increase in OCT CST is compared to a prior lowest measurement. In some embodiments, the subject’s eye health is evaluated by performing optical coherence tomography (OCT) and / or optical coherence tomography angiography (OCT-A).

[0156] In some embodiments, the method of treating a subject’s eye disease comprises evaluating the subject’s eye health and determining that the subject has new or worsening macular hemorrhage due to wAMD activity. In some embodiments, the new or worsening macular hemorrhage due to wAMD activity is determined based on a fundus photograph. In some embodiments, evaluating the subject’s eye health includes taking a fundus photograph of the subject.

[0157] In some embodiments, the individualized dose is administered no more frequently than once every four weeks (+ / - 7 days). In some embodiments the individualized dose is administered 1, 2, 3, 4, 5, 6, or 7 days before or after the at least 4-weeks after the fourth dose.

[0158] Some non-limiting embodiments disclosed herein (with reference to Figure 9) are a method of treating a subject with wAMD (900), the method comprising administering to a subject in need thereof 4 loading doses of an anti-VEGF antibody conjugate at frequency of one loading dose every four weeks (or at Q4W or at QM) (901); and determining, at least 4 weeks (+ / - 7 days) after a last loading dose, a presence or absence of intraretinal fluid (IRF) and / or subretinal fluid (SRF) in an eye of the subject, and / or if macular hemorrhage due to wAMD activity has clinically worsened (902). Further, if there is IRF or SRF in the eye, or thesubject has clinically worsened macular hemorrhage due to wAMD activity, the method can include administering a maintenance dose of the anti-VEGF antibody (903). In some embodiments, a subsequent dose of the anti-VEGF antibody conjugate is administered to the subject if the subject has not received any dose of the anti-VEGF antibody conjugate for, for about, or for at least 24 weeks after the last dose (e.g., the fourth dose, the last loading dose, the last individualized dose, the last maintenance dose, etc.). In some embodiments, the subsequent dose is administered to the subject at a frequency of, or of about, once every 24 weeks, if the subject does not have a decline in eye health after the fourth dose.

[0159] In some embodiments, the decline in eye health is based on the presence of intraretinal fluid (IRF), the presence of subretinal fluid (SRF), and / or new or worsening macular hemorrhage due to wAMD activity. In some embodiments, the method includes evaluating an IRF volume and / or SRF volume in an optical coherence tomography (OCT) scan of the subject to determine that the subject has the presence of intraretinal fluid (IRF) and / or subretinal fluid (SRF), respectively. In some embodiments, determining the presence of IRF and / or SRF in the subject’s eye comprises performing optical coherence tomography (OCT). In some embodiments, the subject is determined to have the presence of intraretinal fluid (IRF) and / or subretinal fluid (SRF) when the amount of fluid in the subject’s eye above predetermined normal levels (for example, around 5-20 nanoliters for IRF at 3mm, or 10-50 nanoliters for SRF at 3mm, or IRF exceeding 5-10 nL at 1mm, or SRF exceeding 10-25 nL at 1mm), e.g., as determined by OCT. The OCT scan can be analyzed using any suitable option. In some embodiments, the OCT scan or image is analyzed by automated image analysis.

[0160] In some embodiments, a vision assessment, as measured by Best Corrected Visual Acuity (BCVA), increases by at least a threshold after administering at least the fourth dose relative to a baseline, wherein the threshold is 1-5 Early Treatment Diabetic Retinopathy Study (ETDRS) letters. In some embodiments, the threshold is 1, 2, 3, 4, or 5 ETDRS letters. In some embodiments, a vision assessment, as measured by Best Corrected Visual Acuity (BCVA), increases by at least 4.5 ETDRS letters after administering at least the fourth dose relative to a baseline.

[0161] In some embodiments, the decline in eye health comprises an increase in central subfield thickness (CST) as measured by optical coherence tomography (OCT) of at least 25-50 microns after the fourth dose, compared to the prior lowest measurement. In someembodiments, the decline in eye health includes an increase in CST of, of about, or of at least 15, 20, 25, 30, 35, 40, 45, 50, 55, or 60 microns, or more, or optionally an increase in CST in a range defined by any two of the preceding values, for example, 15-60, 25-45, 25-40, 25-35, 25-30, 30-35, 30-40, 30-45, 30-50, 35-40, 35-50, 40-50, or 45-40 microns, compared to the prior lowest measurement.

[0162] With reference to FIG. 17, provided herein is a method (1700) of treating eye disease that includes administering (1701) one or more doses (e.g., loading doses, individualized doses, etc.) of an anti-VEGF therapy no more frequently than Q4W (+ / - 7 days) to a subject with a neovascular retinal disease or condition. The method can include evaluating (1702) the subject’s eye health after administering the one or more doses of the anti-VEGF therapy to determine if the subject has a decline in eye health, wherein the evaluating comprises determining a presence or absence of intraretinal fluid (IRF) and / or subretinal fluid (SRF) in an eye of the subject. In some embodiments, evaluating the subject’s eye health for the presence or absence of IRF and / or SRF includes determining an amount or level of IRF and / or SRF in the subject’s eye. In some embodiments, the subject is determined to have the presence of intraretinal fluid (IRF) and / or subretinal fluid (SRF) when the amount of fluid in the subject’s eye above predetermined normal levels (for example, around 5-20 nanoliters for IRF at 3mm, or 10-50 nanoliters for SRF at 3mm, or IRF exceeding 5-10 nL at 1mm, or SRF exceeding 10-25 nL at 1mm), e.g., as determined by OCT. In some embodiments, the one or more doses of the anti-VEGF therapy are one or more loading doses of the anti-VEGF therapy. In some embodiments, the one or more doses of the anti-VEGF therapy are administered at Q4W (+ / - 7 days). In some embodiments, the one or more doses of the anti-VEGF therapy includes 1, 2, 3, 4, 5, 6, or more doses of the anti-VEGF therapy. In some embodiments, the one or more doses of the anti-VEGF therapy includes at least 4 doses (e.g., loading doses) of the anti-VEGF therapy. In some embodiments, the one or more doses of the anti-VEGF therapy includes 4 doses (e.g., loading doses) of the anti-VEGF therapy.

[0163] The method can be performed for any suitable purpose. In some embodiments, the method is performed as a clinical trial for the anti-VEGF therapy and the dosing regimen. In some embodiments, the method is performed to obtain data that is not for submitting to a regulatory agency (e.g., FDA) to obtain market approval for use of the anti- VEGF therapy in treating the eye disease. In some embodiments, the method is performedafter market and / or regulatory approval of use of the anti-VEGF therapy in treating the eye disease. In some embodiments, the method includes administering one or more individualized doses of the anti-VEGF therapy to the subject after administering the one or more doses if the subject has a decline in eye health, wherein the individualized dose is administered no more frequently than Q4W (+ / - 7 days). In some embodiments, the decline in eye health is or includes the presence of intraretinal fluid (IRF) and / or subretinal fluid (SRF), e.g., as described herein.

[0164] Also provided is a method of treating eye disease that includes administering an anti-VEGF therapy to a subject with a neovascular retinal disease or condition according to a treatment regimen, to thereby treat the eye disease. The treatment regimen can include administering one or more doses (e.g., loading doses, individualized doses, etc.) of the anti-VEGF therapy no more frequently than Q4W (+ / - 7 days) to the subject. In some embodiments, the treatment regimen includes administering one or more doses (e.g., loading doses, individualized doses, etc.) of the anti-VEGF therapy at Q4W (+ / - 7 days). In some embodiments, the treatment regimen includes administering 1, 2, 3, 4, 5, 6, or more doses (e.g., loading doses, individualized doses, etc.) of the anti-VEGF therapy. In some embodiments, the treatment regimen includes administering at least 4 doses (e.g., loading doses, individualized doses, etc.) of the anti-VEGF therapy. In some embodiments, the treatment regimen includes administering 4 doses (e.g., loading doses) of the anti-VEGF therapy.

[0165] The treatment regimen can have an efficacy that has been determined by evaluating an eye health of a cohort of patients with the neovascular retinal disease or condition that have been administered one or more doses of the anti-VEGF therapy no more frequently than Q4W (+ / - 7 days), by determining a change in or presence of an intraretinal fluid (IRF) and / or subretinal fluid (SRF) in an eye of each patient of the cohort (or a subset thereof) after administration of the one or more doses of the anti-VEGF therapy. In some embodiments, the efficacy has been determined by evaluating the eye health of the cohort of patients that have been administered one or more doses (e.g., loading doses, individualized doses, etc.) of the anti-VEGF therapy at a frequency of Q4W (+ / - 7 days). In some embodiments, the patients of the cohort have been administered 1, 2, 3, 4, 5, 6, or more doses (e.g., loading doses, individualized doses, etc.) of the anti-VEGF therapy. In some embodiments, the patients of the cohort have been administered at least 4 doses (e.g., loading doses, individualized doses,etc.) of the anti-VEGF therapy. In some embodiments, the patients of the cohort have been administered 4 doses (e.g., loading doses) of the anti-VEGF therapy. In some embodiments, the patients of the cohort have been further administered one or more individualized doses of the anti-VEGF therapy after administration of the one or more doses upon a decline in eye health, wherein the individualized dose is administered no more frequently than Q4W (+ / - 7 days). In some embodiments, the evaluating has been performed no more frequently than Q4W (+ / - 7 days). In some embodiments, the evaluating has been performed no more frequently than Q4W, Q8W or Q12W (+ / - 7 days). In some embodiments, the evaluating has been performed at a frequency of Q4W, Q8W or Q12W (+ / - 7 days).

[0166] In some embodiments, each patient in the cohort has been determined to have the presence of intraretinal fluid (IRF) and / or subretinal fluid (SRF) when the amount of fluid in the patient’s eye was above predetermined normal levels (for example, around 5-20 nanoliters for IRF at 3mm, or 10-50 nanoliters for SRF at 3mm, or IRF exceeding 5-10 nL at 1mm, or SRF exceeding 10-25 nL at 1mm), e.g., as determined by OCT. In some embodiments, the decline in eye health was based on the presence of intraretinal fluid (IRF) (e.g., presence of an amount of IRF above a threshold amount or above a predetermined normal level). In some embodiments, a patient of the cohort has been determined to have the presence of intraretinal fluid (IRF) when the amount of IRF in the patient’s eye was at or above 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nL at 3 mm. In some embodiments, a patient of the cohort has been determined to have the presence of intraretinal fluid (IRF) when the amount of IRF in the patient’s eye was at or above 5 nL at 3 mm. In some embodiments, a patient of the cohort has been determined to have the presence of intraretinal fluid (IRF) when the amount of IRF in the patient’s eye was at or above 20 nL at 3 mm. In some embodiments, a patient of the cohort has been determined to have the presence of intraretinal fluid (IRF) when the amount of IRF in the patient’s eye was at or above 5, 6, 7, 8, 9, or 10 nL at 1 mm. In some embodiments, a patient of the cohort has been determined to have the presence of intraretinal fluid (IRF) when the amount of IRF in the patient’s eye was at or above 5 nL at 1 mm. In some embodiments, a patient of the cohort has been determined to have the presence of intraretinal fluid (IRF) when the amount of IRF in the patient’s eye was at or above 10 nL at 1 mm. In some embodiments, the decline in eye health was based on the presence of subretinal fluid (SRF) (e.g., presence of an amount of SRF above a threshold amount or abovea predetermined normal level). In some embodiments, a patient of the cohort has been determined to have the presence of SRF when the amount of SRF in the patient’s eye was at or above 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 nL at 3 mm. In some embodiments, a patient of the cohort has been determined to have the presence of SRF when the amount of SRF in the patient’s eye was at or above 10 nL at 3 mm. In some embodiments, a patient of the cohort has been determined to have the presence of SRF when the amount of SRF in the patient’s eye was at or above 50 nL at 3 mm. In some embodiments, a patient of the cohort has been determined to have the presence of SRF when the amount of SRF in the patient’s eye was at or above 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nL at 1 mm. In some embodiments, a patient of the cohort has been determined to have the presence of SRF when the amount of SRF in the patient’s eye was at or above 10 nL at 1 mm. In some embodiments, a patient of the cohort has been determined to have the presence of SRF when the amount of SRF in the patient’s eye was at or above 25 nL at 1 mm.

[0167] In some embodiments, an efficacy of the treatment regimen is determined by a clinical trial for the anti-VEGF therapy and the dosing regimen. In some embodiments, an efficacy of the treatment regimen is determined to obtain data that is not for submitting to a regulatory agency (e.g., FDA) to obtain market approval for use of the anti-VEGF therapy in treating the eye disease. In some embodiments, an efficacy of the treatment regimen is determined after market and / or regulatory approval of use of the anti-VEGF therapy in treating the eye disease. In some embodiments, the decline in eye health is or includes the presence of or a change in intraretinal fluid (IRF) and / or subretinal fluid (SRF), e.g., as described herein. In some embodiments, the decline in eye health is or includes a change (e.g., increase) in intraretinal fluid (IRF) and / or subretinal fluid (SRF), e.g., as described herein. In some embodiments, the efficacy of the treatment regimen is or includes an absence of IRF and / or SRF. In some embodiments, the efficacy of the treatment regimen is or includes an amount or level of IRF and / or SRF at or below a threshold amount or a predetermined normal level, as described herein. In some embodiments, the efficacy of the treatment regimen is or includes a time interval until a loss of a therapeutic result of the anti-VEGF therapy (e.g., until an improvement in visual acuity or anatomical measure of eye health (for example, presence of IRF and / or SRF, or CST) due to administration of the anti-VEGF therapy is no longerobserved) after administration of the one or more doses of the anti-VEGF therapy. In some embodiments, the time interval is, is about, or is at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48 months or more. In some embodiments, the efficacy of the treatment regimen is or includes a reduction in CST (e.g., a clinically relevant reduction in CST), as described herein. In some embodiments, the efficacy of the treatment regimen is or includes a lack of an increase in CST (e.g., lack of a clinically relevant increase in CST). In some embodiments, the efficacy of the treatment regimen is or includes an improvement in visual acuity (e.g., BCVA).

[0168] In some embodiments, the method includes administering one or more individualized doses of the anti-VEGF therapy to the subject after administering the one or more doses, upon a decline in eye health. In some embodiments, the method includes evaluating the subject’s eye health after administering the one or more doses to determine if the subject has the decline in eye health, optionally wherein the evaluating comprises evaluating an OCT scan of the subject’s eye. In some embodiments, the method includes evaluating the subject’s eye health after administering the one or more doses to determine if the subject has a second decline in eye health, optionally wherein the evaluating comprises evaluating an OCT scan of the subject’s eye; and administering one or more individualized doses of the anti-VEGF therapy to the subject upon determining that the subject has the second decline in eye health, thereby treating the eye disease. In some embodiments, the efficacy of the treatment regimen (e.g., including administering one or more individualized doses to patients of the cohort) determined by evaluating the eye health of a cohort of patients is recapitulated in treating the subject by administering the anti-VEGF therapy, including administering one or more individualized doses to the subject upon the second decline in eye health determined based on the OCT evaluation (e.g., by a clinician without measuring IRF or SRF).

[0169] In some embodiments, the neovascular retinal disease or condition is selected from: diabetic retinopathy (DR), choroidal neovascularization (CNV), age-related macular degeneration (AMD), diabetic macular edema (DME), pathological myopia, von Hippel-Lindau disease, histoplasmosis of the eye, retinal vein occlusion (RVO), central retinal vein occlusion (CRVO), branch retinal vein occlusion (BRVO), corneal neovascularization, retinal neovascularization, retinopathy of prematurity (ROP), subconjunctival hemorrhage, and hypertensive retinopathy. In some embodiments, the neovascular retinal disease orcondition is or includes wAMD. In some embodiments, the neovascular retinal disease or condition is or includes DR. In some embodiments, the neovascular retinal disease or condition is or includes non-proliferative diabetic retinopathy (NPDR).

[0170] In some embodiments, the anti-VEGF therapy is or includes an anti-VEGF antibody (e.g., anti-VEGF-A antibody) and / or an anti-VEGF antibody conjugate (e.g., anti- VEGF-A antibody conjugate), as described herein. In some embodiments, the anti-VEGF therapy is or includes KSI-301, as described herein. In some embodiments, the anti-VEGF therapy is or includes an anti-VEGF fusion protein. In some embodiments, the anti-VEGF therapy is or includes aflibercept, conbercept, ranibizumab, or bevacizumab.

[0171] In any method of treatment of the present disclosure, in some embodiments, each dose (e.g., a first, second, third, fourth dose, or a loading dose, or an individualized dose, or a maintenance dose) of the anti-VEGF therapy administered to the subject includes a therapeutically effective amount of the anti-VEGF therapy. In some embodiments, each dose (e.g., a first, second, third, fourth dose, or a loading dose, or an individualized dose, or a maintenance dose) of the anti-VEGF therapy administered to the subject includes a therapeutically effective amount of the anti-VEGF antibody conjugate. In some embodiments, each dose (e.g., a first, second, third, fourth dose, or a loading dose, or an individualized dose, or a maintenance dose) of the anti-VEGF therapy administered to the subject includes a therapeutically effective amount of the anti-VEGF antibody conjugate and an unconjugated anti-VEGF antibody. In some embodiments, a therapeutically effective amount of the anti- VEGF therapy includes 1-6 mg of an anti-VEGF antibody (e.g., unconjugated, conjugated to a phosphorylcholine-containing polymer as described herein, or a mixture of both as described herein). In some embodiments, a therapeutically effective amount of the anti-VEGF therapy includes 1-5 mg, 1-4 mg, 1-3 mg, 1-2 mg, 2-5 mg, 2-4 mg, 2-3 mg, 3-4 mg, 3-5, or 4-5 mg of an anti-VEGF antibody (e.g., unconjugated, conjugated to a phosphorylcholine-containing polymer as described herein, or a mixture of both as described herein). In some embodiments, a therapeutically effective amount of the anti-VEGF therapy includes or includes about 5 mg of an anti-VEGF antibody (e.g., unconjugated, conjugated to a phosphorylcholine-containing polymer as described herein, or a mixture of both as described herein). In some embodiments, each dose (e.g., a first, second, third, fourth dose, or a loading dose, or an individualized dose, or a maintenance dose) of the anti-VEGF antibody conjugate comprises 1-6 mg of protein ofthe anti-VEGF antibody conjugate. In some embodiments, the anti-VEGF antibody conjugate comprises 1, 2, 3, 4, 5, or 6 mg of protein of the anti-VEGF antibody conjugate, or a range defined by any two of the preceding values, for example, 1-5 mg, 1-4 mg, 1-3 mg, 1-2 mg, 2- 5 mg, 2-4 mg, 2-3 mg, 3-4 mg, 3-5, or 4-5 mg. In some embodiments, each dose comprises about 5 mg of protein of the anti-VEGF antibody conjugate. In some embodiments, each loading dose of the anti-VEGF antibody conjugate includes the same or about the same amount (e.g., 5 mg) of the anti-VEGF antibody conjugate (e.g., as measured based on the amount of protein). In some embodiments, each individualized or maintenance dose of the anti-VEGF antibody conjugate includes the same or about the same amount (e.g., 5 mg) of the anti-VEGF antibody conjugate (e.g., as measured based on the amount of protein). In some embodiments, each subsequent dose of the anti-VEGF antibody conjugate includes the same or about the same amount (e.g., 5 mg) of the anti-VEGF antibody conjugate (e.g., as measured based on the amount of protein). In some embodiments, each dose (e.g., a first, second, third, fourth dose, or a loading dose, or an individualized dose, or a maintenance dose) includes 1-6 mg total of an anti-VEGF-A antibody conjugate (by weight of the antibody portion) and an unconjugated anti-VEGF-A antibody. In some embodiments, each dose (e.g., a first, second, third, fourth dose, or a loading dose, or an individualized dose, or a maintenance dose) includes 1-5 mg, 1-4 mg, 1-3 mg, 1-2 mg, 2-5 mg, 2-4 mg, 2-3 mg, 3-4 mg, 3-5, or 4-5 mg total of an anti-VEGF-A antibody conjugate (by weight of the antibody portion) and an unconjugated anti-VEGF-A antibody. In some embodiments, each dose (e.g., a first, second, third, fourth dose, or a loading dose, or an individualized dose, or a maintenance dose) includes or includes about 5 mg total of an anti-VEGF-A antibody conjugate (by weight of the antibody portion) and an unconjugated anti-VEGF-A antibody.

[0172] In some embodiments, the anti-VEGF antibody conjugate comprises an anti-VEGF-A antibody and a phosphorylcholine containing polymer, wherein the polymer is covalently bonded to the anti-VEGF-A antibody at a non-native cysteine outside a variable region of the anti-VEGF-A antibody.

[0173] In some embodiments, the anti-VEGF antibody conjugate comprises an anti-VEGF-A antibody and a phosphorylcholine containing polymer, wherein the polymer is covalently bonded to the anti-VEGF-A antibody at a non-native cysteine outside a variable region of the anti-VEGF-A antibody, wherein the anti-VEGF-A antibody comprises a lightchain and heavy chain, said heavy chain comprising an Fc region, wherein the cysteine is in the Fc region of the heavy chain, wherein the antibody conjugate has the following structure:; wherein each heavy chain of the anti-VEGF-A antibody is denoted by the letter H, and each light chain of the anti-VEGF-A antibody is denoted by the letter L; the polymer is bonded to the anti-VEGF-A antibody through a sulfhydryl at C443 according to EU numbering, whichbond is depicted on one of the heavy chains above; PC is, where the curvy line indicates the point of attachment to the rest of the polymer; 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 sequence of the heavy chain comprises SEQ ID NO: 1 or a sequence at least 90% identical thereto, and wherein thesequence of the light chain comprises SEQ ID NO: 2, or a sequence at least 90% identical thereto. In some embodiments, the anti-VEGF antibody conjugate is any one of the antibody conjugates described herein (e.g., KSI-301).

[0174] After receiving the last dose (e.g., the final loading dose, or any dosing that occurred last), the subject may retain a therapeutic result of the anti-VEGF therapy for a sustained period of time without the subject receiving a maintenance or individualized dose of the antibody conjugate. A therapeutic result of the anti-VEGF therapy may include an improvement in one or more of visual acuity, eye health, or retinal health (e.g., retinal thickness, extent of retinal perfusion, etc.) at or around the time of the final loading dose compared to before or at the time of the first loading dose. Any suitable therapeutic result of an anti-VEGF therapy may be used according to methods of the present disclosure. Suitable measures for determining therapeutic results include, e.g., visual acuity, retinal thickness, perfusion in at least one eye, diabetic retinopathy severity score (DRSS), disease activity of the eye disorder, the absence or a decrease in intraretinal fluid (IRF), subretinal fluid (SRF), or any combination thereof.

[0175] In some embodiments, the therapeutic result includes an improvement, or at least a slowed decline, in visual acuity. Visual acuity may be monitored using any suitable method. In some embodiments, the visual acuity is measured by best corrected visual acuity (BCVA) using e.g., ETDRS letters or Snellen chart, etc. In some embodiments, the therapeutic result may include an improvement in BCVA measured by ETDRS letters of 1 letter or more, e.g., 2 letters or more, 3 letters or more, 4 letters or more, 5 letters or more, 6 letters or more, 7 letters or more, 8 letters or more, 9 letters or more, 10 letters or more, 12 letters or more, 15 letters or more, 18 letters or more, 20 letters or more, 22 letters, or more, including 25 letters or more, or by a number within a range defined by any two of the preceding values, compared to pre-treatment. In some embodiments, the therapeutic result may include a reduction in the rate of deterioration of BCVA by at least 10%, e.g., at least 15%, at least 25%, at least 50%, at least 75%, at least 90%, including about 100%, or any percentage in a range defined by any two of the preceding values, over pre-treatment.

[0176] In some embodiments, the therapeutic result includes a reduction, or at least a slowed increase, in retinal thickness (e.g., central subfield thickness). The retinal thickness may be measured using any suitable method, including, but not limited to, optical coherencetomography (OCT) or OCT-A. In some embodiments, the therapeutic result may include a reduction in retinal thickness of about 25 μm or more, e.g., about 30 μm or more, about 40 μm or more, about 50 μm or more, about 75 μm or more, about 100 μm or more, about 125 μm or more, about 150 μm or more, about 175 μm or more, about 200 μm or more, about 225 μm or more, about 250 μm or more, about 275 μm or more, about 300 μm or more, about 325 μm or more, about 350 μm or more, about 375 μm or more, about 400 μm or more, or a reduction within a range defined by any two of the preceding values, compared to pre-treatment. In some embodiments, the therapeutic result may include a reduction in the rate of increase in retinal thickness by at least 10%, e.g., at least 15%, at least 25%, at least 50%, at least 75%, at least 90%, including about 100%, or any percentage in a range defined by any two of the preceding values, over pre-treatment.

[0177] In some embodiments, the therapeutic result includes improved perfusion, or at least a reduction in the rate of expansion of non-perfusion, of the retina. Perfusion may be monitored using any suitable method. Suitable methods include, without limitation, OCT- angiography (OCT-A), fluorescein angiogram or ultrawide-field fluorescein angiogram. The degree of perfusion, or non-perfusion, may be measured using any suitable measure. In some embodiments, non-perfusion area or area of capillary non-perfusion is measured. In some embodiments, an ischemic index is calculated by dividing the non-perfusion area by the total retinal area. In some embodiments, the presence or absence of retinal non-perfusion in retinal quadrants on the angiogram is measured. In some embodiments, the therapeutic result may include a reduction in the area of non-perfusion of at least 10%, e.g., at least 15%, at least 25%, at least 50%, at least 75%, at least 90%, including about 100%, or any percentage in a range defined by any two of the preceding values, over pre-treatment. In some embodiments, the therapeutic result may include a reduction in the rate of progressive non-perfusion of at least 10%, e.g., at least 15%, at least 25%, at least 50%, at least 75%, at least 90%, including about 100%, or any percentage in a range defined by any two of the preceding values, over pre- treatment.

[0178] In some embodiments, the therapeutic result includes improved, or prevented worsening of, diabetic retinopathy severity score (DRSS). In some embodiments, the therapeutic result may include an improved DRSS of 2 steps or more, or 3 steps or more compared to pre-treatment. In some embodiments, the therapeutic result may includepreventing worsening of DRSS by 2 steps or more, or 3 steps or more compared to pre- treatment.

[0179] The therapeutic result is retained if the level of visual acuity or retinal health (e.g., retinal thickness, presence or absence of IRF, or presence or absence of SRF, etc.) does not worsen by more than a predetermined amount compared to the improved level. In some embodiments, the therapeutic result is retained if the level of visual acuity or retinal health does not revert by 10% or more, e.g., 20% or more, 30% or more, 40% or more, 50% or more, 75% or more, 90% or more, including 100% or more to the pretreatment level of visual acuity or retinal health after the last dose (e.g., final loading dose). In some embodiments, the therapeutic result is retained if the rate of change of visual acuity or retinal health does not revert by 10% or more, e.g., 20% or more, 30% or more, 40% or more, 50% or more, 75% or more, 90% or more, including 100% or more to the pretreatment level of the rate of change of visual acuity or retinal health after the last dose (e.g., final loading dose).

[0180] In some embodiments, the therapeutic result includes an improvement in visual acuity. In some embodiments the therapeutic result may be retained if BCVA does not fall by 1 letter or more, 2 letters or more, 3 letters or more, 4 letters or more, 5 letters or more, 6 letters or more, 7 letters or more, 8 letters or more, 9 letters or more, or 10 letters or more from the BCVA score at the time of the final loading dose (e.g., at Week 12 after three monthly loading doses). In some embodiments the therapeutic result may be retained if BCVA does not fall by 1 letter or more, 2 letters or more, 3 letters or more, 4 letters or more, 5 letters or more, 6 letters or more, 7 letters or more, 8 letters or more, 9 letters or more, or 10 letters or more from the BCVA score measured at the last assessment (e.g., 4 weeks ago). In some embodiments the therapeutic result may be retained if BCVA does not fall by 1 letter or more, 2 letters or more, 3 letters or more, 4 letters or more, 5 letters or more, 6 letters or more, 7 letters or more, 8 letters or more, 9 letters or more, or 10 letters or more from the best measured BCVA score, or the average of the 2 best measured BCVA scores, of the subject.

[0181] In some embodiments, the therapeutic result includes a reduction in retinal thickness (e.g., central subfield thickness). In some embodiments, the therapeutic result may be retained if retinal thickness (e.g., central subfield thickness) does not increase by 25 μm or more, 30 μm or more, 40 μm or more, 50 μm or more, 75 μm or more, 100 μm or more, 125 μm or more, or 150 μm or more from the retinal thickness at the time of the last dose (e.g.,final loading dose) (e.g., at Week 12 after three monthly loading doses). In some embodiments, the therapeutic result may be retained if retinal thickness (e.g., central subfield thickness) does not increase by 25 μm or more, 30 μm or more, 40 μm or more, 50 μm or more, 75 μm or more, 100 μm or more, 125 μm or more, or 150 μm or more from the retinal thickness measured at the last assessment (e.g., 4 weeks ago). In some embodiments, a retained therapeutic result includes retinal thickness that is not greater than 150 μm, 125 μm, 100 μm, 75 μm, 50 μm, 40 μm, or 30 μm, compared to the lowest measured retinal thickness of the subject.

[0182] In some embodiments, the therapeutic result includes improved perfusion of the retina. In patients with DR and DME, the retina can have an area of non-perfusion, or absence of blood flow. In some embodiments, non-perfusion is visualized on angiograms. In some embodiments, a therapeutic result of the anti-VEGF antibody conjugate, e.g., KSI-301, administration, according to methods of the present disclosure, includes regression of non- perfusion, or re-perfusion of the retina. In some embodiments, the therapeutic result may be retained if the area of non-perfusion is reduced by 10% or more, e.g., 15% or more, 25% or more, 50% or more, 75% or more, 90% or more, or about 100% relative to the area of non- perfusion at the time of the last dose (e.g., final loading dose). In some embodiments, the therapeutic result may be retained if the area of non-perfusion is not increased by 10% or more, e.g., 15% or more, 25% or more, 50% or more, 75% or more, 90% or more, or about 100% relative to the area of non-perfusion at the time of the last dose (e.g., final loading dose). In some embodiments, the therapeutic result may be retained if the area of non-perfusion is not increased by 10% or more, e.g., 15% or more, 25% or more, 50% or more, 75% or more, 90% or more, or about 100% relative to the area of non-perfusion measured at the last assessment (e.g., 4 weeks ago). In some embodiments, the therapeutic result may be retained if the area of non-perfusion is not increased by 10% or more, e.g., 15% or more, 25% or more, 50% or more, 75% or more, 90% or more, or about 100% relative to the smallest area of non-perfusion measured in the subject.

[0183] In some embodiments, the method includes administering at least four loading doses of the anti-VEGF antibody conjugate (e.g., KSI-301) to the subject, and administering one or more maintenance or individualized doses of the anti-VEGF antibody conjugate, e.g., KSI-301, after the final loading dose. The maintenance or individualized dosesmay be administered to the subject no more frequently than once every 4 weeks, e.g., every 6 weeks, every 8 weeks, every 10 weeks, every 12 weeks, every 14 weeks, every 16 weeks, every 18 weeks, every 20 weeks, or every 24 weeks. Whether to administer a maintenance or individualized dose of the anti-VEGF antibody conjugate may be determined based on the presence of intraretinal fluid (IRF), subretinal fluid (SRF) and or new or clinically worsening macular hemorrhage due to wAMD activity. In some embodiments, patients who do not have the presence of intraretinal fluid (IRF), subretinal fluid (SRF) and or new or clinically worsening macular hemorrhage due to wAMD activity may receive treatment if there is an increase in OCT CST compared to prior lowest measurement. In some embodiments, the presence of new or clinically worsening macular hemorrhage due to wAMD activity is documented with a fundus photograph.

[0184] In some embodiments, the maintenance or individualized dose may be administered based on a predetermined schedule (e.g., a schedule determined before the subject is administered any of the one or more of the loading doses). In some embodiments, the maintenance or individualized dose may be administered based on a predetermined schedule based on the severity of the eye disorder, the subject’s previous response, or lack thereof, to other therapies for the eye disorder, or any other clinically relevant factors associated with the subject. In some embodiments, the maintenance or individualized dose may be administered based on the outcome of one or more assessment tests for ocular health and / or function carried out on the subject during the course of treatment with the anti-VEGF antibody conjugate. In some embodiments, the maintenance or individualized dose may be administered based on the outcome of one or more assessment tests carried out on the subject every 4 or more weeks, e.g., every 6 or more weeks, every 8 or more weeks, every 10 or more weeks, every 12 or more weeks, every 16 or more weeks, every 20 or more weeks, every 24 or more weeks, every 28 or more weeks, every 32 or more weeks, every 36 or more weeks, including every 40 or more weeks. In some embodiments, the maintenance or individualized dose may be administered if one or more assessment tests indicates a diminishment of the therapeutic result of the anti- VEGF therapy that is greater than a predetermined threshold. In some embodiments, the maintenance or individualized dose may be administered if one or more assessment tests does not indicate a diminishment of the therapeutic result of the anti-VEGF therapy or the diminishment is not greater than a predetermined threshold

[0185] As used herein, “Q4W”, “QM” and the like each refers to a dosing schedule, and have the ordinary and customary meaning to one of ordinary skill in the art in view of the present disclosure. The number may indicate the number of the unit of time specified by the subsequent letter. “W” indicates a unit of a week; “M” specifies an interval of a month. Thus, Q4W refers to a dosing interval of 4 weeks, which also includes a dosing interval of one month; Q8W refers to a dosing interval of 8 weeks, which also includes a dosing interval of two months; and so on. QM refers to a dosing interval of one month. Q4W and QM are used interchangeably herein. As used herein, specification of a dosing schedule does not necessarily imply a number of doses beyond two, unless indicated otherwise. In some embodiments, a dosing schedule refers to the dosing schedule for maintenance doses (including the interval between the last loading dose, and the first maintenance dose). A reference to a dosing schedule being “longer” (e.g., “Q12W or longer”) refers to the time interval between doses being longer than that specified (e.g., a dosing interval of 12 weeks or longer).

[0186] In some embodiments, the total number of injections (including loading and maintenance doses) of the anti-VEGF antibody conjugate (e.g., KSI-301) administered to the subject in the first year of treatment is 10 times or less, 9 times or less, 8 times or less, 7 times or less, 6 times or less, 5 times or less, 4 times or less, 3 times or less, 2 times or less, or once, in order to retain the therapeutic result of the anti-VEGF antibody conjugate therapy. As used herein, “first year of treatment” and similar reference to the first “X” years of treatment denotes the time period measured from the first dose (e.g., first loading dose) of the anti-VEGF antibody polymer conjugate administered to the subject under the particular treatment regimen or schedule. In some embodiments, the time period is measured from the first dose (e.g., first loading dose) of the anti-VEGF antibody polymer conjugate administered to a treatment naïve subject (e.g., a subject who has not received an anti-VEGF therapy before being administered the anti-VEGF antibody polymer conjugate of the present disclosure). In some embodiments, the time period is measured from the first dose (e.g., first loading dose) of the anti-VEGF antibody polymer conjugate administered to a subject who has previously been treated with an anti-VEGF therapy (e.g., an anti-VEGF therapy that does not include the anti-VEGF antibody polymer conjugate of the present disclosure) but has not responded to the anti-VEGF therapy. In some embodiments, the total number of injections (including loading and maintenance doses) of the anti-VEGF antibody conjugate (e.g., KSI-301) administered to the subject in thefirst two years of treatment is 10 times or less, 9 times or less, 8 times or less, 7 times or less, 6 times or less, 5 times or less, 4 times or less, 3 times or less, 2 times or less, or once, in order to retain the therapeutic result of the anti-VEGF antibody conjugate therapy. In some embodiments, the total number of injections (including loading and maintenance doses) of the anti-VEGF antibody conjugate (e.g., KSI-301) administered to the subject in the first three years of treatment is 10 times or less, 9 times or less, 8 times or less, 7 times or less, 6 times or less, 5 times or less, 4 times or less, 3 times or less, 2 times or less, or once, in order to retain the therapeutic result of the anti-VEGF antibody conjugate therapy. In any method of the present disclosure, in some embodiments, at least 4 doses (e.g., loading doses and any maintenance or individualized doses) of the anti-VEGF antibody polymer conjugate is administered to the subject in the first year of treatment.

[0187] In some embodiments, the total number of maintenance doses of the anti- VEGF antibody conjugate (e.g., KSI-301) administered to the subject in a one-year period for treatment is 7 times or less, 6 times or less, 5 times or less, 4 times or less, 3 times or less, 2 times or less, once or less, or zero, in order to retain the therapeutic result of the anti-VEGF antibody conjugate therapy. In some embodiments, the total number of maintenance doses of the anti-VEGF antibody conjugate (e.g., KSI-301) administered to the subject in a two-year period for treatment is 7 times or less, 6 times or less, 5 times or less, 4 times or less, 3 times or less, 2 times or less, once or less, or zero, in order to retain the therapeutic result of the anti- VEGF antibody conjugate therapy. In some embodiments, the total number of maintenance doses of the anti-VEGF antibody conjugate (e.g., KSI-301) administered to the subject in a three-year period for treatment is 7 times or less, 6 times or less, 5 times or less, 4 times or less, 3 times or less, 2 times or less, once or less, or zero, in order to retain the therapeutic result of the anti-VEGF antibody conjugate therapy.

[0188] In some embodiments, the total number of injections (including loading and maintenance doses) of the anti-VEGF antibody conjugate (e.g., KSI-301) administered to the subject with wAMD in the first year of treatment is 8 times or less, 7 times or less, 6 times or less, 5 times or less, 4 times or less, 3 times or less, 2 times or less, or once, in order to retain the therapeutic result of the anti-VEGF antibody conjugate therapy. In some embodiments, the total number of injections (including loading and maintenance doses) of the anti-VEGF antibody conjugate (e.g., KSI-301) administered to the subject with wAMD in the first twoyears of treatment is 8 times or less, 7 times or less, 6 times or less, 5 times or less, 4 times or less, 3 times or less, 2 times or less, or once, in order to retain the therapeutic result of the anti- VEGF antibody conjugate therapy. In some embodiments, the total number of injections (including loading and maintenance doses) of the anti-VEGF antibody conjugate (e.g., KSI- 301) administered to the subject with wAMD in the first three years of treatment is 8 times or less, 7 times or less, 6 times or less, 5 times or less, 4 times or less, 3 times or less, 2 times or less, or once, in order to retain the therapeutic result of the anti-VEGF antibody conjugate therapy.

[0189] In some embodiments, the total number of maintenance doses of the anti- VEGF antibody conjugate (e.g., KSI-301) administered to the subject with wAMD in a one- year period for treatment is 4 times or less, 3 times or less, 2 times or less, once or less, or zero, in order to retain the therapeutic result of the anti-VEGF antibody conjugate therapy. In some embodiments, the total number of maintenance doses of the anti-VEGF antibody conjugate (e.g., KSI-301) administered to the subject with wAMD in a two-year period for treatment is 4 times or less, 3 times or less, 2 times or less, once or less, or zero, in order to retain the therapeutic result of the anti-VEGF antibody conjugate therapy. In some embodiments, the total number of maintenance doses of the anti-VEGF antibody conjugate (e.g., KSI-301) administered to the subject with wAMD in a three-year period for treatment is 4 times or less, 3 times or less, 2 times or less, once or less, or zero, in order to retain the therapeutic result of the anti-VEGF antibody conjugate therapy.

[0190] In some embodiments, a first maintenance or individualized dose is administered at a first time after the last loading dose, and a second maintenance or individualized dose is administered at a second period of time after the first maintenance or individualized dose, where no other dose is administered between the last loading dose and the first maintenance or individualized dose, or between the first maintenance or individualized dose and the second maintenance or individualized dose. The second period of time between the first and second subsequent doses may be the same or different from the first period of time between the last loading dose and the first maintenance or individualized dose. In some embodiments, the first time period is 8 weeks or more, e.g., 10 weeks or more, 12 weeks or more, 14 weeks or more, 16 weeks or more, 18 weeks or more, 20 weeks or more, or 24 weeks or more. In some embodiments, the second period of time is longer than the first period oftime by 0 weeks or more, e.g., by 4 weeks or more, by 6 weeks or more, by 8 weeks or more, by 10 weeks or more, by 12 weeks or more, by 16 weeks or more, by 20 weeks or more. The timing for administering the second maintenance or individualized dose may depend on the outcome of one or more assessments for ocular health and / or function of the subject.

[0191] Any suitable amount of the anti-VEGF antibody conjugate, or anti-VEGF protein conjugate (e.g., KSI-301), may be administered to the subject in an individualized or maintenance dose. In some embodiments, the subsequent dose includes about 1 mg or more, e.g., about 1.25 mg or more, about 1.5 mg or more, about 1.75 mg or more, about 2 mg or more, about 2.5 mg or more, about 3 mg or more, about 3.5 mg or more, about 4 mg or more, about 4.5 mg of more, including about 5 mg or more (by weight of the anti-VEGF antibody portion) of the anti-VEGF antibody conjugate. In some embodiments, the subsequent dose includes from about 1 mg to about 10 mg, e.g., about 1 mg to about 7.5 mg, about 1.25 mg to about 5 mg, including about 2 mg to about 5 mg (by weight of the anti-VEGF antibody portion) of the anti-VEGF antibody conjugate.

[0192] As the therapeutic result of the anti-VEGF therapy is retained for a sustained period of time after the last dose (e.g., final loading dose), the subject may not need to receive a dose of the anti-VEGF antibody conjugate, or anti-VEGF protein conjugate (e.g., an aflibercept biopolymer conjugate), while the therapeutic effect lasts. In some embodiments, no further administration of the anti-VEGF antibody conjugate (e.g., KSI-301), or anti-VEGF protein conjugate, is made to the subject within 4 weeks or more, e.g., within 6 weeks or more, within 8 weeks or more, within 10 weeks or more, within 12 weeks or more, within 14 weeks or more, within 16 weeks or more, within 20 weeks or more, within 24 weeks or more, within 28 weeks or more, within 32 weeks or more, within 36 weeks or more, within 40 weeks or more, within 44 weeks or more, within 48 weeks or more, including within 52 weeks or more, after the last dose (e.g., final loading dose).

[0193] Pharmacokinetic studies have been conducted in rabbit which demonstrate that KSI-301 has extended ocular half-life, penetrates ocular tissues well and is distributed to the retina and choroid. In rabbit, KSI-301 has an ocular half-life of approximately 11 days, which is significantly longer than the reported rabbit half-life measured for aflibercept and ranibizumab, which are 4 to 5 and 3 to 4 days, respectively (CovanceStudy 8376321, Park 2016). A series of non-clinical GLP repeat dose (4-week dosing intervals) toxicology studiesin cynomolgus monkeys testing the ocular and systemic safety of KSI-301 have been conducted through 26 weeks (7 intravitreal doses) and 10 weeks (3 intravenous doses), respectively. Results show that KSI-301 was well tolerated up to the maximum dose tested of 5 mg / eye (intravitreal) and 5 mg / kg (intravenous) in the ocular and systemic studies, respectively. Together, data extrapolated from non-clinical PK and toxicology studies indicate that KSI-301 can be safely and effectively dosed in human subjects. In some embodiments, the route of administration is via an intravitreal injection. In some embodiments, an anti-VEGF antibody conjugate (e.g., KSI-301) can be administered every 3 - 4 months, after a loading dose completion, or even less frequently.

[0194] Also provided herein is a method of treating wet age-related macular degeneration (wAMD) that includes: identifying a subject with wAMD; administering a first dose of an anti-VEGF antibody conjugate (e.g., KSI-301) to the subject; administering a second dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the first dose; administering a third dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the second dose; administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the third dose; and administering an individualized dose of the anti-VEGF antibody conjugate to the subject at least 4 weeks (+ / - 7 days) after the fourth dose if the subject has a decline in eye health, wherein the individualized dose is administered no more frequently than, once every four weeks (or no more frequently than Q4W or QM), wherein the first, second, third, fourth doses and the individualized dose each includes about 5 mg of protein of the anti-VEGF antibody conjugate, wherein the anti-VEGF antibody conjugate includes: (1) an anti-VEGF-A antibody that includes: a light chain that has the amino acid sequence set forth in SEQ ID NO: 2; and a heavy chain that has the amino acid sequence set forth in SEQ ID NO :1 (or any of the variants thereof in FIG.10, e.g., SEQ ID NOs: 9, 10, 11, or 12); and (2) a phosphorylcholine containing polymer, wherein the polymer is covalently bonded to the heavy chain of the anti-VEGF-A antibody, where the antibody conjugate has the following structure:where: each heavy chain of the anti-VEGF-A antibody is denoted by the letter H, and each light chain of the anti-VEGF-A antibody is denoted by the letter L; the polymer is bonded to the heavy chain through a sulfhydryl at C443 according to EU numbering, which bond isdepicted on one of the heavy chains above; PC is, where the curvy line indicates the point of attachment to the rest of the polymer; 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%.

[0195] Also provided is a method of treating wet age-related macular degeneration (wAMD) that includes: identifying a subject with wAMD; administering a first dose of an anti- VEGF antibody conjugate (e.g., KSI-301) to the subject; administering a second dose of theanti-VEGF antibody conjugate to the subject about 4 weeks after the first dose; administering a third dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the second dose; administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the third dose; and if the subject has a decline in eye health within about 24 weeks after the fourth dose, administering an individualized dose of the anti-VEGF antibody conjugate, wherein the first, second, third, fourth doses and the individualized dose each includes about 5 mg of protein of the anti-VEGF antibody conjugate, wherein the anti-VEGF antibody conjugate includes: (1) an anti-VEGF-A antibody that includes: a light chain that has the amino acid sequence set forth in SEQ ID NO: 2; and a heavy chain that has the amino acid sequence set forth in SEQ ID NO :1 (or any of the variants thereof in FIG. 10, e.g., SEQ ID NOs: 9, 10, 11, or 12); and (2) a phosphorylcholine containing polymer, wherein the polymer is covalently bonded to the heavy chain of the anti-VEGF-A antibody, where the antibody conjugate has the following structure:where: each heavy chain of the anti-VEGF-A antibody is denoted by the letter H, and each light chain of the anti-VEGF-A antibody is denoted by the letter L; the polymer is bonded to the heavy chain through a sulfhydryl at C443 according to EU numbering, which bond isdepicted on one of the heavy chains above; PC is, where the curvy line indicates the point of attachment to the rest of the polymer; 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%.

[0196] Also provided is a method of treating a subject with wet age-related macular degeneration (wAMD) that includes: administering to a subject in need thereof 4 loading doses of an anti-VEGF antibody conjugate at frequency of one loading dose every four weeks (or atQ4W or at QM); determining, at least 4 weeks (+ / - 7 days) after a last loading dose, a presence or absence of intraretinal fluid (IRF) and / or subretinal fluid (SRF) in an eye of the subject, and / or if macular hemorrhage due to wAMD activity has clinically worsened; and if there is IRF or SRF in the eye, or the subject has clinically worsened macular hemorrhage due to wAMD activity, administering a maintenance dose, wherein the loading doses and the maintenance dose each includes about 5 mg of protein of the anti-VEGF antibody conjugate, wherein the anti-VEGF antibody conjugate includes: (1) an anti-VEGF-A antibody that includes: a light chain that has the amino acid sequence set forth in SEQ ID NO: 2; and a heavy chain that has the amino acid sequence set forth in SEQ ID NO :1 (or any of the variants thereof in FIG. 10, e.g., SEQ ID NOs: 9, 10, 11, or 12); and (2) a phosphorylcholine containing polymer, wherein the polymer is covalently bonded to the heavy chain of the anti-VEGF-A antibody, where the antibody conjugate has the following structure:where: each heavy chain of the anti-VEGF-A antibody is denoted by the letter H, and each light chain of the anti-VEGF-A antibody is denoted by the letter L; the polymer is bonded to the heavy chain through a sulfhydryl at C443 according to EU numbering, which bond isdepicted on one of the heavy chains above; PC is, where the curvy line indicates the point of attachment to the rest of the polymer; 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%.

[0197] Also provided herein is a method of treating diabetic retinopathy, such as non-proliferative diabetic retinopathy. Provided herein is a method of treating diabetic retinopathy (DR), the method comprising: identifying a subject with DR; administering a first dose of an anti-VEGF antibody conjugate to the subject; administering a second dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the first dose; administering a third dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the second dose; administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 12 weeks after the third dose; and administering a fifth dose of the anti-VEGF antibody conjugate to the subject about 24 weeks after the fourth dose. In some embodiments, the method includes administering one or more subsequent doses of the anti-VEGF antibody conjugate to the subject after the fifth dose at or no more frequently than Q24W. In some embodiments, the method includes administering 3 doses of the anti-VEGF antibody conjugate to the subject at Q4W (+ / - 7 days); administering a fourth dose of the anti-VEGF antibody conjugate to the subject 12 weeks (+ / - 7 days) after the third dose; then administering one or more doses of the anti-VEGF antibody conjugate to the subject at Q24W after the third dose. In some embodiments, the DR is non-proliferative diabetic retinopathy (NPDR). In some embodiments, the subject is identified as having moderately severe to severe NPDR. In some embodiments, the identified subject is a treatment naive subject.

[0198] Also provided is a method of treating Diabetic Retinopathy (DR), the method comprising: (a) identifying a treatment naive subject with moderately severe to severe NPDR; (b) administering a first dose of an anti-VEGF antibody conjugate to the subject; (c) administering a second dose of the anti-VEGF antibody conjugate to the subject about 4 weeksafter the first dose; (d) administering a third dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the second dose; (e) administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 12 weeks after the third dose; and (f) administering a fifth dose of the anti-VEGF antibody conjugate to the subject about 24 weeks after the fourth dose, (g) performing (b)-(f) unless the subject develops DME, PDR, and / or ASNV, in which case: (h) administering a fifth dose of an anti-VEGF antibody conjugate to the subject when the subject is identified as having developed DME, PDR, and / or ASNV; (i) administering a sixth dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the fifth dose; (j) administering a seventh dose of the anti-VEGF antibody conjugate to the subject about 12 weeks after the sixth dose; (k) administering an eighth dose of the anti-VEGF antibody conjugate to the subject about 12 weeks after the seventh dose; and (l) administering a ninth dose of the anti-VEGF antibody conjugate to the subject about 12 weeks after the eighth dose.

[0199] In some embodiments, the subject has a diabetic retinopathy severity score (DRSS) from 47 to 53 before administering the first dose. In some embodiments, before administering the first dose the subject has a CST 320 microns and a BCVA ETDRS letter score in the of 69 letters (approximate Snellen equivalent of 20 / 40 or better), or a CST >320 and 350 microns and a BCVA ETDRS letter score of 79 letters (approximate Snellen equivalent of 20 / 25 or better). Indicators for Retreatment

[0200] In some embodiments, subjects in need of retreatment for wAMD can be identified by specific criteria indicating a decline in eye health. In some embodiments, subjects in need of retreatment for wAMD can be identified by an IRF exceeding 5-20 nL or SRF exceeding 10-50 nL at 3mm. In some embodiments, subjects in need of retreatment for wAMD can be identified by an IRF exceeding 5-10 nL or SRF exceeding 10-25 nL at 1mm, or CST- RPE / CST-BM increase above 25-100 m. In some embodiments, subjects in need of retreatment for wAMD can be identified by a CST-RPE / CST-BM increase above 25-100 m. In some embodiments, subjects in need of retreatment for wAMD can be identified by an IRF exceeding 5-20 nL or SRF exceeding 10-50 nL at 3mm or IRF exceeding 5-10 nL or SRF exceeding 10-25 nL at 1mm. In some embodiments, subjects in need of retreatment for wAMD can be identified by new onset of retinal hemorrhage from CNV. In some embodiments, criteriaare assessed after the fourth dose of an anti-VEGF-A antibody conjugate, and the criteria are compared to the prior lowest measurement. Remote monitoring of treatment progress

[0201] In some embodiments, determining a subject’s eye health after administration of an anti-VEGF-A antibody conjugate according to embodiments described herein includes the subject obtaining an image of the subject’s own retina using a suitable imaging device. In some embodiments, a device is provided herein, comprising an imaging device for remote monitoring of treatment progress in a subject wherein the subject has been, is, or will be treated with any of the compositions or methods described herein. In some embodiments, the device can comprise wearable goggles and / or a compact desktop model. In some embodiments, the device can assess retinal fluid volume and / or retinal thickness using traditional and / or machine learning algorithms. In some embodiments, the device can use scanning laser ophthalmoscopy (SLO) and / or optical coherence tomography (OCT) imaging modalities to capture images. In some embodiments, the device can be controlled remotely (i.e., by a physician or other healthcare professional). In some embodiments, the device can provide real-time image viewing and evaluation by a physician or other healthcare professional. In some embodiments, the device provides real-time image viewing and evaluation by a physician or other healthcare professional at the same or a different location as the patient. In some embodiments, the device can detect potential bacterial infections. In some embodiments, the device can be used to detect bacterial infections within the first 24 hours following treatment with any of the compositions or methods described herein. In some embodiments, the device is used daily by a subject to capture images. In some embodiments, the device uploads the captured images to a cloud system for storage and / or analysis. In some embodiments, the device transmits the images and / or processed data to a physician or other healthcare professional. In some embodiments, the device provides diagnostic feedback to the subject regarding the uploaded images. In some embodiments, the physician or other healthcare professional can analyze the images and / or processed data and provide a recommendation regarding further course of treatment. In some embodiments, the subject has wet age-related macular degeneration (AMD). As used herein, “remote” in the context of monitoring treatment progress denotes a location that is not specialized to carry out a medical treatment and / or evaluation of the subject. In some embodiments, a remote location is one thatis not a doctor’s office, a clinic, a hospital, or a medical facility. In some embodiments, a remote location is one where there is no medical practitioner (e.g., doctor, physician, ophthalmologist, nurse, etc.) onsite. In some embodiments, a remote location is the subject’s home or workplace.

[0202] In some embodiments, systems for remote monitoring of treatment progress in a subject, wherein the subject has been, is, or will be treated with any of the compositions or methods described herein are provided herein, comprising, a memory; at least one processor; and at least one non-transitory computer readable medium containing instructions that, when executed by the at least one processor, cause the at least one processor to perform operations comprising: capturing images for assessment of retinal fluid volume and retinal thickness, uploading the captured images to a cloud system, processing the images with traditional and / or machine learning algorithms to generate processed data, sending the processed data to a physician or other health care professional, diagnosing a subject based upon the processed data, providing the diagnosis directly to the subject, detecting a bacterial infection based upon the processed data, or any or all of the steps recited herein. In some embodiments, the subject has wet age-related macular degeneration (AMD).

[0203] In some embodiments, methods for remote monitoring of treatment progress in a subject, wherein the subject has been, is, or will be treated with any of the compositions or methods described herein are provided herein. In some embodiments, the methods comprise capturing images for assessment of retinal fluid volume and retinal thickness. In some embodiments, the methods comprise uploading the captured images to a cloud system. In some embodiments, the methods comprise processing the images with traditional and / or machine learning algorithms to generate processed data. In some embodiments, the methods comprise sending the processed data to a physician or other health care professional. In some embodiments, the methods comprise diagnosing a subject based upon the processed data. In some embodiments, the methods comprise providing the diagnosis directly to the subject. In some embodiments, the methods comprise retreatment of the subject with any of the compositions or methods described herein. In some embodiments, the methods comprise detection of a bacterial infection based upon the processed data. In some embodiments, capturing comprises the use of a device comprising wearable goggles orcompact desktop model. In some embodiments, the subject has wet age-related macular degeneration (AMD). ANTIBODIES AND ANTIBODY CONJUGATES

[0204] Provided herein are anti-VEGF antibodies (including anti-VEGF proteins) and conjugates thereof (e.g., KSI-301). In some embodiments, the antibodies themselves are different from other anti-VEGF agents and provide superior results over other anti-VEGF agents or therapies. In some embodiments, the anti-VEGF antibody conjugate (or a combination of the anti-VEGF antibody conjugate and unconjugated anti-VEGF antibody) displays a surprising superiority over other antibodies and / or the expectation of the activity other antibody conjugates. FIG. 10 displays some embodiments of the amino acid sequence of the antibody portion of KSI-301, including those with a terminal lysine removed. Terminal lysine removal is a post-translational modification in antibodies. The lysine residues at the heavy chain C-terminus of recombinant IgGs can be removed (often to a large extent) during cell culture by carboxypeptidases that are endogenous to CHO host cells. For all antibody sequences recited herein, although specific sequences (which may be longer) 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. Thus, in some embodiments, any of the antibody sequences provided herein may be modified by this lysine clip, which can include the modified version as shown in FIG. 10 (with one or more of the underlined residues being removed). In some embodiments, the lysine in the heavy chain of FIG.10 is removed. In some embodiments, the GK is removed. In some embodiments, the PGK is removed. In some embodiments, the SPGK is removed. As will be appreciated by those in the art, across a population of molecules, the lysine clip may vary and not be complete. Thus, for example, 90, 95, 98, 99, or 100% of the molecules may have one or more of the clip versions, while 10, 5, 2, 1, or down to 0% may be full length (including any range defined between any two of the preceding values). Thus, as used herein, KSI-301 includes any one of and all options for the heavy chain variations outlined in FIG.10. Any reference to the antibody portion of KSI-301 or an amino acid sequence of an anti-VEGF antibody heavy chain is intended to encompass, but not be limited to, a polypeptideexpressed recombinantly from a nucleic acid construct encoding the heavy chain amino acid sequence without any of the post-translational modifications noted above (e.g., encoding the amino acid sequence of SEQ ID NO:1).

[0205] In some embodiments, the anti-VEGF antibody conjugate is KSI-301 (or KSI-301ABC), which is an antibody conjugate comprising: (1) an anti-VEGF-A antibody that includes: a light chain that has the amino acid sequence set forth in SEQ ID NO: 2; and a heavy chain that has the amino acid sequence set forth in SEQ ID NO :1 (or any of the variants thereof in FIG. 10, e.g., SEQ ID NOs: 9, 10, 11, or 12); and (2) a phosphorylcholine containing polymer, wherein the polymer is covalently bonded to the heavy chain of the anti-VEGF-A antibody, where the antibody conjugate has the following structure:wherein: each heavy chain of the anti-VEGF-A antibody is denoted by the letter H, and each light chain of the anti-VEGF-A antibody is denoted by the letter L; the polymer is bonded tothe anti-VEGF-A antibody through a sulfhydryl at C443 according to EU numbering, whichbond is depicted on one of the heavy chains above; PC is, where the curvy line indicates the point of attachment to the rest of the polymer; 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%.

[0206] Historically, conjugating a molecule to a protein often resulted in a decrease in the protein’s binding interaction to its intended target. In some embodiments of the present disclosure, when conjugating to a location that is outside of the active site, the same level of decrease as might have been expected is not necessarily observed. The evidence provided herein shows the opposite effect as to what may have been expected. In some embodiments, and without intending to be limited by theory, the conjugate can be superior to the antibody alone. For example, the interaction of a ligand and its specific receptor is often driven through the stereospecific interaction of the ligand and the receptor, as directed by the interactions of the hydrophilic amino acids on the ligand with the hydrophilic amino acids on the receptor, and water molecules are front and center in those interactions. At the same time, this hydrophilic stereospecificity is further enhanced by de-emphasizing and / or suppressing non- specific hydrophobic interactions that might generally be mediated / created by hydrophobic- to-hydrophobic amino acids.

[0207] In some embodiments, an anti-VEGF antibody conjugate is provided that is capable of blocking at least 90% of an interaction between a VEGF ligand (“VEGFL”) and a VEGF-receptor (“VEGFR”). For example, it can block at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or effectively all of the interaction between VEGFR and VEGFL. In some embodiments, the noted blocking occurs at saturating concentrations. In some embodiments, an anti-VEGF antibody conjugate is provided that blocks at least 95% of an interaction between a VEGF ligand and a VEGF-receptor. An example of such superiority of blocking is the ability of the anti-VEGF antibody bioconjugate (an antibody conjugate provided herein, e.g., KSI-301) to block to a higher degree than Lucentis® (ranibizumab) or Avastin® (bevacizumab) or even the antibody OG1950 (unconjugated). Indeed, this result was unexpected in that while the addition of a polymer to an antibody (to form an antibodyconjugate), could be expected to have some or no detrimental impact on binding / activity of the antibody, it was unexpected that it would actually improve the blocking ability of the antibody in this manner.

[0208] In some embodiments, the antibodies or conjugates thereof inhibit at least 70, 80, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% of the activity and / or interaction between VEGFR and VEGFL. In some embodiments, the IC50value can be 0.1, 1, 2, 3, 4, 5, 6, 7 , 8, 9, 10, 20, 30, 40, 50, 100 nM or less than any one or more of the preceding values. In some embodiments, the KD can be 2x10-13, 1x10-13, 1x10-12, 1x10-11, 1x10-10M or less than any one of the preceding values. In some embodiments, the IC50value can be 1, 5, 10, 20, 30, 40, 50, 60, 7080, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1,000, 1,100, 1,200, 1,300 nM, or less than any one of the preceding values.

[0209] In some embodiments, an anti-VEGF antibody is provided that blocks at least 90% of an interaction between a VEGF ligand and a VEGF-receptor. For example, it can block at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or effectively all of the interaction between VEGFR and VEGFL. As example of such superiority of blocking, is the ability of OG1950 (and antibody provided herein) to block to a higher degree than Lucentis®(ranibizumab) or Avastin®(bevacizumab).

[0210] In some embodiments, other antibodies, such as Lucentis® (ranibizumab) or Avastin® (bevacizumab) can be conjugated to one or more of the polymers as described herein, by one or more of the processes described herein. In some embodiments, any antibody, or fragment thereof, can be conjugated to one or more of the polymers as described herein, for example by one or more of the processes described herein.

[0211] In some embodiments the antibody comprises a heavy chain amino acid variable region that comprises SEQ ID NO: 1 (with or without the C-terminal lysine), and a light chain amino acid variable region that comprises SEQ ID NO: 2. In some embodiments, the antibody is conjugated to one or more of the polymers provided herein. In some embodiments, the conjugated antibody is at least 90% identical to SEQ ID NO: 1 and / or 2. In some embodiments, the antibody contains the 6 CDRs within SEQ ID NO:1 and SEQ ID NO: 2, as well as a point mutation of L443C (EU numbering, or 449C in SEQ ID NO: 1). In some embodiments, the conjugated antibody is at least 90% identical to SEQ ID NO: 1 and / or 2 andincludes the following mutations: L234A, L235A, and G237A (EU numbering), and at least one of the following mutations: Q347C (EU numbering) or L443C (EU numbering).

[0212] In some embodiments, an antibody that binds to VEGF-A is provided. The antibody can comprise: 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: L234A, L235A, and G237A (EU numbering), and at least one of the following mutations: Q347C (EU numbering) or L443C (EU numbering). In some embodiments, the antibody includes a heavy chain that includes 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; and has a sequence at least 90% identical to SEQ ID NO: 1. In some embodiments, the antibody includes a light chain that includes 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; and has a sequence at least 90% identical to SEQ ID NO: 2. In some embodiments, the antibody includes a heavy chain that includes 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; and has a sequence at least 95% identical to SEQ ID NO: 1. In some embodiments, the antibody includes a light chain that includes 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; and has a sequence at least 95% identical to SEQ ID NO: 2. In some embodiments, the antibody includes a heavy chain that includes 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; and has a sequence at least 97% identical to SEQ ID NO: 1. In some embodiments, the antibody includes a light chain that includes 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; and has a sequence at least 97% identical to SEQ ID NO: 2. In some embodiments, the antibody includes a heavy chain that includes 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; and has a sequence at least 98% identical to SEQ ID NO: 1. In some embodiments, the antibody includes a light chain that includes 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 inSEQ ID NO: 2; and has a sequence at least 98% identical to SEQ ID NO: 2. In some embodiments, the antibody includes a heavy chain that includes 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; and has a sequence at least 99% identical to SEQ ID NO: 1. In some embodiments, the antibody includes a light chain that includes 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; and has a sequence at least 99% identical to SEQ ID NO: 2.

[0213] As will be appreciated by one of skill in the art, in light of the present specification, any of the antibodies provided herein can be conjugated to any of the polymers provided herein and / or any antibody provided herein can have a cysteine added such that it allows for site specific conjugation to a polymer.

[0214] “KSI-301” refers to a recombinant, mammalian cell expressed full-length humanized anti-VEGF monoclonal antibody and a bioconjugate thereof, which is covalently conjugated to a branched high molecular weight phosphorylcholine-based biopolymer. In some embodiments, KSI-301 is supplied as a preservative free, sterile, aqueous solution in a single-use glass vial at a concentration of 50 mg / mL (based on antibody mass, e.g., excluding any contribution from the polymer). KSI-301 is an anti-vascular endothelial growth factor (VEGF) biopharmaceutical with an extended ocular half-life. “KSI-301P” denotes the unconjugated anti-VEGF monoclonal antibody in KSI-301 (e.g., OG1950, having the light chain and heavy chain amino acid sequences set forth in FIG. 10). “KSI-301ABC” is a bioconjugate of two intermediates: (1) OG1950 antibody intermediate, a recombinant, full- length humanized, anti-huVEGF A monoclonal antibody (e.g., having the light chain and heavy chain amino acid sequences set forth in FIG. 10), and (2) OG1802 biopolymer intermediate, a phosphorylcholine biopolymer (e.g., as set forth in FIG. 3). The addition of OG1802, an inert biopolymer, increases the size of the biologic, thereby extending the ocular pharmacokinetics (PK) of KSI-301 beyond that of currently approved anti-huVEGF-A therapeutics. Nonclinical studies with KSI-301ABC indicate that it appropriately binds with high affinity to huVEGF-A whose binding to huVEGF Receptors 1 and 2 (huVEGFR) is then inhibited. This in turn abrogates huVEGF-A mediated function.

[0215] “VEGF” or “vascular endothelial growth factor” is a human vascular endothelial growth factor that affects angiogenesis or an angiogenic process. In particular, theterm VEGF means any member of the class of growth factors that (i) bind to a VEGF receptor such as VEGFR-1 (Flt-1), VEGFR-2 (KDR / Flk-1), or VEGFR-3 (FLT-4); (ii) activates a tyrosine kinase activity associated with the VEGF receptor; and (iii) thereby affects angiogenesis or an angiogenic process.

[0216] The VEGF family of factors is made up of five related glycoproteins: VEGF-A (also known as VPE), -B, -C, -D and PlGF (placental growth factor). Of these, VEGF-A is the most well studied and is the target of anti-angiogenic therapy. Ferrara et al, (2003) Nat. Med. 9:669-676. VEGF-A exists as a number of different isotypes which are generated both by alternative splicing and proteolysis: VEGF-A206, VEGF-A189, VEGF-A165, and VEGF-A121. The isoforms differ in their ability to bind heparin and non-signaling binding proteins called neuropilins. The isoforms are all biologically active as dimers.

[0217] The various effects of VEGF are mediated by the binding of a VEGF, e.g., VEGF-A (P15692), -B (P49766), -C (P49767) and –D (Q43915), to receptor tyrosine kinases (RTKs). The VEGF family receptors belong to class V RTKs and each carry seven Ig-like domains in the extracellular domain (ECD). In humans, VEGF binds to three types of RTKs: VEGFR-1 (Flt-1) (P17948), VEGFR-2 (KDR, Flk-1) (P935968) and VEGFR-3 (Flt-4) (P35916). Unless otherwise apparent from the context reference to a VEGF means any of VEGF-A, -B, -C , –D, and PlGF, in any of the natural isoforms or natural variants or induced variants having at least 90, 95, 98 or 99% or 100% sequence identity to a natural form. In some embodiments, such VEGFs are human VEGFs. Likewise reference to a VEGFR means any of VEGR-1, R-2 or R-3, including any natural isoform or natural variant, or an induced variant having at least 90, 95, 98 or 99% or 100% sequence identity to a natural sequences.

[0218] VEGF antagonist therapies have been approved for the treatment of certain cancers and wAMD. Bevacizumab (AVASTIN, Genentech / Roche) is a humanized mouse monoclonal antibody that binds to and neutralizes human VEGF, in particular to all isoforms of VEGF-A and to bioactive proteolytic fragments of VEGF-A. See, e.g., Ferrara N, Hillan KJ, Gerber HP, Novotny W.2004. Discovery and development of bevacizumab, an anti-VEGF antibody for treating cancer. Nat Rev Drug Discov.3(5):391-400. Ranibizumab is an antibody fragment or Fab.

[0219] In some embodiments, an antibody conjugate is presented having an anti- VEGF-A antibody bonded at a cysteine outside a variable region of the antibody to aphosphorylcholine containing polymer, wherein the cysteine has been added via recombinant DNA technology. In some embodiments, the polymer is bonded to a single cysteine. In some embodiments, “added by recombinant DNA technology” means that the cysteine residue replaces a non-cysteine amino acid that occurs in the same position in a known or existing antibody or in a consensus antibody sequence. Thus, for example where the antibody is an IgG1 and the heavy chain possess a leucine at EU position 443, the leucine is replaced via recombinant DNA technology with a cysteine (L443C, EU numbering, or 449C in SEQ ID NO: 1). Correspondingly, the native IgG1 sequence at EU position 347 is Q (glutamine) and the Q is replaced with cysteine via recombinant DNA technology to yield Q347C.

[0220] In some embodiments, the anti-VEGF-A antibody comprises a light chain and a heavy chain where the heavy chain has an Fc region. In some embodiments, the cysteine is in the Fc region and the anti-VEGF-A antibody is an immunoglobulin G (IgG). In some embodiments, the anti-VEGF-A heavy chain has CDRH1: GYDFTHYGMN (SEQ ID NO: 3), CDRH2: WINTYTGEPTYAADFKR (SEQ ID NO: 4), and CDRH3: YPYYYGTSHWYFDV (SEQ ID NO: 5), and position 231 (via sequential counting as in SEQ ID NO: 1) is T, and the anti-VEGF-A light chain has CDRL1: SASQDISNYLN (SEQ ID NO: 6), CDRL2: FTSSLHS (SEQ ID NO: 7), and CDRL3: QQYSTVPWT (SEQ ID NO: 8), and Kabat position 4 is L (FIG. 11). In some embodiments, the anti-VEGF-A heavy chain has CDRH1: GYDFTHYGMN (SEQ ID NO: 3), CDRH2: WINTYTGEPTYAADFKR (SEQ ID NO: 4), and CDRH3: YPYYYGTSHWYFDV (SEQ ID NO: 5), and the anti-VEGF-A light chain has CDRL1: SASQDISNYLN (SEQ ID NO: 6), CDRL2: FTSSLHS (SEQ ID NO: 7), and CDRL3: QQYSTVPWT (SEQ ID NO: 8).

[0221] In some embodiments, the anti-VEGF-A heavy chain isotype is IgG1. In some embodiments, the IgG1 constant domain has one or more mutations relative to an IgG1 constant domain (e.g. constant region of SEQ ID NO: 1) to modulate effector function. In some embodiments, the effector function mutations are one or more of the following: (EU numbering) E233X, L234X, L235X, G236X, G237X, A327X, A330X, and P331X wherein X is any natural or unnatural amino acid. In some embodiments, the mutations are selected from the group consisting of (EU numbering): E233P, L234V, L234A, L235A, G237A, A327G, A330S, and P331S. In some embodiments, the antibody conjugate has the following mutations (EU numbering): L234A, L235A, and G237A.

[0222] In some embodiments, the cysteine residue is in the anti-VEGF-A heavy chain and is Q347C (EU numbering) or L443C (EU numbering). In some embodiments, the cysteine residue is L443C (EU numbering, or 449C in SEQ ID NO: 1). In some embodiments, the sequence of the anti-VEGF-A heavy chain is SEQ ID NO. 1 and the sequence of the anti- VEGF-A light chain is SEQ ID NO.2 (FIG.10).

[0223] In some embodiments, the phosphorylcholine containing polymer comprises 2-(methacryloyloxyethyl)-2'-(trimethylammonium)ethyl phosphate (MPC) monomers as set forth below:.

[0224] Such that the polymer comprises the following repeating units:where n is an integer from 1 to 3000 and the wavy lines indicate the points of attachment between monomer units in the polymer.

[0225] In some embodiments, the polymer has three or more arms, or is synthesized with an initiator comprising 3 or more polymer initiation sites. In some embodiments, thepolymer 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. More preferably, the polymer has 3, 6, or 9 arms, or is synthesized with an initiator comprising 3, 6, or 9 polymer initiation sites. In some embodiments, the polymer has 9 arms, or is synthesized with an initiator comprising 9 polymer initiation sites.

[0226] In some embodiments, the polymer that is added has a molecular weight between about 300,000 and about 1,750,000 Da (SEC-MALs). In some embodiments, the polymer has a molecular weight between about 500,000 and about 1,000,000 Da. In some embodiments, the polymer has a molecular weight of between about 600,000 to about 900,000 Da. In some embodiments, the polymer has a molecular weight of between about 750,000 to about 850,000 Da. In some embodiments, the polymer has a molecular weight of between about 800,000 to about 850,000 Da. In some embodiments, the polymer has a molecular weight of between about 750,000 to about 800,000 Da.

[0227] In some embodiments, any of the antibodies described herein can be further conjugated to a polymer to form a bioconjugate. The molecular weight of the bioconjugate (in total, SEC-MALs) can be between about 350,000 and 2,000,000 Daltons, for example, between about 450,000 and 1,900,000 Daltons, between about 550,000 and 1,800,000 Daltons, between about 650,000 and 1,700,000 Daltons, between about 750,000 and 1,600,000 Daltons, between about 850,000 and 1,500,000 Daltons, between about 900,000 and 1,400,000 Daltons, between about 950,000 and 1,300,000 Daltons, between about 900,000 and 1,000,000 Daltons, between about 1,000,000 and 1,300,000 Daltons, between about 850,000 and 1,300,000 Daltons, between about 850,000 and 1,000,000 Daltons, and between about 1,000,000 and 1,200,000 Daltons.

[0228] In some embodiments, the antibody conjugate is purified. In some embodiments, the polymer is aspect of the antibody conjugate is polydisperse, i.e. the polymer PDI is not 1.0. In some embodiments, the PDI is less than 1.5. In some embodiments, the PDI is less than 1.4. In some embodiments, the PDI is less than 1.3. In some embodiments the PDI is less than 1.2. In some embodiments the PDI is less than 1.1.

[0229] In some embodiments, the antibody conjugate has an anti-VEGF-A immunoglobulin G (IgG) bonded to a polymer, which polymer comprises MPC monomers, wherein the sequence of the anti-VEGF-A heavy chain is SEQ ID NO.1, and the sequence ofthe anti-VEGF-A light chain is SEQ ID NO. 2, and wherein the antibody is bonded only at C449 in SEQ ID NO. 1 to the polymer. In some embodiments, the polymer has 9 arms and has a molecular weight of between about 600,000 to about 1,000,000 Da.

[0230] In some embodiments, the antibody conjugate has an anti-VEGF-A immunoglobulin G (IgG) bonded to a polymer, which polymer comprises MPC monomers, wherein the sequence of the anti-VEGF-A heavy chain is SEQ ID NO.1, and the sequence of the anti-VEGF-A light chain is SEQ ID NO. 2, and wherein the antibody is bonded only at C443 (EU numbering, or 449C in SEQ ID NO: 1) to the polymer. In some embodiments, the polymer has 9 arms and has a molecular weight of between about 600,000 to about 1,000,000 Da.

[0231] In some embodiments, the antibody conjugate has the following structure:wherein: each heavy chain of the anti-VEGF-A antibody is denoted by the letter H, and each light chain of the anti-VEGF-A antibody is denoted by the letter L; the polymer is bonded to the anti-VEGF-A antibody through the sulfhydryl of C449 of SEQID NO: 1, 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; wherein X is a) –OR where R is H, methyl, ethyl, propyl, or isopropyl, b) –H, c) any halogen, including – Br, –Cl, or –I, d) –SCN, or 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 10%. In some embodiments, n1, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different and are integers from 0 to 3000. In some embodiments, n1, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different and are integers from 0 to 500. In some embodiments, X is –OR, where R is a sugar, an aminoalkyl, mono-substituted, poly-substituted or unsubstituted variants of the following residues: saturated C1-C24alkyl, unsaturated C2-C24alkenyl or C2-C24alkynyl, acyl, acyloxy, alkyloxycarbonyloxy, aryloxycarbonyloxy, cycloalkyl, cycloalkenyl, alkoxy, cycloalkoxy, aryl, heteroaryl, arylalkoxy carbonyl, alkoxy carbonylacyl, amino, aminocarbonyl, aminocarboyloxy, nitro, azido, phenyl, hydroxy, alkylthio, arylthio, oxysulfonyl, carboxy, cyano, and halogenated alkyl including polyhalogenated alkyl, --CO-- O--R7, carbonyl --CCO--R7, --CO--NR8R9, --(CH2)n--COOR7, --CO--(CH) n--COOR7, --(CH2) n--NR8R9, ester, alkoxycarbonyl, aryloxycarbonyl, wherein n is an integer from 1 to 6, wherein each R7, R8and R9is separately selected from the group consisting of a hydrogen atom, halogen atom, mono-substituted, poly-substituted or unsubstituted variants of the following residues: saturated C1- C24alkyl, unsaturated C2-C24alkenyl or C2- C24alkynyl, acyl, acyloxy, alkyloxycarbonyloxy, aryloxycarbonyloxy, cycloalkyl, cycloalkenyl, alkoxy, cycloalkoxy, aryl, heteroaryl, arylalkoxy carbonyl, alkoxy carbonylacyl, amino, aminocarbonyl, aminocarboyloxy, nitro, azido, phenyl, hydroxy, alkylthio, arylthio, oxysulfonyl, carboxy, cyano, and halogenated alkyl including polyhalogenated alkyl, a 5-membered ring, and a 6- membered ring.

[0232] In some embodiments, the antibody conjugate has the following structure:wherein: each heavy chain of the anti-VEGF-A antibody is denoted by the letter H, and each light chain of the anti-VEGF-A antibody is denoted by the letter L; the polymer is bonded to the anti-VEGF-A antibody through the sulfhydryl of C443 (EU numbering, or 449C in SEQ ID NO: 1), 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; wherein X is a) –OR where R is H, methyl, ethyl, propyl, or isopropyl, b) –H, c) any halogen, including Br, –Cl, or –I, d) –SCN, or 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, n6, n7, n8 and n9 is 2500 plus or minus 10%. In some embodiments, n1, n2, n3, n4, n5, n6, n7, n8 and n9 are the same or different and are integers from 0 to 3000. In some embodiments, n1, n2,n3, n4, n5, n6, n7, n8 and n9 are the same or different and are integers from 0 to 500. In some embodiments, X is -OR, where R is a sugar, an aminoalkyl, mono-substituted, poly-substituted or unsubstituted variants of the following residues: saturated C1 -C24 alkyl, unsaturated C2 -C24 alkenyl or C2 -C24 alkynyl, acyl, acyloxy, alkyloxycarbonyloxy, aryloxycarbonyloxy, cycloalkyl, cycloalkenyl, alkoxy, cycloalkoxy, aryl, heteroaryl, arylalkoxy carbonyl, alkoxy carbonylacyl, amino, aminocarbonyl, aminocarboyloxy, nitro, azido, phenyl, hydroxy, alkylthio, arylthio, oxysulfonyl, carboxy, cyano, and halogenated alkyl including polyhalogenated alkyl, --CO--O--R7, carbonyl --CCO--R7, --CO--NR8R9, --(CH2)n--COOR7, - -CO--(CH)n--COOR7, --(CH2)n--NR8R9, ester, alkoxycarbonyl, aryloxycarbonyl, wherein n is an integer from 1 to 6, wherein each R7, R8 and R9 is separately selected from the group consisting of a hydrogen atom, halogen atom, mono-substituted, poly-substituted or unsubstituted variants of the following residues: saturated C1- C24 alkyl, unsaturated C2 -C24 alkenyl or C2- C24 alkynyl, acyl, acyloxy, alkyloxycarbonyloxy, aryloxycarbonyloxy, cycloalkyl, cycloalkenyl, alkoxy, cycloalkoxy, aryl, heteroaryl, arylalkoxy carbonyl, alkoxy carbonylacyl, amino, aminocarbonyl, aminocarboyloxy, nitro, azido, phenyl, hydroxy, alkylthio, arylthio, oxysulfonyl, carboxy, cyano, and halogenated alkyl including polyhalogenated alkyl, a 5-membered ring, and a 6-membered ring.

[0233] In some embodiments, the half-life of the anti-VEGF-A antibodies is extended by attachment of a “half-life (“half life”) extending moieties” or “half-life (“half life”) extending groups”. Half-life extending moieties include, without limitation, peptides and proteins which can be expressed in frame with the biological drug of issue (or conjugated chemically depending on the situation) and various polymers which can be attached or conjugated to one or more amino acid side chain or end functionalities such as -SH, -OH, - COOH, -CONH2, -NH2, or one or more N- and / or O-glycan structures. Half-life extending moieties generally act to increase the in vivo circulatory half-life of biologic drugs.

[0234] Non-limiting examples of peptide / protein half-life extending moieties include Fc fusion (Capon DJ, Chamow SM, Mordenti J, et al. Designing CD4 immunoadhesions for AIDS therapy. Nature.1989.337:525-31), human serum albumin (HAS) fusion (Yeh P, Landais D, Lemaitre M, et al. Design of yeast-secreted albumin derivatives for human therapy: biological and antiviral properties of a serum albumin-CD4 genetic conjugate. Proc Natl Acad Sci USA. 1992. 89:1904-08 ), carboxy terminal peptide (CTP) fusion (FaresFA, Suganuma N. Nishimori K, et al. Design of a long-acting follitropin agonist by fusing the C-terminal sequence of the chorionic gonadotropin beta subunit to the follitropin beta subunit. Proc Natl Acad Sci USA. 1992. 89:4304-08), genetic fusion of non-exact repeat peptide sequence (XTEN) fusion (Schellenberger V, Wang CW, Geething NC, et al. A recombinant polypeptide extends the in vivo half-life of peptides and proteins in a tunable manner. Nat Biotechnol.2009.27:1186-90), elastin like peptide (ELPylation) (MCpherson DT, Morrow C, Minehan DS, et al. Production and purification of a recombinant elastomeric polypeptide, G(VPGVG19-VPGV, from Escheriachia coli. Biotechnol Prog. 1992. 8:347-52), human transferrin fusion (Prior CP, Lai C-H, Sadehghi H et al. Modified transferrin fusion proteins. Patent WO2004 / 020405. 2004), proline-alanine-serine (PASylation) (Skerra A, Theobald I, Schlapsky M. Biological active proteins having increased in vivo and / or vitro stability. Patent WO2008 / 155134 A1. 2008), homo-amino acid polymer (HAPylation) (Schlapschy M, Theobald I, Mack H, et al. Fusion of a recombinant antibody fragment with a homo-amino acid polymer: effects on biophysical properties and prolonged plasma half-life. Protein Eng Des Sel. 2007. 20:273-84) and gelatin like protein (GLK) fusion (Huang Y-S, Wen X-F, Zaro JL, et al. Engineering a pharmacologically superior form of granulocyte-colony-stimulating-factor by fusion with gelatin-like protein polymer. Eur J. Pharm Biopharm.2010.72:435-41).

[0235] Non-limiting examples of polymer half-life extending moieties include 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).

[0236] In one embodiment a half-life extending moiety can be conjugated to an antibody via free amino groups of the protein using N-hydroxysuccinimide (NHS) esters.Reagents targeting conjugation to amine groups can randomly react to -amine group of lysines, -amine group of N-terminal amino acids, and -amine group of histidines.

[0237] However, the anti-VEGF-A antibodies disclosed herein have many amine groups available for polymer conjugation. Conjugation of polymers to free amino groups, thus, might negatively impact the ability of the antibody proteins to bind to VEGF.

[0238] In some embodiments, a half-life extending moiety is coupled to one or more free SH groups using any appropriate thiol-reactive chemistry including, without limitation, maleimide chemistry, or the coupling of polymer hydrazides or polymer amines to carbohydrate moieties of the antibody after prior oxidation. In some embodiments maleimide coupling is used. In some embodiments, coupling occurs at cysteines naturally present or introduced via genetic engineering.

[0239] In some embodiments, polymers are covalently attached to cysteine residues introduced into anti-VEGF-A antibodies by site directed mutagenesis. In some embodiments, the cysteine residues are employed in the Fc portion of the antibody. In some embodiments, the sites to introduce cysteine residues into an Fc region are provided in WO 2013 / 093809, US 7,521,541, WO 2008 / 020827, US 8,008,453, US 8,455,622 and US2012 / 0213705, incorporated herein by reference for all purposes. In some embodiments, the cysteine mutations are Q347C (EU numbering) and L443C referring to the human IgG heavy chain by EU numbering.

[0240] In some embodiments, conjugates of antibody and high MW polymers serving as half-life extenders are provided. In some embodiments, a conjugate comprises an antibody that is coupled to a zwitterionic polymer wherein the polymer is formed from one or more monomer units and wherein at least one monomer unit has a zwitterionic group is provided. In some embodiments, the zwitterionic group is phosphorylcholine.

[0241] In some embodiments, one of the monomer units is HEMA-PC. In some embodiments, a polymer is synthesized from a single monomer which is HEMA-PC.

[0242] In some embodiments, some antibody conjugates have 2, 3, or more polymer arms wherein the monomer is HEMA-PC. In some embodiments, the conjugates have 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 polymer arms wherein the monomer is HEMA-PC. In some embodiments, the conjugates have 3, 6 or 9 arms. In some embodiments, the conjugate has 9 arms.

[0243] In some embodiments, polymer-antibody conjugates have a polymer portion with a molecular weight of between 100,000 and 1,500,000 Da. In some embodiments, the conjugate has a polymer portion with a molecular weight between 500,000 and 1,000,000 Da. In some embodiments, the conjugate has a polymer portion with a molecular weight between 600,000 to 800,000 Da. In some embodiments, the conjugate has a polymer portion with a molecular weight between 600,000 and 850,000 Da and has 9 arms. When a molecular weight is given for an antibody conjugated to a polymer, the molecular weight will be the addition of the molecular weight of the protein, including any carbohydrate moieties associated therewith, and the molecular weight of the polymer.

[0244] In some embodiments, an anti-VEGF-A antibody has a HEMA-PC polymer which has a molecular weight measured by Mw of between about 100 kDa and 1650 kDa is provided. In some embodiments, the molecular weight of the polymer as measured by Mw is between about 500 kDa and 1000 kDa. In some embodiments, the molecular weight of the polymer as measured by Mw is between about 600 kDa to about 900 kDa. In some embodiments, the polymer molecular weight as measured by Mw is 750 kDa plus or minus 15%. In some embodiments, the polymer molecular weight as measured by Mw is 800 kDa plus or minus 20%.

[0245] In some embodiments, the polymer is made from an initiator suitable for ATRP having one or more polymer initiation sites. In some embodiments, the polymer initiation site has a 2-bromoisobutyrate site. In some embodiments, the initiator has 3 or more polymer initiation sites. In some embodiments, the initiator has 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 polymer initiation sites. In some embodiments, the initiator has 3, 6 or 9 polymer initiation sites. In some embodiments, the initiator has 9 polymer initiation sites. In some embodiments, the initiator is OG1786 (FIG.1).

[0246] The anti-VEGF-A antibodies can be produced by recombinant expression including (i) the production of recombinant DNA by genetic engineering, (ii) introducing recombinant DNA into prokaryotic or eukaryotic cells by, for example and without limitation, transfection, electroporation or microinjection, (iii) cultivating the transformed cells, (iv) expressing antibody, e.g. constitutively or on induction, and (v) isolating the antibody, e.g. from the culture medium or by harvesting the transformed cells, in order to (vi) obtain purified antibody.

[0247] The anti-VEGF-A antibodies can be produced by expression in a suitable prokaryotic or eukaryotic host system characterized by producing a pharmacologically acceptable antibody molecule. Examples of eukaryotic cells are mammalian cells, such as CHO, COS, HEK 293, BHK, SK-Hip, and HepG2. Other suitable expression systems are prokaryotic (e.g., E. coli with pET / BL21 expression system), yeast (Saccharomyces cerevisiae and / or Pichia pastoris systems), and insect cells.

[0248] A wide variety of vectors can be used for the preparation of the antibodies disclosed herein and are selected from eukaryotic and prokaryotic expression vectors. Examples of vectors for prokaryotic expression include plasmids such as, and without limitation, preset, pet, and pad, wherein the promoters used in prokaryotic expression vectors include one or more of, and without limitation, lac, trc, trp, recA, or araBAD. Examples of vectors for eukaryotic expression include: (i) for expression in yeast, vectors such as, and without limitation, pAO, pPIC, pYES, or pMET, using promoters such as, and without limitation, AOX1, GAP, GAL1, or AUG1; (ii) for expression in insect cells, vectors such as and without limitation, pMT, pAc5, pIB, pMIB, or pBAC, using promoters such as and without limitation PH, p10, MT, Ac5, OpIE2, gp64, or polh, and (iii) for expression in mammalian cells, vectors such as, and without limitation, pSVL, pCMV, pRc / RSV, pcDNA3, or pBPV, and vectors derived from, in one aspect, viral systems such as and without limitation vaccinia virus, adeno-associated viruses, herpes viruses, or retroviruses, using promoters such as and without limitation CMV, SV40, EF-1, UbC, RSV, ADV, BPV, and beta-actin. Method of Conjugating Proteins to Polymers

[0249] In some embodiments, a method is presented of preparing a therapeutic protein-half life extending moiety conjugate having the step of conjugating a therapeutic protein which has a cysteine residue added via recombinant DNA technology to a half-life extending moiety having a sulfhydryl specific reacting group selected from the group consisting of maleimide, vinylsulfones, orthopyridyl-disulfides, and iodoacetamides to provide the therapeutic protein-half life extending moiety conjugate.

[0250] In some embodiments a method of preparing the anti-VEGF antibody conjugate, e.g., OG1953, from OG1950 is provided. The method comprises reducing the OG1950 protein with a 50x molar excess of the TCEP reducing agent. After reduction, theantiobody is oxidized to produce a decapped OG1950 antibody where the inter- and intra- light and heavy chain disulfide bonds naturally occurring in the antibody are formed, but the engineered Cysteine on the heavy chain position L443C (EU numbering, or 449C in SEQ ID NO: 1) remains to be decapped. The OG1950 is then conjugated by adding an excipient and adding 5-10x molar excess of a maleimide biopolymer (e.g., OG1802). The biopolymer links to the OG1950 antibody through a covalent thiolether linkage. After conjugation, the anti- VEGF antibody conjugate, e.g., OG1953, is purified with both unconjugated antibody and polymer removed.

[0251] The protein and process described above can be varied as well. Thus, in some embodiments, a process for preparing a conjugated protein (which need not be an antibody or an anti-VEGF antibody) is provided. The process includes reducing one or more cysteines in a protein to form a decapped protein in a solution. After reducing the one or more cysteines the decapped protein is reoxidized to restore at least one disulfide linkage in the reduced protein while ensuring that an engineered cysteine residue in the protein remains in a free thiol form to form a reoxidized decapped protein in the solution. At least one excipient is then added to the solution. The excipient reduces a polymer induced protein precipitation. After the excipient is added, a polymer is added to the solution, which is conjugated to the reoxidized decapped protein at the engineered cysteine residue to form a conjugated protein.

[0252] In some embodiments, the molar excess of the reducing agent can be altered to any amount that functions. In some embodiments 10, 20, 30, 40, 50, 60, 70, 80, 90x molar excess of the reducing agent (which need not be TCEP in all embodiments) can be employed. In some embodiments, any antibody (therapeutic or otherwise) can be employed. In some embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15x molar excess of a maleimide biopolymer can be employed. In some embodiments, there is an excess of decapped protein to polymer. In some embodiments, the amount of the reduced protein is less than the amount of the polymer. In some embodiments, the amount of the reduced protein is 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10%, 5%, 4%, 3%, 2%, 1% of the amount of the polymer. In some embodiments, 10-15 times as much polymer is used as protein. In some embodiments the amount of the reduced antibody is greater than the amount of the polymer. In some embodiments the amount of the polymer is greater than the amount of the reduced antibody.

[0253] In some embodiments, the purification step is optional.

[0254] In some embodiments, the method of making an antibody conjugate comprises conjugating an anti-VEGF-A antibody to a phosphorylcholine containing polymer. In some embodiments the method comprises the steps of conjugating an anti-VEGF-A antibody to a phosphorylcholine containing polymer. The anti-VEGF-A antibody comprises an amino residue added via recombinant DNA technology. In some embodiments, the added amino acid residue is a cysteine residue. In some embodiments, the cysteine residue is added outside a variable region of the antibody. The cysteine residue can be added to either the heavy chain or light chain of the antibody.

[0255] In some embodiments, the polymer comprises or consists of a phosphorylcholine containing polymer. In some embodiments, the phosphorylcholine containing polymer comprises a sulfhydryl specific reacting group selected from the group consisting of a maleimide, a vinylsulfone, an orthopyridyl-disulfide, and an iodoacetamide. In some embodiments, the sulfhydryl specific reacting group on the phosphorylcholine containing polymer reacts with the cysteine residue on the anti-VEGF-A antibody to make the antibody conjugate.

[0256] In some embodiments, the protein to be conjugated can be an antibody, an antibody protein fusion, or a binding fragment thereof. In some embodiments, the protein is not an antibody but is an enzyme, a ligand, a receptor, or other protein or mutants or variants thereof. In some embodiments, the native protein contains at least one disulfide bond and at least one non-native cysteine.

[0257] In some embodiments, the excipient can be an acid or a base. In some embodiments, the excipient is a detergent, a sugar, or a charged amino acid. In some embodiments, the excipient assists in keeping the protein in solution during the conjugation to the polymer. In some embodiments, the excipient is added to the solution containing the protein, prior to the addition of the polymer to the solution that contains the protein.

[0258] In some embodiments, the reaction occurs under aqueous conditions between about pH 5 to about pH 9. In some embodiments, the reaction occurs between 6.0 and 8.5, between 6.5 and 8.0 or between 7.0 and 7.5.

[0259] In some embodiments, the polymer is conjugated to the protein at 2-37 degrees Celsius. In some embodiments, the conjugation occurs at 0-40 degrees Celsius, 5-35 degrees Celsius, 10-30 degrees Celsius, and 15-25 degrees Celsius.

[0260] In some embodiments, the conjugated proteins described herein can be contacted to an ion exchange medium or hydrophobic interaction chromatography or affinity chromatography medium for purification (to remove the conjugated from the unconjugated). In some embodiments, the ion exchange medium, hydrophobic interaction chromatography, and / or affinity chromatography medium separates the conjugated protein from the free polymer and from the reoxidized decapped protein.

[0261] In some embodiments, the processes described herein and outlined in FIG. 18 involves using an excipient that is capable of facilitating and / or maintaining a solubility system. In some embodiments, the process allows the solution to maintain the solubility of the two components meant to interact. This can include the solubility of the protein and the polymer and then the end conjugate as well. In some embodiments, without the excipient approach, the issue can be that while the protein is soluble, when the biopolymer is added, the solubility of the solution (e.g., protein) drops and it crashes / precipitates out of solution. Of course, when the protein crashes out, it is not available to conjugate efficiently with the biopolymer. Thus, an excipient can be employed to maintain the solubility of the protein in the presence of the biopolymer so the two can couple to form the protein conjugate (or as depicted in FIG.18, an antibody conjugate). This also allows for the solubility of the conjugate to be maintained.

[0262] In some embodiments, the polymers disclosed herein can comprise one or more of the following: a zwitterion, a phosphorylcholine, or a PEG linker bridging a center of a polymer branching point to the maleimide functional group. In some embodiments, any of the polymers provided herein can be added to a protein via the methods provided herein.

[0263] In some embodiments, any of the proteins provided herein can be conjugated to any of the polymers provided herein via one or more of the methods provided herein.

[0264] In some embodiments, the process(es) provided herein allow(s) for larger scale processing to make and purify protein and / or antibody conjugates. In some embodiments, the volume employed is at least 1 liter, for example 1, 10, 100, 1,000, 5,000, 10,000, liters or more. In some embodiments, the amount of the antibody conjugate produced and / or purified can be 0.1, 1, 10, 100, 1000, or more grams.

[0265] In some embodiments, the therapeutic protein may be any of the anti- VEGF-A antibodies described herein having a cysteine residue added via recombinant DNA technology (FIG. 11). In some embodiments, the anti-VEGF antibody heavy chain has the following CDRs: CDRH1: GYDFTHYGMN (SEQ ID NO: 3), CDRH2: WINTYTGEPTYAADFKR (SEQ ID NO: 4), and CDRH3: YPYYYGTSHWYFDV (SEQ ID NO: 5). The heavy chain can also have threonine (T) at position 231 (via sequential counting as in SEQ ID NO. 1). In some embodiments, the anti-VEGF light chain has the following CDRs: CDRL1: SASQDISNYLN (SEQ ID NO: 6), CDRL2: FTSSLHS (SEQ ID NO: 7), and CDRL3: QQYSTVPWT (SEQ ID NO: 8). The anti-VEGF-A light chain can also have leucine (L) at Kabat position 4. In some embodiments, the anti-VEGF antibody heavy chain has the following CDRs: CDRH1: GYDFTHYGMN (SEQ ID NO: 3), CDRH2: WINTYTGEPTYAADFKR (SEQ ID NO: 4), and CDRH3: YPYYYGTSHWYFDV (SEQ ID NO: 5), and the anti-VEGF light chain has the following CDRs: CDRL1: SASQDISNYLN (SEQ ID NO: 6), CDRL2: FTSSLHS (SEQ ID NO: 7), and CDRL3: QQYSTVPWT (SEQ ID NO: 8).

[0266] In some embodiments, the anti-VEGF-A antibody is IgG1. In some embodiments, the heavy chain has one or more mutations to modulate effector function. In some embodiments, the mutations are to one or more of the following amino acid positions (EU numbering): E233, L234, L235, G236, G237, A327, A330, and P331. In some embodiments, the mutations are selected from the group consisting of: E233P, L234V, L234A, L235A, G237A, A327G, A330S and P331S (EU numbering). In some embodiments, the mutations are (EU numbering) L234A, L235A and G237A.

[0267] In some embodiments, the cysteine residue added to the therapeutic protein via recombinant DNA technology should not be involved in Cys-Cys disulfide bond pairing. In this regard, therapeutic proteins may be dimeric. So for example, an intact anti-VEGF-A antibody has two light chains and two heavy chains. If a Cys residue is introduced into the heavy chain for instance, the intact antibody will have two such introduced cysteines at identical positions and the possibility exists that these cysteine residues will form intra-chain disulfide bonds. If the introduced cysteine residues form Cys-Cys disulfide bonds or have a propensity to do so, that introduced Cys residue will not be useful for conjugation. It is knownin the art how to avoid positions in the heavy and light chains that will give rise to intra-chain disulfide pairing. See, e.g., U.S. Patent Application No. 2015 / 0158952.

[0268] In some embodiments, the cysteine residue introduced via recombinant DNA technology is selected from the group consisting of (EU numbering) Q347C and L443C. In some embodiments, the cysteine residue is L443C (EU numbering, or 449C in SEQ ID NO: 1). In some embodiments, the heavy chain the antibody has the amino acid sequence set forth in SEQ ID NO.1 and the light chain has the amino acid sequence of SEQ ID NO.2.

[0269] In some embodiments, the sulfhydral specific reacting group is maleimide.

[0270] In some embodiments, the half-life extending moiety is selected from the group consisting 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 2- methacryloyloxy-2’-ethyltrimethylammoniumphosphate (MPC).

[0271] In some embodiments, the half-life extending moiety is a zwitterionic polymer. In some embodiments, the zwitterion is phosphorylcholine, i.e. a phosphorylcholine containing polymer. In some embodiments, the polymer is composed of MPC units.

[0272] In some embodiments, the MPC polymer has three or more arms. In some embodiments, the MPC polymer has 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 arms. In some embodiments, the MPC polymer has 3, 6, or 9 arms. In some embodiments, the MPC polymer has 9 arms. In some embodiments, the polymer is synthesized with an initiator comprising 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or more polymer initiation sites

[0273] In some embodiments, the MPC polymer has a molecular weight between about 300,000 and 1,750,000 Da. In some embodiments, the MPC polymer has a molecular weight between about 500,000 and 1,000,000 Da or between about 600,000 to 900,000 Da.

[0274] In some embodiments, the method of preparing a therapeutic protein half life-extending moiety conjugate has an additional step of contacting the therapeutic protein with a thiol reductant under conditions that produce a reduced cysteine sulfhydryl group. As discussed above, it is preferable that the cysteine residue added via recombinant DNA technology are unpaired, i.e. are not involved in Cys-Cys intra chain disulfide bonds or are not substantially involved in such bonding. However, Cys residues which are not involved in such Cys-Cys disulfide bonding and are free for conjugation are known to react with free cysteine in the culture media to form disulfide adducts. See, e.g., WO 2009 / 052249. A cysteine so derivatized will not be available for conjugation. To free the newly added cysteine from the disulfide adduct, the protein after purification is treated with a reducing agent, e.g., dithiothreitol. However, such treatment with a reducing agent will reduce all of the cysteine residues in the therapeutic protein, including native cysteines many of which are involved in inter and intra chain Cys-Cys disulfides bonds. The native Cys-Cys disulfides are generally crucial to protein stability and activity and they should be reformed. In some embodiments, all native (e.g., inter and intra) Cys-Cys disulfides are reformed.

[0275] To reform native inter and intra-chain disulfide residues, after reduction to remove the cysteine disulfide adducts, the therapeutic protein is exposed to oxidizing conditions and / or oxidizing agents for a prescribed period of time, e.g., overnight. In some embodiments, ambient air exposure overnight can be used to achieve reformation of the native disulfide bonds. In some embodiments, an oxidizing agent is employed to restore the native disulfides. In some embodiments, the oxiding agent is selected from the group consisting of acqueous CuSO4 and dehydroascorbic acid (DHAA). In some embodiments, the oxidizing agent is DHAA. In some embodiments, the range of DHAA used is in the range of 5-30 equivalents. In some embodiments, the range is 10-20 equivalents. In some embodiments, the range is 15 equivalents.

[0276] In some embodiments, the thiol reductant is selected from the group consisting of: Tris[2-carboxyehtyl]phosphine hydrochloride (TCEP), dithiothreitol (DTT), dithioerythritol (DTE), sodium borohydride (NaBH4), sodium cyanoborohydride (NaCNBH3), -mercaptoethanol (BME), cysteine hydrochloride and cysteine. In some embodiments, the thiol reductant is TCEP.

[0277] In some embodiments, the thiol reductant concentration is between 1 and 100 fold molar excess relative to the therapeutic protein concentration. In some embodiments, the thiol reductant concentration is between 20 to 50 fold molar excess relative to the therapeutic protein concentration. In some embodiments, the thiol reductant is removed following incubation with the therapeutic protein prior to oxidation of the therapeutic protein.

[0278] In some embodiments, the method for conjugating a therapeutic protein to a half-life extending moiety has a further step of purifying the therapeutic protein conjugate after conjugation. In some embodiments, the therapeutic protein conjugate is purified using a technique selected from the group consisting of ion exchange chromatography, hydrophobic interaction chromatography, size exclusion chromatography, and affinity chromatography or combinations thereof.

[0279] In some embodiments, the therapeutic protein conjugate retains at least 20% biological activity relative to unconjugated therapeutic protein. In some embodiments, the therapeutic protein conjugate retains at least 50% biological activity relative to unconjugated therapeutic protein. In some embodiments, the therapeutic protein conjugate retains at least 90% biological activity relative to native therapeutic protein.

[0280] In some embodiments, the therapeutic protein conjugate has an increased half-life relative to unconjugated therapeutic protein. In some embodiments, the therapeutic protein conjugate has at least a 1.5 fold increase in half-life relative to unconjugated therapeutic protein. In some embodiments, the therapeutic protein conjugate has at least a 5 fold increase in half-life relative to unconjugated therapeutic protein.

[0281] In some embodiments, the zwitterionic polymer of the method of conjugating a therapeutic protein to a half-life extending moiety is a radically polymerizable monomer having a zwitterion group and the method has a further step of polymerizing the free radically polymerizable zwitterionic monomer in a polymerization medium to provide a polymer, the medium comprising: the radically polymerizable zwitterionic monomer; a transition metal catalyst M+wherein Mt is a transition metal, q is a higher oxidation state of the metal and q-1 is a lower oxidation state of the metal, wherein the metal catalyst is supplied as a salt of the form Mt(q-1)+X’(q-1) wherein X’ is a counterion or group or the transition metal catalyst is supplied in situ by providing the inactive metal salt at its higher oxidationstate Mtq+X’qtogether with a reducing agent that is capable of reducing the transition metal from the oxidized inactive state to the reduced active state; a ligand; and an initiator.

[0282] To function as an ATRP transition metal catalyst, the transition metal should have at least two readily accessible oxidation states separated by one electron, a higher oxidation state and a lower oxidation state. In ATRP, a reversible redox reaction results in the transition metal catalyst cycling between the higher oxidation state and the lower oxidation state while the polymer chains cycle between having propagating chain ends and dormant chain ends. See, e.g., U.S. Patent No.7,893,173.

[0283] In some embodiments, the radically polymerizable zwitterionic monomer is selected from the group consisting ofwherein R1 is H or C1-6alkyl, ZW is a zwitterion and n is an integer from 1-6.

[0284] In some embodiments, the radically polymerizable monomer iswherein R1 is H or C1-6 alkyl, R2, R3, R4 are the same or different and are H or C1-4alkyl and X and Y are the same or different and are integers from 1-6. In some embodiments, R1, R2, R3 and R4 are each methyl and X and Y are each 2.

[0285] In some embodiments, the radically polymerizable monomer iswherein R1 is H or C1-6alkyl, R2 and R3 are the same or different and are H or C1-4alkyl, R4 is PO4-, SO3- or CO2- and X and Y are the same or different and are integers from 1-6. In some embodiments, R1, R2 and R3 are methyl, R4 is PO4- and X and Y are each 2.

[0286] In some embodiments, the monomer iswherein R1 is H or C1-6alkyl, R2, R3 and R4 are the same or different and are H or C1-4alkyl, R5 is PO4-, SO3- or CO2- and X and Y are the same or different and are integers from 1-6. In some embodiments, R1, R2, R3 and R4 are methyl, R5 is PO4- and X and Y are 2.

[0287] In some embodiments, the transition metal Mt is selected from the group consisting of Cu, Fe, Ru, Cr, Mo, W, Mn, Rh, Re, Co, V, Zn, Au, and Ag. In some embodiments, the metal catalyst is supplied as a salt of the form Mt(q-1)+X’(q-1).is selected from the group consisting of Cu+, Fe2+, Ru2+, Cr2+, Mo2+, W2+, Mn3+, Rh3+, Re2+, Co+, V2+, Zn+, Au+, and Ag+and X’ is selected from the group consisting of halogen, C1-6 alkoxy, (SO4)1 / 2, (PO4)1 / 3, (R7PO4)1 / 2, (R72PO4), triflate, hexafluorophosphate, methanesulfonate, arylsulfonate, CN and R7CO2, where R7 is H or a straight or branched C1-6alkyl group which may be substituted from 1 to 5 times with a halogen. In some embodiments, Mt(q-1)+is Cu+and X’ is Br.

[0288] In some embodiments,is supplied in situ. In some embodiments, Mtq+X’q is CuBr2. In some embodiments, the reducing agent is an inorganic compound. In some embodiments, the reducing agent is selected from the group consisting of a sulfur compound of a low oxidation level, sodium hydrogen sulfite, an inorganic salt comprising a metal ion, a metal, hydrazine hydrate and derivatives of such compounds. In some embodiments, the reducing agent is a metal. In some embodiments, the reducing agent is Cu0.

[0289] In some embodiments, the reducing agent is an organic compound. In some embodiments, the organic compound is selected from the group consisting of alkylthiols, mercaptoethanol, or carbonyl compounds that can be easily enolized, ascorbic acid, acetylacetonate, camphosulfonic acid, hydroxy acetone, reducing sugars, monosaccharides, glucose, aldehydes, and derivatives of such organic compounds.

[0290] In some embodiments, the ligand is selected from the group consisting of 2,2'-bipyridine, 4,4'-Di-5-nonyl-2,2'-bipyridine, 4,4-dinonyl-2,2'-dipyridyl, 4,4',4''-tris(5- nonyl)-2,2':6',2''-terpyridine, N,N,N',N',N''-Pentamethyldiethylenetriamine, 1,1,4,7,10,10- Hexamethyltriethylenetetramine, Tris(2-dimethylaminoethyl)amine, N,N-bis(2- pyridylmethyl)octadecylamine, N,N,N',N'-tetra[(2-pyridal)methyl]ethylenediamine, tris[(2- pyridyl)methyl]amine, tris(2-aminoethyl)amine, tris(2-bis(3-butoxy-3- oxopropyl)aminoethyl)amine, tris(2-bis(3-(2-ethylhexoxy)-3-oxopropyl)aminoethyl)amine and Tris(2-bis(3-dodecoxy-3-oxopropyl)aminoethyl)amine. In some embodiments, the ligand is 2,2’-bipyridine.

[0291] In some embodiments the initiator has the structure: wherein R1’ is a nucleophilic reactive group, R2’ comprises a linker, and R3’ comprises a polymer synthesis initiator moiety having the structurewherein R4’ and R5’ and are the same or different and are selected from the group consisting of alkyl, substituted alkyl, alkylene, alkoxy, carboxyalkyl, haloalkyl, cycloalkyl, cyclic alkyl ether, alkenyl, alkenylene, alkynyl, alkynylene, cycloalkylene, heterocycloalkyl, heterocycloalkylene, aryl, arylene, arylene-oxy, heteroaryl, amino, amido or any combination thereof; Z’ is a halogen, –OR (where R is –H, methyl, ethyl, propyl, or isopropyl), –SCN or – NCS; and s is an integer between 1 and 20.

[0292] In some embodiments, Z’ is Br and R4’ and R5’ are each methyl. In some embodiments, R1’ is selected from the group consisting of -NH2, -OH, and -SH.

[0293] In some embodiments R2’ is alkyl, substituted alkyl, alkylene, alkoxy, carboxyalkyl, haloalkyl, cycloalkyl, cyclic alkyl ether, alkenyl, alkenylene, alkynyl, alkynylene, cycloalkylene, heterocycloalkyl, heterocycloalkylene, aryl, arylene, arylene-oxy, heteroaryl, amino, amido or any combination thereof. In some embodiments, R2’ iswherein A and B are the same or different and are integers from 1-20. In some embodiments, A and B are each 4.

[0294] In some embodiments, R3’ iswherein R6, R7 and R8 are the same or different and are selected from the group consisting ofwherein Z is –OR (where R is –H, methyl, ethyl, propyl, or isopropyl), –SCN, –NCS, –F, –Cl, –Br or –I. In some embodiments, Z is –Br and R6, R7 and R8 are each.

[0295] In some embodiments, the initiator has the structure:wherein A and B are the same or different and are integers from 2 to 12 and Z is any halogen, for example Br. In some embodiments, A and B are each 4.

[0296] In some embodiments, the method further has the step of reacting the nucleophilic reactive group with a maleimide reagent to provide the phosphorylcholine containing polymer having a terminal maleimide. In some embodiments, the maleimide compound is.

[0297] Also provided are pharmaceutical compositions that include an antibody conjugate as provided herein. In some embodiment, a pharmaceutical compositions includes an antibody conjugate as described herein, and a pharmaceutically acceptable excipient (e.g., water, NaCl, normal saline solutions, lactated Ringer’s, normal sucrose, normal glucose, a buffer, and the like). The antibody conjugates and pharmaceutical compositions containing them may be administered in an effective regime for treating or prophylaxis of a patient’s disease including, for instance, administration by oral, intravitreal, intravenous, subcutaneous, intramuscular, intraosseous, intranasal, topical, intraperitoneal, and intralesional administration. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration or routes among others. In therapy or as a prophylactic, the active agent may be administered to an individual as an injectable composition, for example as a sterile aqueous dispersion. In some embodiments the agent is isotonic or substantially isotonic.

[0298] For administration to mammals, and particularly humans, it is expected that the dosage of the active agent is from 0.01 mg / kg body weight, typically around 1 mg / kg. The physician can determine the actual dosage most suitable for an individual which depends on factors including the age, weight, sex and response of the individual, the disease or disorder being treated and the age and condition of the individual being treated. The above dosages are exemplary of the average case. There can, of course, be instances where higher or lower dosages are merited. In some embodiments, the dosage can be 0.5 to 20 mg / eye, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 mg.

[0299] This dosage may be repeated as often as appropriate (e.g., weekly, fortnightly, monthly, once every two months, quarterly, twice a year, yearly). If side effects develop the amount and / or frequency of the dosage can be reduced, in accordance with normal clinical practice. In one embodiment, the pharmaceutical composition may be administered once every one to thirty days. In one embodiment, the pharmaceutical composition may be administered twice every thirty days. In one embodiment, the pharmaceutical composition may be administered once a week.

[0300] The antibodies and pharmaceutical compositions can be employed alone or in conjunction with other compounds, such as therapeutic compounds or molecules, e.g. anti- inflammatory drugs, analgesics or antibiotics. Such administration with other compounds maybe simultaneous, separate or sequential. The components may be prepared in the form of a kit which may comprise instructions as appropriate.

[0301] The antibodies, antibody conjugates, and pharmaceutical compositions disclosed herein can be used for treatment or prophylaxis of disease, particularly the ocular diseases or conditions described herein. Any of the antibodies, antibody conjugates, and pharmaceutical compositions disclosed herein, is for the treatment of a subject in need thereof, e.g., in need of treatment for an ocular disease or condition as described herein, by any of the methods described herein.

[0302] The anti-VEGF antibody conjugates, or anti-VEGF protein conjugates, and pharmaceutical compositions containing them may be formulated for and administered by ocular, intraocular, and / or intravitreal injection, and / or juxtascleral injection, and / or subretinal injection and / or subtenon injection, and / or superchoroidal injection and / or subconjunctival and / or topical administration in the form of eye drops and / or ointment. Such antibodies and compositions can be delivered by a variety of methods, e.g. intravitreally as a device and / or a depot that allows for slow release of the compound into the vitreous, including those described in references such as Intraocular Drug Delivery, Jaffe, Ashton, and Pearson, editors, Taylor & Francis (March 2006). In one example, a device may be in the form of a minipump and / or a matrix and / or a passive diffusion system and / or encapsulated cells that release the compound for a prolonged period of time (Intraocular Drug Delivery, Jaffe, Ashton, and Pearson, editors, Taylor & Francis (March 2006)).

[0303] Formulations for ocular, intraocular or intravitreal administration can be prepared by methods and using ingredients known in the art. A main requirement for efficient treatment is proper penetration through the eye. Unlike diseases of the front of the eye, where drugs can be delivered topically, retinal diseases require a more site-specific approach. Eye drops and ointments rarely penetrate the back of the eye, and the blood-ocular barrier hinders penetration of systemically administered drugs into ocular tissue. Accordingly, usually the method of choice for drug delivery to treat retinal disease, such as AMD and DME, is direct intravitreal injection. Intravitreal injections are usually repeated at intervals which depend on the patient’s condition, and the properties and half-life of the drug delivered.

[0304] Therapeutic antibodies and related conjugates generally are placed into a container having a sterile access port, for example, an intravenous solution bag or vial havinga stopper pierceable by a hypodermic injection needle. Such compositions may also be supplied in the form of pre-filled syringes.

[0305] A “stable” formulation is one in which the protein or protein conjugated to a polymer of other half-life extending moiety therein essentially retains its physical stability and / or chemical stability and / or biological activity upon storage. By “stable” is also meant a formulation which exhibits little or no signs of instability, including aggregation and / or deamidation. For example, the formulations provided may remain stable for at least two year, when stored as indicated at a temperature of 5-8°C. Suitable formulations for an anti-VEGF antibody conjugate of the present disclosure is described in e.g., PCT publication number WO2017117464, which is incorporated by reference herein in its entirety.

[0306] Various analytical techniques for measuring protein stability are available in the art and are reviewed in Peptide and Protein Drug Delivery, 247-301 (Vincent Lee ed., New York, N.Y., 1991) and Jones, 1993 Adv. Drug Delivery Rev. 10: 29-90, for examples. Stability can be measured at a selected temperature for a selected time period. In some embodiments the storage of the formulations is stable for at least 6 months, 12 months, 12- 18 months, or for 2 or more years.

[0307] A protein, such as an antibody or fragment thereof, "retains its physical stability" in a pharmaceutical formulation if it shows no signs of aggregation, precipitation, deamidation and / or denaturation upon visual examination of color and / or clarity, or as measured by UV light scattering or by size exclusion chromatography.

[0308] A protein "retains its chemical stability" in a pharmaceutical formulation, if the chemical stability at a given time is such that the protein is considered to still retain its biological activity. Chemical stability can be assessed by detecting and quantifying chemically altered forms of the protein. Chemical alteration may involve size modification (e.g., clipping), which can be evaluated using size exclusion chromatography, SDS-PAGE and / or matrix-assisted laser desorption ionization / time-of-flight mass spectrometry (MALDI / TOF MS), for examples. Other types of chemical alteration include charge alteration (e.g., occurring as a result of deamidation), which can be evaluated by ion-exchange chromatography, for example. An antibody "retains its biological activity" in a pharmaceutical formulation, if the biological activity of the antibody at a given time is within about 10% (within the errors of the assay) of the biological activity exhibited at the time thepharmaceutical formulation was prepared as determined in an antigen binding assay, for example.

[0309] A protein-polymer conjugate “retains its chemical stability” the chemical bond between the protein and the polymer is maintained intact, e.g., it is not hydrolyzed or otherwise disrupted. The protein part of the conjugate retains its chemical stability as described above.

[0310] By “isotonic” is meant that the formulation of interest has essentially the same osmotic pressure as human blood or the vitreous for intravitreal injections. Isotonic formulations will generally have an osmotic pressure from about 250 to 400 mOsm. Isotonicity can be measured using a vapor pressure or ice-freezing type osmometer, for example.

[0311] As used herein, “buffer” refers to a buffered solution that resists changes in pH by the action of its acid-base conjugate components. In some embodiments, the buffer has a pH from about 3.0 to about 8.0; for example from about 4.5 to 8; or about pH 6 to about 7.5; or about 6.0 to about 7.0, or about 6.5-7.0, or about pH 7.0 to about 7.5; or about 7.1 to about 7.4. A pH of any point in between the above ranges is also contemplated.

[0312] In some embodiments, “PBS” phosphate buffered saline, Tris based buffers and histidine based buffers are used.

[0313] In some embodiments, the PBS buffer is made up of at least Na2HPO4, KH2PO4 and NaCl adjusted so as to provide the appropriate pH. In some embodiments, the buffer may contain other pharmaceutical excipients such as KCl and other salts, detergents and / or preservatives so as to provide a stable storage solution.

[0314] A “preservative” is a compound which can be included in the formulation to essentially reduce bacterial action therein, thus facilitating the production of a multi-use formulation, for example. Examples of potential preservatives include octadecyldimethylbenzyl ammonium chloride, hexamethonium chloride, benzalkonium chloride (a mixture of alkylbenzyldimethylammonium chlorides in which the alkyl groups are long-chain compounds), and benzethonium chloride. Other types of preservatives include aromatic alcohols such as phenol, butyl and benzyl alcohol, alkyl parabens such as methyl or propyl paraben, catechol, resorcinol, cyclohexanol, 3-pentanol, and m-cresol.

[0315] In some embodiments, formulations, to be safe for human use or for animal testing, should have sufficiently low levels of endotoxin. “Endotoxin” is lipopolysaccharide(LPS) derived from the cell membrane of Gram-negative bacteria. Endotoxin is composed of a hydrophilic polysaccharide moiety covalently linked to a hydrophobic lipid moiety (lipid A). Raetz CR, Ulevitch RJ, Wright SD, Sibley CH, Ding A, Nathan CF. 1991. Gram-negative endotoxin: an extraordinary lipid with profound effects on eukaryotic signal transduction. FASEB J. 5(12):2652-2660. Lipid A is responsible for most of the biological activities of endotoxin, i.e., its toxicity. Endotoxins are shed in large amount upon bacterial cell death as well as during growth and division. They are highly heat-stable and are not destroyed under regular sterilizing conditions. Extreme treatments with heat or pH, e.g., 180-250°C and over 0.1 M of acid or base must be used (Petsch D, Anspach F. 2000. Endotoxin removal from protein solutions. J Biotechnol. 76: 97-119). Such conditions of course would be highly detrimental to biological drugs.

[0316] In the biotech and pharmaceutical industries, it is possible to find endotoxin during both production processes and in final products. As bacteria can grow in nutrient poor media, including water, saline and buffers, endotoxins are prevalent unless precautions are taken. Endotoxin injection into an animal or human causes a wide variety of pathophysiological effects, including endotoxin shock, tissue injury and even death. Ogikubo Y, Ogikubo Y, Norimatsu M, Noda K, Takahashi J, Inotsume M, Tsuchiya M, Tamura Y. 2004. Evaluation of the bacterial endotoxin test for quantifications of endotoxin contamination of porcine vaccines. Biologics 32:88-93.

[0317] Pyrogenic reactions and shock are induced in mammals upon intravenous injection of endotoxin at low concentrations (1 ng / mL) (Fiske JM, Ross A, VanDerMeid RK, McMichael JC, Arumugham. 2001. Method for reducing endotoxin in Moraxella catarrhalis UspA2 protein preparations. J Chrom B. 753:269-278). The maximum level of endotoxin for intravenous applications of pharmaceutical and biologic product is set to 5 endotoxin units (EU) per kg of body weight per hour by all pharmacopoeias (Daneshiam M, Guenther A, Wendel A, Hartung T, Von Aulock S. 2006. In vitro pyrogen test for toxic or immunomodulatory drugs. J Immunol Method 313:169-175). EU is a measurement of the biological activity of an endotoxin. For example, 100 pg of the standard endotoxin EC-5 and 120 pg of endotoxin from Escherichia coli O111:B4 have activity of 1 EU (Hirayama C, Sakata M. 2002. Chromatographic removal of endotoxin from protein solutions by polymer particles. J Chrom B 781:419-432). Meeting this threshold level has always been a challengein biological research and pharmaceutical industry (Berthold W, Walter J. 1994. Protein Purification: Aspects of Processes for Pharmaceutical Products. Biologicals 22:135-150; Petsch D, Anspach FB.2000. Endotoxin removal from protein solutions. J Biotech 76:97-119).

[0318] The presence of endotoxin in drugs to be delivered via intravitreal injection is of particular concern. Intravitreal injection of drug (penicillin) was first performed in 1945 by Rycroft. Rycroft BW.1945. Penicillin and the control of deep intra-ocular infection. British J Ophthalmol 29 (2): 57-87. The vitreous is a chamber where high level of drug can be introduced and maintained for relatively long periods of time. The concentration of drug that can be achieved via intravitreal injection far exceeds what can be generated by topical administration or by systemic administration (e.g. intravenous).

[0319] One of the most dangerous complications potentially arising from intravitreal injections is endophthalmitis. Endophthalmitis falls into two classes: infectious and sterile. Infectious endophthalmitis is generally cause by bacteria, fungi or parasites. The symptoms of infectious endophthalmitis include severe pain, loss of vision, and redness of the conjunctiva and the underlying episclera. Infectious endophthalmitis requires urgent diagnosis and treatment. Possible treatments include intravitreal injection of antibiotics and pars plana vitrectomy in some cases. Enucleation may be called for to remove a blind and painful eye. See, e.g., Christy NE, Sommer A. 1979. Antibiotic prophylaxis of postoperative endophthalmitis. Ann Ophthalmol 11 (8): 1261–1265.

[0320] Sterile endophthalmitis in contrast does not involve an infectious agent and can be defined as the acute intraocular inflammation of the vitreous cavity that resolves without the need of intravitreal antibiotics and / or vitreoretinal surgery. If a vitreous microbiological study has been done, it needs to be negative culture proven to sustain a diagnosis of sterile endophthalmitis. Marticorena J, Romano V, Gomez-Ulla F. 2012 “Sterile Endophthalmitis after Intravitreal Injections” Med Inflam. 928123.

[0321] It has been observed that intravitreal injection of biological drugs contaminated with endotoxin can result in sterile endophthalmitis. Marticorena, et al. Bevacizumab (Avastin) is approved by the Food and Drug Administration for the treatment of glioblastoma and of metastatic colorectal cancer, advanced nonsquamous non-small-cell lung cancer and metastatic kidney cancer. Bevacizumab is also widely used off label as a treatment for wAMD. Bevacizumab comes from the manufacturer as a 100 mg / 4 ml. This solutioncannot be directly used for intravitreal injection and should be compounded by a pharmacist. Clusters of sterile endophthalmitis have been observed and are theorized to be cause by inadvertent contamination of bevacizumab by endotoxin by the compounding pharmacist.

[0322] Given the dire clinical results of intravitreal injection of endotoxin, the total amount of endotoxin that can be given to a patient via intravitreal dosing is highly limited. In some embodiments, a solution having an antibody or antibody-conjugate is provided having an endotoxin level that does not exceed 5.0 EU / ml. In some embodiments, the endotoxin level does not exceed 1.0 EU / ml. In some embodiments, the endotoxin level does not exceed 0.5 EU / ml. In some embodiments, the endotoxin level does not exceed 0.2 EU / ml. In some embodiments, the endotoxin level does not exceed 2, 1, 0.5, 0.2, 0.1, 0.09, 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02 or 0.01 EU / ml.

[0323] Two commonly used FDA-approved tests for the presence of endotoxin are the rabbit pyrogen test and Limulus Amoebodyte Lysate (LAL) assay (Hoffman S, et al.2005. International validation of novel pyrogen tests based on human monocytoid cells J. Immunol. Methods 298:161-173; Ding JL, Ho BA. 2001. New era in pyrogen testing. Biotech. 19:277- 281). The rabbit pyrogen test was developed in the 1920s and involves monitoring the temperature rise in a rabbit injected with a test solution. However, use of the rabbit pyrogen test has greatly diminished over the years due to expense and long turnaround time. Much more common is the LAL test. LAL is derived from the blood of a horseshoe crab and clots upon exposure to endotoxin.

[0324] One of the simplest LAL assays is the LAL gel-clot assay. Essentially, the LAL clotting assay is combined with a serial dilution of the sample in question. Formation of the gel is proportional to the amount of endotoxin in the sample. Serial dilutions are prepared from the sample and each dilution assayed for its ability to form LAL gel. At some point a negative reaction is contained. The amount of endotoxin in the original sample can be estimated from the dilution assay.

[0325] Other LAL tests have also been developed, including the turbidimetric LAL assay (Ong KG, Lelan JM, Zeng KF, Barrett G, Aourob M, Grimes CA.2006. A rapid highly- sensitive endotoxin detection system. Biosensors and Bioelectronics 21:2270-2274) and the chromogenic LAL assay (Haishima Y, Hasegawa C, Yagami T, Tsuchiya T, Matsuda R, Hayashi Y. 2003. Estimation of uncertainty in kinetic-colorimetric assay of bacterialendotoxins. J Pharm Biomed Analysis. 32:495-503). The turbidimetric and chromogenic assays are much more sensitive and quantitative than the simple gel-clot dilution assay.

[0326] In some embodiments a method of reducing the amount of endotoxin in a composition having an antibody disclosed herein is provided. The method having the steps of contacting the composition with an affinity chromatography resin that binds to the antibody; eluting the antibody from the affinity chromatography resin to form an affinity chromatography eluent having the antagonist; contacting the affinity chromatography eluent with an ion-exchange resin that binds the antibody; and eluting the antibody from the ion- exchange resin, wherein the antibody eluted from the ion-exchange resin is substantially free from endotoxin.

[0327] The above method for reducing the amount of endotoxin, or other method or process recited herein, can be performed in the order described in the steps above or it can optionally be performed by varying the order of the steps or even repeating one or more of the steps. In one embodiment, the method of reducing the amount of endotoxin in a composition is performed in the order of the described steps. In some embodiments, the affinity chromatography resin contacting, washing and eluting steps are repeated in the same order more than one time before contacting the affinity chromatography eluent with the ion exchange resin. The method can also include a filtering step using, for example, a 0.1 micron, 0.22 micron, or 0.44 micron filter, that can be performed on either one or more of the eluents removed after each resin binding step.

[0328] In certain instances, the steps of contacting the composition with affinity chromatography resin, washing and eluting the antibody from the affinity chromatography resin can be repeated more than one time before contacting the first eluent with an ion- exchange resin. In one embodiment, the affinity chromatography resin comprises a recombinant Protein A (“rProteinA”) resin. One example of a suitable recombinant Protein A resin is rProteinA Sepharose FF® resin (Amersham, Piscataway, N.J.). In another embodiment, a suitable affinity chromatography resin would comprise a protein G chromatography resin. In other embodiments, a suitable affinity chromatography resin comprises a mixed Protein A / Protein G resin. In other embodiments, a suitable affinity chromatography resin comprises a hydrophobic charge induction resin that comprises a 4-mercaptoethylpyridine ligand such as a MEP HyperCel® resin (BioSepra, Cergy, Saint Christophe, France).

[0329] In some embodiments, the ion exchange resin comprises an anion-exchange resin. As will be known by the person skilled in the art, ion exchangers may be based on various materials with respect to the matrix as well as to the attached charged groups. For example, the following matrices may be used, in which the materials mentioned may be more or less cross-linked: MacroCap Q (GE Healthcare Biosciences, Piscataway, NJ), agarose based (such as Sepharose CL-6B®, Sepharose Fast Flow® and Sepharose High Performance ®), cellulose based (such as DEAE Sephacel®), dextran based (such as Sephadex®), silica based and synthetic polymer based. For the anion exchange resin, the charged groups, which are covalently attached to the matrix, may, for example, be diethylaminoethyl, quaternary aminoethyl, and / or quaternary ammonium. In some embodiments the anion-exchange resin comprises a quaternary amine group. An exemplarily anion-exchange resin that has a quaternary amine group for binding the anti-M-CSF antibody is a Q Sepharose® resin (Amersham, Piscataway, N.J.).

[0330] In other aspects, if the endotoxin levels are higher than desired after subjecting the composition to the aforementioned anion-exchange chromatography step, the composition may in the alternative be subjected to a cation exchange resin. In some embodiments, any endotoxin in the composition should have a differential binding to the ion- exchange resin than the protein in question to allow purification of the protein from the endotoxin. In this regard, endotoxin is negatively charged and will generally bind to an anion exchange resin. If both the protein and the endotoxin bind to the anion exchange resin, purification of one from the other may be effectuated by using a salt gradient to elute the two into different fractions. The relative binding of the protein to a particular resin may also be effected by changing the pH of the buffer relative to the pI of the protein. In some embodiments, cation-exchange chromatography is the sole ion-exchange chromatography employed.

[0331] In some embodiments, if the endotoxin levels are too high after the anion exchange resin, the composition may be further subjected to a second ion-exchange step, for example, by contacting the compositions with a cation exchange resin and followed by a wash step, then elution from the ion-exchange resin. In some embodiments, the cation exchangeresin comprises a sulfonic group for binding. Exemplary cation exchange resins are SP Sepharose® resin FF (Amersham, Piscataway, N.J.) Poros XS (CEX) (Life Technology, Grand Island, New York).

[0332] In some embodiments, after the solution of antibody protein is produced having the specified level of endotoxin, there are a number of steps prior to final formulation of the protein. In some embodiments, a half-life extending moiety is conjugated to the protein. The conjugate is then formulated into a final drug formulation which is injected into the patients. In some embodiments, the conjugate is again purified on an ion-exchange resin which can be a cation-exchange resin. In other embodiments, the protein is formulated. In all cases, normal laboratory procedures should be employed to prevent the introduction of endotoxin contaminants into the protein sample or into the protein-polymer conjugate. Formulations

[0333] In any method of treating an eye disorder (e.g., wAMD, DR, etc.) provided herein, in some embodiments, the method includes administering to the subject a therapeutic formulation or pharmaceutical composition containing the dose (e.g., loading dose, subsequent dose, etc.) of the anti-VEGF antibody conjugate as provided herein. The pharmaceutical composition or therapeutic formulation can include any suitable components, as disclosed herein. In some embodiments, the therapeutic formulation or pharmaceutical composition includes a mixture of the conjugated and unconjugated form of the anti-VEGF antibody, as described herein (e.g., mixture of KSI-301ABC and KSI-301P). In some embodiments, the therapeutic formulation or pharmaceutical composition includes: an unconjugated anti-VEGF antibody, with or without the non-native cysteine in the Fc region, wherein the unconjugated anti-VEGF antibody is not covalently attached to a phosphorylcholine-containing polymer; and the anti-VEGF antibody conjugate, wherein the unconjugated anti-VEGF antibody is present in the formulation in a range of 10-60% (e.g., 15-25%) of a total molar amount of the anti-VEGF antibody conjugate and the unconjugated anti-VEGF antibody, wherein the total molar amount is the sum of the molar amount of the anti-VEGF antibody conjugate and the molar amount of the unconjugated anti-VEGF antibody. In some embodiments, the unconjugated anti-VEGF antibody is present in the formulation at about 20% of the total molar amount of the anti-VEGF antibody conjugate and the unconjugated anti-VEGF antibody.

[0334] 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%, 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.

[0335] 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%, 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 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.

[0336] 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%, 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 of the conjugate and the second protein. In some embodiments, the polymer is a phosphorylcholine-containing polymer.

[0337] 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%, etc.) of a total molar amount of the conjugate and the second protein, wherein the total molar amountcomprises 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.

[0338] In some embodiments, the formulation or composition comprises a mixture, by mass weight concentration, of 5% to 10% unconjugated protein (e.g. unconjugated antibody) with the remainder comprising the conjugated protein (e.g., conjugated antibody). 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) 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) 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 compositioncomprises a mixture of the two proteins (e.g., antibodies) 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) by mass weight concentration of 5% to 30% 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 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) 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) by mass weight concentration of 5% to 55% unconjugated protein (e.g., unconjugated antibody) with the remainder comprising the conjugated protein.

[0339] In some embodiments, the composition or formulation comprises a mixture of the two proteins (e.g., antibodies) by mass weight concentration of 5% to 55% unconjugated protein (e.g., unconjugated antibody) with the remainder comprising the conjugated protein (e.g., the conjugated antibody). 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) by mass weight concentration of 5% to 65% 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., 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) 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 some embodiments, the composition comprises a mixture of the two proteins (e.g., antibodies) 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) by mass weight concentration of 5% to 90% unconjugated protein (e.g., unconjugated antibody) with the remainder comprising the conjugated protein.

[0340] 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.

[0341] 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 weightconcentration of the proteins (e.g., excluding any contribution of a polymer that may be conjugated thereto) in the mixture.

[0342] 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%, etc.).

[0343] 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%, etc.), 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.

[0344] 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%, etc.), 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 weightconcentration of the first protein and the second protein in the composition, at a pH within 0.5 pH units of the pI of the second protein. 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 within 0.5 pH units of the pI of the second protein.

[0345] 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 5-90%, 15-25%, 25-35%, etc.).

[0346] 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%, 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%, 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.

[0347] 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. Insome 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., unco...

Claims

WHAT IS CLAIMED IS:

1. A method of treating wet age-related macular degeneration (wAMD), the method comprising: identifying a subject with wAMD; administering a first dose of an anti-VEGF antibody conjugate to the subject; administering a second dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the first dose; administering a third dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the second dose; administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the third dose; and administering an individualized dose of the anti-VEGF antibody conjugate to the subject at least 4 weeks (+ / - 7 days) after the fourth dose if the subject has a decline in eye health, wherein the individualized dose is administered no more frequently than about once every four weeks, optionally wherein one or more subsequent doses of the anti-VEGF antibody conjugate are administered to the subject no less frequently than about once every 24 weeks after the fourth and / or the last dose.

2. The method of claim 1, comprising administering a subsequent dose of the anti- VEGF antibody conjugate to the subject if the subject has not received any dose of the anti- VEGF antibody conjugate for at least 24 weeks after the last dose.

3. A method of treating wAMD, the method comprising: identifying a subject with wAMD; administering a first dose of an anti-VEGF antibody conjugate to the subject; administering a second dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the first dose; administering a third dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the second dose; administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the third dose;evaluating the subject’s eye health no more frequently than about once every four weeks after the fourth dose; and if the subject has a decline in eye health upon evaluating, administering an individualized dose of the anti-VEGF antibody conjugate to the subject, optionally wherein one or more subsequent doses of the anti-VEGF antibody conjugate are administered to the subject no less frequently than about once every 24 weeks after the fourth and / or the last dose.

4. A method of treating wAMD, the method comprising: identifying a subject with wAMD; administering a first dose of an anti-VEGF antibody conjugate to the subject; administering a second dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the first dose; administering a third dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the second dose; administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the third dose; evaluating the subject’s eye health no more frequently than about once every four weeks (or no more frequently than Q4W or QM) after the fourth dose; and administering an individualized dose to the subject based on the subject’s evaluated eye health, optionally wherein one or more subsequent doses of the anti- VEGF antibody conjugate are administered to the subject no less frequently than about once every 24 weeks after the fourth and / or the last dose.

5. A method of treating wAMD, the method comprising: identifying a subject with wAMD; administering a first dose of an anti-VEGF antibody conjugate to the subject; administering a second dose of the anti-VEGF antibody conjugate to the subject 4 weeks after the first dose; and administering a third dose of the anti-VEGF antibody conjugate to the subject 4 weeks after the second dose;administering a fourth dose of the anti-VEGF antibody conjugate to the subject 4 weeks after the third dose; and if the subject has a decline in eye health within about 24 weeks after the fourth dose, administering an individualized dose of the anti-VEGF antibody conjugate, optionally wherein one or more subsequent doses of the anti-VEGF antibody conjugate are administered to the subject no less frequently than about once every 24 weeks after the fourth and / or the last dose.

6. The method of claim 5, wherein the individualized dose is administered no more frequently than about once every 4 weeks (or no more frequently than Q4W or QM).

7. A method of treating eye disease, the method comprising: identifying a subject with a neovascular eye disease; administering a first dose of an anti-VEGF antibody conjugate to the subject; administering a second dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the first dose; administering a third dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the second dose; administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the third dose; administering an individualized dose of the anti-VEGF antibody conjugate to the subject at least 4 weeks (+ / - 7 days) after the fourth dose if the subject has a decline in eye health, wherein the individualized dose is administered no more frequently than about once every four weeks (or no more frequently than Q4W or QM).

8. The method of claim 7, comprising evaluating the subject’s eye health after the fourth dose to determine if the subject has the decline in eye health.

9. The method of claim 7, wherein the neovascular eye disease is wAMD.

10. The method of claim 1, wherein the decline in eye health is based on the presence of intraretinal fluid (IRF), the presence of subretinal fluid (SRF), and / or new or worsening macular hemorrhage due to wAMD activity.

11. The method of claim 10, wherein (a) the subject is determined to have the presence of intraretinal fluid (IRF) when the amount of fluid in an OCT IRF volume is above a predetermined normal level, optionally wherein the predetermined normal level is 5-20 nanoliters over a 3 mm diameter circle, or 5-10 nanoliters over a 1 mm diameter circle, and (b) the subject is determined to have the presence of subretinal fluid (SRF) when the amount of fluid in an OCT SRF volume is above a predetermined normal level, optionally wherein the predetermined normal level is 10-50 nanoliters over a 3 mm diameter circle, or 10-25 nanoliters over a 1 mm diameter circle.

12. The method of claim 1, wherein the decline in eye health comprises an increase in retina thickness, optionally wherein the retina thickness is based on an optical coherence tomography central subfield thickness (OCT CST).

13. The method of claim 12, wherein the decline in eye health comprises an increase in OCT CST compared to a previous measurement of OCT CST, optionally wherein the increase in OCT CST is compared to a prior lowest measurement.

14. The method of claim 1, comprising evaluating the subject’s eye health by performing optical coherence tomography (OCT) and / or optical coherence tomography angiography (OCT-A).

15. The method of claim 1, comprising evaluating the subject’s eye health and determining that the subject has new or worsening macular hemorrhage due to wAMD activity, optionally wherein the new or worsening macular hemorrhage due to wAMD activity is determined based on a fundus photograph.

16. The method of claim 1, comprising administering the individualized dose no more frequently than once every four weeks (+ / - 7 days).

17. A method of treating a subject with wAMD, the method comprising: administering to a subject in need thereof 4 loading doses of an anti-VEGF antibody conjugate at frequency of one loading dose every four weeks (or at Q4W or at QM); determining, at least 4 weeks (+ / - 7 days) after a last loading dose, a presence or absence of intraretinal fluid (IRF) and / or subretinal fluid (SRF) in an eye of the subject, and / or if macular hemorrhage due to wAMD activity has clinically worsened; and if there is IRF or SRF in the eye, or the subject has clinically worsened macular hemorrhage due to wAMD activity, administering a maintenance dose.

18. The method of claim 17, wherein determining the presence or absence of IRF and / or SRF in the subject’s eye comprises performing optical coherence tomography (OCT).

19. The method of claim 17, wherein the subject is determined to have the presence of intraretinal fluid (IRF) and / or subretinal fluid (SRF) when the amount of fluid in the subject’s eye is above predetermined normal levels, optionally the predetermined normal levels are around 5-20 nanoliters for IRF at 3mm, or 10-50 nanoliters for SRF at 3mm, or IRF exceeding 5-10 nL at 1mm, or SRF exceeding 10-25 nL at 1mm.

20. The method of claim 1, wherein a vision assessment as measured by Best Corrected Visual Acuity (BCVA) increases by at least 1 ETDRS letters or by at least a threshold after administering at least the fourth dose relative to a baseline, wherein the threshold is 1-5 Early Treatment Diabetic Retinopathy Study (ETDRS) letters.

21. The method of claim 1, wherein the decline in eye health comprises an increase in central subfield thickness (CST) as measured by optical coherence tomography (OCT) of at least 20-50 microns after the fourth dose, compared to the prior lowest measurement.

22. A method of treating diabetic retinopathy (DR), the method comprising:identifying a subject with DR; administering a first dose of an anti-VEGF antibody conjugate to the subject; administering a second dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the first dose; administering a third dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the second dose; administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 12 weeks after the third dose; and administering a fifth dose of the anti-VEGF antibody conjugate to the subject about 24 weeks after the fourth dose.

23. The method of claim 22, comprising administering one or more subsequent doses of the anti-VEGF antibody conjugate to the subject after the fifth dose at or no more frequently than Q24W.

24. The method of claim 22, wherein the DR is non-proliferative diabetic retinopathy (NPDR).

25. The method of claim 22, wherein the subject is identified as having moderately severe to severe NPDR.

26. The method of claim 22, wherein the identified subject is a treatment naïve subject.

27. A method of treating diabetic retinopathy (DR), the method comprising: (a) identifying a treatment naive subject with moderately severe to severe NPDR; (b) administering a first dose of an anti-VEGF antibody conjugate to the subject; (c) administering a second dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the first dose; (d) administering a third dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the second dose; (e) administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 12 weeks after the third dose; and (f) administering a fifth dose of the anti-VEGF antibody conjugate to the subject about 24 weeks after the fourth dose,(g) performing (b)-(f) unless the subject develops DME, PDR, and / or ASNV, in which case: (h) administering a fifth dose of an anti-VEGF antibody conjugate to the subject when the subject is identified as having developed DME, PDR, and / or ASNV; (i) administering a sixth dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the fifth dose; (j) administering a seventh dose of the anti-VEGF antibody conjugate to the subject about 12 weeks after the sixth dose; (k) administering an eighth dose of the anti-VEGF antibody conjugate to the subject about 12 weeks after the seventh dose; and (l) administering a ninth dose of the anti-VEGF antibody conjugate to the subject about 12 weeks after the eighth dose.

28. The method of claim 22, wherein the subject has a diabetic retinopathy severity score (DRSS) from 47 to 53 before administering the first dose.

29. The method of claim 22, wherein before administering the first dose the subject has a CST 320 microns and a BCVA ETDRS letter score in the Study Eye of 69 letters (approximate Snellen equivalent of 20 / 40 or better), or a CST >320 and 350 microns and a BCVA ETDRS letter score in the Study Eye of 79 letters (approximate Snellen equivalent of 20 / 25 or better).

30. The method of claim 1, wherein each dose of the anti-VEGF antibody conjugate comprises 1-6 mg of protein of the anti-VEGF antibody conjugate, optionally wherein each dose comprises about 5 mg of protein of the anti-VEGF antibody conjugate.

31. The method of claim 1, wherein the anti-VEGF antibody conjugate comprises an anti-VEGF-A antibody and a phosphorylcholine-containing polymer, wherein the polymer is covalently bonded to the anti-VEGF-A antibody at a non-native cysteine outside a variable region of the anti-VEGF-A antibody.

32. The method of claim 1, wherein the anti-VEGF antibody conjugate is administered intravitreally.

33. The method of claim 1, wherein the anti-VEGF antibody conjugate comprises an anti-VEGF-A antibody and a phosphorylcholine containing polymer, wherein the polymer is covalently bonded to the anti-VEGF-A antibody at a non-native cysteine outside a variable region of the anti-VEGF-A antibody, wherein the anti-VEGF-A antibody comprises a light chain and heavy chain, said heavy chain comprising an Fc region, wherein the cysteine is in the Fc region of the heavy chain, wherein the antibody conjugate has the following structure:; wherein: each heavy chain of the anti-VEGF-A antibody is denoted by the letter H, and each light chain of the anti-VEGF-A antibody is denoted by the letter L;the polymer is bonded to the anti-VEGF-A antibody through a sulfhydryl at C443 according to EU numbering, which bond is depicted on one of the heavy chains above; PC is, where the curvy line indicates the point of attachment to the rest of the polymer; 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%, optionally wherein the sequence of the heavy chain comprises SEQ ID NO: 1 or a sequence at least 90% identical thereto, and wherein the sequence of the light chain comprises SEQ ID NO: 2, or a sequence at least 90% identical thereto.

34. A method of treating wet age-related macular degeneration (wAMD) comprising: identifying a subject with wAMD; administering a first dose of an anti-VEGF antibody conjugate (e.g., KSI-301) to the subject; administering a second dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the first dose; administering a third dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the second dose; administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the third dose; and administering an individualized dose of the anti-VEGF antibody conjugate to the subject at least 4 weeks (+ / - 7 days) after the fourth dose if the subject has a decline in eye health, wherein the individualized dose is administered no more frequently than, once every four weeks (or no more frequently than Q4W or QM), wherein the first, second, third, fourth doses and the individualized dose each comprises about 5 mg of protein of the anti-VEGF antibody conjugate, wherein the anti-VEGF antibody conjugate comprises:(1) an anti-VEGF-A antibody that includes: a light chain that has the amino acid sequence set forth in SEQ ID NO: 2; and a heavy chain that has the amino acid sequence set forth in SEQ ID NO :1 (or any of the variants thereof in FIG.10, e.g., SEQ ID NOs: 9, 10, 11, or 12); and (2) a phosphorylcholine containing polymer, wherein the polymer is covalently bonded to the heavy chain of the anti-VEGF-A antibody, where the antibody conjugate has the following structure:wherein: each heavy chain of the anti-VEGF-A antibody is denoted by the letter H, and each light chain of the anti-VEGF-A antibody is denoted by the letter L; the polymer is bonded to the heavy chain through a sulfhydryl at C443 according to EU numbering, which bond is depicted on one of the heavy chains above;PC is, where the curvy line indicates the point of attachment to the rest of the polymer; 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%.

35. A method of treating wet age-related macular degeneration (wAMD) comprising: identifying a subject with wAMD; administering a first dose of an anti-VEGF antibody conjugate (e.g., KSI-301) to the subject; administering a second dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the first dose; administering a third dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the second dose; administering a fourth dose of the anti-VEGF antibody conjugate to the subject about 4 weeks after the third dose; and if the subject has a decline in eye health within about 24 weeks after the fourth dose, administering an individualized dose of the anti-VEGF antibody conjugate, wherein the first, second, third, fourth doses and the individualized dose each comprises about 5 mg of protein of the anti-VEGF antibody conjugate, wherein the anti-VEGF antibody conjugate comprises: (1) an anti-VEGF-A antibody that includes: a light chain that has the amino acid sequence set forth in SEQ ID NO: 2; and a heavy chain that has the amino acid sequence set forth in SEQ ID NO :1 (or any of the variants thereof in FIG.10, e.g., SEQ ID NOs: 9, 10, 11, or 12); and (2) a phosphorylcholine containing polymer, wherein the polymer is covalently bonded to the heavy chain of the anti-VEGF-A antibody, where the antibody conjugate has the following structure:wherein: each heavy chain of the anti-VEGF-A antibody is denoted by the letter H, and each light chain of the anti-VEGF-A antibody is denoted by the letter L; the polymer is bonded to the heavy chain through a sulfhydryl at C443 according to EU numbering, which bond is depicted on one of the heavy chains above; PC is, where the curvy line indicates the point of attachment to the rest of the polymer; 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%.

36. A method of treating a subject with wet age-related macular degeneration (wAMD) comprising: administering to a subject in need thereof 4 loading doses of an anti-VEGF antibody conjugate at frequency of one loading dose every four weeks (or at Q4W or at QM); determining, at least 4 weeks (+ / - 7 days) after a last loading dose, a presence or absence of intraretinal fluid (IRF) and / or subretinal fluid (SRF) in an eye of the subject, and / or if macular hemorrhage due to wAMD activity has clinically worsened; and if there is IRF or SRF in the eye, or the subject has clinically worsened macular hemorrhage due to wAMD activity, administering a maintenance dose, wherein the loading doses and the maintenance dose each comprises about 5 mg of protein of the anti-VEGF antibody conjugate, wherein the anti-VEGF antibody conjugate comprises: (1) an anti-VEGF-A antibody that includes: a light chain that has the amino acid sequence set forth in SEQ ID NO: 2; and a heavy chain that has the amino acid sequence set forth in SEQ ID NO :1 (or any of the variants thereof in FIG.10, e.g., SEQ ID NOs: 9, 10, 11, or 12); and (2) a phosphorylcholine containing polymer, wherein the polymer is covalently bonded to the heavy chain of the anti-VEGF-A antibody, where the antibody conjugate has the following structure:wherein: each heavy chain of the anti-VEGF-A antibody is denoted by the letter H, and each light chain of the anti-VEGF-A antibody is denoted by the letter L; the polymer is bonded to the heavy chain through a sulfhydryl at C443 according to EU numbering, which bond is depicted on one of the heavy chains above; PC is, where the curvy line indicates the point of attachment to the rest of the polymer; 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%.

37. The method of claim 1, wherein the decline in eye health comprises an IRF exceeding 5-20 nL or a SRF exceeding 10-50 nL at 3mm, after the fourth dose.

38. The method of claim 1, wherein the decline in eye health comprises an IRF exceeding 5-10 nL or a SRF exceeding 10-25 nL at 1mm, or a CST-RPE (internal limiting membrane to retinal pigment epithelium) / CST-BM (internal limiting membrane to Bruch's membrane) increase above 25-100 m, after the fourth dose.

39. The method of claim 1, wherein the decline in eye health comprises a CST- RPE / CST-BM increase above 25-100 m, after the fourth dose.

40. The method of claim 1, wherein the decline in eye health comprises an IRF exceeding 5-20 nL or a SRF exceeding 10-50 nL at 3mm or an IRF exceeding 5-10 nL or a SRF exceeding 10-25 nL at 1mm, after the fourth dose.

41. The method of claim 1, wherein the decline in eye health comprises new onset of retinal hemorrhage from choroidal neovascularization (CNV).

42. The method of claim 1, wherein the diabetic retinopathy is non-proliferative diabetic retinopathy (NPDR).

43. The method of claim 1, wherein the anti-VEGF-A antibody conjugate is tarcocimab tedromer.

44. The method of claim 1, wherein the administering comprises administering to the subject a therapeutic formulation comprising: an unconjugated anti-VEGF-A antibody comprising the fusion protein with or without the non-native cysteine in the Fc region, wherein the unconjugated anti-VEGF-A antibody is not covalently attached to a phosphorylcholine-containing polymer; and the anti-VEGF-A antibody conjugate, wherein the unconjugated anti-VEGF-A antibody is present in the formulation in a range of 10-60% of a total molar amount of the anti-VEGF-A antibody conjugate and the unconjugated anti-VEGF-A antibody, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated anti-VEGF- A antibody.

45. The method of claim 44, wherein the unconjugated anti-VEGF-A antibody is present in the therapeutic formulation at about 20% of the total molar amount of the anti- VEGF-A antibody conjugate and the unconjugated anti-VEGF-A antibody.

46. The method of claim 44, wherein the total amount of the anti-VEGF-A antibody conjugate (by weight of the antibody portion) and the unconjugated anti-VEGF-A antibody in each dose is 1-6 mg, optionally the total amount of the anti-VEGF-A antibody conjugate (by weight of the antibody portion) and the unconjugated anti-VEGF-A antibody in each dose is or is about 5 mg.

47. A method of treating eye disease, comprising: administering one or more doses of an anti-VEGF therapy no more frequently than Q4W (+ / - 7 days) to a subject with a neovascular retinal disease or condition; and evaluating the subject’s eye health after administering the one or more doses of the anti-VEGF therapy to determine if the subject has a decline in eye health, wherein the evaluating comprises determining a presence or absence of intraretinal fluid (IRF) and / or subretinal fluid (SRF) in an eye of the subject.

48. The method of claim 47, comprising administering one or more individualized doses of the anti-VEGF therapy to the subject after administering the one or more doses if the subject has a decline in eye health, wherein the individualized dose is administered no more frequently than Q4W (+ / - 7 days).

49. A method of treating eye disease, comprising administering an anti-VEGF therapy to a subject with a neovascular retinal disease or condition according to a treatment regimen, to thereby treat the eye disease, wherein the treatment regimen comprises administering one or more doses of the anti- VEGF therapy no more frequently than Q4W (+ / - 7 days) to the subject, and wherein an efficacy of the treatment regimen has been determined by evaluating an eye health of a cohort of patients with the neovascular retinal disease or condition that have been administered one or more doses of the anti-VEGF therapy no more frequently than Q4W (+ / - 7 days), by determining a change in an intraretinal fluid (IRF) and / or subretinal fluid (SRF) in an eye of each patient of the cohort after administration of the one or more doses of the anti- VEGF therapy.

50. The method of claim 49, wherein the patients if the cohort have been further administered one or more individualized doses of the anti-VEGF therapy after administration of the one or more doses upon a decline in eye health, wherein the individualized dose is administered no more frequently than Q4W (+ / - 7 days).

51. The method of claim 49, wherein the efficacy of the treatment regimen comprises a time interval until a loss of a therapeutic result of the anti-VEGF therapy after administration of the one or more doses of the anti-VEGF therapy.

52. The method of claim 49, comprising administering one or more individualized doses of the anti-VEGF therapy to the subject after administering the one or more doses, upon a decline in eye health.

53. The method of claim 52, comprising evaluating the subject’s eye health after administering the one or more doses to determine if the subject has the decline in eye health, optionally wherein the evaluating comprises evaluating an OCT scan of the subject’s eye.

54. The method of claim 50, comprising: evaluating the subject’s eye health after administering the one or more doses to determine if the subject has a second decline in eye health, optionally wherein the evaluating comprises evaluating an OCT scan of the subject’s eye; and administering one or more individualized doses of the anti-VEGF therapy to the subject upon determining that the subject has the second decline in eye health, thereby treating the eye disease.

55. The method of claim 49, wherein the decline in eye health comprises an IRF exceeding 5-20 nL or a SRF exceeding 10-50 nL at 3mm.

56. The method of claim 49, wherein the decline in eye health comprises an IRF exceeding 5-10 nL or a SRF exceeding 10-25 nL at 1mm, or a CST-RPE (internal limiting membrane to retinal pigment epithelium) / CST-BM (internal limiting membrane to Bruch's membrane) increase above 25-100 m.

57. The method of claim 49, wherein the decline in eye health comprises a CST- RPE / CST-BM increase above 25-100 m.

58. The method of claim 49, wherein the decline in eye health comprises an IRF exceeding 5-20 nL or a SRF exceeding 10-50 nL at 3mm or an IRF exceeding 5-10 nL or a SRF exceeding 10-25 nL at 1mm.

59. The method of claim 49, wherein the decline in eye health comprises new onset of retinal hemorrhage from choroidal neovascularization (CNV).

60. The method of claim 49, wherein the evaluating is performed no more frequently than Q4W (+ / - 7 days).

61. The method of claim 49, wherein the neovascular retinal disease or condition is selected from: diabetic retinopathy (DR), choroidal neovascularization (CNV), wet age-related macular degeneration (AMD), diabetic macular edema (DME), pathological myopia, von Hippel-Lindau disease, histoplasmosis of the eye, retinal vein occlusion (RVO), central retinal vein occlusion (CRVO), branch retinal vein occlusion (BRVO), corneal neovascularization, retinal neovascularization, retinopathy of prematurity (ROP), subconjunctival hemorrhage, and hypertensive retinopathy, optionally wherein the neovascular retinal disease or condition is wAMD or DR.

62. The method of claim 61, wherein the DR is non-proliferative diabetic retinopathy (NPDR).

63. The method of claim 49, wherein the anti-VEGF therapy comprises an anti- VEGF antibody or an anti-VEGF fusion protein.

64. The method of claim 49, wherein the anti-VEGF therapy comprises aflibercept, conbercept, ranibizumab, or bevacizumab.

65. The method of claim 49, wherein the anti-VEGF therapy comprises an anti- VEGF antibody conjugate.

66. The method of claim 65, wherein the anti-VEGF antibody conjugate comprises an anti-VEGF-A antibody and a phosphorylcholine containing polymer, wherein the polymer is covalently bonded to the anti-VEGF-A antibody at a non-native cysteine outside a variable region of the anti-VEGF-A antibody.

67. The method of claim 65, wherein the anti-VEGF antibody conjugate comprises an anti-VEGF-A antibody and a phosphorylcholine containing polymer, wherein the polymer is covalently bonded to the anti-VEGF-A antibody at a non-native cysteine outside a variable region of the anti-VEGF-A antibody, wherein the anti-VEGF-A antibody comprises a light chain and heavy chain, said heavy chain comprising an Fc region, wherein the cysteine is in the Fc region of the heavy chain, wherein the antibody conjugate has the following structure:; wherein: each heavy chain of the anti-VEGF-A antibody is denoted by the letter H, and each light chain of the anti-VEGF-A antibody is denoted by the letter L; the polymer is bonded to the anti-VEGF-A antibody through a sulfhydryl at C443 according to EU numbering, which bond is depicted on one of the heavy chains above; PC is, where the curvy line indicates the point of attachment to the rest of the polymer; 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%,optionally wherein the sequence of the heavy chain comprises SEQ ID NO: 1 or a sequence at least 90% identical thereto, and wherein the sequence of the light chain comprises SEQ ID NO: 2, or a sequence at least 90% identical thereto.

68. The method of claim 62, wherein the anti-VEGF-A antibody conjugate is tarcocimab tedromer.

69. The method of claim 62, wherein administering the one or more doses of the anti-VEGF therapy to the subject comprises administering to the subject a therapeutic formulation comprising: an unconjugated anti-VEGF-A antibody comprising the fusion protein with or without the non-native cysteine in the Fc region, wherein the unconjugated anti-VEGF-A antibody is not covalently attached to a phosphorylcholine-containing polymer; and the anti-VEGF-A antibody conjugate, wherein the unconjugated anti-VEGF-A antibody is present in the formulation in a range of 10-60% of a total molar amount of the anti-VEGF-A antibody conjugate and the unconjugated anti-VEGF-A antibody, wherein the total molar amount is the sum of the molar amount of the fusion protein conjugate and the molar amount of the unconjugated anti-VEGF- A antibody.

70. The method of claim 69, wherein the unconjugated anti-VEGF-A antibody is present in the therapeutic formulation at about 20% of the total molar amount of the anti- VEGF-A antibody conjugate and the unconjugated anti-VEGF-A antibody.

71. The method of claim 69, wherein the total amount of the anti-VEGF-A antibody conjugate (by weight of the antibody portion) and the unconjugated anti-VEGF-A antibody in each dose is 1-6 mg, optionally the total amount of the anti-VEGF-A antibody conjugate (by weight of the antibody portion) and the unconjugated anti-VEGF-A antibody in each dose is or is about 5 mg.

72. The method of claim 47, wherein the anti-VEGF therapy is administered intravitreally.

Images