Dendrimer conjugate compositions for intracellular delivery of antibodies and antibody fragments

Abstract

Dendrimer compositions and methods thereof for intracellular delivery of antibodies and antibody fragments are described for treatment of  ocular diseases, inflammatory disorders, and neurological disorders.

Description

 Important Note: The system was unable to import the complete document, including the compounds and tables. Therefore, these sections have been removed from the imported document. Please refer to the complete document attached under (Other Attachments) for the full content.DENDRIMER CONJUGATE COMPOSITIONS FOR INTRACELLULAR DELIVERY OF ANTIBODIES AND ANTIBODY FRAGMENTSCROSS REFERENCE TO RELATED APPLICATIONSThe application claims benefit of and priority to U.S. Provisional Application No. 63 / 625,737, filed January 26, 2024, which is hereby incorporated by reference in its entirety.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCHThis invention was made with government support under NS093416 and AG063831 awarded by the National Institutes of Health. The government has certain rights in the invention.REFERENCE TO SEQUENCE LISTINGThe Sequence Listing XML submitted as a file named “JHUC_17787_PCT_ST26.xml,” created on January ‎27, ‎2025, and having a size of 30,108 bytes is hereby incorporated by reference pursuant to 37 C.F.R. § 1.834(c)(1).FIELD OF THE INVENTIONThis invention is generally in the field of antibody drug formulations, specifically dendrimer-antibody conjugates for selective delivery to the intracellular targets in the CNS and other sites of disease.BACKGROUND OF THE INVENTIONThe delivery of exogeneous cargo into live cells is an important aspect of diagnostic and therapeutic agent drug delivery. Antibodies are highly specific for cell surface and extracellular targets. Antibodies exert high binding specificity to target moieties and low off target effects. The large size of antibodies facilitates the longer circulating time in the body and long-term activity. Antibodies provide superior selectivity, long term plasma pharmacokinetics (PK) and low toxicity.However, the major limitation for further expanding this therapeutic modality lies in the difficulty in achieving intracellular delivery. As estimated more than 20% of the proteome, the entire complement of proteins that is or can be expressed by a cell, tissue, or organism, are located inside the cells (www.proteinatlas.org / humanproteome / cell / cytosol) with limited accessibility of antibody therapeutics. These include oncogenic proteins, components of different signal transduction pathways, cell metabolism regulators and different enzymes. Compared to RNA based approaches such as siRNA, intracellular antibodies can inhibit the protein immediately, resulting in efficacy within hours instead of days as is the case for RNA or DNA based therapeutics. The major obstacle for intracellular antibody delivery is the cell membrane and the endosomal membrane upon receptor mediated internalization.Thus, it is the object of the present invention to provide a means for selective delivery of antibodies, or fusion proteins or fragments thereof, to the intracellular compartment of target cells, more specifically to the target cells at the site of pathology.It is a further object of the present invention to provide compositions and methods for delivery antibodies, or fusion proteins or fragments thereof, to their intracellular targets.It is also an object of the present invention to provide formulations with improved bioavailability, pharmacokinetics, and increased selectivity of delivery, and reduced side effects.SUMMARY OF THE INVENTIONComposition including dendrimers conjugated to at least one antibody or fragment thereof having an intracellular target are described. Typically, the antibody or fragment / fusion protein thereof is covalently conjugated by a covalent link between a modified or unmodified surface group or interior groups of the dendrimer, including and disulfide, ester, amide, or ether linkage. In some embodiments, the covalent link between the dendrimer and antibody or fragment / fusion protein thereof is not cleaved following administration prior to the delivery of the dendrimer-antibody conjugates to the target cells. In some embodiments, the dendrimer is a polyamidoamine (PAMAM) dendrimer of generation 1 – generation 9, with between greater than 40 and 100% of the surface groups being hydroxylated. In other embodiments, the dendrimer is a generation 2 – generation 7 PAMAM dendrimer, modified by a sugar moiety, wherein the sugar group is selected from the group consisting of glucose, galactose, mannose, and fructose. In further embodiments, the dendrimer is a dendritic polymer, optionally a hyperbranched polymer. Preferably, the dendrimer is a PAMAM dendrimer of generation 2 – generation 7, with greater than 50% of the functional groups in the form of hydroxyl (OH) groups. In further preferred embodiments, the dendrimer is a glucose dendrimer, with a central core of dipentaerythritol and one or more branching units of monosaccharide glucose and ethylene glycol, with >10 surface glucose moieties, for example, the dendrimer is a glucose dendrimer of generation 1, 2, or 3. In some embodiments, the antibody or fragment thereof is a humanized or chimeric antibody, or a fragment, variant, or fusion protein thereof such asa single chain antibody, single chain variable fragment (scFv), disulfide linked Fvs (sdFv), Fab, Fab', F(ab')2, Fv, and single domain antibody fragment (sdAb). In one embodiment, the antibody or fragment thereof is a fusion protein of extracellular components of VEGF receptor (VEGFR)-1 and VEGFR-2 fused to the Fc portion of IgG such as in the case of aflibercept. In further embodiments, the antibody or fragment thereof is adalimumab, alemtuzumab, cetuximab, daclizumab, infliximab, etanercept, efalizumab natalizumab, rituximab, ranibizumab, bevacizumab, palivizumab, trastuzumab, natalizumab, omalizumab, canakinumab, ustekinumab, tocilizumab, daclizumab, basiliximab, muromonab-cd3, eculizumab, certolizumab pegol, golimumab, Ibritumomab tiuxetan, gemtuzumab ozogamicin, tositumomab-l131, panitumumab, ofatumumab, motavizumab, ga101, and aflibercept. The antibody selectively binds an intracellular target. In some embodiments, the antibody or fragment thereof binds specifically to one or more of Tau antibody, CTA-4. PD-1, PD-L1, CD20. In other embodiments, the antibody or fragment thereof binds specifically to one or more proteins involved in NLRP3 inflammasome activation including NLRP3, Caspases, RIP1-3, FADD, MLKL, Interleukins and pro-interleukins. In further embodiments, the antibody or fragment thereof binds specifically to one or more intracellular inflammatory signaling proteins of JAK-STAT, Nrf2, STAT 1, STAT 3, STAT 6, MAPK, MEK, ERK1 / 2, and HIF1a. In one embodiment, the antibody or fragment thereof binds specifically to TRIM21 intracellularly. In some cases, the composition further comprises one or more therapeutic or diagnostic agents.Pharmaceutical formulations of the composition of dendrimers conjugated to at least one antibody or fragment thereof, and one or more pharmaceutically acceptable carrier or excipients, are also described. In some embodiments, the formulation is formulated for systemic or local administration. In some embodiments, the formulation is formulated for parenteral or enteral administration, for example, for intramuscular, intraperitoneal, intravenous, or subcutaneous injection, or for intravitreal administration. Methods of administering the dendrimer-antibody conjugates to a subject in need thereof are also provided. The compositions can be administered to the subject via intranasal, intravenous, oral, subcutaneous, sublingual, intramuscular, topical, intraperitoneal, intrathecal, subchoroidal, intracranial, or intravitreal administration. In some embodiments, the subject in need thereof has one or more of ocular diseases, inflammatory disorders, neurological disorders, and cancer, such as arthritis, diabetic nephropathy, renal interstitial disease, fibrosis, multiple sclerosis, irritable bowel syndrome, Crohn’s disease or other autoimmune disorder. In some embodiments, the subject in need thereof has one or more diseases or disorders of the eye such as diabetic eye disease, symptomatic vitreomacular adhesion / vitreomacular traction (sVMA / VMT), wet (neovascular) age-related macular degeneration (AMD), and dry AMD; or one or more retinal and choroidal vascular diseases such as consisting of AMD, retinopathy of prematurity, diabetic macular edema, retinal vein occlusion, retinopathy associated with toxicity of chemotherapy. Generally, in the case of ocular diseases, the dendrimer-antibody conjugates are administered in an amount effective to reduce and / or inhibit the number or activities of activated microglia and macrophages in the retina and / or the choroid in the eye of a subject in need thereof. In other embodiments, the dendrimer-antibody conjugates are effective to reduce and / or inhibit the expression and / or activities of VEGF and / or VEGFR in the activated microglia, activated macrophages, and / or retinal pigment epithelial (RPE) cells in the diseased retina and / or choroid. In preferred embodiments, the dendrimer-antibody conjugates are administered in a dosage effective to decrease expression of one or more pro-inflammatory cytokines such as TNF-α, interleukin-1β (IL-1β), and interferon-γ (IFN-γ). In further preferred embodiments, the dendrimer-antibody conjugates are in an amount effective to reduce and / or inhibit abnormal vascular permeability and leakage, and / or neoangiogenesis in the eye of a subject in need thereof. In cases where the subject in need thereof has one or more neurological disorders, the dendrimer-antibody conjugates are administered in an amount effective to target one or more types neurons associated with the site of pathology. In some embodiments, the subject in need thereof has Huntington disease, synucleinopathies, Alzheimer’s disease, prion disease, or amyotrophic lateral sclerosis (ALS). Typically, the composition or formulation is administered once or twice every month, every two months, every three months, every four months, every five months, or every six months, or less frequently. The antibody or fragment thereof is selectively delivered to one or more reactive immune cells such as reactive microglia, macrophages, astrocytes, retinal pigmental epithelial cells in a disease or disorder-affected region of a subject in need thereof by virtue of the selective uptake of the dendrimer. In one preferred embodiment hydroxyl-terminated PAMAM dendrimers are conjugated to the antibody or antibody fragment, which is effective to selectively deliver the antibody or antibody fragment to the intracellular compartment of the one or more reactive immune cells. In preferred embodiments, the antibody or fragment thereof is retained within the one or more reactive immune cells for over 1, 2, 3, 4, 5, 6, or 7 days. Examples demonstrate delivery of aflibercept.and HUMIRA®.Selection of the dendrimer is used to target the antibody or fragment thereof to one or more neurons or reactive immune cells such as reactive microglia, macrophages, astrocytes, retinal pigmental epithelial cells in a disease or disorder-affected region of a subject in need thereof. Preferably, the dendrimer-antibody conjugates include glucose dendrimers, and the antibody or fragment thereof conjugated thereto binds to a target in an intracellular compartment of the one or more neurons and / or reactive immune cells. In preferred embodiments, the antibody or fragment thereof is retained within the neurons and / or reactive immune cells for over 1, 2, 3, 4, 5, 6, or 7 days.BRIEF DESCRIPTION OF THE DRAWINGSFIGs. 1A-1D are reaction schemes showing stepwise synthesis of G1-glucose (FIG. 1A), stepwise synthesis of G2-glucose (FIG. 1B), synthesis of functionalized Cy5-D-PEG4-TCO (FIG. 1C), and synthesis of Cy5-G2-Glucose-antibody conjugate (FIG. 1D). FIG. 2 is a schematic representation showing hydroxyl PAMAM dendrimers, glucose dendrimers, or glucose-functionalized PAMAM dendrimers, with ligand functionalization, that are conjugated to antibodies and antibody-like molecules. FIG. 3 is a synthesis scheme of functionalized Cy5-D-PEG4-TCO. The hydroxyl PAMAM dendrimer generation 6 (PAMAM-G6-OH) was treated with 4-tert-butoxycarbonylamino)butyric acid (Boc-protected GABA) linker, 2 and the resulted product, 3 was deprotected using dichloromethane (DCM) / trifluoroacetic acid (TFA) (4:1). The product, 4 was labeled with Cy5 fluorophore using Cy5 N-hydroxysuccinimide (NHS) ester and the resulted intermediate, 5 was conjugated with trans-cyclooctene (TCO) linker, PEG4-TCO to obtain functionalized Cy5-D-PEG4-TCO 6. The subscripted numbers in the formulas indicate the number of GABA BOC, PEG4-TCO, or flurophore attached per dendrimer.FIG. 4A-4D are charecterization of functionalized Cy5-D-PEG4-TCO and synthetic intermediates.1H NMR (DMSO-d6, 500 MHz) characterization of dendrimer conjugates, D-GABA-Boc, D-GABA-NH2, Cy5-D, Cy5-D-PEG4-TCO (in DMSO-d6 and D2O) showing the appearance or disaapreance of characteristic signals (FIG. 4A). (C) MALDI-TOF spectra of PAMAM-G6-OH (FIG. 4B), Cy5-D (FIG. 4C), and Cy5-D-PEG4-TCO (FIG. 4D). The degree of conjugation in each step of the synthesis was calculated based on the 1H-NMR and change of molecular weights measured by MALDI-TOF / MS.FIG. 5 is a synthesis scheme of Cy5-D-aflibercept conjugate. Modification of aflibercept fusion protein for conjugation with dendrimer. Aflibercept was first substituted with PEGylated tetrazine using NHS-PEG2-Tz reagent to form aflibercept-Tz (Compound 7), then Compound 7 was reacted with Cy5-D-PEG4-TCO (Compound 6) to obtain the final product Cy5-D-aflibercept, Compound 9.FIGs. 6A-6B are MALDI-TOF spectra showing peak at mass of 111449 Da for free aflibercept (FIG. 6A) and peak at 180319 Da for Cy5-D-aflibercept (FIG. 6B).FIG. 7 is a bar graph showing quantification of CNV area at day 7 and day 14 mice treated with PBS, aflibercept, and D-aflibercept based on micrographs of CNV lesions (isolectin green) in the RPE-choroid flat mounts. **p <0.01 (n=5).FIGs. 8A-8C are bar graphs showing relative expression level of VEGF (FIG. 8A), IL-1 (FIG. 8B), and TGF (FIG. 8C) determined using RT-qPCR in choroid tissues on day 7 and 14 after intravitreal administration of D-aflibercept and aflibercept 40 µg / µL (1 dose at day 0). p˂0.05, n=6 eyes per group, One-way ANOVA.FIGs.9A-9C are graphs showing the dose response curve showing binding towards VEGFA (λ450nm) over Log[concentration] (nm) of free aflibercept and D-aflibercept (FIG. 9A), standard curve showing optical density over a range of concentrations of human VEGF (pg / mL) in ELISA culture supernatant (FIG. 9B), and bar graph showing free VEGF quantification (optical density) in culture supernatant treated with aflibercept, D-aflibercept, or control (FIG. 9C). ARPE-19 cells were incubated 16 h in 1% FBS containing cell culture media and treated with 10 ng / µL of aflibercept and D-aflibercept, as protocols VEGF quantification was determined following manufacture’s protocols after 24 h treatment for D-aflibercept (208±), aflibercept (97±) and control samples (879±) pg / mL.FIG. 10A and 10B are fundus fluorescein angiography images and quantification of fluorescein leakage at Day 7 (10A) and Day 14 (10B) after laser-induced CNV. Intravitreal injection of PBS, Free-Eylea or D-Eylea was performed one-day post laser CNV. n=5-7 (n represents average fluorescence intensity of 3 burns from a single eye).FIG. 11A-11C: Cellular uptake of Cy5-D-Adalimumab. Representative confocal microscopy images of Cy5-D-Adalimumab cell uptake intoBV-2 cells after 24 h treatment, cell nucleus (Blue), dendrimer (Red) (FIG. 11A); Percent Cy5 positive cells in Cy5-D-Adalimumab treated cells compared to BV-2 cells using FACS study (FIG. 11B); merged FIG.11A and FIG. 11B (FIG. 11C).FIG. 12A-12D are confocal microscopy images and graphs of Cy5-D-Adalimumab cell uptake intoBV-2 cells after 24 h treatment, cell nucleus (Blue), dendrimer (Red). FIG. 12A and 12B show no treatment and FIG. 12C and 12D shows percent Cy5 positive cells in Cy5-D-Adalimumab treated cells compared to BV-2 cells as determined by FACS.FIG. 13A-13C and 13D-13F are graphs of the D-Adalimumab and free Adalimumab binding affinities to key targets in the Tumor necrosis factor alpha (TNF-α), quantification of TNF-α levels with LPS or without LPS in THP-1 cells, THP-1 monocytes and THP-1 derived macrophages with LPS (100 ng / mL) / without LPS treated with varying concentrations of D-Adalimumab and Adalimumab.FIG. 14 is a bar graph showing results from trial 2 suppression of laser-Iinduced CNV formation. CNV lesions z in the RPE-choroid flat mounts at post laser day 7 and day 14 from mice treated with aflibercept, D-aflibercept, and saline, respectively, were imaged. Shown are quantification from micrographs of CNV area day 7 and (left) and day 14 (right) mice treated with aflibercept, D-aflibercept, and saline (n=5).FIG. 15A and FIG. 15B are bar graphs showing Fundus fluorescein angiography images and quantification of fluorescein leakage from fundus fluorescein angiography images at D7 (FIG. 15A) and D14 (FIG. 15B) after laser-induced CNV. Intravitreal injection of PBS, Free-aflibercept, or D-aflibercept was performed one day post laser CNV. n=5-7 (n represents the average fluorescence intensity of 3 burns from a single eye). FIG. 16 is a schematic representation of engineered dendrimer nanoparticles for targeted intracellular delivery of antibody in AMD model.FIG. 17A and FIG. 17B depictingsize measurements using DLS of the hydrodynamic diameter of aflibercept is 9.0±0.7 nm, Cas9 9.5±1.1 nm, and D-aflibercept 12.2±0.7 nm.FIGs. 18A-18D show in vitro characterization of aflibercept and D-aflibercept using LPS-stimulated HMC3 cells. FIG. 18A shows extracellular VEGFA reduction by ELISA. FIG. 18B shows intracellular VEGFA reduction by ELISA (VEGF quantification was determined following manufacture protocols). FIGs. 18C and 18D show Cy5-aflibercept and Cy5-D-aflibercept cell internalization by FACS.FIGs. 19A-19Dshow in vitro characterization of aflibercept and D-aflibercept using LPS-stimulated ARPE-19 cells. FIG. 19A show extracellular VEGFA reduction by ELISA. FIG. 19B show intracellular VEGFA reduction by ELISA (VEGF quantification was determined following manufacture protocols ). FIGs. 19C and 19D show Cy5-aflibercept and Cy5-D-aflibercept cell internalization by FACS.FIG. 20 is a schematic of a synthesis scheme for functionalized Cy5-D-PEG4-TCO.FIG. 21 is a synthesis scheme for Cy5-D-Adalimumab conjugate.FIG. 22 shows cellular uptake of Cy5-D-Adalimumab quantification of percent Cy5 positive cells in Cy5-D-Adalimumab treated cells compared to BV-2 cells using FACS.FIGs. 23A-23E show targeting delivery of HD-ADA in THP-1 monocytes.FIG. 23A is an experimental timeline. FIGs. 23B and 23C are bar graphs showing percentage of Cy5+ cells and MFI from in vitro studies after treating activated or resting cells with HD-Cy5 and HD-ADA-Cy5 (0, 0.1 and 1 ug / ml), respectively. FIGs. 23D and 23F are representative data from non-treatment and HD-ADA-Cy5 (1 µg / ml) groups, respectively, were plotted as FSC-H vs Cy5 and single parameter histograms. Data in this figure correspond to FACs studies and are shown as mean ± SEM.FIGs. 24A-24C show efficacy studies of HD-ADA in THP-1 monocytes. FIG. 24A is an experimental timeline. FIG. 24B shows TNF-α levels (pg / ml) observed in cell lysates (intracellular protein content) and FIG. 24C shows supernatants (extracellular protein content). Data in this figure correspond to in vitro studies after treating activated or resting cells with ADA and HD-ADA (0, 0.1 and 1 ug / ml) and are shown as mean ± SEM.FIGs. 25A-25C shows efficacy studies of Adalimumab and HD-Adalimumab in HMC3 cells. FIG. 25A is an experimental timeline. FIG. 25B and 25C shows TNF-α levels (pg / mL) observed in supernatants (extracellular protein) and cell lysates (intracellular protein) following treatment of activated or resting cells with Adalimumab and HD-Adalimumab (0.1, 1, and 10 µg / mL). Data are presented as mean ± SEM.FIGs. 26A and 26B show cellular internalization of Cy5-Adalimumab and Cy5-D-Adalimumab at 4 hours and 24 hours, evaluated by FACS and confocal microscopy at 20 µg / mL in HMC3 cell lines.DETAILED DESCRIPTION OF THE INVENTIONI. DefinitionsThe term “antibody” refers to natural or synthetic antibodies that bind a target antigen. The term includes polyclonal and monoclonal antibodies. In addition to intact immunoglobulin molecules, also included in the term “antibodies” are fragments or polymers of those immunoglobulin molecules, and human or humanized versions of immunoglobulin molecules that bind the target antigen.The term “specifically binds” refers to the binding of an antibody to its cognate antigen (for example DNA) while not significantly binding to other antigens. Preferably, an antibody “specifically binds” to an antigen with an affinity constant (Ka) greater than about 105 mol–1 (e.g., 106 mol–1, 107 mol–1, 108 mol–1, 109 mol–1, 1010 mol–1, 1011 mol–1, and 1012 mol–1 or more) with that second molecule.The term “monoclonal antibody” or “MAb” refers to an antibody obtained from a substantially homogeneous population of antibodies, i.e., the individual antibodies within the population are identical except for possible naturally occurring mutations that may be present in a small subset of the antibody molecules.The term "therapeutic agent" refers to an agent that can be administered to treat one or more symptoms of a disease or disorder. The term “diagnostic agent” generally refers to an agent that can be administered to reveal, pinpoint, and define the localization of a pathological process. The diagnostic agents can label target cells that allow subsequent detection or imaging of these labeled target cells. The term "therapeutically effective amount" refers to an amount of the therapeutic agent that, when incorporated into and / or onto dendrimers, produces some desired effect at a reasonable benefit / risk ratio applicable to any medical treatment. The effective amount may vary depending on such factors as the disease or condition being treated, the particular targeted construct being administered, the size of the subject, or the severity of the disease or condition. One of ordinary skill in the art may empirically determine the effective amount of a particular compound without necessitating undue experimentation. In some embodiments, the term “effective amount” refers to an amount of a therapeutic agent or prophylactic agent to reduce or diminish the symptoms of one or more diseases. The terms “inhibit” or “reduce” in the context of inhibition, mean to reduce, or decrease in activity and quantity. This can be a complete inhibition or reduction in activity or quantity, or a partial inhibition or reduction. Inhibition or reduction can be compared to a control or to a standard level. Inhibition can be 5, 10, 25, 50, 75, 80, 85, 90, 95, 99, or 100%. For example, dendrimer compositions including one or more inhibitors may inhibit or reduce the activity and / or quantity of diseased neurons by about 10%, 20%, 30%, 40%, 50%, 75%, 85%, 90%, 95%, or 99% from the activity and / or quantity of the same cells in equivalent tissues of subjects that did not receive, or were not treated with the dendrimer compositions. In some embodiments, the inhibition and reduction are compared at levels of mRNAs, proteins, cells, tissues, and organs. For example, an inhibition and reduction in the rate of neural loss, in the rate of decrease of brain weight, or in the rate of decrease of hippocampal volume, as compared to an untreated control subject. The term “treating” or “preventing” mean to ameliorate, reduce or otherwise stop a disease, disorder or condition from occurring or progressing in an animal which may be predisposed to the disease, disorder and / or condition but has not yet been diagnosed as having it; inhibiting the disease, disorder or condition, e.g., impeding its progress; and relieving the disease, disorder, or condition, e.g., causing regression of the disease, disorder and / or condition. Treating the disease or condition includes ameliorating at least one symptom of the particular disease or condition, even if the underlying pathophysiology is not affected, such as treating the pain of a subject by administration of an analgesic agent even though such agent does not treat the cause of the pain. Desirable effects of treatment include decreasing the rate of disease progression, ameliorating, or palliating the disease state, and remission or improved prognosis. For example, an individual is successfully “treated” if one or more symptoms associated with cancer are mitigated or eliminated, including, but are not limited to, reducing and / or inhibiting rate of tumor cell proliferation / growth, increasing the quality of life of those suffering from the disease, decreasing the dose of other medications required to treat the disease, delaying the progression of the disease, and / or prolonging survival of individuals.The phrase "pharmaceutically acceptable" or “biocompatible” refers to compositions, polymers, and other materials and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio. The phrase "pharmaceutically acceptable carrier" refers to pharmaceutically acceptable materials, compositions, or vehicles, such as a liquid or solid filler, diluent, solvent, or encapsulating material involved in carrying or transporting any subject composition, from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of a subject composition and not injurious to the patient.The term “biodegradable” generally refers to a material that will degrade or erode under physiologic conditions to smaller units or chemical species that are capable of being metabolized, eliminated, or excreted in vivo. The degradation time is a function of composition and morphology. The term "dendrimer" includes, but is not limited to, a molecular architecture with an interior core, interior layers, or "generations" of repeating units regularly attached to this initiator core, and an exterior surface of terminal groups attached to the outermost generation.The term “functionalize” means to modify a compound or molecule in a manner that results in the attachment of a functional group or moiety. For example, a molecule may be functionalized by the introduction of a molecule that makes the molecule a strong nucleophile or strong electrophile. The term "targeting moiety" refers to a moiety that localizes to or away from a specific location. The moiety may be, for example, a protein, nucleic acid, nucleic acid analog, carbohydrate, or small molecule. The location may be a tissue, a particular cell type, a subcellular compartment, or a molecule such as a receptor. The term "prolonged residence time" refers to an increase in the time required for an agent to be cleared from a patient's body, or organ or tissue of that patient. In certain embodiments, "prolonged residence time" refers to an agent that is cleared with a half-life that is 10%, 20%, 50% or 75% longer than a standard of comparison such as a comparable agent without conjugation to a delivery vehicle such as a dendrimer. In certain embodiments, "prolonged residence time" refers to an agent that is cleared with a half-life of 2, 5, 10, 20, 50, 100, 200, 500, 1000, 2000, 5000, or 10000 times longer than a standard of comparison such as a comparable agent without a dendrimer that specifically target specific cell types.The terms "incorporated" and "encapsulated" refer to incorporating, formulating, or otherwise including an agent into and / or onto a composition that allows for release, such as sustained release, of such agent in the desired application. The agent or other material can be incorporated into a dendrimer, by binding to one or more surface functional groups of such dendrimer (by covalent, ionic, or other binding interaction), by physical admixture, by enveloping the agent within the dendritic structure, and / or by encapsulating the agent inside the dendritic structure.The term “central nervous system” (“CNS”) includes the brain and spinal cord. As used herein, peripheral nervous system (“PNS”) refers to the nerves other than in the brain and spinal cord.II.CompositionsDendrimer conjugates of antibodies or fragments / fusion proteins thereof which are selectively delivered to one or more target cells by virtue of the dendrimer composition have been developed. Generally, dendrimer-antibody conjugates provide selective delivery of antibodies, or fusion proteins or fragments thereof, to the intracellular compartment of target cells, more specifically to the target cells at the site of pathology. In preferred embodiments, the antibody and / or antibody fragment thereof specifically binds one or more intracellular targets within the target cells. Exemplary target cells include, but not limited to, brain cells such as microglia, astrocytes, and / or neurons, for example, those within the site of pathology in the brain or the CNS; and / or cells in the peripheral nervous system, such as peripheral neurons and macrophages. The microglia and / or astrocytes to which the antibody and / or antigen-binding fragment thereof is delivered may be activated or inactive microglia and / or astrocytes.In some embodiments, when the antibody and / or fragment / fusion protein thereof binds to the target, the antibody and / or fragment / fusion protein thereof remains conjugated to the dendrimer. In these embodiments, following binding, the antibody and / or antigen-binding fragment thereof may be released from the dendrimer or remain conjugated to the dendrimer as an intact dendrimer-antibody conjugate. In some embodiments, the antibody and / or fragment / fusion protein thereof is released from the dendrimer after entry into the intracellular compartment of the target cell.A.DendrimersIn some embodiments, dendrimers are prepared using methods in which the dendrimer is assembled from a multifunctional core, which is extended outward by a series of reactions. A multifunctional core moiety allows stepwise addition of branching units (i.e., generations) around the core. Exemplary chemical structures suitable for use as core moieties are shown in Table 1 below. Additional examples of structures suitable for use as core moieties include monosaccharide, disaccharides, trisaccharides, oligosaccharides, or azide-, alkyne-modified moieties thereof. In some embodiments, the core moiety is chitosan. Thus, azide-modified chitosan, or alkyne-modified chitosan are suitable for conjugating to branching units using click chemistry. In some embodiments, the core moiety is ethylenediamine, or tetra(ethylene oxide). In some embodiments, the core moiety is dipentaerythritol. Table 1. Structural representation of various building blocks (cores, branching units, surface functional groups, monomers) for the synthesis of dendrimers.  2.Branching Units The dendrimers generally have one or more inner repeating branched arms or branching units, formed of amides, hydroxyl, and amine groups known as generations attached radially to the core. The chemical structures listed in Table 1, are also suitable as building blocks to form the branching units of the dendrimer. For example, the branching units of the dendrimers can be formed of dipentaerythritol, monosaccharide, disaccharides, trisaccharides, oligosaccharides, or azide- , alkyne-modified moieties thereof, or a combination thereof. Other examples of chemical structures that are suitable for forming the branching units of the dendrimers disclosed herein include, but are not limited to, sugar moieties, such as glucose, galactose, mannose, and fructose, and alkylene glycol, such as ethylene glycol, and combinations thereof. In some embodiments, the branching unit is chitosan. Thus, azide- modified chitosan, or alkyne-modified chitosan are suitable for conjugating to the core moiety or additional same or different branching units using click chemistry. In some embodiments, the branching unit is methyl acrylate or ethylenediamine, or a combination thereof. In some embodiments, the branching unit is polyethylene glycerol linear or branched. In some embodiments, the branching unit is a copolymer of an alkylene glycol (such as ethylene glycol) and a sugar moiety, such as glucose, galactose, mannose, and / or fructose.In some embodiments, the branching units are hyper-monomers i.e., ABn building blocks. Exemplary hyper-monomers include AB4, AB5, AB6, AB7, AB8 building blocks. Hyper-monomer strategy drastically increases the number of available end groups. 3.Surface Functional Groups The dendrimers typically have a plurality of surface functional groups or molecules. The terms “surface functional groups” and “terminal groups” are used interchangeably herein. Exemplary functional groups include primary amine end groups, hydroxyl end groups, carboxylic acid end groups, acetamide end groups, sugar molecules, oligo- or poly-alkylene glycols, and thiol end groups. In some embodiments, the desired terminal functional groups may be introduced through conjugation methods applied to the core and branching units.The surface functional groups in the dendrimers can include hydroxyl groups, such as those found in PAMAM dendrimers, generation 2 PEG dendrimers (e.g., see Structure I), or terminal glucose units in dendrimers with glucose-based branching units (e.g., see Structures III and IV). These functional groups may also be modified or added through various conjugation techniques. Alternatively, the dendrimers can be formed of glucose and oligoethylene glycol building blocks. Structures III and IV depict exemplary generation 1 and generation 2 glucose dendrimers, respectively. In these forms, the dendrimers can have one or more surface functional groups, e.g., hydroxyl (-OH), amine, acetamide, or carboxyl groups, located on the dendrimer periphery. The surface density of the peripheral functional groups is at least functional group per square nanometer (nm²). For example, the surface density of surface functional groups, e.g. hydroxyl groups, carbohydrate group, etc., may exceed 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 groups per nm². In some embodiments, the surface density of the surface functional groups e.g., hydroxyl groups may be in a range from between about 1 and about 50 or more groups per nm². For example, volumetric density values for surface functional groups, such as hydroxyl groups, can range from about 1 to about 10 groups per nm³, from about 4 to about 50 groups per nm³, from about 5 and about 30 groups per nm³, or from about 10 and about 20 groups per nm³, preferably, from about 5 to about 20 groups per nm², while having a molecular weight of between about 500 Da and about 10 kDa.In some embodiments, the quantity of surface functional groups, such as hydroxyl groups, is at least about 40%, about 50%, or more of the dendrimer. In some embodiments, the surface functional groups can be further modified by conjugating at least 40% of the dendrimer with carbohydrate molecules and / or polyalkylene glycols, such as polyethylene glycols. In these embodiments, the surface density of terminal carbohydrate moieties or polyalkylene glycols can have the same ranges as described above for hydroxyl groups. 1 The dendrimers include an effective number of terminal groups e.g., hydroxyl groups and / or amine groups for targeting specific tissues and / or cell types such as the CNS cells and PNS cells as generally described above. For example, the dendrimer can specifically target a particular tissue region and / or cell type without a targeting moiety and the active agent conjugated thereto bind directly to a receptor on the surface and / or interior of target neural and / or glial cells. For delivery to the brain, hydroxyl PAMAM dendrimers, PAMAM dendrimers with surface modifications incorporating sugar moieties (where over 10% of the surface groups are modified with sugars, preferably glucose), and glucose dendrimers made from glucose building blocks are preferred. Constructs with a total molecular weight of less than 30,000 Da are preferred for brain delivery, whereas constructs exceeding 50,000 Da are preferable for confinement to peripheral circulation.When dendrimers incorporate sugar moieties at their termini, such as glucose, the terminal hydroxyl groups may either be part of the sugar moieties, represent additional hydroxyl groups not included in the sugar moieties, or a combination of both. In some embodiments, all terminal hydroxyl groups are derived from the terminal sugar moieties.a.Hydroxyl-terminated Dendrimers 2 In some embodiments, dendrimers include a plurality of hydroxyl groups. Examples of dendrimers containing high-density hydroxyl groups include commercially available polyester dendritic polymers such as hyperbranched 2,2-Bis(hydroxyl-methyl)propionic acid polyester polymers such as hyperbranched bis-MPA polyester with 64 hydroxyl groups (generation 4) and dendritic polyglycerols.3 In some embodiments, dendrimers with high-density hydroxyl groups contain oligoethylene glycol (OEG). For example, a generation 2 OEG dendrimer (D2-OH-60) can be synthesized using highly efficient and robust chemical reactions like Cu(I)-catalyzed alkyne–azide click chemistry and photo-catalyzed thiol-ene click chemistry. These methods, employing orthogonal hypermonomer and hypercore strategies as described in for example WO 2019 / 094952, facilitate the creation of dense polyol dendrimers at low generation levels with minimal reaction steps. In some embodiments, the dendrimer backbone has non-cleavable polyether bonds, preventing in vivo disintegration and enabling elimination as single, non-biodegradable entities. 4Structure I. A generation two (G2) oligo ethylene glycol-like dendrimerThe range for the surface density of hydroxyl (-OH) on the periphery of the dendrimer is generally described above. The amount of the surface hydroxyl groups on the periphery of the dendrimer is preferably greater than 35%, at least 40%, at least 50%, more than 40%, more than 50%, or in a range from more than 40% to 100%. In some embodiments, the dendrimers may have a fraction of the hydroxyl groups exposed on the outer surface, with the others in the interior core of the dendrimers. b.Dendrimers Modified with Carbohydrates In some embodiments, the dendrimers contain one or more carbohydrate molecules at the termination. These terminal carbohydrate molecules can be prepared by conjugating one or more surface functional groups of dendrimers, such as amine groups, carboxyl groups, or hydroxyl groups, with one or more carbohydrate molecules. In preferred embodiments, the dendrimers, prior to carbohydrate conjugation, are hydroxyl-terminated dendrimers such as hydroxyl-terminated PAMAM dendrimers and one or more of the hydroxyl groups are conjugated with one or more carbohydrate molecules. In some embodiments, the carbohydrate moieties used to modify one or more surface functional groups of the dendrimers are monosaccharides. Exemplary monosaccharides suitable for modifying the dendrimers include glucose, glucosamine, galactose, mannose, fructose, dehydroascorbic acid, urate, myo-inositol. In some embodiments, the dendrimers are conjugated to glucose and thus contain glucose as terminal moieties / molecules. In some embodiments, hydroxyl-terminated dendrimers are modified with one or more glucose moieties to the dendrimer (“D-Glu”). In some embodiments, the dendrimers are conjugated to one or more monosaccharides other than glucose, such as galactose, mannose, and / or fructose. For example, the carbohydrate moieties are oligosaccharides which terminate in one or more monosaccharides including glucose, glucosamine, mannose, fructose, thus, exposing these sugar moieties on the surface.In some embodiments, the dendrimers are conjugated to glucose and / or glucosamine moieties. For example, the dendrimer can be a glucose dendrimer. Glucose dendrimers or glucose-modified dendrimers are capable of achieving selective uptake by target cells. In some embodiments, the dendrimers are conjugated to one or more carbohydrate moieties that exhibit affinity for and suitability in binding specific transporters. These carbohydrate moieties, or sugar-based components, can facilitate transport via various glucose transporters, such as GLUT1, GLUT2, GLUT3, GLUT4, GLUT5, GLUT6, GLUT7, GLUT8, GLUT9, GLUT10, GLUT11, GLUT12, GLUT13, and GLUT14. Alternatively, the dendrimers are conjugated to one or more oligosaccharides terminating in glucose and / or glucosamine, i.e., glucose and / or glucosamine moieties are exposed on the surface of the dendrimer conjugates suitable for binding to one or more of the GLUTs. The range for the surface density of carbohydrate moieties / molecules such as monosaccharides, e.g., glucose, on the periphery of the dendrimers, is same as generally described above. In preferred embodiments, the surface density of carbohydrate molecules is between about 1 and about 50, between about 5 and about 20, per nm2, with each carbohydrate moiety having a molecular weight of between about 100 Da and about 1000 Da. In these embodiments, i.e., one or more surface functional groups of the dendrimer are modified to introduce one or more sugar moieties / molecules at termination, the terminal hydroxyl groups may be part of the terminal sugar moieties / molecules or extra hydroxyl groups that are not modified with sugar moieties / molecules and thus are not part of the sugar moieties / molecules, or a combination thereof.In some embodiments, carbohydrate molecules such as monosaccharides, e.g., glucose, are present in an amount by weight that is between about 1% and 40% of the total weight of the glycosylated dendrimer, for example, between about 2% and 20%, between about 5% and 15%, or between 9 % and 12 % of the total weight of the glycosylated dendrimer. For example, in some embodiments, the carbohydrate moieties are present in an amount that is about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% of the total weight of the glycosylated dendrimer following conjugation. In some embodiments, conjugation of carbohydrate molecules through one or more surface functional groups occurs via about 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, or 25% of the total available surface functional groups, preferably hydroxyl groups, of the dendrimers prior to the conjugation. In other embodiments, the conjugation of carbohydrate molecules occurs on less than 5%, less than 10%, less than 15%, less than 20%, less than 25%, less than 30%, less than 35%, less than 40% of total available surface functional groups of the dendrimers prior to the conjugation.c.Dendrimers Modified with Polyalkylene Glycol In some embodiments, the dendrimers contain one or more polyalkylene glycols at the termination. These terminal polyalkylene glycols can be prepared by conjugating one or more of surface functional groups of the dendrimers, such as hydroxyl groups, with a polyalkylene glycol, such as PEG. In some embodiments, the dendrimers, prior to conjugation, are hydroxyl-terminated dendrimers such as hydroxyl-terminated PAMAM dendrimers and at least a portion of the surface hydroxyl groups are conjugated with PEG. The range for the surface density of polyalkylene glycols, such as PEG, on the periphery of the dendrimers, is the same as generally described for surface groups above. In preferred embodiments, the surface density of polyalkylene glycols such as PEG is between about 1 and about 50, between about 5 and about 20, per nm2 (number of surface polyalkylene glycols / surface area in nm2) while having a molecular weight of between about 100 Da and about 10 kDa. In some embodiments, the polyalkylene glycol molecules such as PEG can be present in an amount by weight that is between about 1% and 40% of the total weight of the pegylated dendrimer, for example, between about 2% and 20%, between about 5% and 15%, or between 9 % and 12 % of the total weight of the pegylated dendrimer. For example, in some embodiments, the polyalkylene glycol molecules, such as PEG, are present in an amount that is about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% of the total weight of the pegylated dendrimer following conjugation. In some embodiments, conjugation of polyalkylene glycol molecules such as PEG through one or more surface functional groups of the dendrimer occurs via about 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, or 25% of the total available surface functional groups, preferably hydroxyl groups, of the dendrimers prior to the conjugation. In other embodiments, the conjugation of polyalkylene glycol molecules such as PEG occurs on less than 5%, less than 10%, less than 15%, less than 20%, less than 25%, less than 30%, less than 35%, less than 40% of total available surface functional groups of the dendrimers prior to the conjugation.4.Exemplary Dendrimers a. Monosaccharide-based DendrimersDendrimer compositions that can selectively accumulate inside neurons, particularly in the nucleus of injured and / or hyperactive neurons, referred to as “glucose dendrimers” have been developed. These dendrimers can also accumulate at a high level inside activated microglia. However, compared to hydroxyl dendrimers which primarily accumulate in microglia, these dendrimers primarily go to neurons. Glucose dendrimers are described in U.S.S.N. 63 / 327,610 “Dendrimer Compositions for Targeted Delivery of Therapeutics to Neurons” by The Johns Hopkins University, inventors Kannan Rangaramanujam, Rishi Sharma, Anjali Sharma, Sujatha Kannan, Nirnath Sah, Mira Sachdeva, and Siva P. Kambhampati filed April 5, 2022.Glucose dendrimers generally contain (a) a central core, (b) one or more branching units, wherein the branching units are monosaccharide glucose-based branching units, optionally with a linker conjugated thereto; and optionally (c) one or more therapeutic, prophylactic and / or diagnostic agents. In some embodiments, the monosaccharide dendrimers have a hypercore (e.g., dipentaerythritol) and one or more monosaccharide branching units. In some embodiments, the hypercore is dipentaerythritol and the monosaccharide branching unit is glucose-based branching unit. In further embodiments, spacer molecules can also be alkyl (CH2)n–hydrocarbon-like units. The monosaccharide branching units may be conjugated to the dendrimer core or to the prior generation of monomers via linkers, such as polyethylene glycol (PEG) chains. For example, glucose-based branching units can be used in some embodiments. In some embodiments, the branching units may include PEG and / or alkyl chain linkers between different dendrimer generations. For example, glucose layers may be connected via PEG linkers and triazole rings. In some embodiments, the branching units are the same for each generation of dendrimers generated from the core. Therefore, for example, the branching units can be glucose-based branching units for generating generation 1 dendrimers and generation 2 dendrimers. In a preferred embodiment, the branching units are β-D-Glucopyranoside tetraethylene glycol azide having the following structure, or peracetylated derivatives thereof. Structure II: β-D-Glucopyranoside tetraethylene glycol azideIn some embodiments, dendrimers are made of glucose and oligoethylene glycol building blocks. Exemplary glucose dendrimers are shown in Structures III and IV. Some exemplary glucose dendrimers include a generation 1 glucose dendrimer having 24 hydroxyl (-OH) end groups, a generation 2 glucose dendrimer having 96 hydroxyl (-OH) end groups, a generation 3 glucose dendrimer having 396 hydroxyl (-OH) end groups, and generation 4 glucose dendrimer having 1584 hydroxyl (-OH) end groups. For example, the glucose dendrimer is a generation 2 glucose-based dendrimer that has 24 glucose molecules at the periphery and 6 embedded glucose molecules in the backbone held together by PEG segments.In some embodiments, the glucose dendrimer is a generation 1, generation 2, generation 3, generation 4, generation 5, or generation 6 dendrimer. In one embodiment, the dendrimer is a generation 1 dendrimer having the following structure:  Structure III: a generation 1 glucose dendrimer In a preferred embodiment, the dendrimer is a generation 2 dendrimer having the following structure:  Structure IV: a generation 2 glucose dendrimer b.PAMAM DendrimersThe term “PAMAM dendrimer” refers to poly(amidoamine) dendrimer, which may contain different cores, with amidoamine building blocks, and can have carboxylic, amine, acetamide, and / or hydroxyl terminations of any generation including, but not limited to, generation 1 PAMAM dendrimers, generation 2 PAMAM dendrimers, generation 3 PAMAM dendrimers, generation 4 PAMAM dendrimers, generation 5 PAMAM dendrimers, generation 6 PAMAM dendrimers, generation 7 PAMAM dendrimers, generation 8 PAMAM dendrimers, generation 9 PAMAM dendrimers, or generation 10 PAMAM dendrimers. In some embodiment, the dendrimers are generation (“G”) 4, 5 or 6 dendrimers. In some embodiments, the PAMAM dendrimers have hydroxyl terminations.Generally, the complete architecture of dendrimers can be distinguished into the inner core moiety followed by radially attached branching units (i.e., generations) which are further decorated with chemical functional groups carrying desired terminal groups at the exterior surface of the dendrimers. In some embodiments, the dendrimers are in nanoparticle form and are described in detail in U.S. Published Application Nos. US 2011 / 0034422, US 2012 / 0003155, and US 2013 / 0136697. For example, the molecular weight of the dendrimers can be varied to prepare polymeric nanoparticles that form particles having properties, such as drug release rate, optimized for specific applications. In some embodiments, different variations of dendrimers may be used as a delivery vehicle to conjugate and deliver one or more active agents, including, but not limited to, dendrons and tectodendrimers. Dendrons are dendritic wedges that comprise one type of functionality at the core (functional groups, f=1) and another at the periphery (f=8, 16, 32, etc.). Tectodendrimers are generally composed of a central dendrimer with multiple dendrimers attached at its periphery. Preferred dendrimers are hydroxyl PAMAMs, for example, a generation 1-9 hydroxyl PAMAM (such as generation 6 hydroxyl PAMAM) and / or a glucose dendrimer, for example a generation 1-5 glucose dendrimer (such as generation 2 glucose dendrimer (GD2) composed of 24 glucose molecules and containing 96 surface hydroxyl groups).B.Antibodies and Fragments Thereof, NanobodiesThe dendrimer compositions typically include one or more antibodies, nanobodies or antibody fragments / fusion proteins thereof. In preferred embodiments, one or more antibodies or antibody fragments / fusion proteins to be conjugated to the dendrimers are targeted to one or more intracellular targets. In further embodiments, the antibodies or antibody fragments / fusion proteins retain or enhance their therapeutic efficacy when delivered intracellularly by the dendrimers compared to the unconjugated antibodies or antibody fragments / fusion proteins. Exemplary antibodies that can be used include whole immunoglobulin (i.e., an intact antibody) of any class, fragments thereof, and synthetic proteins containing at least the antigen binding variable domain of an antibody. The variable domains differ in sequence among antibodies and are used in the binding and specificity of each particular antibody for its particular antigen. However, the variability is not usually evenly distributed through the variable domains of antibodies. It is typically concentrated in three segments called complementarity determining regions (CDRs) or hypervariable regions both in the light chain and the heavy chain variable domains. The more highly conserved portions of the variable domains are called the framework (FR). The variable domains of native heavy and light chains each include four FR regions, largely adopting a beta-sheet configuration, connected by three CDRs, which form loops connecting, and in some cases forming part of, the beta-sheet structure. The CDRs in each chain are held together in close proximity by the FR regions and, with the CDRs from the other chain, contribute to the formation of the antigen binding site of antibodies. Also disclosed are variants and fragments of antibodies which have bioactivity. The fragments, whether attached to other sequences or not, include insertions, deletions, substitutions, or other selected modifications of particular regions or specific amino acids residues, provided the activity of the fragment is not significantly altered or impaired compared to the non-modified antibody or antibody fragment. The term antibody refers to natural or synthetic polypeptides that bind a target antigen. The term includes polyclonal and monoclonal antibodies, including intact antibodies and functional (e.g., antigen-binding) antibody fragments, including Fab fragments, F(ab')2 fragments, Fab' fragments, Fv fragments, recombinant IgG (rlgG) fragments, single chain antibody fragments, including single chain variable fragments (scFv), and single domain antibodies (e.g., sdAb, sdFv, nanobody) fragments. The term encompasses genetically engineered and / or otherwise modified forms of immunoglobulins, such as intrabodies, peptibodies, chimeric antibodies, fully human antibodies, humanized antibodies, and heteroconjugate antibodies, multispecific, e.g., bispecific antibodies, diabodies, triabodies, and tetrabodies, tandem di-scFv, tandem tri-scFv. The term also encompasses intact or full-length antibodies, including antibodies of any class or subclass, including IgG and sub classes thereof, IgM, IgE, IgA, and IgD.The antibody can be an IgM, IgE, IgA, IgD, or IgG optionally an IgG1, IgG2, IgG3, or IgG4.Exemplary fragments and fusions include, but are not limited to, single chain antibodies, single chain variable fragments (scFv), disulfide linked Fvs (sdFv), Fab', F(ab')2, Fv, and single domain antibody fragments (sdAb).A single chain variable fragment can be created by fusing together the variable domains of the heavy and light chains using a short peptide linker, thereby reconstituting an antigen binding site on a single molecule. Single-chain antibody variable fragments (scFvs) in which the C-terminus of one variable domain is tethered to the N-terminus of the other variable domain via a linker have been developed without significantly disrupting antigen binding or specificity of the binding. The linker is chosen to permit the heavy chain and light chain to bind together in their proper conformational orientation. The linker is usually rich in glycine for flexibility, and typically also includes serine or threonine for solubility. The linker can link, for example, the N-terminus of the VH with the C-terminus of the VL, or vice versa. scFv can also be created directly from subcloned heavy and light chains derived from a hybridoma. The scFv can retain, improve, or increase the specificity of the original immunoglobulin, while removing of the constant regions and introducing the linker.Divalent single chain variable fragments can be engineered by linking two scFvs. This can be done by producing a single peptide chain with two VH and two VL regions, yielding a di-scFvs referred to as a tandem di-scFv. ScFvs can also be designed with linker peptides that are too short for the two variable regions to fold together (about five amino acids), forcing scFvs to dimerize and form a divalent single chain variable fragment referred to as a diabody. Diabodies have been shown to have dissociation constants up to 40-fold lower than corresponding scFvs, indicating that they have a much higher affinity to their target. Even shorter linkers (one or two amino acids) lead to the formation of trimers (triabodies or tribodies). Tetrabodies have also been produced and have been shown to exhibit an even higher affinity to their targets than diabodies.Exemplary molecules that include two or more single chain variable fragments (scFv) including, but are not limited to, divalent-scFv (di-scFv), trivalent-scFv (tri-scFv), multivalent-scFv (multi-scFv), diabodies, triabodies, tetrabodies, etc., of scFvs.The antibody can be a humanized or chimeric antibody, or a fragment, variant, or fusion protein thereof. Methods for humanizing non-human antibodies are well known in the art. Generally, a humanized antibody has one or more amino acid residues introduced into it from a source that is non-human. These non-human amino acid residues are often referred to as “import” residues, which are typically taken from an “import” variable domain. Antibody humanization techniques generally involve the use of recombinant DNA technology to manipulate the DNA sequence encoding one or more polypeptide chains of an antibody molecule. In some embodiments, the antibody is modified to alter its half-life. In some embodiments, it is desirable to increase the half-life of the antibody so that it is present in the circulation or at the site of treatment for longer periods of time. In other embodiments, the half-life of the antibody is decreased to reduce potential side effects. Antibody fragments are expected to have a shorter half-life than full size antibodies. Other methods of altering half-life are known and can be used in the described methods. For example, antibodies can be engineered with Fc variants that extend half-life, e.g., using XTEND™ antibody half-life prolongation technology (Xencor, Monrovia, CA). Exemplary Antibodies andFragments / Fusion ProteinsIn some embodiments, the antibody to be conjugated to a dendrimer is a commercially available antibody. Some exemplary commercially available antibodies are listed in Table 2.  Table 2: Exemplary clinically available antibodies In some embodiments, the antibody to be conjugated to a dendrimer is a monoclonal antibody for one or more immunological indications. Some exemplary monoclonal antibodies are listed in Table 3. Other suitable exemplary antibodies are listed in Tables 4 and 5. Table 3: Monoclonal antibodies in FDA review or approved as treatments for immunological indications. Table 4: Selected therapeutic antibodies against new targets. Table 5: Second generation antibodies. In some embodiments, the antibody to be conjugated to a dendrimer is an antineoplastic monoclonal antibody. Exemplary antineoplastic monoclonal antibodies are listed in Table 6. Table 6: FDA approved antineoplastic monoclonal antibodies. In some preferred embodiments, the antibody or antigen binding fragment thereof conjugated to the dendrimers is a recombinant protein targeting VEGF pathways. In one embodiment, the antibody or antigen binding fragment thereof conjugated to the dendrimers is aflibercept. Aflibercept is a recombinant protein constituted of two recognition domains: one of VEGFR-1 and one of VEGFR-2. These two domains are merged with the Fc fragment of an immunoglobulin. This molecule has the ability to bind to VEGF-A, VEGF-B, and PlGF. In one preferred embodiment, the antibody or antigen binding fragment thereof conjugated to the dendrimers has the following amino acid sequence with disulfide bridge: 30-79, 124-185, 246-306, 352-410, 211-211', 214-214' (SEQ ID NO:1): SDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGMTKKNSTFVRVHEKDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO:1)In another preferred embodiment, the antibody or antigen binding fragment thereof conjugated to the dendrimers is bevacizumab or ranibizumab. For example, in some forms, the antibody or antigen binding fragment thereof conjugated to the dendrimers is ranibizumab. Exemplary light chain and heavy chain sequences for ranibizumab is represented below by SEQ ID NO:2 and SEQ ID NO:3. Ranibizumab light chain sequence:DIQLTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKVLIYFTSSLHSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYSTVPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO:2)Ranibizumab heavy chain sequence:EVQLVESGGGLVQPGGSLRLSCAASGYDFTHYGMNWVRQAPGKGLEWVGWINTYTGEPTYAADFKRRFTFSLDTSKSTAYLQMNSLRAEDTAVYYCAKYPYYYGTSHWYFDVWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHL (SEQ ID NO:3)In some forms, the antibody or antigen binding fragment thereof conjugated to the dendrimers is bevacizumab. Exemplary light chain and heavy chain sequences for bevacizumab is represented below by SEQ ID NO:4 and SEQ ID NO:5 respectively. . Bevacizumab light chain sequence:DIQMTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKVLIYFTSSLHSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYSTVPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO:4)Bevacizumab heavy chain sequence:EVQLVESGGGLVQPGGSLRLSCAASGYTFTNYGMNWVRQAPGKGLEWVGWINTYTGEPTYAADFKRRFTFSLDTSKSTAYLQMNSLRAEDTAVYYCAKYPHYYGSSHWYFDVWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO:5). In some forms, the antibody or antigen binding fragment thereof conjugated to the dendrimers is Ofatumumab. Exemplary light chain and heavy chain sequences for Ofatumumab is represented below by SEQ ID NO:6 and SEQ ID NO:7 respectively. . Ofatumumab light chain sequence:EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPITFGQGTRLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNR(SEQ ID NO:6).Ofatumumab heavy chain sequence:EVQLVESGGGLVQPGRSLRLSCAASGFTFNDYAMHWVRQAPGKGLEWVSTISWNSGSIGYADSVKGRFTISRDNAKKSLYLQMNSLRAEDTALYYCAKDIQYGNYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPGSSKSTSGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEP (SEQ ID NO:7).In some forms, the antibody or antigen binding fragment thereof conjugated to the dendrimers is Ibritumomab tiuxetan. Exemplary light chain and heavy chain sequences for Ibritumomab tiuxetan is represented below by SEQ ID NO:8 and SEQ ID NO:9 respectively. . Ibritumomab tiuxetan light chain sequence:QIVLSQSPAILSASPGEKVTMTCRASSSVSYMHWYQQKPGSSPKPWIYAPSNLASGVPARFSGSGSGTSYSLTISRVEAEDAATYYCQQWSFNPPTFGAGTKLELKRADAAPTVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFN(SEQ ID NO:8).Ibritumomab tiuxetan heavy chain sequence:QAYLQQSGAELVRPGASVKMSCKASGYTFTSYNMHWVKQTPRQGLEWIGAIYPGNGDTSYNQKFKGKATLTVDKSSSTAYMQLSSLTSEDSAVYFCARVVYYSNSYWYFDVWGTGTTVTVSAPSVYPLAPVCGDTTGSSVTLGCLVKGYFPEPVTLTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVTSSTWPSQSITCNVAHPASSTKVDKKIEPRGPTIKPCPPCKCPAPNLLGGPSVFIFPPKIKDVLMISLSPIVTCVVVDVSEDDPDVQISWFVNNVEVHTAQTQTHREDYNSTLRVVSALPIQHQDWMSGKEFKCKVNNKDLPAPIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMVTDFMPEDIYVEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLRVEKKNWVERNSYSCSVVHEGLHNHHTTKSFSR (SEQ ID NO:9).In some forms, the antibody or antigen binding fragment thereof conjugated to the dendrimers is Rituximab. Exemplary light chain and heavy chain sequences for Rituximab is represented below by SEQ ID NO:10 and SEQ ID NO:11 respectively. . Rituximab light chain sequence:QIVLSQSPAILSASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO:10).Rituximab heavy chain sequence:QVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARSTYYGGDWYFNVWGAGTTVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKAEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO:11).In some forms, the antibody or antigen binding fragment thereof conjugated to the dendrimers is Cetuximab. Exemplary light chain and heavy chain sequences for Cetuximab is represented below by SEQ ID NO:12 and SEQ ID NO:13 respectively. . Cetuximab light chain sequence:DILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNNNWPTTFGAGTKLELKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO:12).Cetuximab heavy chain sequence:QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYDYEFAYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO:13). In some forms, the antibody or antigen binding fragment thereof conjugated to the dendrimers is adalimumab. Exemplary light chain and heavy chain sequences for adalimumab is represented below by SEQ ID NO:14 and SEQ ID NO:15 respectively. . adalimumab light chain sequence:DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO:14).adalimumab heavy chain sequence:EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO:15).In some forms, the fusion protein or binding fragment thereof conjugated to the dendrimers is Etanercept. An exemplary sequence for Etanercept is represented below by SEQ ID NO:16. LPAQVAFTPYAPEPGSTCRLREYYDQTAQMCCSKCSPGQHAKVFCTKTSDTVCDSCEDSTYTQLWNWVPECLSCGSRCSSDQVETQACTREQNRICTCRPGWYCALSKQEGCRLCAPLRKCRPGFGVARPGTETSDVVCKPCAPGTFSNTTSSTDICRPHQICNVVAIPGNASMDAVCTSTSPTRSMAPGAVHLPQPVSTRSQHTQPTPEPSTAPSTSFLLPMGPSPPAEGSTGDEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQ ID NO:16).In some forms, the antibody or antigen binding fragment thereof conjugated to the dendrimers is Efalizumab. Exemplary light chain variable region and heavy chain variable region sequences for Efalizumab is represented below by SEQ ID NO:17 and SEQ ID NO:18 respectively. . Efalizumab light chain sequence:DIQMTQSPSSLSASVGDRVTITCRASKTISKYLAWXQQKPGKAPKLLIYSGSTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHNEYPLTFGQGTKVEIKR(SEQ ID NO:17).Efalizumab heavy chain sequence:EVQLVESGGGLVQPGGSLRLSCAASGYSFTGHWMNWVRQAPGKGLEWVGIMIHPSDSETRYNQKFKDIRFTISVDKSKNTLYLQMNSLRAEDTAVYYCARIGIYFYGTTYFDYIWGQGTLVTVSSK (SEQ ID NO:18).In some forms, the fusion protein or binding fragment thereof conjugated to the dendrimers is Aflibercept. An exemplary sequence for Aflibercept is represented below by SEQ ID NO:19. SDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEKDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO:19).In some forms, the antibody or antigen binding fragment thereof conjugated to the dendrimers is Ranibizumab. Exemplary light chain and heavy chain sequences for Ranibizumab is represented below by SEQ ID NO:20 and SEQ ID NO:21 respectively. . Ranibizumab light chain sequence:DIQLTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKVLIYFTSSLHSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYSTVPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO:20).Ranibizumab heavy chain sequence:EVQLVESGGGLVQPGGSLRLSCAASGYDFTHYGMNWVRQAPGKGLEWVGWINTYTGEPTYAADFKRRFTFSLDTSKSTAYLQMNSLRAEDTAVYYCAKYPYYYGTSHWYFDVWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHL (SEQ ID NO:21).In some forms, the antibody or antigen binding fragment thereof conjugated to the dendrimers is Daclizumab. Exemplary light chain and heavy chain sequences for Daclizumab is represented below by SEQ ID NO:22 and SEQ ID NO:23 respectively. . Daclizumab Humanized Anti-CD25 Light Chain 1 sequence:DIQMTQSPSTLSASVGDRVTITCSASSSISYMHWYQQKPGKAPKLLIYTTSNLASGVPARFSGSGSGTEFTLTISSLQPDDFATYYCHQRSTYPLTFGSGTKVEVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNR(SEQ ID NO:22).Daclizumab Humanized Anti-CD25 Heavy Chain 1 sequence:QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYRMHWVRQAPGQGLEWIGYINPSTGYTEYNQKFKDKATITADESTNTAYMELSSLRSEDTAVYYCARGGGVFDYWGQGTTLTVSSGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKAEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO:23).In some forms, the antibody or antigen binding fragment thereof conjugated to the dendrimers is Trastuzumab. Exemplary light chain and heavy chain sequences for Trastuzumab is represented below by SEQ ID NO:24 and SEQ ID NO:25 respectively. . Trastuzumab Anti-HER2 Light chain (1 and 2) sequence:DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO:24).Trastuzumab Anti-HER2 Heavy chain (1 and 2) sequence:EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO:25). NanobodiesIn some forms, the antibody or antibody fragment conjugated to the dendrimer is not a whole antibody, but more compact recognition domains such as a nanobody or a sybody. As used herein, non-antibody protein include proteins that is not a whole antibody which is a multidomain protein. By contrast, a nanobody, also known as a single domain antibody, is an antibody fragment containing a single monomeric variable antibody domain e.g., the antigen-specific domain (VHH) of antibodies obtained from camelids and dromedaries. Like a whole antibody, a nanobody is able to bind selectively to a specific antigen. With a molecular weight of only 12–15 kDa, single-domain antibodies are much smaller than common antibodies (150–160 kDa) which are composed of two heavy protein chains and two light chains, and are also smaller than Fab fragments (~50 kDa, one light chain and half a heavy chain) or single-chain variable fragments (scFv, ~25 kDa fusion of two variable domains, one from a light and one from a heavy chain).Camelids naturally produce one class of antibodies composed only of heavy chains in which the target recognition module is composed of a single variable domain (VHH or Nb). (De Meyer, et al., 2014). The nanobody offers unique properties, simpler structural conformation and high stability in a range of different conditions. The VHH of a camelid antibody can be obtained by genetic engineering to yield a small protein having high affinity for a target, resulting in a low molecular weight antibody-derived protein known as a “camelid nanobody”. See U.S. Patent No. 5,759,808; which is incorporated by reference herein in its entirety. Engineered libraries of camelids antibodies and antibody fragments are commercially available, for example, from Ablynx, Ghent, Belgium. As with other antibodies of non-human origin, an amino acid sequence of a camelid antibody can be altered recombinantly to obtain a sequence that more closely resembles a human sequence, i.e., the camelid nanobody can be “humanized”.Camelid nanobodies have low molecular weights approximately one-tenth that of a human IgG molecule and the protein has a physical diameter of only a few nanometers. One consequence of the small size is the ability of camelid nanobodies to bind to antigenic sites that are functionally invisible to larger antibody proteins, i.e., camelid nanobodies are useful as reagents detect antigens that are otherwise cryptic using classical immunological techniques.An exemplary nanobody that can be conjugated to the dendrimer is ozoralizumab. Ozoralizumab is a trivalent, bispecific nanobody compound, which recognizes and binds Tumor Necrosis Factor (TNF) α receptor, thereby acting as a TNF inhibitor. Exemplary anti-TNFα nanobodies are provided in U.S. Patent No. 9,067,991B2, which is incorporated herein in its entirety.C.Coupling Agents and SpacersDendrimer-antibody conjugates can be formed from one or more active agents covalently conjugated or non-covalently attached to a dendrimer. In preferred embodiments, the one or more active agents are covalently conjugated to the dendrimer. Optionally, the antibody or antibody fragment is conjugated to the dendrimer via one or more spacers. The term "spacer" includes chemical moieties and functional groups used for linking an active agent to the dendrimer. The spacer can be either a single chemical entity or two or more chemical entities linked together. The spacer can include any small chemical entity, peptide or polymers having sulfhydryl, thiopyridine, succinimidyl, maleimide, vinylsulfone, carbonate, etc. In some embodiments, the spacer via which the active agent is conjugated to the dendrimer contains different linkages such as disulfide, ester, carbonate, carbamate, thioester, hydrazine, hydrazides, ether, and amide linkages. The spacer between a dendrimer and an active agent can be designed to provide a releasable or non-releasable form of the dendrimer conjugate in vivo. In some embodiments, the conjugation between active agent and dendrimer is via an appropriate spacer that contains an ester bond between the active agent and the dendrimer. In some embodiments, one or more spacers between a dendrimer and active agents can provide desired and effective release kinetics in vivo. These spacers may contain cleavable linkages (e.g., ester, disulfide, phosphodiester, triglycyl peptide, and hydrazine) or non-cleavable linkages (e.g., amide, ether, and amino alkyl). The conjugation between active agents and dendrimers can be performed using reaction known in the art, such as click chemistry, acid-amine coupling, Steglich esterification, etc. In some embodiments, the conjugation between active agent and dendrimer is via a spacer that contain disulfide, ester, ether, thioester, carbamate, carbonate, hydrazine, ether, or amide linkages, or a combination thereof. In some embodiments, the conjugation between active agent and dendrimer is via an appropriate spacer that contain an ester linkage or an amide linkage between the agent and the dendrimer depending on the desired release kinetics of the agent. The spacer can be chosen from among a class of compounds terminating in sulfhydryl, thiopyridine, succinimidyl, maleimide, vinylsulfone and carbonate group. The spacer can include thiopyridine terminated compounds such as dithiodipyridine, N-Succinimidyl 3-(2-pyridyldithio)-propionate (SPDP), Succinimidyl 6-(3-[2-pyridyldithio]-propionamido)hexanoate LC-SPDP or Sulfo-LC-SPDP. The spacer can also include peptides wherein the peptides are linear or cyclic essentially having sulfhydryl groups such as glutathione, homocysteine, cysteine and its derivatives, arg-gly-asp-cys (RGDC), cyclo(Arg-Gly-Asp-d-Phe-Cys) (c(RGDfC)), cyclo(Arg-Gly-Asp-D-Tyr-Cys), cyclo(Arg-Ala-Asp-d-Tyr-Cys). The spacer can be a mercapto acid derivative such as 3 mercapto propionic acid, mercapto acetic acid, 4 mercapto butyric acid, thiolan-2-one, 6 mercaptohexanoic acid, 5 mercapto valeric acid and other mercapto derivatives such as 2 mercaptoethanol and 2 mercaptoethylamine. The spacer can be thiosalicylic acid and its derivatives, (4-succinimidyloxycarbonyl-methyl-alpha-2-pyridylthio)toluene, (3-[2-pyridithio]propionyl hydrazide, The spacer can have maleimide terminations wherein the spacer includes polymer or small chemical entity such as bis-maleimido diethylene glycol and bis-maleimido triethylene glycol, Bis-Maleimidoethane, bismaleimidohexane. The spacer can include vinylsulfone such as 1,6-Hexane-bis-vinylsulfone. The spacer can include thioglycosides such as thioglucose. The spacer can be reduced proteins such as bovine serum albumin and human serum albumin, any thiol terminated compound capable of forming disulfide bonds. The spacer can include polyethylene glycol having maleimide, succinimidyl, and / or thiol terminations.D.Dendrimer-Antibody ConjugatesAntibodies are being developed for many clinical applications, typically targeted against extracellular, membrane, or secreted proteins. Most antibodies against intracellular targets cannot be used within living cells, because of challenges with delivering them into cells. To overcome the existing challenges, non-viral, plasma stable delivery systems based on covalent conjugates of antibodies with dendrimers are developed. The dendrimer-conjugates of antibodies or antibody fragments showed intracellular delivery of these antibodies or antibody fragments as well as in vivo efficacy in the Examples.Dendrimer-antibody conjugates can be formed of an antibody or an antibody fragment thereof covalently conjugated or non-covalently attached to a dendrimer, a dendritic polymer, or a hyperbranched polymer. Generally, covalent conjugates are preferred.In some embodiments, the antibody or antibody fragment is covalently conjugated to one or more terminal groups of the dendrimer such as terminal hydroxyl groups. In some embodiments, dendrimer conjugates include an antibody, or an antibody fragment thereof, conjugated to the dendrimer via one or more spacers. The spacer between a dendrimer and an antibody or an antibody fragment thereof can be designed to provide a releasable or non-releasable form of the dendrimer conjugate in vivo. For example, the spacer can be cleavable or contain a chemical linkage that is cleavable, for example, by exposure to the intracellular compartments of target neural and / or glial cells or upon binding to the receptor on the surface or in the interior of the target neural and / or glial cells in vivo. Examples of cleavable linkages that can be used in a spacer of the dendrimer-antibody conjugates include, esterase sensitive ester bond, glutathione sensitive disulfide bond, phosphatase-sensitive phosphodiester bond, oligopeptide such as triglycyl peptide linker capable of lysosomal release, acid cleavable hydrazine linkage etc. In some embodiments, the spacer between a dendrimer and active agents can provide desired and effective release kinetics in vivo. In some embodiments, the spacer between the dendrimer and the active agent can be non-cleavable or contain a chemical linkage that is non-cleavable, such as amide, ether, and amino alkyl linkages.Generally, the spacer between the dendrimer and the antibody (or an antibody fragment thereof) has a length sufficient for the antibody conjugated thereto to reach and bind to the target on the surface and / or inside of the target cell. For example, the spacer between the dendrimer and antibody has a length in a range from 50 Da to 2000 Da, depending on the release kinetics desired, and the receptor binding flexibility desired. The length of the spacer can vary, depending on the location of the target receptor (for example, on the cell surface, in the cytoplasm of the cell, or in an intercellular compartment of the cell) and / or density of the receptor when located on the cell surface. In some embodiments, the conjugation of dendrimers increases the half-life of the antibody. In preferred embodiments, the conjugation of dendrimers retains the biological function of the antibody or antibody fragment thereof (e.g., binding affinity). The dendrimer can be a generation 2, generation 3, generation 4, generation 5, generation 6, generation 7, generation 8, and up to generation 10. In some embodiments, the dendrimer is conjugated to one or more antibodies or antibody fragments thereof via spacers containing cleavable (ester, disulfide, phosphodiester, triglycyl peptide, and hydrazine) or non-cleavable (amide, ether, and amino alkyl) linkages. The optimal loading will necessarily depend on many factors, including the choice of antibody or antibody fragment, dendrimer structure and size, and tissues to be treated. In some embodiments, the antibody or antibody fragment is conjugated in a ratio of protein to dendrimer of between 2:1 and 1:2 for the larger generation dendrimers, i.e., generation four or higher in the case of PAMAM dendrimers. In one embodiment, the antibody or antibody fragment is conjugated in a ratio of protein to dendrimer of 1:1.In preferred embodiments, dendrimer complexes retain an effective amount of surface functional groups for targeting to target cells, whilst conjugated to an effective amount of antibody or antibody fragment for treat, prevent, and / or image the disease or disorder.Typically, the dendrimer-antibody conjugates have a hydrodynamic volume in the nanometer range. For example, in some embodiments, the glucose dendrimer- antibody conjugates including one or more antibody or fragment thereof conjugated to the dendrimer have a diameter of between about 5 nm and about 500 nm, inclusive, or between about 10 nm and about 200 nm, inclusive, between about 15 nm and about 100 nm, inclusive, depending upon the generation of dendrimer, the number of antibody molecules loaded. In some embodiments, a dendrimer-antibody conjugate including an antibody or antigen binding fragment thereof conjugated to the dendrimer has a diameter effective to penetrate tissue and to retain in the intracellular location of the target cells for a prolonged period of time. An exemplary dendrimer-antibody conjugate is represented by Formula (I). The dendrimer of the exemplary conjugate contains surface hydroxyl groups, wherein one or more of the surface hydroxyl groups are conjugate to one or more active agents via one or more spacers as shown in Formula (I) below. Formula (I)wherein D can be a generation 2 to generation 10 dendrimer, such as any one of those described above, for example, PAMAM or a glucose-based dendrimer; each occurrence of L can be any suitable chemical moiety appropriate for providing tailored drug release and receptor binding; Y can be a bond or a linkage selected from secondary amides (-CONH-), tertiary amides (-CONR-), sulfonamide (-S(O)2-NR-), secondary carbamates (-OCONH-; -NHCOO-), tertiary carbamates (-OCONR-; -NRCOO-), carbonate (-O-C(O)-O-), ureas (-NHCONH-; -NRCONH-; -NHCONR-, -NRCONR-), carbinols (-CHOH-, -CROH-), disulfide groups, phosphodiester group (), hydrazino group, hydrazones, hydrazides, ester (-C(O)-O-), ether (-O-), and oligopeptide (e.g., triglycyl peptide), wherein R is an alkyl group, an aryl group, or a heterocyclic group; each occurrence of X can be an antibody or fragment / fusion protein thereof, wherein a functional group of X (such as an amino group including primary amino, secondary amino, or tertiary amino group; a carboxylic group; or a hydroxyl group) forms a portion of linkage Y; n can be an integer from 1 to 10; and m can be an integer from 16 to 4096. The dendrimer can be PAMAM or a glucose dendrimer, which is 100% hydroxyl. m and n depend on the size of the dendrimer D. For example, a G4 or G6 PAMAM dendrimer can have a n=1. This range is also appropriate for binding and internalization.The oxygen atom shown in Formula (I) is from the surface functional group of the dendrimer, such as a surface hydroxyl group, where the surface hydroxyl group may or may not be part of a terminal sugar moiety / molecule (e.g., glucose). Although not illustrated in Formula (I), one or more hydroxyl groups of the dendrimer that are not conjugated to active agents may be modified with one or more carbohydrates and / or polyalkylene glycols, such as PEG. In some embodiments, each occurrence of L can be represented by -A’-L1-B’-L2-, wherein A’ can be a carbonyl (-C(O)-) or a bond (including single, double, and triple bonds, for example a single bond); B’ can be a bond (including single, double, and triple bonds, for example a single bond), an amide, an ester, an ether, a thiol, a dithiol, an aryl, a heteroaryl, a polyaryl, a heteropolyaryl, or a heterocyclic; and L1 and L2 can be independently a bond, an alkylene, a heteroalkylene, an aryl, an aralkyl, an ether, a polyether, a thiol, a dithiol, a thiolether, a polythioether, an oligopeptide, a polypeptide, an oligo(alkylene glycol), or a polyalkylene glycol, or L1 and L2 can be independently composed of a combination of these groups, such as a combination of alkylene and polyether, a combination of alkylene and thiol or dithiol, a combination of alkylene and oligopeptide, a combination of alkylene, polyether, and thiol or dithiol, or a combination of polyether and thiol or dithiol. In some embodiments, B’ can be a bond (including single, double, and triple bonds, for example a single bond), an amide group, or a heterocyclic group, such as a triazole group. In some embodiments, L1 can be a bond; an alkylene, such as a C1-C10 alkylene, a C1-C8 alkylene, a C1-C6 alkylene, a C1-C5 alkylene, a C1-C4 alkylene, or a C1-C3 alkylene; or an oligo- or poly-(alkylene glycol), such as where p is an integer from 1 to 20, from 1 to 18, from 1 to 16, from 1 to 14, from 1 to 12, from 1 to 10, from 1 to 8, from 1 to 6, from 1 to 5, from 1 to 4, from 1 to 3, or 1 or 2. In some embodiments, L2 can be a bond; an alkylene, such as a C1-C10 alkylene, a C1-C8 alkylene, a C1-C6 alkylene, a C1-C5 alkylene, a C1-C4 alkylene, or a C1-C3 alkylene; an oligo- or poly-(alkylene glycol), such as where p is an integer from 1 to 20, from 1 to 18, from 1 to 16, from 1 to 14, from 1 to 12, from 1 to 10, from 1 to 8, from 1 to 6, from 1 to 5, from 1 to 4, from 1 to 3, or 1 or 2; an oligo- or poly-peptide, such as a triglycyl peptide; a thiol; or a dithiol; or L2 is composed of a combination of two or more of alkylene, oligo- or poly-(alkylene glycol), oligo- or poly-peptide, thiols, and dithiols. For example, L2 is represented by , where p, q, r, s, t, and u are independently an integer from 0 to 10, from 0 to 8, from 0 to 6, from 0 to 5, from 0 to 4, from 0 to 3, or from 0 to 2, such as 0, 1, or 2; and G’ is a thiol, a dithiol, an oligo-peptide such as a triglycyl peptide, or a poly-peptide. In some embodiments, Y is a linkage that is minimally cleavable in vivo. In some embodiments, Y is a linkage that is cleavable in vivo. In some embodiments, Y is an amide (-CONH-), an ester (-C(O)-O-), an ether (-O-), a phosphodiester, or a disulfide group. In some embodiments, L and Y are both a single bond, and D is directly conjugated to X (an antibody or fragment / fusion protein thereof) via an ether linkage. In some embodiments, D is a generation 2 PAMAM dendrimer, a generation 3 PAMAM dendrimer, a generation 4 PAMAM dendrimer, a generation 5 PAMAM dendrimer, a generation 6 PAMAM dendrimer, a generation 2 glucose dendrimer, a generation 3 glucose dendrimer, a generation 4 glucose dendrimer, a generation 5 glucose dendrimer, or a generation 6 glucose dendrimer. More specific exemplary dendrimer-antibody conjugates are shown in the Examples below.III.Methods of Making Dendrimer ConjugatesA.Methods of Making DendrimersDendrimers can be prepared via a variety of chemical reaction steps. Dendrimers are usually synthesized according to methods allowing controlling their structure at every stage of construction. The dendritic structures are mostly synthesized by two main different approaches: divergent or convergent.In some embodiments, dendrimers are prepared using divergent methods, in which the dendrimer is assembled from a multifunctional core, which is extended outward by a series of reactions, commonly a Michael reaction. The strategy involves the coupling of monomeric molecules that possess reactive and protective groups with the multifunctional core moiety, which leads to stepwise addition of generations around the core followed by removal of protecting groups. For example, PAMAM-NH2 dendrimers are first synthesized by coupling N-(2-aminoethyl) acryl amide monomers to an ammonia core. In other embodiments, dendrimers are prepared using convergent methods, in which dendrimers are built from small molecules that end up at the surface of the sphere, and reactions proceed inward, building inward, and are eventually attached to a core.Many other synthetic pathways exist for the preparation of dendrimers, such as the orthogonal approach, accelerated approaches, the Double-stage convergent method or the hypercore approach, the hypermonomer method or the branched monomer approach, the Double exponential method; the Orthogonal coupling method or the two-step approach, the two monomers approach, AB2–CD2 approach. In some embodiments, the core of the dendrimer, one or more branching units, one or more spacers, and / or one or more surface functional groups can be modified to allow conjugation to further functional groups (branching units, spacers, surface functional groups, etc.), monomers, and / or agents via click chemistry, employing one or more Copper-Assisted Azide-Alkyne Cycloaddition (CuAAC), Diels-Alder reaction, thiol-ene and thiol-yne reactions, and azide-alkyne reactions (Arseneault M et al., Molecules. 2015 May 20;20(5):9263-94). In some embodiments, pre-made dendrons are clicked onto high-density hydroxyl polymers. ‘Click chemistry’ involves, for example, the coupling of two different moieties (e.g., a core group and a branching unit; or a branching unit and a surface functional group) via a 1,3-dipolar cycloaddition reaction between an alkyne moiety (or equivalent thereof) on the surface of the first moiety and an azide moiety (e.g., present on a triazine composition or equivalent thereof), or any active end group such as, for example, a primary amine end group, a hydroxyl end group, a carboxylic acid end group, a thiol end group, etc.) on the second moiety. In some embodiments, dendrimer synthesis replies upon one or more reactions such as thiol-ene click reactions, thiol-yne click reactions, CuAAC, Diels-Alder click reactions, azide-alkyne click reactions, Michael Addition, epoxy opening, esterification, silane chemistry, and a combination thereof. In some embodiments, methods involve one or more protection and deprotection steps of the function groups (e.g., hydroxyl groups) on the central core, branching units, and / or therapeutic, prophylactic or diagnostic agents to facilitate addition of branching units to generate desired dendrimer molecules, or addition of therapeutic, prophylactic or diagnostic agents to generate desired dendrimer conjugates. In the case of hydroxyl groups, they may be protected by formation of an ether, an ester, or an acetal. Other exemplary protection groups include Boc and Fmoc. Any existing dendritic platforms can be used to make dendrimers of desired functionalities, i.e., with a high-density of surface hydroxyl groups by conjugating high-hydroxyl containing moieties such as 1-thio-glycerol or pentaerythritol. Exemplary dendritic platforms such as polyamidoamine (PAMAM), poly (propylene imine) (PPI), poly-L-lysine, melamine, poly (etherhydroxylamine) (PEHAM), poly (esteramine) (PEA) and polyglycerol can be synthesized and explored. Dendrimers also can be prepared by combining two or more dendrons. Dendrons are wedge-shaped sections of dendrimers with reactive focal point functional groups. Many dendron scaffolds are commercially available. They come in 1, 2, 3, 4, 5, and 6th generations with, respectively, 2, 4, 8, 16, 32, and 64 reactive groups. In certain embodiments, one type of agents is linked to one type of dendron and a different type of agent is linked to another type of dendron. The two dendrons are then connected to form a dendrimer. The two dendrons can be linked via click chemistry i.e., a 1,3-dipolar cycloaddition reaction between an azide moiety on one dendron and alkyne moiety on another to form a triazole linker.Exemplary methods of making dendrimers are described in detail in International Patent Publication Nos. WO2009 / 046446, WO2015168347, WO2016025745, WO2016025741, WO2019094952, and U.S. Patent No. 8,889,101. 1. Methods of Making Glucose dendrimersIn some embodiments, glucose-based dendrimers are prepared using divergent methods, in which the dendrimer is assembled from a multifunctional core, which is extended outward by a series of reactions. The strategy involves the coupling of monomeric molecules that possesses reactive and protective groups with the multifunctional core moiety, which leads to stepwise addition of generations around the core followed by removal of protecting groups. In some embodiments, glucose dendrimers are synthesized by coupling AB4 peracetylated β-D glucose-PEG4-azide monomers to hexapropargylated core as shown in FIG. 8A. In preferred embodiments, the hypercore is prepared from dipentaerythritol, for example by performing propargylation of dipentaerythritol to achieve the hexa-propargylated core.In some embodiments, the branching units are hypermonomers i.e., ABn building blocks. Exemplary hypermonomers include AB3, AB4, AB5, AB6, AB7, AB8 building blocks. Hypermonomer strategy drastically increases the number of available end groups. An exemplary hypermonomer is AB4 orthogonal hypermonomer including one azide functional group and four allyl groups prepared from dipentaerythritol with five allyl groups reacted with mono tosylated triethylene glycol azide. In some embodiments, the branching unit is polyethylene glycerol linear or branched. Other monomers include disaccharides and oligosaccharides, as well as saccharides such as fructose, lactose, and sucrose. a.Synthesis of AB4 building blockExemplary synthesis methods of hypermonomer AB4 are described below. In some embodiments, the hypermonomer AB4 is based on glucose molecules. In preferred embodiments, the hypermonomer AB4 is conjugated to a polyethylene glycerol, for example, tetraethylene glycol (PEG4). In one embodiment, the hypermonomer AB4 is peracetylated β-D-Glucopyranoside tetraethylene glycol azide.In some embodiments, the synthesis of glucose-OAc-TEG-OTs involves the following steps: a solution of peracetylated β-D-glucopyranoside (10g, 25.6mmol) was dissolved in 50mL of anhydrous dichloromethane (DCM) followed by addition of 2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethyl 4-methylbenzenesulfonate (6.2g, 17.9mmol) and the reaction mixture was cooled to 0°C. Boron trifluoride diethyl etherate (2.5 eq.) was added and the reaction was allowed to come to room temperature. The reaction was monitored with the help of TLC and quenched after 5hrs by the addition of saturated sodium bicarbonate solution at 0°C. After 10 minutes of stirring, DCM (300mL) was added and the organic layer was washed with saturated sodium bicarbonate solution 3 times until the effervescence was quenched. The reaction mixture was dried over sodium sulfate, filtered, and evaporated under reduced pressure. The crude product was purified by combiflash chromatography using ethyl acetate / hexanes (70:30) mixture as eluents. The desired compound was achieved in 60% yield. Structure of glucose-OAc-TEG-OTs is shown below: Structure IIIn some embodiments, the synthesis of glucose-OAc-TEG-N3 involves the following steps: a solution of glucose-OAc-TEG-OTs (6g, 8.8 mmoles) is dissolved in 40 mL of anhydrous DMF followed by the addition of sodium azide (2eq) and the reaction mixture is heated to 50 oC for overnight. Upon completion, the reaction mixture is filtered and DMF is evaporated. Once dried, the crude reaction mixture is passed through combiflash using ethyl acetate:hexane (70:30) as eluent. Structure of glucose-OAc-TEG-N3 is shown below: Structure IIIIn some embodiments, the synthesis of glucose-OH-TEG-N3 involves the following steps: the peracetylated β-D-Glucopyranoside tetraethylene glycol azide is dissolved in anhydrous methanol and sodium methoxide is added to adjust the pH around 8.5-9. The reaction is stirred overnight at room temperature, then diluted with methanol and pH is adjusted with Amberlist IR-120+ around 6-7. The reaction mixture is separated by filtration and the solvent removed by rotary evaporation. The structure of glucose-OH-TEG-N3 is shown below. Structure Vb.Synthesis of Glucose DendrimersIn some embodiments, glucose dendrimers are synthesized by coupling AB4 peracetylated β-D glucose-PEG4-azide monomers to hexapropargylated core. In preferred embodiments, the hexapropargylated core is linked to AB4 β-D-glucose-PEG4-azide building block (2) via click reaction to obtain generation 1 dendrimer.In some embodiments, generation one dendrimer D1-Glu6-OAc24 is prepared according to the following: Hexapropargylated compound (0.5g, 1mmoles) and an azido derivative ((4.1g, 7.4mmoles) 1.2 eq. per acetylene) are suspended in a 1:1 mixture of DMF and water in a 20mL microwave vial equipped with a magnetic stir bar. CuSO4·5H2O (5mol% / acetylene, 75mg) and sodium ascorbate (5mol% / acetylene, 60mg) dissolved in the minimum amount of water are added. The reaction is irradiated in a microwave at 50 °C for 6 h. The reaction mixture is dialyzed against DMF followed by water dialysis containing EDTA. The EDTA is further removed by extensive water dialysis. The product is lyophilized to obtain D1-Glu6-OAc24. Structure of D1-Glu6-OAc24 is shown below. Structure VI In some embodiments, generation one dendrimer D1-Glu6-OH24 is prepared according to the following: the peracetylated generation 1 glucose dendrimer (1g, 0.26mmoles) is dissolved in anhydrous methanol and sodium methoxide is added to adjust the pH to around 8.5-9. The reaction is stirred overnight at room temperature, then diluted with methanol and pH is adjusted with AMBERLIST® IR-120+ around 6-7. The reaction mixture is separated by filtration and the solvent removed by rotary evaporation, followed by water dialysis. Structure of generation one glucose dendrimer, D1-Glu6-OH24, is shown in Structure III. In some embodiments, generation one glucose dendrimer D1-Glu6-OH24 is propargylated to provide D1-Acetylene24 according to the following: D1-GLu6-OH24 (2 g, 0.721 mmol) was dissolved in anhydrous dimethylformamide (DMF, 50 mL) by sonication. Sodium hydride [60% dispersion in mineral oil] (951 mg, 39.65 mmol) is slowly added in portions at 0°C to the solution with stirring. The solution is stirred for an addition 15 minutes at 0°C. This is followed by the addition of propargyl bromide (3.85 mL, 34.608 mmol, 80% w / w solution in toluene) at 0°C and the stirring is continued at room temperature for another 6h. The reaction mixture is quenched with ice and water, filtered, and dialyzed against DMF, followed by the water dialysis to afford D1-acetylene24. Structure of D1-acetylene24 is shown below. Structure VII In some embodiments, generation one dendrimer D1-acetylene24 is further reacted with AB4 β-D-glucose-PEG4-azide to provide generation 2 dendrimer with 24 glucose molecules containing 96 surface hydroxyl groups. An exemplary generation two dendrimer D2-Glu24-OAc96 is prepared according to the following: D1-acetylene dendrimer24 (0.5g, 0.13 mmoles) and glucose-OAc-TEG-azide (2.2g, 4mmoles) are suspended in a 1:1 mixture of DMF and water in a 20 mL microwave vial equipped with a magnetic stir bar. To this CuSO4·5H2O (5mol% / acetylene, 5mg) and sodium ascorbate (5mol% / acetylene, 10mg) dissolved in the minimum amount of water are added. The reaction is irradiated in a microwave at 50 °C for 8 h. Upon completion, the reaction mixture is dialyzed against DMF followed by water dialysis containing EDTA. The EDTA is further removed by extensive water dialysis. The product is lyophilized to obtain D2-Glu24-OAc96.In some embodiments, generation two dendrimer D2-Glu24-OH96 is prepared according to the following: the peracetylated generation 2 glucose dendrimer D2-Glu24-OH96 is dissolved in anhydrous methanol and sodium methoxide is added to adjust the pH around 8.5-9.0. The reaction is stirred overnight at room temperature, then diluted with methanol and pH is adjusted with AMBERLIST® IR-120+ around 6-7. The reaction mixture is filtered to remove the resin and the filtrate is evaporated by rotary evaporation followed by water dialysis to obtain the product as off-white solid.Structure of generation two glucose dendrimer, D2-Glu24-OH96, is shown in Structure IV. In some embodiments, generation two dendrimer D2-Glu24-OH96 is propargylated at one or more terminal hydroxyl groups suitable for further conjugation to one or more therapeutic, prophylactic or diagnostic agents. In some embodiments, one or more terminal hydroxyl groups of generation two dendrimer D2-Glu24-OH96 is propargylated according to the following: D2-Glu24-OH96 (5b) (200 mg, 0.016 mmol) is dissolved in anhydrous dimethylformamide (DMF, 10 mL) by sonication. To this stirring solution, sodium hydride [60% dispersion in mineral oil] (22 mg, 0.934 mmol) is slowly added in portions at 0°C. The solution is additionally stirred for 15 minutes at 0°C. This is followed by the addition of propargyl bromide (18.0 µL, 80% w / w solution in toluene) at 0°C and the stirring is continued at room temperature for another 6h. The solvent is evaporated using V10 evaporator system and the crude product is purified by passing through PD10 SEPHADEX® G25 M column. The aqueous solution is lyophilized to afford the product as off-white solid.In some embodiments, one or more fluorescent dyes such as infrared fluorescent Cy5 dyes are conjugated to dendrimer. In one embodiment, Cy5-D2-Glu24-OH96 (compound 7 of FIG. 1B) is prepared according to the following: Compound 6 (200 mg, 0.016 mmol) and Cy5 azide (20.7 mg, 0.02 mmol) are suspended in a 1:1 mixture of DMF and water in a 25mL round bottom flask equipped with a magnetic stir bar. To this, CuSO4·5H2O (5mol% / acetylene, 0.3 mg) and sodium ascorbate (10mol% / acetylene, 0.5 mg) dissolved in the minimum amount of water are added. The reaction is stirred at room temperature for 24 h. Upon completion, the DMF is evaporated using V10 and the purification is performed using PD10 Sephadex G25 M column. The aqueous solution is lyophilized to afford the product as blue solid. In some embodiments, the total hydroxyl groups for further conjugation to active agents including therapeutic and / or diagnostic agents are about 1-30, 2-20, or 5-10 out of total 96 available hydroxyl groups of the exemplary generation 2 dendrimer with 24 glucose molecules containing 96 surface hydroxyl groups. B.Methods of Making Dendrimer-Antibody ConjugatesMethods for conjugating agents with dendrimers are generally known in the art and for example, as described in U.S. Published Application Nos. US 2011 / 0034422, US 2012 / 0003155, and US 2013 / 0136697.In some embodiments, one or more antibodies or fragment / fusion proteins thereof (collectively referred to as antibodies) are covalently attached to the dendrimers. In some embodiments, the antibodies are attached to the dendrimer via a spacer that is designed to be non-cleavable in vivo. In some embodiments, the antibodies are attached to the dendrimer via a spacer that is designed to be cleaved in vivo. For example, the spacer can be designed to be cleaved hydrolytically, enzymatically, or combinations thereof, so as to provide for the sustained release of the antibodies in vivo. The chemical structure of the spacer can also be selected in view of the desired release rate of the antibodies. In some embodiments, the conjugation between the antibody and dendrimer is via one or more of disulfide, ester, ether, phosphodiester, triglycyl peptide, hydrazine, amide, or amino alkyl linkages. In some embodiments, the conjugation between the antibody and dendrimer is via an appropriate spacer that provides an ester bond or an amide bond between the agent and the dendrimer depending on the desired release kinetics of the agent. In some cases, an ester or disulfide bond is introduced for releasable form of agents. In other cases, an amide or amino alkyl bond is introduced for non-releasable form of antibodies. Spacers generally contain one or more organic functional groups. Examples of suitable organic functional groups contained in the spacers include secondary amides (-CONH-), tertiary amides (-CONR-), sulfonamide (-S(O)2-NR-), secondary carbamates (-OCONH-; -NHCOO-), tertiary carbamates (-OCONR-; -NRCOO-), carbonate (-O-C(O)-O-), ureas (-NHCONH-; -NRCONH-; -NHCONR-, -NRCONR-), carbinols (-CHOH-, -CROH-), disulfide groups, hydrazones, hydrazides, ethers (-O-), and esters (-COO-, –CH2O2C-, CHRO2C-), wherein R is an alkyl group, an aryl group, or a heterocyclic group. In general, the identity of the one or more organic functional groups within the spacer is chosen in view of the desired release rate of the antibodies. In addition, the one or more organic functional groups can be selected to facilitate the covalent conjugation of the antibodies to the dendrimers. In some embodiments, the conjugation between the antibody and dendrimer is via an appropriate spacer that provides a disulfide bridge between the antibody and the dendrimer. In some embodiments, the dendrimer-antibody conjugates are capable of rapid release of the antibody in vivo by thiol exchange reactions, under the reduced conditions found in body. In certain embodiments, the spacer contains one or more of the organic functional groups described above in combination with a linking group. The linking group can be composed of any assembly of atoms, including oligomeric and polymeric chains; for example, the total number of atoms in the linking group is between 3 and 200 atoms, between 3 and 150 atoms, between 3 and 100 atoms, or between 3 and 50 atoms. Examples of suitable linking groups include alkyl groups, heteroalkyl groups, alkylaryl groups, oligo- and polyethylene glycol chains, and oligo- and poly(amino acid) chains. Variation of the linking group provides additional control over the release of the agents in vivo. In embodiments where the spacer includes a linking group, one or more organic functional groups will generally be used to connect the linking group to both the antibodies and the dendrimers. Reactions and strategies useful for the covalent conjugation of agents to dendrimers are known in the art. See, for example, March, “Advanced Organic Chemistry,” 5th Edition, 2001, Wiley-Interscience Publication, New York) and Hermanson, “Bioconjugate Techniques,” 1996, Elsevier Academic Press, U.S.A. Appropriate methods for the covalent conjugation of a given agent can be selected in view of the linking moiety desired, as well as the structure of the agents and dendrimers as it relates to compatibility of functional groups, protecting group strategies, and the presence of labile bonds.The optimal loading will necessarily depend on many factors, including the choice of antibody (or fragment / fusion protein thereof), dendrimer structure and size, and tissues to be treated. In one embodiment, the antibody is conjugated to the dendrimer at a ratio of 1:1. However, optimal loading for any given agent, dendrimer, and site of target can be identified by routine methods, such as those described. In preferred embodiments, the conjugation of dendrimers does not significantly reduce the biological function of the antibody or antibody fragment thereof (e.g., binding affinity). More specific methods for preparing exemplary dendrimer-antibody conjugates are described in the Examples below.III.Pharmaceutical FormulationsPharmaceutical compositions including dendrimer-antibody conjugates may be formulated in a conventional manner using one or more physiologically acceptable carriers.The composition, method of, and relative amounts for the formulation is dependent upon the route of administration chosen. The compositions can be stored lyophilized in single use vials for rehydration immediately before use. Other means for rehydration and administration are known to those skilled in the art.Representative excipients include solvents, diluents, pH modifying agents, preservatives, antioxidants, suspending agents, wetting agents, viscosity modifiers, tonicity agents, stabilizing agents, and combinations thereof. Suitable pharmaceutically acceptable excipients are preferably selected from materials which are generally recognized as safe (GRAS) and may be administered to an individual without causing undesirable biological side effects or unwanted interactions. The compositions are preferably formulated in dosage unit form for ease of administration and uniformity of dosage. The phrase "dosage unit form" refers to a physically discrete unit of conjugate appropriate for the patient to be treated. It will be understood, however, that the total single administration of the compositions will be decided by the attending physician within the scope of sound medical judgment. The therapeutically effective dose can be estimated initially either in cell culture assays or in animal models, usually mice, rabbits, dogs, or pigs. The animal model is also used to achieve a desirable concentration range and route of administration. Such information should then be useful to determine effective doses and routes for administration in humans. Therapeutic efficacy and toxicity of conjugates can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., ED50 (the dose is therapeutically effective in 50% of the population) and LD50 (the dose is lethal to 50% of the population). The dose ratio of toxic to therapeutic effects is the therapeutic index, and is expressed as the ratio, LD50 / ED50. Pharmaceutical compositions which exhibit large therapeutic indices are preferred. The data obtained from cell culture assays and animal studies can be used in formulating a range of dosages for human use.In certain embodiments, the compositions are administered locally, for example, by injection directly into a site to be treated. ITypically, local administration causes an increased localized concentration of the compositions, which is greater than that which can be achieved by systemic administration. In some embodiments, the compositions are administered via intravitreal administration or intracerebral injection. The compositions of dendrimer-antibody conjugates can be administered parenterally. The phrases "parenteral administration" and "administered parenterally" are art-recognized terms and include modes of administration other than enteral and topical administration. The dendrimer-antibody compositions or formulations thereof can be administered orally, intranasally, subcutaneously, intraperitoneally, or intramuscularly. For liquid formulations, pharmaceutically acceptable carriers may be, for example, aqueous or non-aqueous solutions, suspensions, emulsions, or oils. Parenteral vehicles (for subcutaneous, intravenous, intraarterial, or intramuscular injection) include, for example, sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's and fixed oils. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, and injectable organic esters such as ethyl oleate. Aqueous carriers include, for example, water, alcoholic / aqueous solutions, cyclodextrins, emulsions or suspensions, including saline and buffered media. The dendrimers can also be administered in an emulsion, for example, water in oil. Formulations suitable for parenteral administration can include antioxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient, and aqueous and nonaqueous sterile suspensions that can include suspending agents, solubilizers, thickening agents, stabilizers, and preservatives. Intravenous vehicles can include fluid and nutrient replenishers, electrolyte replenishers such as those based on Ringer's dextrose. In general, water, saline, aqueous dextrose and related sugar solutions, and glycols such as propylene glycols or polyethylene glycols are preferred liquid carriers, particularly for injectable solutions. Injectable pharmaceutical carriers for injectable compositions are well-known to those of ordinary skill in the art (see, e.g., Pharmaceutics and Pharmacy Practice, J.B. Lippincott Company, Philadelphia, PA, Banker and Chalmers, eds., pages 238250 (1982), and ASHP Handbook on Injectable Drugs, Trissel, 15th ed., pages 622630 (2009)).Formulations for convection enhanced delivery (“CED”) include solutions of low molecular weight sales and sugars such as mannitol.IV.Methods of UseThe dendrimer composition, in particular glucose dendrimers, selectively targets neurons, which play a key role in the pathogenesis of many disorders and conditions including neurodevelopmental, neurodegenerative diseases, and brain cancer. In preferred embodiments, the dendrimer composition, in particular glucose dendrimers, selectively target or enriched within neurons, preferably within the nucleus of neurons of injured / hyperactive neurons. In further embodiments, the dendrimer compositions enable targeted editing of a specific cell in the body, including reactive immune cells, including reactive microglia, macrophages, astrocytes, retinal pigment epithelial (RPE cells), enabled by the ability of the hydroxyl PAMAM dendrimers to target these cells.Thus, the dendrimer compositions are administered in a dosage unit amount effective to treat or alleviate conditions associated with the pathological conditions of neurons, reactive microglia, macrophages, astrocytes, and / or retinal pigment epithelial (RPE cells). Generally, by targeting these cells, the dendrimers deliver an effective amount of antibody composition to specifically bind to its intracellular target to achieve a therapeutic result. Methods of treating one or more diseases, disorders, and injury including ocular diseases, neurological disorders, and cancers using the dendrimer-antibody compositions are described. In some embodiments, the dendrimer complexes are used to treat one or more ocular diseases such as exudative form of age-related macular degeneration (AMD). The methods typically include administering a subject in a need thereof an effective amount of a composition including an antibody and one or more dendrimers conjugated thereto. Methods of reducing and / or inhibiting the number or activities of activated microglia and macrophages in the retina and / or the choroid in the eye of a subject in need thereof are provided. IMethods of reducing and / or inhibiting the expression and / or activities of VEGF and / or VEGFR in the activated microglia, activated macrophages, and / or retinal pigment epithelial (RPE) cells in the diseased retina and / or choroid are also described. In some embodiments, the compositions are administered via an intravitreal or subchoroidal route. Methods of reducing and / or inhibiting one or more pro-inflammatory cytokines (e.g., TNF-α, interleukin-1β (IL-1β), or interferon-γ (IFN-γ)) secreted by the activated microglia and macrophages in the diseased retina and / or choroid are also described. In some embodiments, treatment using an effective amount of the compositions leads to a decrease in expression of one or more pro-inflammatory cytokines secreted by the activated microglia and macrophages in the diseased retina and / or choroid.Methods of reducing and / or inhibiting abnormal vascular permeability and leakage, and / or neoangiogenesis in the eye of a subject in need thereof are also described. In some embodiments, treatment using an effective amount of the compositions leads to a decrease in vascular permeability, leakage, and / or neoangiogenesis.The compositions are suitable for treatment of a variety of diseases, disorders, and injury including ocular diseases, inflammatory disorders, neurological disorders, and cancer. The compositions and methods are also suitable for prophylactic use. For example, the compositions may be administered to a patient in need thereof to ameliorate, treat or prevent one or more symptoms related with an inflammatory disease, neurodegenerative disease, or cancer. In one example, the dendrimer compositions are suitable for treating one or more diseases and conditions in the eye, the brain, and the nervous system, particularly those associated with pathological activation of neurons, microglia and / or astrocytes. The compositions and methods are also suitable for prophylactic use.The compositions and methods are suitable for treatment one or more diseases or disorders of the eye. The compositions and methods are suitable for alleviating one or more symptoms associated with one or more diseases or disorder of the eye, for example, discomfort, pain, dryness, excessive tearing, injuries, infections, burns, and gradual loss of vision.In some embodiments, the eye disorder to be treated is a back of the eye disease such as diabetic eye disease, symptomatic vitreomacular adhesion / vitreomacular traction (sVMA / VMT), and wet (neovascular) or dry AMD (age-related macular degeneration). In some embodiments, the eye disorder to be treated is one or more retinal and choroidal vascular diseases (e.g., AMD, retinopathy of prematurity, diabetic macular edema, retinal vein occlusion, retinopathy associated with toxicity of chemotherapy e.g., MEK retinopathy). In other embodiments, the eye disorder to be treated is an inflammatory disease of the eye, i.e., diseases of the eye associated with inflammation of the tissues of the eye, including, for example, AMD, retinitis pigmentosa, optic neuritis, sarcoid, retinal detachment, temporal arteritis, retinal ischemia, arteriosclerotic retinopathy, hypertensive retinopathy, retinal artery blockage, retinal vein blockage, diabetic retinopathy, macular edema, Stargardt disease (also known as Stargardt macular dystrophy or juvenile macular degeneration), geographic atrophy, neuromyelitis optica, and also including angiogenic diseases including, for example, retinal neovascularization and choroidal neovascularization. Other conditions can also result in inflammation and / or angiogenesis in the eye, for example, infection, sickle cell disease, hypotension, etc. Further examples of eye disorders that may be treated include amoebic keratitis, fungal keratitis, bacterial keratitis, viral keratitis, onchorcercal keratitis, bacterial keratoconjunctivitis, viral keratoconjunctivitis, corneal dystrophic diseases, Fuchs' endothelial dystrophy, meibomian gland dysfunction, anterior and posterior blepharitis, conjunctival hyperemia, conjunctival necrosis, cicatrical scaring and fibrosis, punctate epithelial keratopathy, filamentary keratitis, corneal erosions, thinning, ulcerations and perforations, Sjogren's syndrome, Stevens-Johnson syndrome, autoimmune dry eye diseases, environmental dry eye diseases, corneal neovascularization diseases, post-corneal transplant rejection prophylaxis and treatment, autoimmune uveitis, infectious uveitis, anterior uveitis, posterior uveitis (including toxoplasmosis), pan-uveitis, inflammatory disease of the vitreous or retina, endophthalmitis prophylaxis and treatment, macular edema, macular degeneration, age-related macular degeneration, proliferative and non-proliferative diabetic retinopathy, hypertensive retinopathy, an autoimmune disease of the retina, primary and metastatic intraocular melanoma, other intraocular metastatic tumors, open angle glaucoma, closed angle glaucoma, pigmentary glaucoma and combinations thereof. Other disorders include injury, burn, or abrasion of the cornea, cataracts and age related degeneration of the eye or vision associated therewith.In preferred embodiments, the eye disorder to be treated is age-related macular degeneration (AMD). Age-related macular degeneration (AMD) is a neurodegenerative, neuroinflammatory disease of the macula, which is responsible for central vision loss. The pathogenesis of age-related macular degeneration involves chronic neuroinflammation in the choroid (a blood vessel layer under the retina), the retinal pigment epithelium (RPE), a cell layer under the neurosensory retina, Bruch's membrane, and the neurosensory retina, itself. Choroidal neovascularization (CNV) is major pathological feature of neovascular AMD and defined by excessive growth of blood vessels from the choroid into subretinal space. The condition primarily affects the macula, the region of the retina responsible for central vision, while peripheral or side vision remains normal.The dendrimer-conjugated anti-VEGF antibodies (e.g., dendrimer-aflibercept) are administered at a frequency of once a month or less, once every two months or less, once every three months or less, once every four months or less, once every five months or less, once every six months or less,once every seven months or less,once eight months or less,once every nine months or less, once every ten months or less, once every eleven months or less,once every year or less, frequent. The compositions and methods are suitable for the treatment of symptoms associated with neurological and neurodegenerative diseases.Neurodegenerative diseases are chronic progressive disorders of the nervous system that affect neurological and behavioral function and involve biochemical changes leading to distinct histopathologic and clinical syndromes (Hardy H, et al., Science. 1998;282:1075–9). Abnormal proteins resistant to cellular degradation mechanisms accumulate within the cells. The pattern of neuronal loss is selective in the sense that one group gets affected, whereas others remain intact. Often, there is no clear inciting event for the disease. The diseases classically described as neurodegenerative are Alzheimer's disease, Huntington's disease, and Parkinson's disease.Passive immunotherapeutics including exogenous antibodies and antibody fragments, have many applications in the treatment of central nervous system (CNS) disorders (Table 3), including Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, and brain cancer. However, the lack of sufficient brain delivery of antibodies is the major limiting factor in development of novel antibody therapeutics for CNS disorders. Delivery of therapeutic antibodies or other macromolecules is challenging due to the restrictive properties of blood brain barrier (BBB), with about only 0.1% of circulating antibodies estimated to reach the brain at steady-state concentration. This limited amount of antibody uptake in brain may be due too whether antibodies cross the BBB or if the trace amount of antibody found in brain is due to blood contamination (Yu, Y. J.; Watts, R. J., Neurotherapeutics 2013, 10 (3), 459-72). Table 3. Monoclonal antibodies and therapies and current clinical studies regarding their use in neurologic Disease (Novak, J. C., Lovett-Racke, A. E., Racke, M. K., Archives of Neurology 2008, 65 (9), 1162-1165).  In preferred embodiments, the compositions and methods are effective in treating, and / or alleviating one or more symptoms of neuroinflammation associated with a neurological or neurodegenerative disease or disorder or central nervous system disorder. The methods typically include administering to the subject an effective amount of the composition to increase cognition or reduce a decline in cognition, increase a cognitive function or reduce a decline in a cognitive function, increase memory or reduce a decline in memory, increase the ability or capacity to learn or reduce a decline in the ability or capacity to learn, or a combination thereof.Neurodegeneration refers to the progressive loss of structure or function of neurons, including death of neurons. For example, the compositions and methods can be used to treat one or more symptoms of subjects with a disease or disorder such as Parkinson’s Disease (PD) and PD-related disorders, Huntington’s Disease (HD), Amyotrophic Lateral Sclerosis (ALS), Alzheimer’s Disease (AD) and other dementias, Prion Diseases such as Creutzfeldt-Jakob Disease, Corticobasal Degeneration, Frontotemporal Dementia, HIV-Related Cognitive Impairment, Mild Cognitive Impairment, Motor Neuron Diseases (MND), Spinocerebellar Ataxia (SCA), Spinal Muscular Atrophy (SMA), Friedreich's Ataxia, Lewy Body Disease, Alpers’ Disease, Batten Disease, Cerebro-Oculo-Facio-Skeletal Syndrome, Corticobasal Degeneration, Gerstmann-Straussler-Scheinker Disease, Kuru, Leigh's Disease, Monomelic Amyotrophy, Multiple System Atrophy, Multiple System Atrophy With Orthostatic Hypotension (Shy-Drager Syndrome), Multiple Sclerosis (MS), Neurodegeneration with Brain Iron Accumulation, Opsoclonus Myoclonus, Posterior Cortical Atrophy, Primary Progressive Aphasia, Progressive Supranuclear Palsy, Vascular Dementia, Progressive Multifocal Leukoencephalopathy, Dementia with Lewy Bodies (DLB), Lacunar syndromes, Hydrocephalus, Wernicke-Korsakoff’s syndrome, post-encephalitic dementia, cancer and chemotherapy-associated cognitive impairment and dementia, and depression-induced dementia and pseudodementia. In some embodiments, the subject has an excitotoxicity disorder. Excitotoxicity is a process through which nerve cells become damaged because they are overstimulated. A number of conditions are linked with excitotoxicity including stroke, traumatic brain injury, multiple sclerosis, amyotrophic lateral sclerosis, Alzheimer's disease, and spinal injuries. Damage to the nerve cells results in corresponding neurological symptoms which can vary depending on which cells are damaged and how extensive the damage is. Once damaged, nerve cells cannot be repaired, and the patient can experience permanent impairment. A number of drugs have been developed and used in an attempt to interrupt, influence, or temporarily halt the glutamate excitotoxic cascade toward neuronal injury. One strategy is the “upstream” attempt to decrease glutamate release. Thus, in some embodiments, the dendrimer complexes include one or more active agent for treating excitotoxicity disorder. In some embodiments, the subject has a nervous system disorder or is in need of neuroprotection. Exemplary conditions and / or subjects include, but are not limited to, subjects having had, subjects with, or subjects likely to develop or suffer from a stroke, a traumatic brain injury, a spinal cord injury, Post-Traumatic Stress syndrome, or a combination thereof.In some embodiments, the compositions and methods are administered to a subject in need thereof in an effective amount to reduce, or prevent one or more molecular or clinical symptoms of a neurodegenerative disease, or one or more mechanisms that cause neurodegeneration. The formulations can be administered directly or indirectly to the brain using methods of administration known in the art. In one embodiment, the compositions or formulations thereof are directly injected to the brain via intracranial administration by convection enhanced delivery (“CED”).In some embodiments, compositions of dendrimers conjugated or complexed with one or more antibodies and / or additional therapeutic or diagnostic agents are administered to a subject having a proliferative disease, such as a benign or malignant tumor. In some embodiments, the subjects to be treated have been diagnosed with stage I, stage II, stage III, or stage IV cancer.The term cancer refers specifically to a malignant tumor. In addition to uncontrolled growth, malignant tumors exhibit metastasis. In this process, small clusters of cancerous cells dislodge from a tumor, invade the blood or lymphatic vessels, and are carried to other tissues, where they continue to proliferate. In this way a primary tumor at one site can give rise to a secondary tumor at another site.The compositions and methods are useful for treating subjects having benign or malignant tumors by delaying or inhibiting the growth of a tumor in a subject, reducing the growth or size of the tumor, inhibiting or reducing metastasis of the tumor, and / or inhibiting or reducing symptoms associated with tumor development or growth.The dendrimer-antibody compositions are administered to a subject in a therapeutically effective amount. The term “effective amount” or “therapeutically effective amount” means a dosage sufficient to treat, inhibit, or alleviate one or more symptoms of the disorder being treated or to otherwise provide a desired pharmacologic and / or physiologic effect. The precise dosage will vary according to a variety of factors such as the severity and location of the disorder or injury and / or methods of administration, and are known to those skilled in the art. A pharmaceutical composition including a therapeutically effective amount of the dendrimer-antibody compositions and a pharmaceutically acceptable diluent, carrier or excipient is described. The actual effective amounts of dendrimer complex can vary according to factors including the specific active agent administered, the particular composition formulated, the mode of administration, and the age, weight, condition of the subject being treated, as well as the route of administration and the disease or disorder. Dosage forms of the pharmaceutical composition including the dendrimer compositions are also provided. “Dosage form” refers to the physical form of a dose of a therapeutic compound, such as a capsule or vial, intended to be administered to a patient. The term “dosage unit” as used herein refers to the amount of the therapeutic compounds to be administered to a patient in a single dose. The optimal dose is selected from the safety and efficacy results of each tested dose for each drug in patients. In general, the timing and frequency of administration will be adjusted to balance the efficacy of a given treatment or diagnostic schedule with the side-effects of the given delivery system. Exemplary dosing frequencies include continuous infusion, single and multiple administrations such as hourly, daily, or weekly dosing. Higher doses may be given initially to load the patient with drug and maximize uptake in the diseased tissues (e.g., eye). After the loading dose, patients may receive a maintenance dose. In some embodiments, dosages are administered once daily, or less frequently, for example, every other day, two days, three days, four days, five days, or six days to a human. In some embodiments, dosages are administered only about once a week, every two weeks, every three weeks, or every four weeks. In some embodiments, dosages are administered about once or twice every month, every two months, every three months, every four months, every five months, or every six months, or less frequent. In a preferred embodiment, dosages are administered once every four weeks or less frequently, as needed.The dendrimer-antibody compositions can be administered in combination with one or more additional therapeutically active agents, which are known to be capable of treating conditions or diseases discussed above.The therapeutic result of the dendrimer complex compositions including one or more active agents can be compared to a control. Suitable controls are known in the art and include, for example, untreated cells or an untreated subject. The dendrimer complex compositions can be administered alone or in combination with one or more additional active agent(s), as part of a therapeutic or prophylactic treatment regime. The dendrimer-antibody compositions can be administered on the same day, or a different day than the second active agent. For example, compositions including dendrimer complex compositions can be administered on the first, second, third, or fourth day, or combinations thereof. The present invention will be further understood by reference to the following non-limiting examples.ExamplesExample 1: Synthesis of Antibody-Dendrimer conjugatesMethods and MaterialsFIG. 1A shows stepwise synthesis of G1-glucose. The hexapropagylated core 1, was treated treated with AB4 building block (β-Glucose-PEG4-azide), 2under clasical click regents (CuAAC click reaction), catalytic amount ofcopper sulfate pentahydrate (CuSO4.5H2O) and sodium ascorbate in DMF:H2O(1:1) to produce G1-glucose-24-OAc, 3. Then compound 3 was treated under typical Zemplén conditions (to remove acetate groups) to obtain the desired product 4 (G1-glucose).FIG. 1B shows stepwise synthesis of G2-glucose. G1-glucose dendrimer 4 was treated with sodium hydride (60% dispersion in mineral oil) for 15 min at 0 ˚C and then treated with Propargyl bromide (80% w / w solution in toluene). The reaction was stirred at room temperature for 8 h to form compound 5. The compound 5 next treated with AB4 building block (β-Glucose-PEG4-azide), 2under clasical click regents (CuAAC click reaction), catalytic amount ofcopper sulfate pentahydrate (CuSO4.5H2O) and sodium ascorbate in DMF:H2O(1:1) to produce G2-glucose-96-OAc, 6. And then compound 6 was reacted under typical Zemplén conditions to obtain the desired product 7(G2-glucose).FIG. 1C shows synthesis of functionalized Cy5-D-PEG4-TCO. G2-glucose dendrimer was treated with sodium hydride (60% dispersion in mineral oil) at 0˚C and addition of propagyl bromide at O˚C-RT for 8 h to form compound 8. The resulted product 8 was reacted with azido-PEG2-amine (9) to form product 10. The product, 10 was labeled with Cy5 fluorophore and the resulted intermediate, 11 was conjugated with Pegylated trans-cyclooctene (TCO) to obtain functionalized Cy5-D-PEG4-TCO (13).FIG. 1D shows synthesis of Cy5-G2-Glucose-antibody conjugate. Me-Tz attached antibody, 14 and TCO attached G2-Glucose dendrimer 13 were reacted under highly specific inverse Diels-alder click reaction (IEDDA) to form Cy5-G2-Glucose-antibody conjugate 16.FIG. 2 is a schematic representation showing hydroxyl PAMAM dendrimers, glucose dendrimers, or glucose-functionalized PAMAM dendrimers, with ligand functionalization, that are conjugated to antibodies and antibody-like molecules. Example 2: Targeted intracellular delivery of aflibercept fusion protein using hydroxyl PAMAM dendrimer mediated nano-constructsAge related macular degeneration (AMD) causes progressive degeneration of the photoreceptor-dense macula and affects fine visual activity like reading, facial recognition and drivingMaterials and MethodsA covalently conjugated hydroxyl PAMAM dendrimer-aflibercept system was made and then tested Generation 6 hydroxyl PAMAM dendrimer (D6-OH) was used, and conjugation was carried out using a trans-cyclooctene-tetrazine (TCO-Tz) strain-promoted click chemistry strategy under catalyst-free, mild conditions. The PEGylated trans-cyclooctene (PEG4-TCO) labeled hydroxyl PAMAM dendrimer and PEGylated tetrazine(PEG2-Tz) labeled aflibercept were reacted under physiological conditions to produce dendrimer-aflibercept (D-aflibercept). Without the guidance of any targeting ligand, intravenously administrated D-aflibercept appeared to accumulate and internalize at the site of neuro-inflammation and long-term retention in activated macrophages in the disease-affected area. Delivering aflibercept to key pathological cells via dendrimer provided significant improvement in efficacy compared to aflibercept alone. While, dendrimer-mediated internalization of aflibercept improved the trapping of VEGF in its origin (Fig 16). Materials and MethodsCovalently conjugated hydroxyl PAMAM dendrimer-aflibercept conjugates were made using a generation six hydroxyl PAMAM dendrimer (D6-OH). Conjugation was conducted using trans-cyclooctene-tetrazine (TCO-Tz) strain-promoted click chemistry under catalyst free, mild conditions. The PEGylated trans-cyclooctene (PEG4-TCO) labeled hydroxyl PAMAM dendrimer and PEGylated tetrazine (PEG2-Tz) labeled aflibercept were reacted under physiological conditions to produce dendrimer-aflibercept (D-aflibercept). Biomolecules, chemicals, and reagentsUnless stated otherwise, all the reactions were performed in flame-dried glassware under a positive pressure of nitrogen using dry solvents. Commercial grade reagents and anhydrous solvents were purchased from chemical suppliers and used without further purification. 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC.HCl), N, N-diisoprpylethylamine (DIPEA), 4-(dimethylamino)pyridine (DMAP) trifluoracetic acid (TFA), γ-(Boc-amino) butyric acid (Boc-GABA-OH), anhydrous dichloromethane (DCM), N, Nʹ-dimethylformamide (DMF) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Cyanine 5 (Cy5)-mono-NHS ester was purchased from Amersham Bioscience-GE Healthcare. Trans-cyclooctene (TCO) was purchased from AAT bioquest, Inc. Tetrazine (Tz) precursors were purchased from BroadPharm. Deuterated solvents dimethylsulfoxide (DMSO-d6), water (D2O), and Chloroform (CDCl3) were purchased from Cambridge Isotope Laboratories Inc. (Andover, MA). Ethylenediamine-core polyamidoamine (PAMAM) dendrimer, generation 6.0, hydroxy surface (G6-OH; diagnostic grade; consisting of 256 hydroxyl end-groups), methanol solution (13.75% w / w) was purchased from Dendritech Inc. (Midland, MI, USA). Dialysis membranes were purchased from Spectrum Laboratories Inc. (Rancho Dominguez, CA, USA). Instrumentation for characterization of intermediates and productsProton nuclear magnetic resonance (1H NMR) spectra were recorded on a Bruker 500 MHz spectrometer at ambient temperatures and analyzed using MestReNova software. 1H NMR chemical shifts were reported as δ using residual solvent as an internal standard (DMSO-d6, 2.50), and (D2O, 4.79 ppm). Analytical high-performance liquid chromatography (HPLC) was performed using a Shimadzu LC-AD HPLC system equipped with a variable wavelength absorbance detector and a C18 reverse phase column (Waters, BEH300 5 µm, 19×250 mm). The eluents were monitored at 210 nm using a photodiode array (PDA) detector, and fluorescently labeled conjugate was monitored at both 650 and 210 nm using fluorescence and PDI detectors, respectively. HPLC elution was carried out with a 40 min linear gradient of 0%-90% HPLC grade acetonitrile (CH3CN) in water containing 0.1% TFA, maintaining the flow rate at 1.0 mL / min. Dendrimer-aflibercept conjugates and intermediates were purified using Fast Protein Liquid Chromatography (FPLC) system (AKTA FPLC, 280 nm single wave length detector) equipped with SUPEROSETM 6 Increase 10 / 300 GL size exclusion column (minimum 500 µL sample volume) at the flow rate 0.5 mL / min and analyzed using UNICORN start 1.1 software. The sample elution was conducted with PBS pH 7.4(1×). Synthesis of compoundsSynthesis of D-GABABoc(Compound3)A solution of PAMAM G6-OH (Compound 1in FIG. 1) (2.00 g, 0.03 mmol) in anhydrous DMF (15 mL) was treated with Boc-GABA-OH (0.14 g, 0.68 mmol), DMAP (0.16 g, 0.82 mmol) and stirred at room temperature for 5 minutes. Then EDC.HCI (0.92 g, 0.75 mmol) was added in portions to the reaction mixture over the period of 5 minutes. The reaction mixture was stirred at room temperature for 48 hours. The crude product was diluted with DMF and was dialyzed against DMF using a 3.5 kDa MW cut-off cellulose dialysis tubing for 12 hours followed by water for 24 hours. The aqueous solution was lyophilized to yield desired product 3 (FIG. 1) as a hygroscopic white solid (1.95 g, 95%). 1H NMR (500 MHz, DMSO-d6) 8.10-7.70 (m, internal amide H), 6.60 (s, GABA amide H, 10H), 4.74 (s, surface OH, 213H), 3.99 (s, ester linked H, 22H) 3.39 (t, J = 5.0 Hz, dendrimer –CH2), 3.40-3.35 (m, dendrimer CH2), 3.11 (m, dendrimer-CH2), 2.89 (m, dendrimer CH2), 2.73-2.65(m, dendrimer CH2), 2.45(m, dendrimer-CH2), 2.21(m, dendrimer CH2), 1.64-1.59 (m, GABA linker-CH2, 25H), 1.36 (s, Boc group, 85H). HPLC C18 retention time 19 min.Synthesis of D-GABA-NH2(Compound4)The Boc protected GABA linker containing PAMAM G6-OH (Compound 3 in FIG. 1) (500 mg, 0.008 mmol) was treated with TFA / DCM (3:4) solvent mixture. The reaction was stirred at room temperature for 12 hours, then diluted with methanol (MeOH) to remove excess of TFA (Trifluoroacetic acid), and concentrated in vacuo. The crude product was directly used for the next step without any further purification. 1H NMR (500 MHz, DMSO-d6) δ 8.50-7.75 (m, internal amide H), 5.50-4.50 (broad s, surface -OH), 4.00 (s, ester linked H), 3.50-2.25(m, dendrimer-CH2), 1.93-1.59 (m, GABA linker-CH2). Synthesis of Cy5-D (Compound5)A solution of compound 4 (FIG. 1) (574 mg, 0.01 mmol) in DMF (5 mL) was treated with DIPEA to adjust pH of the reaction mixture (~7.0-7.5). Then the reaction was treated with Cy5-NHS ester (17.4 mg, 0.023 mmol, 1.2 eq) and stirred at room temperature (rt) for 12 hours. It was then dialysed against DMF using 3.5 kDa for 12 hours followed by against water for 24 hours. The aqueous layer was frozen and lyophilized to yield desired product 5 (FIG. 1) as blue solid (yield 85%).1H NMR (500 MHz, DMSO-d6) δ 8.25-7.75 (m, internal amide H), 7.30 (s, Cy5 H), 7.10 (s Cy5 H), 6.70 (s, GABA amide H), 6.50 (m Cy5 H), 6.25 (m Cy5 H), 4.75 (s, surface OH, 226H), 4.00 (m, ester CH2), 3.50-2.00 (m, dendrimer CH2), 1.64-1.59 (s, 31H), 1.25 (s, 66H), 0.8 (s, 21H). HPLC C18 retention time (MeCN in H2O with 0.1% TFA, linear gradient, 40 min). HPLC C18 retention time: 17.5 min.Synthesis of Cy5-D-PEG4-TCO (Compound 6)A solution of compound 5 (FIG. 1) (48 mg, 0.0008 mmol) in DMF (5 mL) was treated with DIPEA to adjust pH of the reaction mixture (~7.0-7.5). The reaction was treated with TCO-PEG4-NHS ester (4 mg, 0.0080 mmol) and stirred the reaction mixture at rt for 12 h. It was then dialysed against DMF 12 h followed by against water for 24 h. The aqueous layer was frozen and lyophilized to yield desired product as blue solid (yield 55%). 1H NMR (500 MHz, DMSO-d6) δ 8.14-7.73 (m, internal amide H), 7.35 (m, Cy5 H), 7.25 (m, Cy5 H), 7.05 (m, Cy5 H), 6.6 (m, Cy5 H), 6.3 (m, Cy5 H), 6.83 (s, GABA amide H), 5.65-5.50 (m, TCO H), 5.45-5.35 (m, TCO H), ( 4.74 (s, surface OH, H), 4.01-3.39 (t, J = 5.0 Hz, ester –CH2), 3.50-2.00 (m, dendrimer CH2), 1.9 (s, 24H), 1.6 (s, 80H), 1.2 (s, 126H), 0.8 (s, 80H). HPLC C18 retention time: 19.5 min.Ultrafiltration and SEC chromatographyAfter each step of the synthesis, the excess small molecules reagents and byproducts and buffer exchange were performed by Amicon ultrafiltration using 15 mL, 30 kDa MWCO units (for ≥2 mg samples) or 0.5 mL, 30 kDa MWCO units (for ≤2 mg samples). Sample preparation and MALDI-TOF analysisPAMAM dendrimer conjugates:The MALDI matrix 2’, 4’,6’-Trihydroxyacetophenone monohydrate (THAP) (10 mg) was dissolved in 1mL of Acetonitrile in water (1:1) with 0.1% trifluoroacetic acid). Then 2 µL of PAMAM dendrimer was deposited on the MALDI sample plate. The matrix (2 µL of the 10 mg / mL) was deposited on the air-dried sample and allowed it to air dry for 10-20 min. The MALDI-TOF MS analysis was performed in a reflective-positive mode. AfliberceptThe Aflibercept fusion protein used for this study was manufactured by Regeneron Pharmaceuticals. Cell lineThe ARPE-19 cells, an immortal human RPE cell line, was used for this study. Cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, ATCC, Manassas, VA) supplemented with 10% (v / v) heat-inactivated fetal bovine serum (FBS, Invitrogen Corp., Carlsbad, CA), 1% penicillin / streptomycin (P / S, Invitrogen Corp., Carlsbad CA). Cells were maintained at 37˚C and 5% CO2 under humidified atmosphere.In vitro evaluation of the delivery strategyStudy of time dependent uptake: ARPE-19 cells were seeded in glass-bottom culture dishes and grown for 24-48 h to 70-80% confluency. Cells were treated with Cy5 fluorescently labeled aflibercept and aflibercept-conjugated Cy5-labeled dendrimer (Cy5-D-aflibercept, compound 9in FIG. 3) in DMEM supplemented with 1% P / S (serum free media). The cells were then washed with PBS (×3) and fixed in 5% formalin solution. Cells were incubated and confocal microscopic images were taken by Zeiss Axiovert 200 system equipped with an LSM 510-Meta confocal module. Image acquisition parameters were kept constant during the imaging. Images were processed by Zen 2011 software (Zeiss). AnimalsMale six- to eight-week-old C57BL / 6J mice (Envigo, Indianapolis, IN) were used for the experiments. Mice were treated in accordance with the Association for Research in Vision and Ophthalmology Statement on the care and use of Animals in Ophthalmic and Vision Research. All protocols were reviewed and approved by the Animal Care and Use Committee of the Johns Hopkins University School of Medicine. Induction of CNV by laser photocoagulation and drug treatmentLaser photocoagulation-induced rupture of Bruch’s membrane was used to generate CNV (choroidal neovascularization; Gong, Y. et al. PLoS One 10, e0132643, (2015)). The 6–8-week-old C57BL / 6J mice were anesthetized with ketamine hydrochloride (100 mg / Kg body weight) and pupils were dilated with 1% tropicamide (Alcon Labs, Inc., Fort Worth, TX). After full dilation of pupils, lubricating eye drops (Alcon Laboratories) were applied to the cornea. The fundus was viewed with an imaging camera, and laser photocoagulation was induced by the image-guided laser system (Micron III, Phoenix Research Laboratories, Pleasanton, CA). Four laser burns were equally induced around the optic nerve in each eye by Argon laser (532 nm, spot size of 50 μm, duration of 70 ms, power levels of 240mW) was performed in the 9, 12, and 3 o’clock position. A vaporization bubble was created at each laser spot, which indicated successful laser photocoagulation. Only burns that produced bubbles without vitreous hemorrhage were included in the study. For Real-time PCR measurements of cytokines, ten laser burns were applied per eye. Immediately after the laser photocoagulation, intravitreal injection of 1 μl of D-aflibercept (40 μg / μl), aflibercept (40 μg / μl) or PBS was conducted by 31-gauge Hamilton syringe.Fundus Fluorescein Angiography (FFA) FFA was performed with the Retinal Imaging Microscope (Micron III, Phoenix Research Laboratories, Pleasanton, CA) 7 days or 14 days after laser photocoagulation. Mice were anesthetized, pupils dilated, and intra-peritoneal injected with fluorescein (Altaire, Aquebogue, NY). Fluorescent fundus images were taken 5 minutes after fluorescein injection. Retinal pigment epithelium / Choroid / Sclera Flat-mount, Imaging and Quantification Mice were sacrificed 7 days after laser photocoagulation. Eyes were immediately enucleated and fixed with 4% paraformaldehyde (Sigma-Aldrich, St. Louis, MO) in PBS for 1 hour at room temperature. The posterior eyecups containing the retinal pigment epithelium / choroid / sclera were dissected, radially sectioned, and incubated with 0.1% Triton X-100 (Thermo Fisher Scientific, Tewksbury, MA) in phosphate buffered saline (PBS, Life Technologies, Grand Island, NY) for 1 hour at room temperature. After overnight incubation at 4 °C with IBA-1 antibodies (ionized calcium binding adaptor molecule 1) (1:500, Wako Chemicals, Richmond, VA), flat mounts were incubated with a second antibody Cy3 (1:500, Life Technologies, Grand Island, NY) and Isolectin B4 (IB4, 1:100, Thermo Fisher, MA) at 4 °C overnight. After washing with PBS three times, the posterior eye cups were flat-mounted onto slides in Fluoromount-G mounting medium (Thermo Fisher Scientific, Tewksbury, MA). The fluorescent images were taken with the inverted fluorescence microscope (Zeiss, Peabody, MA). The areas of CNV lesions were quantified by image analysis with Image-J software (National Institutes of Health, Bethesda, MD) by two observers masked with respect to experimental groups.CNV vascular leakageFFA was performed with the Retinal Imaging Microscope (Micron III, Phoenix Research Laboratories, Pleasanton, CA) 7 or 14 days after laser photocoagulation. Mice were anesthetized, pupils dilated, and intra-peritoneal injected with fluorescein (0.1mL, 10% wt / vol.). Fluorescent fundus images were taken 5 minutes and 10 minutes after fluorescein injection. The fluorescent intensity of CNV lesions was quantified using ImageJ (National Institute of Health, Bethesda, MD), and the difference in fluorescence intensity between 10- and 5-minute images was calculated as CNV vascular leakage.ELISA-based VEGF binding assessmentThe sandwich ELISA-based assay was used to confirm the binding of D-aflibercept on VEGF. The VEGF-coated plate was prepared by incubating a pre-blocked nickel-coated 96-well plate (Thermo Scientific, cat. No. 15442) with His-tagged recombinant human VEGF (Prospec, cat. No. CYT496) diluted in PBST (PBS containing 0.1% TWEEN 20) at 1 µg / mLin 100 µL per well. The plate was incubated 1h at room temperature and washed 3 times with PBST (200 L / well each). Then the serially diluted (3-fold dilution) aflibercept or D-aflibercept in PBST was added to each well and incubated overnight at 4 °C. The plate was washed again 3 times with PBST (200 l / well each), and to each well was added HRP-conjugated anti-human secondary antibody (1 / 5000 dilution; Thermo Scientific, cat. No. A18811) diluted in PBST. The plate was developed by adding 1-step slow TMB-ELISA substrate (100 µL / well) and incubating for 15-30 minutes. The chromogenic reaction was stopped by the addition of 1N sulfuric acid (Thermo Scientific, cat. No. N600, 100 µL / well) and the absorbance of each well was read at 450 nm using a microplate reader (BioTek instruments).Quantification of VEGFA by ELISAHMC3 cells were seeded in 96-well plates at a density of 10,000 cells per well and cultured in standard conditions (37°C, 5% CO₂). The cells were pre-treated with 100 ng / mL lipopolysaccharide (LPS, derived from *Escherichia coli* 0127: B8, Lot#125M4091V) (Sigma-Aldrich, St. Louis, MO) for 3 hours to induce an inflammatory response. Following LPS pre-activation, the cells were treated with varying concentrations of Eylea and D-Eylea (conjugated Eylea) at 50, 10, and 1 µg / mL, respectively, for 12 hours. After treatment, the medium was aspirated, and the cells were gently washed with warm PBS. Fresh culture medium containing 100 ng / mL LPS was added to maintain the inflammatory environment, and the cells were incubated for an additional 1, 2, 3, and 4 days. At each time point (1-, 2-, 3-, and 4-days post-treatment), the culture medium was collected from the wells, centrifuged at 5,000 rpm for 5 minutes at 4°C, and the supernatant was stored at -80°C until further analysis. The effect of aflibercept and D-aflibercept was evaluated by measuring the secreted vascular endothelial growth factor (VEGF) levels in the collected media. VEGF concentrations were quantified using a Human VEGF ELISA kit (Bio Legend, San Diego, CA) according to the manufacturer’s protocol. Absorbance at 450 nm was measured using a microplate reader with a wavelength correction at 570 nm. Untreated (resting) cells and cells treated with LPS alone served as control conditions for the experiment.RT-PCR for cytokine expressionGene expression was quantified by quantitative RT-PCR in CNV model 7 and 14 days after laser induction. Choroids were dissected in RNase free conditions. Total RNA was isolated with Trizol reagent (Life Technologies, Grand Island, NY). 3 μg of total RNA was reverse-transcribed to cDNA using high-capacity cDNA reverse transcription system (Life Technologies, Grand Island, NY) according to the manufacturer's instructions. Real-time PCR was performed with SYBR Green Master Mix (Life Technologies, Grand Island, NY) and the following primers: GAPDH, VEGF, IL-1, TGF-β.Statistical analysisData are presented as mean ± SEM and analyses were performed in GraphPad Prism (Version 7: La Jolla, CA). Statistical significance of the data was analyzed using the one-way ANOVA with a Fisher’ LSD post hoc test was performed using SPSS software 25.0 (IBM, Armonk, NY). P<0.05 was considered as statistically significant. ResultsSynthesis and characterization of dendrimer conjugated Aflibercept (EYLEA®) and efficacy in laser CNV model for Age related macular degenerationCy5-D-PEG4-TCO. Cy5-D-PEG4-TCO conjugate was synthesized using PAMAM-G6-OH (D6-OH) dendrimer (FIG. 3). Generation 6 (G6) PAMAM dendrimer has 256 free hydroxyl groups (D6-OH) available on the surface for further conjugations. After each reaction step, the products were purified via dialysis in DMF for 24 h to eliminate small molecule impurities followed by water dialysis to remove DMF. 1H NMR (in DMSO-d6) comparison of intermediates and the final conjugates from top to bottom (FIG. 4A) and analytical reverse phase-High-performance liquid chromatography (HPLC) traces confirmed the product formation by appearance and disappearance of peaks and showing shifts in the retention times, respectively.Molecular weights (Figures4B-4D) of all intermediates and final components were determined by matrix-assisted laser desorption / ionization-time of flight (MALDI-TOF). The synthesis of compound 1 was carried out using D6-OH in methanol (13.75% w / w), which was dried under reduced pressure, followed by the dissolution in water and lyophilization. The lyophilized mono-functionalized D6-OH was functionalized with Boc protected amine by treatment of 4-tert-butoxycarbonylamino)butyric acid (Boc-GABA-OH) under N-(3-dimethylaminopropyl)-N´-ethylcarbodiimide hydrochloride (EDC.HCl) and 4-(dimethylamino) pyridine (4-DMAP) in DMF for 36 h at room temperature to yield the Boc protected bifunctional dendrimer product. The crude dendrimer was dialyzed by 3.5kDa membrane against ultrapure water for 24 h followed by lyophilization. 1H NMR of dendrimer 3 depicted the appearance of tert-butyl protons of Boc group at 1.3 ppm as a singlet along with GABA methylene protons at  1.6 ppm. The peak at  3.9 ppm is for the methylene protons of the dendrimer next to hydroxyl groups once converted to ester and amidic protons from GABA linker also appeared at  6.8 ppm. Then the Boc groups were deprotected under mild acidic conditions using trifluoroacetic acid (TFA) in dichloromethane (DCM) 1:4 to obtain bi-functionalized dendrimer. The excess TFA was removed by co-evaporation with methanol and resulted crude product was used for next step without further purification. The complete disappearance of Boc protons was confirmed by 1H NMR while no ester hydrolysis was observed under this condition (FIG. 3A). The total number of amine groups was maintained at ~10. Then bifunctional dendrimer was treated with fluorescent dye Cy5 to yield dendrimer 4 with~1-2successful Cy5 attachment at dendrimer surface. 1H NMR showed the appearance of Cy5 signals in the aromatic region (FIG. 3A) and HPLC retention time was shifted from 19.0 to 17.5 min confirming the product formation. After Cy5 attachment rest of amine groups were reacted with trans-cyclooctene containing, hetero-bi functional (NHS-PEG4-TCO) linker. This hetero-bi functional linker is used to form a chemical bonding between dendrimer and aflibercept. Analytical HPLC was conducted to measure the purity of products (FIG. 3B). The sizes of components were measured by dynamic light scattering (DLS) using Zetasizer. The mean hydrodynamic diameter of both D-aflibercept (12.22±0.74 nm) and aflibercept (9.04 ±0.74 nm) was measured (FIG. 17A and FIG. 17B). Upon conjugation of aflibercept to dendrimer-aflibercept there was significant increase in size. Synthesis and characterization of D-aflibercept conjugateThe aflibercept fusion protein was functionalized with terminal tetrazine (Tz) while D6-OH was functionalized with trans-cyclooctene (TCO) for click reaction (FIG. 5). The aflibercept (2 mg in 500 µL of PBS) was treated with Tz-PEG2-NHS ester (20 mol equiv in 10-20 µL of anhydrous DMSO) and incubated for 1h. Initially, the reaction mixture became cloudy and cleared after a few minutes. The excess Tz-PEG2-NHS and byproducts were removed by ultrafiltration. The product 8 was further purified by equipped with size-exclusion column chromatography (SEC). The number of Tz groups attached per aflibercept was determined based on the change in the molecular weight measured by MALDI-TOF mass spectrometer. The number of groups attached is denoted as the subscripted number. The TCO-PEG4 attached dendrimer 6(5.2 mg in 500 µL PBS)wasreacted with 8via trans-cyclooctene-tetrazine (TCO-Tz) to afford crude product 9. The resulting crude product was purified by ultrafiltration, and the product was further purified by Fast Protein Liquid Chromatography (FPLC) system (AKTA FPLC, 280 nm single wavelength detector) equipped with SEC column (FIG. 6A). The TCO-Tz click reaction used herein is fast, quantitative and releases no toxic byproducts. At low biomolecule concentrations (less than < 5 µM) TCO-Tz works well compared to strain-promoted alkyne-azide cycloaddition SPAAC and Cu(I) catalyzed azide-alkyne cycloaddition (CuAcc). The TCO-Tz “click” reaction proceeds via an inverse-electron demand Diels-Alder reaction (IEDDA) between a TCO and a Tz, followed by retro Diels-Alder reaction eliminating N2to form a dihydropyridazine bond. In contrast to a regular Diels-Alder reaction, where an electron diene reacts with an electron-poor dienophile, in an inverse-electron-demand Diels-Alder reaction, an electron-rich dienophile reacts with an electron poor diene. The TCO as a precursor gave tremendous rate difference compared to cis-cyclooctene and other cyclic alkenes. The high reactivity is related to a crown confirmation adapted by TCO, which is lower in energy than the ‘half-chair’ confirmation of cis form. The chemo selective TCO-Tz ligation possess ultrafast kinetics (>800 M-1s-1) unmatched by any other biorthogonal ligation pair. The click ligation was performed at near neutral pH, aqueous condition at room temperature. The ultrafast kinetics, selectivity, and long-term aqueous stability make TCO-Tz the ideal pair in low concentration aflibercept-dendrimer coupling reactions. MALDI-TOF MS, and analytical HPLC traces were used to confirm the synthesis of all the intermediates and final dendrimer aflibercept conjugates (FIGs.6A-6B). D-aflibercept was analyzed using a standard SDS-PAGE (4-12% gradient gel) followed by Coomassie blue staining for the protein visualization on the gel (data not shown). D-aflibercept mediated suppression of laser-Induced CNV formation.Generally, the VEGF upregulation can happen in response to ischemia, hypoxia, inflammation, and trauma (Croll, S. D., Goodman, J. H. & Scharfman, H. E. in Recent Advances in Epilepsy Research (eds Devin K. Binder & Helen E. Scharfman) 57-68 (Springer US, 2004)). In AMD the accumulation of oxidized lipids in Bruch’s membrane and ischemic component due to atrophy triggers VEGF upregulation. The well-established laser-induced mouse CNV model (C57BL / 6) was used to evaluate efficacy of the dendrimer conjugate (Shah, J Vis Exp, e53502 (2015)). The targeted laser injury to RPE and Bruche’s membrane induces angiogenesis, modeling the characteristic pathology observed in neovascular AMD. Four laser spots were created, and mice were treated one day after the laser injury.The efficacy of a single intravitreal dose of D-aflibercept (40 µg / µL) was evaluated (n=6 eyes) on mice with laser induced CNV, which reflects the pathological end stage of wet AMD. The suppression of CNV lesions were measured separately from two individuals (FIG. 8B and FIG. 8C). The CNV lesions were assessed by fluorescein angiography (FFA). The mice with laser CNV created on day 0- and one-day post laser CNV, were treated with D-aflibercept and free aflibercept administrated intravitreally. On day 7, animals (n=6) were sacrificed, the Choroidal flat mounts were stained using lectin and CNV areas were assessed using fluorescein angiography in a blinded manner. Intravitreal free aflibercept and D-aflibercept demonstrated comparable effect in CNV reduction in 7 days. However, at 14 days after a single 40 µg / µL dose of D-aflibercept, CNV area was significantly reduced. CNV area analysis on day 14 (FIG. 7 and FIG. 14) shows >50% reduction compared to free aflibercept, indicating sustainable pharmacodynamic effect upon only single dose injection while free aflibercept shows short-term efficacy in this model.The vascular leakage was assessed by fundus fluorescein angiography (FFA). Intravitreal injection of free-aflibercept and D-aflibercept showed comparable effect in suppressing vascular leakage (p<0.05) at the lesion sites at day 7 when compared with PBS-treated control (Fig. 15A). The reduction in the leakage was sustained up to day 14 (FIG. 15B). (Of note, due to spontaneous reduction (~25%) in vascular leakage in PBS-treated group from healing of the laser lesions, the statistical significances between PBS- and Free-aflibercept / D-aflibercept-treated groups were lost at day 14).Intravitreal D-aflibercept therapy attenuates choroidal inflammation.D-aflibercept therapy produced a significant reduction in multiple pro-inflammatory cytokines (TNF-a, IL-6, and VEGF) compared to free aflibercept (FIGs.8A-8C). The expression of pro-angiogenic factor VEGF was significantly lower after treatment of both aflibercept and D-aflibercept group compared to control group (p=0.031 and p=0.034, respectively). Compared to day 7 on day 14, treatment of D-aflibercept significantly suppressed the expression of VEGF than aflibercept group (p=0.046) and control group (p=0.019) (FIG.8A). Inflammatory cytokines IL-1 was significantly suppressed in D-aflibercept treatment group compared to control group on day 7 (p=0.03) and day 14 (p=0.047). Similarly, the expression of TGF-β was also significantly increased in D-aflibercept group compared to control group on both time points (p=0.040 and p=0.001, respectively). The intravitreally administrated dendrimer-aflibercept localized in the activated macrophages at CNV lesionsOn day 1 after laser CNV, the intravitreally administered dendrimer-aflibercept was assessed for localization at CNV lesions. The intravitreal injected Cy5-D-aflibercept and Cy5-aflibercept demonstrated specific localization to the CNV area (data not shown). The choroidal macrophages in CNV lesions were positive for dendrimer, confirming targeting of inflammatory cells in CNV. Cy5-D-aflibercept was found only in the CNV bulb area. The Cy5-D-aflibercept was still detected in CNV lesions after 21 days whereas free aflibercept is completely cleared by day 7.Delivering aflibercept to the key pathological cells via dendrimer provides significant improvement in efficacy in CNV reduction compared to free afliberceptIntercellular trafficking of aflibercept to the key pathological cells is important. This study showed delivery of aflibercept into cytosol using a dendrimer conjugate (data not shown).The D-aflibercept and free aflibercept binding affinities to key targets in the VEGFA, PGFRA and quantification of VEGF levels under serum starvation of ARPE-19 cellsThe binding properties of D-aflibercept and aflibercept to VEGFA wereassessed usingELISA. As shown in FIGs.9A-9C, D-aflibercept and aflibercept bound human VEGF in dose dependent manner with calculated KD values of 49 pM and 129 pM, respectively (FIG. 9A). KD values were calculated using the GraphPad / Prism software. The amount of unbound VEGF was quantified by ELISA (FIGs.9B-9C). A reduction of free VEGF (serum starvation of ARPE-19 cells) as early as after 24 h of treatment was quantified and free VEGF was reduced by 76% and 89%in the presence of 10 ng / µLD-aflibercept and similar concentrations of aflibercept, respectively (FIG. 9C). The difference in VEGF reduction by D-aflibercept likely depends on its cell internalization capability compared to free aflibercept or may be related to cell viability (D-aflibercept shows better cell viability compared to aflibercept). At 24 h, D-aflibercept is mostly localized in cytosol. However, this fast internalization of D-aflibercept is a disadvantage in early time points to trap extracellular VEGF.The quantification of VEGF levels under LPS-activated ARPE-19 and HMC3 cells.VEGF on LPS-activated HMC3 (human microglia) (FIG. 18A-18D) and ARPE-19 (differentiated) cells (FIG. 19A-19D) were also quantified. The cells were pre-treated with 100 ng / mL lipopolysaccharide (300 endotoxin units) for 3 hours to induce an inflammatory response. Following this pre-activation, the cells were treated with varying concentrations of aflibercept and D-aflibercept at 50, 10, and 1 µg / mL for 12 hours and replenished with LPS-containing media. The effect of aflibercept and D-aflibercept in VEGF reduction was determined up to 4 days of continued stimulation with LPS using the Human VEGF ELISA assay. On the fourth day of the experiment, D-aflibercept exhibited a notable reduction in VEGF levels in both the supernatant and lysate when compared to cells treated with aflibercept. This effect was consistent across all tested concentrations, indicating that D-aflibercept may be more effective at suppressing VEGF production than its counterpart. This is in line with the superior cell internalization of D-aflibercept compared to free aflibercept (FIG. 18C and 18D, and FIG. 19A and 19D). In vitro delivery of D-aflibercept / aflibercept using ARPE-19 cellsIn vitro delivery of D-aflibercept into ARPE-19 cells was assessed. The time dependent uptake was conducted using both Cy5-D-aflibercept and Cy5-aflibercept and cells were incubated for 7 days. The cells were washed prior to imaging at each time point and the treatment media reapplied after each imaging session at least 96 h. At 96 h the treatment media was completely removed and replenished with regular media. As early as 30 min after treatment, D-aflibercept showed intracellular accumulation and within 24 h showed significant accumulation. Although, aflibercept did not accumulate in ARPE 19 cells to any appreciable degree, at 24 h aflibercept seemed to completely wash away. However, even though the level of internalization is not appreciable, aflibercept accumulation is related to the involvement of Fc-receptor, neonatal FcRn, in the uptake and intracellular trafficking of the VEGF-antagonist aflibercept. It was postulated that specific delivery of D-aflibercept to the cytoplasm is a crucial factor for the success of D-aflibercept therapy compared to aflibercept. Cy5-D-aflibercept (Alexa 488) conjugate were synthesized and in vitro localization was assessed. The ARPE-19 cells were treated with Cy5-D-afflibercept (Alexa 488), and cells were incubated for 24 h. FIG. 10A and 10B show fundus fluorescein angiography images and quantification of fluorescein leakage at (A) Day 7 and (B) Day 14 after laser-induced CNV. Intravitreal injection of PBS, Free-Eylea or D-Dylea was performed one-day post laser CNV. n=5-7 (n represents average fluorescence intensity of 3 burns from a single eye).Cy5-D-aflibercept showed intracellular accumulation and dendrimer Cy5 (red) signal is colocalized with aflibercept Alexa 488 (green) signal. The colocalization of Cy5 and Cy3 signal is confirmed by orange signal in the merged panel.Discussion and ConclusionAMD is the leading cause of irreversible central vision loss in the adult population. Even though there is a multifactorial etiology for AMD, the vascular endothelial growth factor is a crucial mediator of pathological angiogenesis. The anti-VEGF agent Aflibercept has shown greater efficacy in suppressing neovascularization. However, it requires chronic intravitreal (IVT) injections to maintain visual function, which poses a significant burden on patients. A dendrimer-aflibercept conjugation strategy was developed in the present study based on mild Cu-free TCO-Tz click reaction conditions via a tunable synthetic route using simple purification and advanced characterization techniques. Interestingly, chemical conjugation does not impair the activity of the aflibercept fusion protein, confirmed by the VEGF dose-response curve. Most importantly, unlike other antibody / antibody fragments or antibody-like molecules, D-aflibercept localized in cells of interest, macrophages, and retained for an extended time. This allows D-aflibercept to neutralize or re-localize the protein of interest VEGF / PIGF. Also, D-aflibercept may gain entry to the macrophages and then binds intracellular VEGF, the complex could be recognized by the Tripartite motif containing-21 (TRIM2), a protein that contains the highest affinity IgG (Fc)binding domain of any mammalian protein and a ubiquitin ligase domain and targeted to degradation by the proteasome. In the standard mouse laser CNV model, dendrimer-aflibercept led to significant CNV suppression over 21 days performing better efficacy than intravitreal aflibercept (positive control). Intravitreal D-aflibercept conjugates specifically target the key pathological cells and macrophages and are retained in the CNV lesions for a longer period (28 days), thus providing significant improvement in efficacy compared to free aflibercept. The enhanced anti-neovascularization ability of D-aflibercept compared to aflibercept further stressed the involvement of inflammation in the progression of choroidal neovascularization. D-aflibercept can regulate the levels of inflammatory cytokines (e.g., IL-1, TGF-β) in the target cells, thus significantly attenuating intracellular VEGF production and reducing CNV area. Further, the single dose of D-aflibercept containing ~ 40 µg / µL of aflibercept retained > 28 days in the bleb area stressed our dendrimer-based delivery system's significantly improved pharmacodynamic effect. The study herein highlights the potential of the covalently conjugated dendrimer-aflibercept platform in advancing the clinical translation as therapeutic for AMD.Thus, a D-aflibercept nano-construct was created in this study using a Cu-free trans-cyclooctene-tetrazine click chemistry strategy for treating AMD. The dendrimer-aflibercept conjugates affect the VEGF suppression in CNV lesions in the eye model. The dendrimer-aflibercept conjugates localized in macrophages within the CNV lesion, and unlike aflibercept, which has extracellular VEGF binding properties, D-aflibercept internalized to cells of interest and localizes in the cytoplasm as confirmed by in vivo / in vitro analysis. Interestingly, intravitreal injection of D-aflibercept led to significant therapeutic efficacy in CNV suppression over 14 days in mouse laser CNV model performing better than intravitreal aflibercept control. Most importantly, dendrimer-based intracellular delivery of aflibercept seems to attenuate VEGF / PDGF activity at the protein level. This strategy has potential to clinically deliver aflibercept into intracellular targets that will extend therapeutic efficacy in AMD. Example 3:Dendrimer-Adalimumab (HUMIRA®) conjugateAdalimumab is a tumor necrosis factor blocker. Adalimumab is a recombinant human IgG1 monoclonal antibody created using phage display technology resulting in an antibody with human derived heavy and light chain variable regions and human IgG1:k constant regions. Adalimumab is produced by recombinant DNA technology in a mammalian cell (Chinese Hamster Ovary (CHO)) expression system and is purified by a process that includes specific viral inactivation and removal steps. It consists of 1330 amino acids and has a molecular weight ofapproximately 148 kilodaltons. HUMIRA® is a tumor necrosis factor (TNF) blocker indicated for: Rheumatoid Arthritis (RA); Juvenile Idiopathic Arthritis (JIA); Psoriatic Arthritis (PsA); Ankylosing Spondylitis (AS); Crohn’s Disease (CD); Ulcerative Colitis (UC); Plaque Psoriasis (Ps); Hidradenitis Suppurativa (HS); and Uveitis (UV). The recommended subcutaneous dosage of HUMIRA® for adult patients with rheumatoid arthritis (RA), psoriatic arthritis (PsA), or ankylosing spondylitis (AS) is 40 mg administered every other week. Methotrexate (MTX), other non-biologic DMARDS, glucocorticoids, nonsteroidal antiinflammatory drugs (NSAIDs), and / or analgesics may be continued during treatment with HUMIRA®. In the treatment of RA, some patients not taking concomitant MTX may derive additional benefit from increasing the dosage of HUMIRA® to 40 mg every week or 80 mg every other week. Materials and MethodsA hydroxyl-terminated poly(amidoamine), PAMAM dendrimer-conjugated Adalimumab (D-Adalimumab) was constructed using trans-cyclooctene-tetrazine (TCO-TZ) click chemistry strategy. The click reaction was performed at near neutral pH, aqueous conditions at room temperature. MALDI-TOF MS, and analytical HPLC traces were used to confirm the synthesis of all the intermediates and final dendrimer adalimumab conjugates. D-Adalimumab was further analyzed using a standard SDS-PAGE (4-12% gradient gel) followed by Coomassie blue staining for the protein visualization on the gel. The chemical structure of Cy5-D-Adalimumab is  The efficient cellular delivery of Cy5-D-Adalimumab was studied by confocal microscopy. C57 / BL6 derived microglial (BV-2) cells were treated with Cy5-D-Adlimumab. The D-Adalimumab and free Adalimumab binding affinities to key targets in the Tumor necrosis factor alpha (TNF-α) and quantification of TNF-α levels with LPS / without LPS in THP-1 cells were then measured. Human derivedTHP-1Monocytes and Macrophages were incubated 3 h with LPS (100 ng / mL) / without LPS containing cell culture media and treated with varying concentrations of Adalimumab and D-Adalimumab. TNF-α quantification was conducted using Human TNF-α ELISA kit following manufacture’s protocols. A) Cell lysate B) Cell supernatant.Synthesis of functionalized Cy5-D-PEG4-TCOFIG. 20 shows the synthesis of functionalized Cy5-D-PEG4-TCO. The hydroxyl PAMAM dendrimer generation 6 (PAMAM-G6-OH) was treated with 4-tert-butoxycarbonylamino)butyric acid (Boc-protected GABA) linker, 2 and the resulted product, 3 was deprotected using dichloromethane (DCM) / trifluoroacetic acid (TFA) (4:1). The product, 4 was labeled with Cy5 fluorophore using Cy5 N-hydroxysuccinimide (NHS) ester and the resulted intermediate, 5 was conjugated with trans-cyclooctene (TCO) linker, PEG4-TCO to obtain functionalized Cy5-D-PEG4-TCO 6. The subscripted numbers in the formulas indicate the number of GABA BOC, PEG4-TCO, or flurophore attached per dendrimer.Synthesis of Cy5-D-Adalimumab conjugate FIG. 21 is a schematic of the synthesis of Cy5-D-Adalimumab conjugate. Modification of Adalimumab antibody for conjugation with dendrimer. Adalimumab was first substituted with PEGylated tetrazine using NHS-PEG2-Tz reagent to form aflibercept-Tz 7, then 7 was reacted with Cy5-D-PEG4-TCO, 6 to obtain the final product Cy5-D-aflibercept, 8.The Adalimumab antibody was functionalized with terminal tetrazine (Tz), while D6-OH was functionalized with trans-cyclooctene (TCO) for click reaction (Scheme 1 (FIG. 20) and Scheme 2 (FIG. 21)). The Adalimumab (1 mg in 250 µL of PBS) was treated with Tz-PEG2-NHS ester (20 mol equiv in 10-20 µL of anhydrous DMSO) and incubated for 1h. The excess Tz-PEG2-NHS and byproducts were removed by ultrafiltration. Product 8 was further purified by being equipped with size-exclusion column chromatography (SEC). The number of Tz groups attached per aflibercept was determined based on the change in the molecular weight measured by the MALDI-TOF mass spectrometer. The number of groups attached is denoted as the subscripted number. The TCO-PEG4 attached dendrimer 6 wasreacted with 7 via trans-cyclooctene-tetrazine (TCO-Tz) to afford crude product 8. The resulting crude product was purified by ultrafiltration, and the product was further purified by a Fast Protein Liquid Chromatography (FPLC) system (AKTA FPLC, 280 nm single wavelength detector) equipped with SEC column. The TCO-Tz “click” reaction proceeds via an inverse-electron demand Diels-Alder reaction (IEDDA) between a TCO and a Tz, followed by retro Diels-Alder reaction eliminating N2 to form a dihydropyridazine bond. D-adalimumab was further analyzed using a standard SDS-PAGE (4-12% gradient gel) followed by Coomassie blue staining for the protein visualization on the gel (data not shown). Confocal microscopy further demonstrated the cellular uptake of Cy5-D-Adalimumab cell uptake into BV-2 cells after 24 h treatment (FIG. 22). The efficient cellular delivery of Cy5-D-Adalimumab was studied by confocal microscopy. The C57 / BL6 derived microglial (BV-2) cells were treated with Cy5-D-Adlimumab. After 24 h incubation, dendrimer-Adalimumab (red fluorescence) showed significant accumulation in BV-2 cells. (FIG. 22). Thus the results demonstrate improved uptake of the dendrimer-adalimumab in BV-2 microglia cells.ResultsThe binding affinities of D-Adalimumab and free Adalimumab to key targets in the Tumor necrosis factor alpha (TNF-α), and the TNF-α levels with LPS or without LPS in THP-1 cells were quantified..Representative confocal microscopy images and graphs of Cy5-D-Adalimumab cell uptake intoBV-2 cells after 24 h treatment, cell nucleus (Blue), dendrimer (Red) are shown in FIG. 12A-12D. FIG. 12A and 12B show no treatment and FIG. 12C and 12D shows percent Cy5 positive cells in Cy5-D-Adalimumab treated cells compared to BV-2 cells as determined by FACS.FIG. 13A-13C and 13D-13F are graphs of the THP-1 monocytes and THP-1 derived macrophages with LPS (100 ng / mL) / without LPS treated with varying concentrations of D-Adalimumab and Adalimumab.After 24 h incubation, dendrimer-Adalimumab (red fluorescence) showed significant accumulation in BV-2 cells.SummaryIn a proof-of-concept study, the efficacy of the dendrimer-TNFa antibody conjugate was assessed by measuring the TNF-α level in the cell lysate (FIG. 12A, 12B) and cell supernatant (FIG. 12C, 12D), using Human TNF-α ELISA kit. The LPS treated groups showed significant upregulation of TNF-α levels compared to non-treated groups. Both 1 ng / µL and 0.1 ng / µL Dendrimer-Adalimumab samples showed significant reduction of TNF-α levels in cell lysate and cell supernatant samples compared to free Adalimumab groups, indicating that dendrimer-delivery enhances the efficacy of the antibody. In vivo, the dendrimer-antibody conjugate target specific cells, and produces additional advantages of injured-cell targeted intracellular delivery of antibodies. Given that the glucose dendrimer can target neuronal cells (neurons, ganglion cells, photoreceptors) and reactive inflammatory cells, this provides a means for greatly improved delivery of intracellular antibodies to a variety of cells.Example 4:Improved uptake and efficacy of dendrimer-adalimumab in THP-1 human monocytesStudies have suggested that monocytes in patients with RA express and secrete more TNFα (Li et al). The THP1 monocyte cell line has a higher expression of transmembrane TNFα when compared to healthy, normal monocytes (Li et al). Therefore, this cell line is a relevant cell line to test for adalimumab cellular uptake. Materials and Methods:Cell culture.To evaluate the efficacy of HD-ADA, cell studies were performed. THP-1 human acute monocyte leukemia cells were obtained from the American type culture collection (ATCC) and cultured in RPMI-1640 medium (Cat# A1049101, ThermoFischer Scientific, USA) supplemented with 10% fetal bovine serum (Cat#F2442, Millipore Sigma, USA), 1% Antibiotic / antimycotic solution (Penicilin G, Streptomycin, Amphotericin B - Cat# SV30079.01, Cytiva, HyClone Laboratories, USA) and 0.05 mM 2-mercaptoethanol (Sigma-Aldrich, USA) in 5% CO2 at 37°C. The morphological features of cells in suspension and adherent cells were evaluated under a light microscope. Images were taken through a Motic AE31E inverted microscope with a PLAN LWD 20× / 0.3 Ph1 objective and equipped with a Moticam 3.0 MP camera. Experimental design.Cells were seeded at seeded 1x106 cells / ml into a 12 well plate and then differentiated into macrophage by stimulation with 100 nM PMA for 24 h followed by a 24-hour rest period without PMA. To evaluate the monocyte population, same conditions as macrophages were kept except that the cells were exposed to VEH. In order to mimic the pro-inflammatory milieu that drives ADA prescription, macrophages, as well as monocytes, were exposed to LPS stimulus for 4 h (100 ng / mL; Sigma Aldrich). Non-LPS-exposed cells were used as control. After that, treatments (ADA or HD-ADA) were added accordingly to each well at the following concentrations 0, 0.1 or 1 ug / ml for 4 h. After that, a wash out step was performed and complete medium was added to every well. 18-20 h later, samples were collected for biochemical studies. To evaluate the preventive effect of the treatment, non-LPS-exposed cells were exposed to the treatments for 4 h and a final washout step followed by the addition of LPS stimulus was performed. Samples (cell lysates and supernatants) were collected 18-20 h later. To evaluate the prolonged effect of the treatment, cells were exposed to LPS for 4 h followed by treatments for another 4 h. A final washout step followed by the addition of LPS (100 ng / ml) was included in the experimental design and samples (cell lysates and supernatants) were also collected 18-20 h later. Studies in monocytes and macrophages were run in parallel at the same time and three independent experiments were run for each cell lineage. Experimental design is shown in each figure.Cell lysates. To evaluate intracellular efficacy, cells were washed twice with 1X cold PBS to remove residual medium. 100 ul of 1X cold cell lysis buffer (Cat# 9803, Cell signaling, USA) was added to lyse cells under nondenaturing conditions. The lysing procedure was performed according to the manufacturer instructions. Briefly, cells exposed to the lysis buffer were incubated on ice for 5 min. Samples were centrifuged at 14,000 g during 10 min at 4°C. Supernatant was removed to assess TNF-α.ELISA for TNFα measurement.Cell lysates and supernatants from THP-1 monocytes and THP-1-derived macrophages were analyzed for TNF-α by human TNF-α ELISA kit (Cat#KHC3014 and Cat#KHC3011, ThermoFisher Scientific, USA) according to the manufacturers’ protocols and analyzed in FlexStation 3 plate reader through SoftMax® Pro7.1 software. Total protein content was determined by using PierceTM BCA protein assay kit (Cat#23225, ThermoFischer Scientific, USA) and same concentration of total proteins for each sample was used to performed the TNF-α measurement through ELISA assay. Data reported correspond to 100 ug of total proteins in 100 ul of sample seeded in the ELISA plate. Flow cytometry.Similar experimental design as described above was performed except that the cells were treated with Cy5-HD or Cy5-HD-ADA at the same concentrations (0, 0.1, 1 ug / ml). Cells were exposed to a solution of trypsin 0.05% (1X, Cat#SH30236.01; Cytiva, USA) for 10 min at 37°C, collected in a centrifuge tube and washed twice with PBS 1X. Then 10% formalin for 15 min was used for cell fixation. To avoid clusters and wall attachment to the tube during centrifugation, EDTA (Corning, USA) was added to the cell suspension. Cells were analyzed using a FACs cytometer (BD® LSR II Flow Cytometer, USA); and at least 10,000 events were included in the analysis. Data were analyzed by FlowJoX software. THP-1 monocytes and THP-1-derived macrophages were gated by forward scatter – area (FSC-A) and side scatter – area (SSC-A) to exclude debris and then doublets were excluded by performing a forward scatter height (FSC-H) vs forward scatter area (FSC-A) density plot. Finally, single cells were plotted as FSC-H vs Cy5 signal. The percentage of Cy5+ cells and median fluorescence intensity (MFI) for the single cells was calculated.  ResultsIncreased uptake of Dendrimer-adalimumab (D-Ada) in THP1 monocytes while dendrimer alone does not localize in monocytesFlow cytometry of THP1 monocytes exposed to D-CY5 or D-Ada-Cy5 showed that D-Cy5 did not localize in the monocytes. However, D-Ada-Cy5 was rapidly taken up into the THP-1 monocytes both with and without LPS stimulation. Since THP1 cells normally have high expression of transmembrane TNFα (indicating a pro-inflammatory state at baseline), the increased uptake and binding was observed even without LPS stimulation. A dose response was observed in the uptake with higher dose of D-Ada showing greater uptake as demonstrated by mean fluorescence intensity. Close to 100% of cells take up some D-Ada-Cy5 (FIG. 23A) but the extent of uptake is dose dependent (FIG. 23C). Since HD-Ada-Cy5 and HD-Cy5 were exposed only for 4 hours and was then washed out and fluorescence was measured after ~18 hours in the cells, the fluorescence intensity is only indicative of intracellular D-Ada-Cy5. This result indicates that Adalimumab conjugation leads to localization of dendrimer in THP1 monocytes that is not seen with dendrimer alone. D-Ada is effective in decreasing intracellular TNFα levels while Adalimumab alone does not in THP1 monocytes. Adalimumab acts by binding and inhibiting soluble TNFα (sTNFα) that is secreted and to a lesser extent also transmembrane TNFα (tmTNFα) (Li et al; Mease et al,; Oberoi et al). However, antibodies are not internalized and can only affect intracellular TNFα by indirect effects on sTNFα and tmTNFα (Zamora-Atenza et al). It was observed that D-Ada is better than Adalimumab alone at these doses in inhibiting the secreted sTNFα (FIG. 24C), and importantly, D-Ada inhibits intracellular TNFα in the cell lysate while free adalimumab at these doses does not (FIG. 24B). Since this is only after a 4 hour exposure to D-Ada and the treatment is removed and cell lysate tested after ~18 hours, this indicates the effect of D-Ada that has internalized and is acting intracellularly for a prolonged period. This demonstrates that not only does D-Ada deliver the Antibody intracellularly, it is also functional and demonstrates efficacy in inhibiting TNFα levels inside the cell. Example 5:Improved uptake of dendrimer-adalimumab in HMC3 human microglia cellsMaterials and MethodsCell linesHMC3, a human microglia cell line, was used for this study. Cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM, ATCC, Manassas, VA) supplemented with 10% (v / v) heat-inactivated fetal bovine serum (FBS, Invitrogen Corp., Carlsbad, CA) and 1% penicillin / streptomycin (P / S, Invitrogen Corp., Carlsbad, CA). Cells were maintained at 37°C and 5% CO₂ in a humidified atmosphere.Quantification of TNF-α by ELISAHMC3 cells were seeded in 96-well plates at a density of 10,000 cells per well and cultured under standard conditions (37°C, 5% CO₂). To induce an inflammatory response, the cells were pre-treated with 100 ng / mL lipopolysaccharide (LPS, derived from Escherichia coli O127:B8, Lot#125M4091V) (Sigma-Aldrich, St. Louis, MO) for 3 hours. After LPS pre-activation, cells were treated with varying concentrations of Adalimumab and HD-Adalimumab (10, 1, and 0.1 µg / mL) for 12 hours. The medium was aspirated after treatment, and the cells were gently washed with warm PBS. Fresh culture medium containing 100 ng / mL LPS was added to maintain the inflammatory environment, and cells were incubated for 1, 3, and 4 days. At each time point, the culture medium was collected and centrifuged at 5,000 rpm for 5 minutes at 4°C, and the supernatant was stored at -80°C until analysis. TNF-α levels in the collected media were quantified using a Human TNF-α ELISA kit (Cat#KHC3014 and Cat#KHC3011, ThermoFisher Scientific, USA), following the manufacturer's protocol. Absorbance was measured at 450 nm using a microplate reader, with a wavelength correction at 570 nm. Untreated (resting) cells and cells treated with LPS alone were controls.Flow Cytometry and Confocal MicroscopyHMC3 cells were seeded in 35 mm petri plates at a density of 100 cells per well and cultured under standard conditions (37°C, 5% CO₂). Cells were treated with Cy5-Adalimumab or Cy5-HD-Adalimumab (20 µg / mL). After 12 hours of incubation, the cells were washed three times with PBS (pH 7.4). The cells were then stained with DAPI (10 µg / mL) for 5 minutes, followed by three washes with PBS (pH 7.4). Confocal microscopy was used to visualize the cells. For flow cytometry, cells were detached using 0.05% trypsin (Cat#SH30236.01; Cytiva, USA), collected in centrifuge tubes, and washed twice with PBS. Cells were fixed with 10% formalin for 15 minutes, and centrifugation was performed for 5 minutes at 2000 RPM. Cells were analyzed using a BD® LSR II Flow Cytometer (USA); at least 10,000 events were included in the analysis. Data were processed using FlowJoX software. The percentage of Cy5+ cells and the median fluorescence intensity (MFI) were calculated.Results and DiscussionEvaluation of the in vitro efficacy of Adalimumab and HD-AdalimumabHMC3 cells are a suitable and reliable in vitro model for evaluating immune response modulation under activated and resting conditions. HD-Adalimumab therapy significantly reduced pro-inflammatory cytokine (TNF-α) levels compared to free Adalimumab. TNF-α expression was notably lower in the Adalimumab and HD-Adalimumab-treated groups than in the control. From day 3 to day 4, HD-Adalimumab treatment suppressed TNF-α expression more effectively than free Adalimumab and the control group (FIG. 25A and 25B).Evaluation of the in vitro cellular uptake of Cy5-Adalimumab and Cy5-HD-AdalimumabA more significant increase in Cy5 cells was observed in those treated with Cy5-HD-Adalimumab (20 µg / mL), compared to Cy5-Adalimumab or untreated cells, regardless of LPS stimulation. This trend was consistent with MFI results after LPS activation, indicating higher cellular uptake of Cy5-HD-Adalimumab compared to Cy5-Adalimumab for flow cytometry and confocal microscopy. Our data demonstrate a significant increase in MFI following treatment with Cy5-HD-ADA at 20 µg / mL (non-treatment vs. Cy5-HD-Adalimumab, and Cy5-Adalimumab vs. Cy5-HD-Adalimumab: p < 0.01) (FIG. 26A and 26B).Example 6:In vivo targeting and intracellular delivery of antibody to microglia in the brain by dendrimer-adalimumab, upon systemic administration in a rabbit CP modelAdalimumab (monoclonal antibody targeting human TNF-α) was used for this proof-of-concept study demonstrating delivery of the antibody to cells in the brain. Dendrimer conjugated with Cy5 and adalimumab was injected intravenously to newborn rabbit kits with neuroinflammation and brain injury. Brains were collected after 24 hours, sectioned and stained with Iba1 for identifying microglia in the brain. Adalimumab was detected by immunohistochemistry using anti-adalimumab tagged with Alexa-488 (data not shown). Since adalimumab is a human antibody, it will not bind rabbit TNF-α. Imaging data shows strong co-localization of D-Ada-Cy5 in Iba1 stained microglia indicating that D-Ada-Cy5 is inside microglial cells (data not shown). The presence of adalimumab inside the cell is confirmed by staining directly for the adalimumab with Anti-Ada Ab (data not shown). The merged image shows co-localization of D-Ada-Cy5 with Anti-Ada Ab and Iba1, indicating that systemically administered D-ada crosses the blood brain barrier intact and is delivered into microglial cells. Antibody delivery to the brain is often difficult to achieve since antibodies do not usually cross the BBB (blood brain barrier). In the healthy brain, the BBB is not impaired and this large dendrimer-antibody conjugate will not be transported. In the injured brain, in the areas of pathology, the BBB is impaired, and the conjugate is transported. The results here relate to the fact that the dendrimer is able to transport, even a large antibody, 2-3 times its size (in molecular weight), into the specific injured cells, preferred by the dendrimer. This has implications for targeted treatment of neuroinflammatory and neurodegenerative diseases such as multiple sclerosis, ALS, Parkinson’s disease, Alzheimer’s, seizures, neuromyelitis optica, glioblastoma, peripheral neuropathies, CNS complications of CRS (cytokine response syndrome) etc. Modifications and variations of the present invention will be apparent to those skilled in the art and are intended to come within the scope of the appended claims. Important Note: The system was unable to import the complete document, including the compounds and tables. Therefore, these sections have been removed from the imported document. Please refer to the complete document attached under (Other Attachments) for the full content. 

Claims

1. A composition comprising dendrimers covalently conjugated to at least one antibody, nanobody or fragment thereof binding to an intracellular target.

2. The compositions of claim 1, where the antibody or fragment / fusion protein thereof is covalently conjugated by a covalent link between a modified or unmodified surface group or interior groups of the dendrimer, such as a disulfide, ester, amide, or ether linkage.

3. The composition of any of claims 1or 2, wherein the dendrimer is a polyamidoamine (PAMAM) dendrimer of generation 1 – generation 9, with between greater than 40 and 100% of the surface groups being hydroxylated.

4. The composition of claim 1, wherein the dendrimer is a generation 2 – generation 7 PAMAM dendrimer, modified by a sugar moietyselected from the group consisting of glucose, galactose, mannose, and fructose.

5. The compositions of claim 3, wherein the dendrimer is a glucose dendrimer of generation 1, 2, or 3.

6. The compositions of claim 1, wherein the dendrimer is a glucose dendrimer, with a central core of dipentaerythritol and one or more branching units of monosaccharide glucose and ethylene glycol, with >10 surface glucose moieties.

7. The composition of any of claims 1-6, wherein the antibody or fragment thereof is a single chain antibody, single chain variable fragment (scFv), disulfide linked Fvs (sdFv), Fab, Fab', F(ab')2, Fv, and single domain antibody fragment (sdAb) or nanobody.

8. The composition of any of claims 1-7, wherein the antibody or fragment thereof binds specifically to one or more of Tau antibody, CTLA-4. PD-1, PD-L1, and CD20.

9. The composition of any of claims 1-7, wherein the antibody or fragment thereof binds specifically to one or more proteins involved in NLRP3 inflammasome activation selected from the group consisting of NLRP3, Caspases, RIP1-3, FADD, MLKL, Interleukins, and pro-interleukins.

10. The composition of any of claims 1-7, wherein the antibody or fragment thereof binds specifically to one or more intracellular inflammatory signaling proteins selected from the group consisting of JAK-STAT, Nrf2, STAT 1, STAT 3, STAT 6, MAPK, MEK, ERK1 / 2, and HIF1a.

11. The composition of any of claims 1-7, wherein the antibody or fragment thereof binds specifically to TRIM21.

12. A pharmaceutical formulation comprising the composition of any one of claims 1-11, and one or more pharmaceutically acceptable carrier or excipientsfor systemic or local administration.

13. A method of administering the composition or formulation of any one of claims 1-12 to a subject in need thereof having one or more of ocular diseases, inflammatory disorders, neurological disorders, and cancer.

14. The method of claim 13, wherein the subject in need thereof hasone or more diseases or conditions selected from the group consisting of arthritis, diabetic nephropathy, renal interstitial disease, fibrosis, multiple sclerosis, irritable bowel syndrome and Crohn’s disease, autoimmune disorders, traumatic brain injury, acute pain, chronic inflammatory pain, neuropathic pain, or glaucoma, inflammatory disorders.

15. The method of claim 13, wherein the subject in need thereof has one or more diseases or disorders of the eye selected from the group consisting of diabetic eye disease, symptomatic vitreomacular adhesion / vitreomacular traction (sVMA / VMT), wet (neovascular) age-related macular degeneration (AMD), and dry AMD.

16. The method of claim 13, wherein the subject in need thereof has one or more retinal and choroidal vascular diseases selected from the group consisting of AMD, retinopathy of prematurity, diabetic macular edema, retinal vein occlusion, and retinopathy associated with toxicity of chemotherapy.

17. The method of claim 13, wherein the composition or formulation is administered in an amount effective to reduce and / or inhibit the number or activities of activated microglia and macrophages in the retina and / or the choroid in the eye of a subject in need thereof.

18. The method of claim 13, wherein the composition or formulation is administered in an amount effective to decrease expression of one or more pro-inflammatory cytokines selected from the group consisting of TNF-α, interleukin-1β (IL-1β), and interferon-γ (IFN-γ). or 19. The method of claim 13, wherein the subject in need thereof has one or more neurological disorders selected from the group consisting of Huntington disease, synucleinopathies, Alzheimer’s disease, prion disease, and amyotrophic lateral sclerosis (ALS), and wherein the dendrimer-antibody conjugates are in an amount effective to target one or more neuronal cells (including ganglion cells), peripheral neurons, and photoreceptors associated with the site of pathology.

20. The method of claim 13 wherein the subject in need thereof has rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, plaque psoriasis, hidradenitis suppurativa, uveitis, or juvenile idiopathic arthritis.   

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