Processes for the filtration or purification of viral particles

Abstract

Provided for herein are methods of purifying and / or concentrating viral vectors, methods of using the same to treat diseases, such as cancer or autoimmune diseases, and methods of producing sterile compositions, such as, sterile pharmaceutical compositions, wherein the methods can perform steps after passing the viral vector through a protein coated sterile filter.

Description

DOCKET NO: INH-024WO PATENT PROCESSES FOR THE FILTRATION OR PURIFICATION OF VIRAL PARTICLES RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application Ser. No. 63 / 482,617 filed February 01, 2023, and U.S. Provisional Application Ser. No. 63 / 510,391 filed June 27, 2023, each of which are hereby incorporated by reference in their entirety. REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY

[0002] The instant application contains a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. Said XML copy, created on January 20, 2024, is named “INH-024WO_SL” and is 97,699 bytes in size. FIELD

[0003] Embodiments provided herein relate to the process of production of viral vectors, involving filtration. BACKGROUND

[0004] Sterile filtration is typically performed at the end of large biological molecule (protein, monoclonal antibody, viral vector) processing to remove bioburden and produce sterile drug substance. Many sterile filters are available commercially, with variations in pore size, material of construction, surface charge of the membrane, pore morphology, and filter membrane format. Any of the variables could affect recovery of the product of interest across the filter. Loss of product across sterile filtration has been reported for viruses due to physical retention of the product, related to either size and / or charge. Investigation of sterile filter types and feed concentration to maximize recovery of a particular product of interest have been reported. The present invention addressed these needs and others. BRIEF SUMMARY

[0005] In some embodiments, a method of purifying a viral vector or a method producing a concentrated sterilized solution are provided, the methods comprising the steps of clarifying a solution comprising cell culture media and the viral vector; filtering the clarified solution comprising the viral vector through a first chromatographic filter to produce a filtered clarified solution comprising the viral vector; passing the filtered clarified solution comprising the viral vector through a protein coated sterile membrane to produce a sterilized solution IPTS / 126938130.1DOCKET NO: INH-024WO PATENT comprising the viral vector; and concentrating the sterilized solution comprising the viral vector to produce a concentrated sterilized solution comprising the viral vector.

[0006] In some embodiments, a protein solution comprises one or more component selected from a protein, 2-Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS), histidine, phosphate, HEPES ((4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), sodium chloride, or combinations thereof.

[0007] In some embodiments, the protein solution comprises about 0.1% w / v to about 3.0% w / v of a protein, about 20 mM 2-Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS), and about 100 mM to about 400 mM sodium chloride.

[0008] In some embodiments, the protein coated sterile membrane has a positive charge prior to being coated with a protein.

[0009] In some embodiments, the membrane is selected from a polyethersulfone (PES) membrane, a nylon membrane, or a polyvinylidene fluoride (PVDF) membrane.

[0010] In some embodiments, the method comprises the steps of digesting DNA in a solution comprising the viral vector to produce a first digested solution comprising the viral vector; clarifying the digested solution comprising the viral vector; filtering the clarified, digested solution comprising the viral vector through a first chromatographic filter to produce a filtered, clarified, digested solution comprising the viral vector; digesting DNA in the filtered, clarified, digested solution comprising the viral vector to produce a filtered, clarified, twice- digested solution comprising viral vector; filtering the filtered, clarified, twice-digested solution comprising the viral vector through a second chromatographic filter to produce a twice-filtered, clarified, twice-digested solution comprising viral vector; passing the twice- filtered, clarified, twice-digested solution comprising the viral vector through a protein coated sterile membrane to produce a sterilized solution comprising the viral vector; concentrating the sterilized solution comprising the viral vector to produce a concentrated sterilized solution comprising the viral vector.

[0011] In some embodiments, the protein solution comprises one or more component selected from a protein, 2-Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS), histidine, phosphate, HEPES ((4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), sodium chloride, or combinations thereof.

[0012] In some embodiments, the protein solution comprises about 0.1% w / v to about 3.0% w / v of a protein, about 20 mM 2-Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS), about 100 mM to about 400 mM sodium chloride, and a pH of about 6.0 to about 8.0. -2- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT

[0013] In some embodiments, the protein coated sterile membrane has a positive charge prior to being coated with a protein.

[0014] In some embodiments, the membrane is selected from a polyethersulfone (PES), a nylon membrane, or a polyvinylidene fluoride (PVDF) membrane. BRIEF DESCRIPTION OF FIGURES

[0015] FIG.1 illustrates a viral vector production process.

[0016] FIG.2 illustrates the recovery of a viral vector from PES with and without pre- use flush with protein.

[0017] FIG.3 illustrates the recovery of a viral vector from an uncharged sterilefilter.

[0018] FIG.4 illustrates the impact of pre-coating on a charged sterile filter.

[0019] FIG.5 illustrates the impact of pre-coating on a neutral sterile filter.

[0020] FIG.6 illustrates that both HSA and recombinant HSA may be used as a sterile filter pre-coating protein solution.

[0021] FIG.7A and FIG.7B illustrate crystal structures of VSV-G bound to LDL-R.

[0022] FIG. 8A and FIG. 8B illustrate the effect of adding negatively charged amino acids to the VSV-G:LDL-R binding interface on native tropism and fusogenicity.

[0023] FIG.9 illustrates an alignment of the ectodomains of different VSV-G proteins from different strains. DETAILED DESCRIPTION

[0024] Unless otherwise defined, scientific and technical terms used herein have the meanings that are commonly understood by those of ordinary skill in the art. In the event of any latent ambiguity, definitions provided herein take precedent over any dictionary or extrinsic definition. Unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. The use of “or” means “and / or” unless stated otherwise. The use of the term “including,” as well as other forms, such as “includes” and “included,” is not limiting.

[0025] Generally, nomenclature used in connection with cell and tissue culture, molecular biology, immunology, microbiology, genetics and protein and nucleic acid chemistry and hybridization described herein is well-known and commonly used in the art. The methods and techniques provided herein are generally performed according to conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout the present specification unless otherwise indicated. -3- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT Enzymatic reactions are performed according to manufacturer’s specifications, as commonly accomplished in the art or as described herein. The nomenclatures used in connection with, and the laboratory procedures and techniques of, analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein are those well-known and commonly used in the art.

[0026] That the disclosure may be more readily understood, select terms are defined below.

[0027] As used herein, the terms “a” or “an” means that “at least one” or “one or more” unless the context clearly indicates otherwise.

[0028] As used herein, the term “about” means that the numerical value is approximate and small variations would not significantly affect the practice of the disclosed embodiments. Where a numerical limitation is used, unless indicated otherwise by the context, “about” means the numerical value can vary by ±10% and remain within the scope of the disclosed embodiments. Additionally, where a phrase recites “about x to y,” the term “about” modifies both x and y and can be used interchangeably with the phrase “about x to about y” unless context dictates differently.

[0029] As used herein, the term “individual” or “subject,” or “patient” used interchangeably, means any animal, including mammals, such as mice, rats, other rodents, rabbits, dogs, cats, swine, cattle, sheep, horses, or primates, such as humans.

[0030] As used herein, the terms “comprising” (and any form of comprising, such as “comprise”, “comprises”, and “comprised”), “having” (and any form of having, such as “have” and “has”), “including” (and any form of including, such as “includes” and “include”), or “containing” (and any form of containing, such as “contains” and “contain”), are inclusive or open-ended and do not exclude additional, unrecited elements or method steps. Any step or composition that uses the transitional phrase of “comprise” or “comprising” can also be said to describe the same with the transitional phase of “consisting of” or “consists.”

[0031] As used herein, the term “pre-flushed” or “pre-coated”, or any variation of these terms such as “pre-flushing” or “pre-coating” describes a filter, wherein a solution has been passed through the filter before the filter is used with a viral vector solution.

[0032] As used herein, the phrase “viral vector recovery” is defined as the amount of viral vector post-filtration compared to the amount of viral vector pre-filtration expressed as a percentage.

[0033] As used herein, the phrase “purified viral vector” is defined as viral vector which has been purified from cell culture harvest using at least one chromatography step. -4- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT

[0034] As used herein, “non-concentrated” viral vector is a viral vector in a solution that has not undergone a concentration step during its production. As used herein, a “non- concentrated” viral vector does not have a limit on the concentration of viral particles therein, but rather, unless explicitly state otherwise, solely denotes whether a solution comprising a viral vector has undergone a concentration step or not.

[0035] As used herein, the term “contacting” means bringing together of two elements in an in vitro system or an in vivo system. For example, “contacting” virus or vector described herein with an individual or patient or cell includes the administration of the virus to an individual or patient, such as a human, as well as, for example, introducing a compound into a sample containing a cellular or purified preparation containing the cell.

[0036] As used herein, the term “fused” or “linked” when used in reference to a protein having different domains or heterologous sequences means that the protein domains are part of the same peptide chain that are connected to one another with either peptide bonds or other covalent bonding. The domains or section can be linked or fused directly to one another or another domain or peptide sequence can be between the two domains or sequences and such sequences would still be considered to be fused or linked to one another. In some embodiments, the various domains or proteins provided for herein are linked or fused directly to one another or a linker sequences, such as the glycine / serine sequences described herein link the two domains together.

[0037] A “disease” in an animal is a state of health wherein the animal cannot maintain homeostasis, and wherein if the disease is not ameliorated then the animal’s health continues to deteriorate. In contrast, a “disorder” in an animal is a state of health in which the animal is able to maintain homeostasis, but in which the animal’s state of health is less favorable than it would be in the absence of the disorder. Left untreated, a disorder does not necessarily cause a further decrease in the animal’s state of health.

[0038] “Effective amount” or “therapeutically effective amount” are used interchangeably herein, and refer to an amount of a compound, formulation, material, or composition, as described herein effective to achieve a particular biological result or provides a therapeutic or prophylactic benefit. Such results may include, but are not limited to an amount that when administered to a mammal, causes a detectable level of immune cell activation compared to the immune cell activation detected in the absence of the composition. The immune response can be readily assessed by a plethora of art-recognized methods. The skilled artisan would understand that the amount of the composition administered herein varies and can be readily determined based on a number of factors such as the disease or condition being -5- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT treated, the age and health and physical condition of the mammal being treated, the severity of the disease, the particular compound being administered, and the like.

[0039] “Encoding” refers to the inherent property of specific sequences of nucleotides in a polynucleotide, such as a gene, a cDNA, or an mRNA, to serve as templates for synthesis of other polymers and macromolecules in biological processes having either a defined sequence of nucleotides (i.e., rRNA, tRNA and mRNA) or a defined sequence of amino acids and the biological properties resulting therefrom. Thus, a gene encodes a protein if transcription and translation of mRNA corresponding to that gene produces the protein in a cell or other biological system. Both the coding strand, the nucleotide sequence of which is identical to the mRNA sequence and is usually provided in sequence listings, and the non-coding strand, used as the template for transcription of a gene or cDNA, can be referred to as encoding the protein or other product of that gene or cDNA.

[0040] “Expression vector” refers to a vector comprising a recombinant polynucleotide comprising expression control sequences operatively linked to a nucleotide sequence to be expressed. An expression vector comprises sufficient cis-acting elements for expression; other elements for expression can be supplied by the host cell or in an in vitro expression system. Expression vectors include all those known in the art, such as cosmids, plasmids (e.g., naked or contained in liposomes) and viruses (e.g., Sendai viruses, lentiviruses, retroviruses, adenoviruses, and adeno-associated viruses) that incorporate the recombinant polynucleotide.

[0041] As used herein, the phrase “ex vivo” in reference to a cell being transduced, transfected or transformed ex vivo, refers to a cell being transduced, transfected or transformed outside of the subject, that is with the cells being removed from the subject before such cells are transduced, transfected or transformed.

[0042] As used herein, “filter” refers to any substance through which a solution or composition is passed to remove portions (e.g.) of the solution or composition. Therefore, “filter” is not meant to be limited to a membrane filter, but also includes any substance of any thickness through with the solution or composition is passed. Accordingly, the term “filter” may include, but not be limited to, membranes, such as PES, nylon, or PVDF membranes; membrane chromatography units such as Sartobind Q, Mustang Q, and the like; or stationary phases such as an ion exchange stationary phase (e.g. cross linked polymer resin such as divinylbenzene cross-linked polystyrene), affinity stationary phase (e.g. nickel resin, streptavidin resin, glutathione conjugated resin, protein A or protein G conjugated resin, and the like), hydrophobic stationary phase (e.g. silica resin bonded with butyl, phenyl, ether, amide, or propyl ligands), size exclusion stationary phase (e.g. silica resin with appropriate -6- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT diameter and pore size), or any combination thereof (i.e. multimodal chromatography). In some embodiments, the “filter” is a resin, such as a chromatography resin as provided for herein. Similarly, the term “chromatographic filter” refers to any substance that may be used for chromatographic separation of a solution or composition. Therefore, “chromatographic filter” is not meant to be limited to a membrane filter, but also includes any substance of any thickness through with the solution or composition is passed. Accordingly, the term “chromatographic filter” may include, but not be limited to, membranes, such as PES, nylon, or PVDF membranes; membrane chromatography units such as Sartobind Q, Mustang Q, and the like; or stationary phases such as an ion exchange stationary phase (e.g. cross linked polymer resin such as divinylbenzene cross-linked polystyrene), affinity stationary phase (e.g. nickel resin, streptavidin resin, glutathione conjugated resin, protein A or protein G conjugated resin, and the like), hydrophobic stationary phase (e.g. silica resin bonded with butyl, phenyl, ether, amide, or propyl ligands), size exclusion stationary phase (e.g. silica resin with appropriate diameter and pore size), or any combination thereof (i.e. multimodal chromatography).

[0043] “Identity” as used herein refers to the subunit sequence identity between two polymeric molecules such as between two nucleic acid or amino acid molecules, such as, between two polynucleotide or polypeptide molecules. When two amino acid sequences have the same residues at the same positions; e.g., if a position in each of two polypeptide molecules is occupied by an Arginine, then they are identical at that position. The identity or extent to which two amino acid or two nucleic acid sequences have the same residues at the same positions in an alignment is often expressed as a percentage. The identity between two amino acid or two nucleic acid sequences is a direct function of the number of matching or identical positions; e.g., if half of the positions in two sequences are identical, the two sequences are 50% identical; if 90% of the positions (e.g., 9 of 10), are matched or identical, the two amino acids sequences are 90% identical.

[0044] By "substantially identical" is meant a polypeptide or nucleic acid molecule exhibiting at least 50% identity to a reference amino acid sequence (for example, any one of the amino acid sequences described herein) or nucleic acid sequence (for example, any one of the nucleic acid sequences described herein). In some embodiments, such a sequence is at least 60%, 80% or 85%, or 90%, 95% or even 99% identical at the amino acid level or nucleic acid to the sequence used for comparison. Other percentages of identity in reference to specific sequences are described herein. -7- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT

[0045] Sequence identity can be measured / determined using sequence analysis software (for example, Sequence Analysis Software Package of the Genetics Computer Group, University of Wisconsin Biotechnology Center, 1710 University Avenue, Madison, Wis. 53705, BLAST, BESTFIT, GAP, or PILEUP / PRETTYBOX programs). Such software matches identical or similar sequences by assigning degrees of homology to various substitutions, deletions, and / or other modifications. Conservative substitutions typically include substitutions within the following groups: glycine, alanine; valine, isoleucine, leucine; aspartic acid, glutamic acid, asparagine, glutamine; serine, threonine; lysine, arginine; and phenylalanine, tyrosine. In an exemplary approach to determining the degree of identity, a BLAST program may be used, with a probability score between e3 and e100 indicating a closely related sequence. In some embodiments, sequence identity is determined by using BLAST with the default settings.

[0046] To the extent embodiments provided for herein, include compositions comprising various proteins, these proteins may, in some instances, comprise amino acid sequences that have sequence identity to the amino acid sequences disclosed herein. Therefore, in certain embodiments, depending on the particular sequence, the degree of sequence identity is preferably greater than 50% (e.g.60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more) to the SEQ ID NOs disclosed herein. In addition to these percentages, other percentages of identity are provided for herein. Identity between polypeptides can be determined by the Smith-Waterman homology search algorithm as implemented in the MPSRCH program (Oxford Molecular), using an affine gap search with parameters gap open penalty – 12 and gap extension penalty = 1. These proteins may, compared to the disclosed proteins, include one or more (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) conservative amino acid replacements i.e. replacements of one amino acid with another which has a related side chain. Genetically-encoded amino acids are generally divided into four families: (1) acidic i.e. aspartate, glutamate; (2) basic i.e. lysine, arginine, histidine; (3) non polar i.e. alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan; and (4) uncharged polar i.e. glycine, asparagine, glutamine, cysteine, serine, threonine, tyrosine. Phenylalanine, tryptophan, and tyrosine are sometimes classified jointly as aromatic amino acids. In general, Substitution of single amino acids within these families does not have a major effect on the biological activity. The proteins may have one or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) single amino acid deletions relative to the disclosed protein sequences. The proteins may also include one or more (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) insertions (e.g. each of 1, 2, 3, 4 or 5 amino acids) relative to the disclosed protein sequences. -8- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT

[0047] As used herein, the phrase “in vivo” in reference to a cell being transduced, transfected or transformed in vivo, refers to a cell being transduced, transfected or transformed in the subject without the cells being removed from the subject before such cells are transduced, transfected or transformed.

[0048] “Isolated” means altered or removed from the natural state. For example, a nucleic acid or a peptide naturally present in a living animal is not “isolated,” but the same nucleic acid or peptide partially or completely separated from the coexisting materials of its natural state is “isolated.” An isolated nucleic acid or protein can exist in substantially purified form, or can exist in a non-native environment such as, for example, a host cell.

[0049] A “lentivirus” as used herein refers to a genus of the Retroviridae family that is able to infect non-dividing cells. Non-limiting examples of lentiviruses are HIV, SIV, and FIV. Vectors or viral-like particles derived from lentiviruses can be used to transduce cells and deliver genes or other molecules and have them expressed in a cell either in vitro, ex-vivo, or in vivo.

[0050] By the term “modified” as used herein, is meant a changed state or structure of a molecule or cell as provided herein. Molecules may be modified in many ways, including chemically, structurally, and functionally, such as mutations, substitutions, insertions, or deletions (e.g. internal deletions truncations). Cells may be modified through the introduction of nucleic acids or the expression of heterologous proteins.

[0051] By the term “modulating,” as used herein, is meant mediating an increase or decrease in the level of a response in a subject compared with the level of a response in the subject in the absence of a treatment or compound, and / or compared with the level of a response in an otherwise identical but untreated subject. The term encompasses perturbing and / or affecting a native signal or response thereby mediating a beneficial therapeutic response in a subject, such as, a human.

[0052] Unless otherwise specified, a “nucleotide sequence encoding an amino acid sequence” includes all nucleotide sequences that are degenerate versions of each other and that encode the same amino acid sequence. The phrase nucleotide sequence that encodes a protein or an RNA may also include introns to the extent that the nucleotide sequence encoding the protein may in some version contain an intron(s).

[0053] The term “oligonucleotide” typically refers to short polynucleotides. It will be understood that when a nucleotide sequence is represented by a DNA sequence (i.e., A, T, C, G), this also provides the corresponding RNA sequence (i.e., A, U, C, G) in which “U” replaces “T.” -9- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT

[0054] “Parenteral” administration of a composition includes, e.g., subcutaneous (s.c.), intravenous (i.v.), intramuscular (i.m.), or intrasternal injection, intrathecal, or infusion techniques.

[0055] The term “polynucleotide” as used herein is defined as a chain of nucleotides. Furthermore, nucleic acids are polymers of nucleotides. Thus, the terms “nucleic acids” and “polynucleotides” as used herein are interchangeable. As used herein polynucleotides include, but are not limited to, all nucleic acid sequences which are obtained by any methods available in the art, including, without limitation, recombinant methods, i.e., the cloning of nucleic acid sequences from a recombinant library or a cell genome, using cloning technology and PCR, and the like, and by synthetic means.

[0056] As used herein, the terms “peptide,” “polypeptide,” and “protein” are used interchangeably, and refer to a compound comprised of a plurality of amino acid residues covalently linked by peptide bonds. As used herein, the term refers to both short chains, which also commonly are referred to in the art as peptides, oligopeptides and oligomers, for example, and to longer chains, which generally are referred to in the art as proteins, of which there are many types. “Polypeptides” include, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, fusion proteins, among others. The polypeptides include natural peptides, recombinant peptides, synthetic peptides, or a combination thereof.

[0057] The term “pseudotyped” or “pseudotyped viral particle”, as used herein, refers to a viral particle bearing glycoproteins derived from other viruses having envelopes or a viral vector encoding envelope glycoproteins from a virus that is different from the parental virus. The host range of the vector particles can thus be expanded or altered depending on the type of cell surface receptor used by the glycoprotein. For example, a virus can be pseudotyped with a VSV-G mutant protein as provided for herein.

[0058] By the term “specifically binds,” as used herein with respect to an antibody, is meant an antibody which recognizes a specific antigen, but does not substantially recognize or bind other molecules in a sample. For example, an antibody that specifically binds to an antigen from one species may also bind to that antigen from one or more species. But, such cross- species reactivity does not itself alter the classification of an antibody as specific. In another example, an antibody that specifically binds to an antigen may also bind to different allelic forms of the antigen. However, such cross reactivity does not itself alter the classification of an antibody as specific. In some instances, the terms “specific binding” or “specifically binding,” can be used in reference to the interaction of an antibody, a protein, or a peptide with -10- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT a second chemical species, to mean that the interaction is dependent upon the presence of a particular structure (e.g., an antigenic determinant or epitope) on the chemical species; for example, an antibody recognizes and binds to a specific protein structure rather than to proteins generally. If an antibody is specific for epitope “A”, the presence of a molecule containing epitope A (or free, unlabeled A), in a reaction containing labeled “A” and the antibody, will reduce the amount of labeled A bound to the antibody. In some embodiments, the targeting moieties described herein that can be used to target the viral particles comprising the mutant VSV-G protein, or other viral structural proteins used to pseudotype a virus, can specifically bind to their target.

[0059] The term “therapeutic” as used herein means a treatment and / or prophylaxis. A therapeutic effect is obtained by suppression, remission, or eradication of a disease state.

[0060] The term “transfected” or “transformed” or “transduced” as used herein refers to a process by which exogenous nucleic acid is transferred or introduced into a cell. A “transfected” or “transformed” or “transduced” cell is one which has been transfected, transformed or transduced with exogenous nucleic acid. The cell includes the primary subject cell and its progeny. In some embodiments, the transfection, transformation, or transduction is performed or occurs in vivo.

[0061] A “vector” is a composition of matter which comprises an isolated nucleic acid encoding a protein or a peptide. Numerous vectors are known in the art including, but not limited to, linear polynucleotides, plasmids, DNA, and RNA. Examples of viral vectors include, but are not limited to, Sendai viral vectors, adenoviral vectors, adeno-associated virus vectors, retroviral vectors, lentiviral vectors, and the like.

[0062] A “carrier” or “delivery vehicle” includes viral particles, viruses, polylysine compounds, and liposomes, which facilitate transfer of nucleic acid into cells. A carrier or delivery vehicle can also be used to deliver a protein or peptide to a cell.

[0063] Ranges: throughout this disclosure, various aspects of the embodiments can be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. -11- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT This applies regardless of the breadth of the range. Unless otherwise explicitly stated to the contrary, a range that is disclosed also includes the endpoints of the range.

[0064] Without being bound to any particular theory, standard viral purification techniques tend to result in a loss of viral product during filtration. This is especially the case when the viral product is concentrated prior to sterile filtration, as is standard practice. Passing a pre-flush protein solution through the sterile filter prior to filtration of the viral product results in increased viral yields, possibly due to the protein solution preventing the viral product from becoming trapped in the filter due to charge interactions. Further, performing sterile filtration prior to concentrating the viral product was also found to increase product yields. Methods of Purifying Viral Vectors and / or Producing Concentrated Sterilized Solutions Comprising Viral Vectors

[0065] In some embodiments, methods of purifying a viral vector or producing a concentrated sterilized solution comprising a viral vector are provided. In some embodiments, the methods comprise clarifying a solution comprising cell culture media and the viral vector; filtering the clarified solution comprising the viral vector through a first chromatographic filter to produce a filtered clarified solution comprising the viral vector; passing the filtered clarified solution comprising the viral vector through a protein-coated sterile membrane to produce a sterilized solution comprising the viral vector; and concentrating the sterilized solution comprising the viral vector to produce a concentrated sterilized solution comprising the viral vector.

[0066] In some embodiments, the methods comprise clarifying a solution comprising cell culture media and the viral vector; filtering the clarified solution comprising the viral vector through a first chromatographic filter to produce a filtered clarified solution comprising the viral vector; concentrating the filtered clarified solution comprising the viral vector to produce a concentrated solution comprising the viral vector; and passing the concentrated solution comprising the viral vector through a protein-coated sterile membrane to produce a concentrated sterilized solution comprising the viral vector.

[0067] In some embodiments, the methods further comprises the step of mixing the concentrated sterilized solution comprising the viral vector with one or more sterile, pyrogen- free buffers and / or excipients to produce a sterile composition comprising the viral vector. In some embodiments, the concentrated sterilized solution comprising the viral vector is mixed with one pyrogen-free buffer and / or excipient. In some embodiments, the concentrated sterilized solution comprising the viral vector is mixed with more than one pyrogen-free buffer -12- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT and / or excipient. In some embodiments, the method of purifying a viral vector comprises the steps of clarifying a solution comprising cell culture media and the viral vector, filtering the clarified solution comprising the viral vector through a first chromatographic filter to produce a filtered clarified solution comprising the viral vector, passing the filtered clarified solution comprising the viral vector through a protein-coated sterile membrane to produce a sterilized solution comprising the viral vector, concentrating the sterilized solution comprising the viral vector to produce a concentrated sterilized solution comprising the viral vector, and mixing the concentrated sterilized solution comprising the viral vector with one or more sterile, pyrogen- free buffers and / or excipients to produce a sterile composition comprising the viral vector. In some embodiments, the concentrated sterilized solution comprising the viral vector is mixed with one pyrogen-free buffer and / or excipient. In some embodiments, the concentrated sterilized solution comprising the viral vector is mixed with more than one pyrogen-free buffer and / or excipient.

[0068] In some embodiments, the methods further comprise the step of collecting the media from a cell culture producing the viral vector prior to clarifying the solution. In some embodiments, the media collected from a cell culture producing the viral vector comprises the viral vector.

[0069] In some embodiments, the filtering of the clarified solution through the first chromatographic filter comprises filtration of the clarified solution via capture chromatography. In some embodiments, the capture chromatography is ion exchange chromatography, affinity chromatography, hydrophobic chromatography, size exclusion chromatography, or multimodal chromatography. In some embodiments, the capture chromatography is ion exchange chromatography. In some embodiments, the capture chromatography is affinity chromatography. In some embodiments, the capture chromatography is hydrophobic chromatography. In some embodiments, the capture chromatography is size exclusion chromatography. In some embodiments, the capture chromatography is multimodal chromatography.

[0070] In some embodiments, the methods further comprise the step of filtering the filtered clarified solution comprising the viral vector through a second chromatographic filter to produce a twice-filtered clarified solution comprising the viral vector. In some embodiments, the second filtering step is performed prior to passing filtered clarified solution through the protein coated sterile membrane. In some embodiments, the twice-filtered clarified solution comprising the viral vector is passed through the protein coated sterile membrane. -13- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT

[0071] In some embodiments, the filtering of the first filtered solution through the second chromatographic filter comprises filtration of the first filtered solution via polishing chromatography. In some embodiments, the polishing chromatography is ion exchange chromatography, hydrophobic interaction chromatography, size exclusion chromatography, or multimodal chromatography. In some embodiments, the polishing chromatography is ion exchange chromatography. In some embodiments, the polishing chromatography is hydrophobic interaction chromatography. In some embodiments, the polishing chromatography is size exclusion chromatography. In some embodiments, the polishing chromatography is multimodal chromatography.

[0072] In some embodiments, the protein coated sterile membrane comprises a sterile membrane pre-flushed with a protein solution. In some embodiments, a sterile membrane is pre-flushed with a protein solution to produce the protein coated sterile membrane. In some embodiments, the method further comprises the step of pre-flushing a sterile membrane with a protein solution to produce a protein coated sterile membrane. In some embodiments, the method further comprises the step of coating a sterile membrane with a protein solution to produce a protein coated sterile membrane.

[0073] In some embodiments, the method comprises passing the filtered clarified solution comprising the viral vector through a protein coated sterile membrane to produce a sterilized solution comprising the viral vector, wherein the protein coated sterile membrane has a pore size of about 0.01µm to about 0.45µm. In some embodiments, the protein coated sterile membrane has a pore size of about 0.01µm, about 0.10µm, about 0.11µm, about 0.12µm, about 0.13µm, about 0.14µm, about 0.15µm, about 0.16µm, about 0.17µm, about 0.18µm, about 0.19µm, about 0.2µm, about 0.21µm, about 0.22µm, about 0.23µm, about 0.24µm, about 0.25µm, about 0.26µm, about 0.27µm, about 0.28µm, about 0.29µm, about 0.30µm, about 0.31µm, about 0.32µm, about 0.33µm, about 0.34µm, about 0.35µm, about 0.36µm, about 0.37µm, about about 0.38µm, about 0.39µm, about 0.40µm, about 0.41µm, about 0.42µm, about 0.43µm, about 0.44µm, or about 0.45µm, or any value or range in-between. In some embodiments, the protein coated sterile membrane has a pore size of 0.01µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.10µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.11µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.12µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.13µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.14µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.15µm. In some -14- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT embodiments, the protein coated sterile membrane has a pore size of 0.16µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.17µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.18µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.19µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.20µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.21µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.22µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.23µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.24µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.25µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.26µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.27µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.28µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.29µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.30µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.31µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.32µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.33µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.34µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.35µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.36µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.37µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.38µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.39µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.40µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.41µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.42µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.43µm. In some embodiments, the protein coated sterile membrane has a pore size of 0.44µm. In some embodiments, protein coated sterile membrane has a pore size of 0.45µm.

[0074] In some embodiments, the protein coated sterile membrane does not significantly retain the viral vector, such as when the solution comprising the viral vector is passed through the membrane. In some embodiments, the recovery of viral vector from the solution comprising the viral vector is at least about 60%, 61%, 62%, 63%, 64%, 65%, 66%, -15- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 60%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 61%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 62%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 63%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 64%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 65%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 66%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 67%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 68%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 69%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 70%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 71%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 72%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 73%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 74%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 75%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 76%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 77%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 78%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 79%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 80%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 81%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 82%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 83%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 84%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 85%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 86%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 87%. In some embodiments, the recovery of the solution comprising the viral vector is at least about -16- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT 88%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 89%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 90%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 91%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 92%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 93%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 94%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 95%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 96%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 97%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 98%. In some embodiments, the recovery of the solution comprising the viral vector is at least about 99%. In some embodiments, the recovery of the solution comprising the viral vector is about 100%.

[0075] In some embodiments, the protein solution comprises one or more components selected from a protein, 2-Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS), histidine, phosphate, HEPES ((4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), sodium chloride, or combinations thereof. In some embodiments, the protein solution comprises a protein. In some embodiments, the protein solution comprises 2-Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS). In some embodiments, the protein solution comprises histidine, phosphate. In some embodiments, the protein solution comprises HEPES ((4-(2-hydroxyethyl)-1- piperazineethanesulfonic acid). In some embodiments, the protein solution comprises sodium chloride. In some embodiments, the protein solution comprises a protein and one or more components selected from 2-Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS), histidine, phosphate, HEPES ((4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), sodium chloride, or combinations thereof. In some embodiments, the protein solution comprises a protein and 2- Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS). In some embodiments, the protein solution comprises a protein and histidine, phosphate. In some embodiments, the protein solution comprises a protein and HEPES ((4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid). In some embodiments, the protein solution comprises a protein and sodium chloride.

[0076] In some embodiments, the protein solution comprises about 0.1% to about 3% w / v of a protein or any value or range therein. In some embodiments, the protein solution comprises about 0.2% to about 3% w / v of a protein. In some embodiments, the protein solution comprises about 0.3% to about 3% w / v of a protein. In some embodiments, the protein solution -17- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT comprises about 0.4% to about 3% w / v of a protein. In some embodiments, the protein solution comprises about 0.5% to about 3% w / v of a protein. In some embodiments, the protein solution comprises about 0.6% to about 3% w / v of a protein. In some embodiments, the protein solution comprises about 0.7% to about 3% w / v of a protein. In some embodiments, the protein solution comprises about 0.8% to about 3% w / v of a protein. In some embodiments, the protein solution comprises about 0.9% to about 3% w / v of a protein. In some embodiments, the protein solution comprises about 1% to about 3% w / v of a protein. In some embodiments, the protein solution comprises about 1.2% to about 3% w / v of a protein. In some embodiments, the protein solution comprises about 1.4% to about 3% w / v of a protein. In some embodiments, the protein solution comprises about 1.6% to about 3% w / v of a protein. In some embodiments, the protein solution comprises about 1.8% to about 3% w / v of a protein. In some embodiments, the protein solution comprises about 2% to about 3% w / v of a protein. In some embodiments, the protein solution comprises about 2.2% to about 3% w / v of a protein. In some embodiments, the protein solution comprises about 2.4% to about 3% w / v of a protein. In some embodiments, the protein solution comprises about 2.6% to about 3% w / v of a protein. In some embodiments, the protein solution comprises about 2.8% to about 3% w / v of a protein.

[0077] In some embodiments, the protein solution comprises about 0.1% to about 3% w / v of a protein. In some embodiments, the protein solution comprises about 0.1% to about 2.8% w / v of a protein. In some embodiments, the protein solution comprises about 0.1% to about 2.6% w / v of a protein. In some embodiments, the protein solution comprises about 0.1% to about 2.4% w / v of a protein. In some embodiments, the protein solution comprises about 0.1% to about 2.2% w / v of a protein. In some embodiments, the protein solution comprises about 0.1% to about 2% w / v of a protein. In some embodiments, the protein solution comprises about 0.1% to about 1.8% w / v of a protein. In some embodiments, the protein solution comprises about 0.1% to about 1.6% w / v of a protein. In some embodiments, the protein solution comprises about 0.1% to about 1.4% w / v of a protein. In some embodiments, the protein solution comprises about 0.1% to about 1.2% w / v of a protein. In some embodiments, the protein solution comprises about 0.1% to about 1.0% w / v of a protein. In some embodiments, the protein solution comprises about 0.1% to about 0.9% w / v of a protein. In some embodiments, the protein solution comprises about 0.1% to about 0.8% w / v of a protein. In some embodiments, the protein solution comprises about 0.1% to about 0.7% w / v of a protein. In some embodiments, the protein solution comprises about 0.1% to about 0.6% w / v of a protein. In some embodiments, the protein solution comprises about 0.1% to about 0.5% w / v of a protein. In some embodiments, the protein solution comprises about 0.1% to about -18- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT 0.4% w / v of a protein. In some embodiments, the protein solution comprises about 0.1% to about 0.3% w / v of a protein. In some embodiments, the protein solution comprises about 0.1% to about 0.2% w / v of a protein.

[0078] In some embodiments, the protein solution comprises about 0.1% w / v, about 0.2% w / v, about 0.3% w / v, about 0.4% w / v, about 0.5% w / v, about 0.6% w / v, about 0.7% w / v, about 0.8% w / v, about 0.9% w / v, about 1.0% w / v, about 1.2% w / v, about 1.4% w / v, about 1.6% w / v, about 1.8% w / v, about 2.0% w / v, about 2.2% w / v, about 2.4% w / v, about 2.6% w / v, about 2.8% w / v, or about 3.0% w / v of a protein, or any value therein. In some embodiments, the protein solution comprises about 0.1% w / v of a protein. In some embodiments, the protein solution comprises about 0.2% w / v of a protein. In some embodiments, the protein solution comprises about 0.3% w / v of a protein. In some embodiments, the protein solution comprises about 0.4% w / v of a protein. In some embodiments, the protein solution comprises about 0.5% w / v of a protein. In some embodiments, the protein solution comprises about 0.6% w / v of a protein. In some embodiments, the protein solution comprises about 0.7% w / v of a protein. In some embodiments, the protein solution comprises about 0.8% w / v of a protein. In some embodiments, the protein solution comprises about 0.9% w / v of a protein. In some embodiments, the protein solution comprises about 1.0% w / v of a protein. In some embodiments, the protein solution comprises about 1.2% w / v of a protein. In some embodiments, the protein solution comprises about 1.4% w / v of a protein. In some embodiments, the protein solution comprises about 1.6% w / v of a protein. In some embodiments, the protein solution comprises about 1.8% w / v of a protein. In some embodiments, the protein solution comprises about 2.0% w / v of a protein. In some embodiments, the protein solution comprises about 2.2% w / v of a protein. In some embodiments, the protein solution comprises about 2.4% w / v of a protein. In some embodiments, the protein solution comprises about 2.6% w / v of a protein. In some embodiments, the protein solution comprises about 2.8% w / v of a protein. In some embodiments, the protein solution comprises about 3.0% w / v of a protein.

[0079] In some embodiments, the protein solution comprises about 100mM to about 400mM sodium chloride, or any value or range therein. In some embodiments, the protein solution comprises about 110mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 120mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 130mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 140mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 150mM to about 400mM -19- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT sodium chloride. In some embodiments, the protein solution comprises about 160mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 170mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 180mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 190mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 200mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 210mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 220mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 230mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 240mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 250mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 260mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 270mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 280mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 290mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 300mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 310mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 320mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 330mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 340mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 350mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 360mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 370mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 380mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 390mM to about 400mM sodium chloride.

[0080] In some embodiments, the protein solution comprises about 100mM to about 400mM sodium chloride, or any value or range therein. In some embodiments, the protein solution comprises about 100mM to about 390mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 380mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 370mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 360mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 350mM -20- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 340mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 330mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 320mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 310mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 300mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 290mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 280mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 270mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 260mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 250mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 240mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 230mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 220mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 210mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 200mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 190mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 180mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 170mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 160mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 150mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 140mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 130mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 120mM sodium chloride. In some embodiments, the protein solution comprises about 100mM to about 110mM sodium chloride.

[0081] In some embodiments, the protein solution comprises about 100mM, about 110mM, about 120mM, about 130mM, about 140mM, about 150mM, about 160mM, about 170mM, about 180mM, about 190mM, about 200mM, about 210mMm, about 220mM, about 230mM, about 240mM, about 250mM, about 260mM, about 270mM, about 280mM, about 290mM, about 300mM, about 310mM, about 320mM, about 330mM, about 340mM, about 350mM, about 360mM, about 370mM, about 380mM, about 390mM, or about 400mM sodium chloride. In some embodiments, the protein solution comprises about 100mM sodium chloride. -21- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT In some embodiments, the protein solution comprises about 110mM sodium chloride. In some embodiments, the protein solution comprises about 120mM sodium chloride. In some embodiments, the protein solution comprises about 130mM sodium chloride. In some embodiments, the protein solution comprises about 140mM sodium chloride. In some embodiments, the protein solution comprises about 150mM sodium chloride. In some embodiments, the protein solution comprises about 160mM sodium chloride. In some embodiments, the protein solution comprises about 170mM sodium chloride. In some embodiments, the protein solution comprises about 180mM sodium chloride. In some embodiments, the protein solution comprises about 190mM sodium chloride. In some embodiments, the protein solution comprises about 200mM sodium chloride. In some embodiments, the protein solution comprises about 210mM sodium chloride. In some embodiments, the protein solution comprises about 220mM sodium chloride. In some embodiments, the protein solution comprises about 230mM sodium chloride. In some embodiments, the protein solution comprises about 240mM sodium chloride. In some embodiments, the protein solution comprises about 250mM sodium chloride. In some embodiments, the protein solution comprises about 260mM sodium chloride. In some embodiments, the protein solution comprises about 270mM sodium chloride. In some embodiments, the protein solution comprises about 280mM sodium chloride. In some embodiments, the protein solution comprises about 290mM sodium chloride. In some embodiments, the protein solution comprises about 300mM sodium chloride. In some embodiments, the protein solution comprises about 310mM sodium chloride. In some embodiments, the protein solution comprises about 320mM sodium chloride. In some embodiments, the protein solution comprises about 330mM sodium chloride. In some embodiments, the protein solution comprises about 340mM sodium chloride. In some embodiments, the protein solution comprises about 350mM sodium chloride. In some embodiments, the protein solution comprises about 360mM sodium chloride. In some embodiments, the protein solution comprises about 370mM sodium chloride. In some embodiments, the protein solution comprises about 380mM sodium chloride. In some embodiments, the protein solution comprises about 390mM sodium chloride. In some embodiments, the protein solution comprises about 300mM sodium chloride.

[0082] In some embodiments, the protein solution comprises about 5mM to about 50mM 2-Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS), or any value or range therein. In some embodiments, the protein solution comprises about 10mM to about 50mM TRIS. In some embodiments, the protein solution comprises about 15mM to about 50mM TRIS. In -22- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT some embodiments, the protein solution comprises about 20mM to about 50mM TRIS. In some embodiments, the protein solution comprises about 25mM to about 50mM TRIS. In some embodiments, the protein solution comprises about 30mM to about 50mM TRIS. In some embodiments, the protein solution comprises about 35mM to about 50mM TRIS. In some embodiments, the protein solution comprises about 40mM to about 50mM TRIS. In some embodiments, the protein solution comprises about 45mM to about 50mM TRIS.

[0083] In some embodiments, the protein solution comprises about 5mM to about 50mM 2-Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS), or any value or range therein. In some embodiments, the protein solution comprises about 10mM to about 45mM TRIS. In some embodiments, the protein solution comprises about 10mM to about 40mM TRIS. In some embodiments, the protein solution comprises about 10mM to about 35mM TRIS. In some embodiments, the protein solution comprises about 10mM to about 30mM TRIS. In some embodiments, the protein solution comprises about 10mM to about 25mM TRIS. In some embodiments, the protein solution comprises about 10mM to about 20mM TRIS. In some embodiments, the protein solution comprises about 10mM to about 15mM TRIS.

[0084] In some embodiments, the protein solution comprises about 5mM, about 10mM, about 15mM, about 20mM, about 25mM, about 30mM, about 35mM, about 40mM, about 45mM, or about 50mM 2-Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS). In some embodiments, the protein solution comprises about 5mM TRIS. In some embodiments, the protein solution comprises about 10mM TRIS. In some embodiments, the protein solution comprises about 15mM TRIS. In some embodiments, the protein solution comprises about 20mM TRIS. In some embodiments, the protein solution comprises about 25mM TRIS. In some embodiments, the protein solution comprises about 30mM TRIS. In some embodiments, the protein solution comprises about 35mM TRIS. In some embodiments, the protein solution comprises about 40mM TRIS. In some embodiments, the protein solution comprises about 45mM TRIS. In some embodiments, the protein solution comprises about 50mM TRIS.

[0085] In some embodiments, the protein solution comprises about 5mM to about 100mM histidine buffer or any value or range therein. In some embodiments, the protein solution comprises about 10mM to about 100mM histidine buffer. In some embodiments, the protein solution comprises about 15mM to about 100mM histidine buffer. In some embodiments, the protein solution comprises about 20mM to about 100mM histidine buffer. In some embodiments, the protein solution comprises about 25mM to about 100mM histidine buffer. In some embodiments, the protein solution comprises about 30mM to about 100mM histidine buffer. In some embodiments, the protein solution comprises about 35mM to about -23- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT 100mM histidine buffer. In some embodiments, the protein solution comprises about 40mM to about 100mM histidine buffer. In some embodiments, the protein solution comprises about 45mM to about 100mM histidine buffer. In some embodiments, the protein solution comprises about 50mM to about 100mM histidine buffer. In some embodiments, the protein solution comprises about 55mM to about 100mM histidine buffer. In some embodiments, the protein solution comprises about 60mM to about 100mM histidine buffer. In some embodiments, the protein solution comprises about 65mM to about 100mM histidine buffer. In some embodiments, the protein solution comprises about 70mM to about 100mM histidine buffer. In some embodiments, the protein solution comprises about 75mM to about 100mM histidine buffer. In some embodiments, the protein solution comprises about 80mM to about 100mM histidine buffer. In some embodiments, the protein solution comprises about 85mM to about 100mM histidine buffer. In some embodiments, the protein solution comprises about 90mM to about 100mM histidine buffer. In some embodiments, the protein solution comprises about 95mM to about 100mM histidine buffer.

[0086] In some embodiments, the protein solution comprises about 5mM to about 100mM histidine buffer or any value or range therein. In some embodiments, the protein solution comprises about 5mM to about 95mM histidine buffer. In some embodiments, the protein solution comprises about 5mM to about 90mM histidine buffer. In some embodiments, the protein solution comprises about 5mM to about 85mM histidine buffer. In some embodiments, the protein solution comprises about 5mM to about 80mM histidine buffer. In some embodiments, the protein solution comprises about 5mM to about 75mM histidine buffer. In some embodiments, the protein solution comprises about 5mM to about 70mM histidine buffer. In some embodiments, the protein solution comprises about 5mM to about 65mM histidine buffer. In some embodiments, the protein solution comprises about 5mM to about 60mM histidine buffer. In some embodiments, the protein solution comprises about 5mM to about 55mM histidine buffer. In some embodiments, the protein solution comprises about 5mM to about 50mM histidine buffer. In some embodiments, the protein solution comprises about 5mM to about 45mM histidine buffer. In some embodiments, the protein solution comprises about 5mM to about 40mM histidine buffer. In some embodiments, the protein solution comprises about 5mM to about 35mM histidine buffer. In some embodiments, the protein solution comprises about 5mM to about 30mM histidine buffer. In some embodiments, the protein solution comprises about 5mM to about 25mM histidine buffer. In some embodiments, the protein solution comprises about 5mM to about 20mM histidine buffer. In some embodiments, the protein solution comprises about 5mM to about 15mM histidine buffer. -24- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT In some embodiments, the protein solution comprises about 5mM to about 10mM histidine buffer.

[0087] In some embodiments, the protein solution comprises about 5mM, about 10mM, about 15mM, about 20mM, about 25mM, about 30mM, about 35mM, about 40mM, about 45mM, about 50mM, about 55mM, about 60mM, about 65mM, about 70mM, about 75mM, about 80mM, about 85mM, about 90mM, about 95mM, or about 100mM histidine buffer, or any value therein. In some embodiments, the protein solution comprises about 5mM histidine buffer. In some embodiments, the protein solution comprises about 10mM histidine buffer. In some embodiments, the protein solution comprises about 15mM histidine buffer. In some embodiments, the protein solution comprises about 20mM histidine buffer. In some embodiments, the protein solution comprises about 25mM histidine buffer. In some embodiments, the protein solution comprises about 30mM histidine buffer. In some embodiments, the protein solution comprises about 35mM histidine buffer. In some embodiments, the protein solution comprises about 40mM histidine buffer. In some embodiments, the protein solution comprises about 45mM histidine buffer. In some embodiments, the protein solution comprises about 50mM histidine buffer. In some embodiments, the protein solution comprises about 55mM histidine buffer. In some embodiments, the protein solution comprises about 60mM histidine buffer. In some embodiments, the protein solution comprises about 65msM histidine buffer. In some embodiments, the protein solution comprises about 70mM histidine buffer. In some embodiments, the protein solution comprises about 75mM histidine buffer. In some embodiments, the protein solution comprises about 80mM histidine buffer. In some embodiments, the protein solution comprises about 85mM histidine buffer. In some embodiments, the protein solution comprises about 90mM histidine buffer. In some embodiments, the protein solution comprises about 95mM histidine buffer. In some embodiments, the protein solution comprises about 100mM histidine buffer.

[0088] In some embodiments, the protein solution has a pH of about 5.5 to about 8.5, or any value or range therein. In some embodiments, the protein solution has a pH of about 6.0 to about 8.5. In some embodiments, the protein solution has a pH of about 6.5 to about 8.5. In some embodiments, the protein solution has a pH of about 7.0 to about 8.5. In some embodiments, the protein solution has a pH of about 7.5 to about 8.5. In some embodiments, the protein solution has a pH of about 8.0 to about 8.5. In some embodiments, the protein solution has a pH of about 6.0 to about 8.0. -25- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT

[0089] In some embodiments, the protein solution has a pH of about 5.5 to about 8.5. In some embodiments, the protein solution has a pH of about 5.5 to about 8.0. In some embodiments, the protein solution has a pH of about 5.5 to about 7.5. In some embodiments, the protein solution has a pH of about 5.5 to about 7.0. In some embodiments, the protein solution has a pH of about 5.5 to about 6.5. In some embodiments, the protein solution has a pH of about 5.5 to about 6.0.

[0090] In some embodiments, the protein solution has a pH of about 5.5, about 6.0, about 6.5, about 6.6, about 6.7, about 6.8, about 6.9, about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7, about 7.8, about 7.9 about 8.0, or about 8.5, or any value therein. In some embodiments, the protein solution has a pH of about 5.5. In some embodiments, the protein solution has a pH of about 6.0. In some embodiments, the protein solution has a pH of about 6.5. In some embodiments, the protein solution has a pH of about 6.6. In some embodiments, the protein solution has a pH of about 6.7. In some embodiments, the protein solution has a pH of about 6.8. In some embodiments, the protein solution has a pH of about 6.9. In some embodiments, the protein solution has a pH of about 7.0. In some embodiments, the protein solution has a pH of about 7.1. In some embodiments, the protein solution has a pH of about 7.2. In some embodiments, the protein solution has a pH of about 7.3. In some embodiments, the protein solution has a pH of about 7.4. In some embodiments, the protein solution has a pH of about 7.5. In some embodiments, the protein solution has a pH of about 7.6. In some embodiments, the protein solution has a pH of about 7.7. In some embodiments, the protein solution has a pH of about 7.8. In some embodiments, the protein solution has a pH of about 7.9. In some embodiments, the protein solution has a pH of about 8.0. In some embodiments, the protein solution has a pH of about 8.5.

[0091] In some embodiments, the protein solution comprises about 0.1% w / v to about 3.0% w / v of a protein, about 5mM to about 50mM 2-Amino-2-(hydroxymethyl)propane-1,3- diol (TRIS), and about 100mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 0.1% w / v of a protein, about 20mM 2-Amino-2- (hydroxymethyl)propane-1,3-diol (TRIS), and about 100mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 1% w / v of a protein, about 20mM 2-Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS), and about 100mM to about 400mM sodium chloride.

[0092] In some embodiments, the protein solution comprises about 0.1% w / v to about 3.0% w / v of a protein, about 5mM to about 50mM 2-Amino-2-(hydroxymethyl)propane-1,3- diol (TRIS), and about 100mM to about 400mM sodium chloride. In some embodiments, the -26- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT protein solution comprises about 0.1% w / v to about 3.0% w / v of a protein, about 20mM 2- Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS), about 100mM to about 400mM sodium chloride, and has a pH of about 6.0 to about 8.0. In some embodiments, the protein solution comprises about 0.1% w / v of a protein, about 20mM 2-Amino-2-(hydroxymethyl)propane-1,3- diol (TRIS), about 100mM to about 400mM sodium chloride, and has a pH of about 6.0 to about 8.0. In some embodiments, the protein solution comprises about 1% w / v of a protein, about 20mM 2-Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS), about 100mM to about 400mM sodium chloride, and has a pH of about 6.0 to about 8.0.

[0093] In some embodiments, the protein solution comprises about 0.1% w / v to about 3.0% w / v of a protein, about 5mM to about 100mM histidine buffer, and about 100mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 0.1% w / v of a protein, about 5mM to about 100mM histidine buffer, and about 100mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 1% w / v of a protein, about 5mM to about 100mM histidine buffer, and about 100mM to about 400mM sodium chloride.

[0094] In some embodiments, the protein solution comprises about 0.1% w / v to about 3.0% w / v of a protein, about 5mM to about 100mM histidine buffer, and about 100mM to about 400mM sodium chloride. In some embodiments, the protein solution comprises about 0.1% w / v to about 3.0% w / v of a protein, about 5mM to about 100mM histidine buffer, about 100mM to about 400mM sodium chloride, and has a pH of about 6.0 to about 8.0. In some embodiments, the protein solution comprises about 0.1% w / v of a protein, about 5mM to about 100mM histidine buffer, about 100mM to about 400mM sodium chloride, and has a pH of about 6.0 to about 8.0. In some embodiments, the protein solution comprises about 1% w / v of a protein, about 5mM to about 100mM histidine buffer, about 100mM to about 400mM sodium chloride, and has a pH of about 6.0 to about 8.0.

[0095] In some embodiments, the protein solution comprises any protein that has an isoelectric point from about 5.0 to about 8.0, or any value or range therein. In some embodiments, the protein has an isoelectric point from about 5.0 to about 8.0. In some embodiments, the protein has an isoelectric point from about 5.2 to about 8.0. In some embodiments, the protein has an isoelectric point from about 5.4 to about 8.0. In some embodiments, the protein has an isoelectric point from about 5.6 to about 8.0. In some embodiments, the protein has an isoelectric point from about 5.8 to about 8.0. In some embodiments, the protein has an isoelectric point from about 6.0 to about 8.0. In some embodiments, the protein has an isoelectric point from about 6.2 to about 8.0. In some -27- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT embodiments, the protein has an isoelectric point from about 6.4 to about 8.0. In some embodiments, the protein has an isoelectric point from about 6.6 to about 8.0. In some embodiments, the protein has an isoelectric point from about 6.8 to about 8.0. In some embodiments, the protein has an isoelectric point from about 7.0 to about 8.0. In some embodiments, the protein has an isoelectric point from about 7.2 to about 8.0. In some embodiments, the protein has an isoelectric point from about 7.4 to about 8.0. In some embodiments, the protein has an isoelectric point from about 7.6 to about 8.0. In some embodiments, the protein has an isoelectric point from about 7.8 to about 8.0. In some embodiments, the protein has an isoelectric point from about 5.0 to about 7.8. In some embodiments, the protein has an isoelectric point from about 5.0 to about 7.6. In some embodiments, the protein has an isoelectric point from about 5.0 to about 7.4. In some embodiments, the protein has an isoelectric point from about 5.0 to about 7.2. In some embodiments, the protein has an isoelectric point from about 5.0 to about 7.0. In some embodiments, the protein has an isoelectric point from about 5.0 to about 6.8. In some embodiments, the protein has an isoelectric point from about 5.0 to about 6.6. In some embodiments, the protein has an isoelectric point from about 5.0 to about 6.4. In some embodiments, the protein has an isoelectric point from about 5.0 to about 6.2. In some embodiments, the protein has an isoelectric point from about 5.0 to about 6.0. In some embodiments, the protein has an isoelectric point from about 5.0 to about 5.8. In some embodiments, the protein has an isoelectric point from about 5.0 to about 5.6. In some embodiments, the protein has an isoelectric point from about 5.0 to about 5.4. In some embodiments, the protein has an isoelectric point from about 5.0 to about 5.2. In some embodiments, the protein has an isoelectric point of about 5.0, 5.2, 5.4, 5.6, 5.8, 6.0, 6.2, 6.4, 6.6, 6.8, 7.0, 7.2, 7.4, 7.6, 7.8, 8.0, or any value in between. In some embodiments, the protein has an isoelectric point of about 5.0. In some embodiments, the protein has an isoelectric point of about 5.1. In some embodiments, the protein has an isoelectric point of about 5.2. In some embodiments, the protein has an isoelectric point of about 5.3. In some embodiments, the protein has an isoelectric point of about 5.4. In some embodiments, the protein has an isoelectric point of about 5.5. In some embodiments, the protein has an isoelectric point of about 5.6. In some embodiments, the protein has an isoelectric point of about 5.7. In some embodiments, the protein has an isoelectric point of about 5.8. In some embodiments, the protein has an isoelectric point of about 5.9. In some embodiments, the protein has an isoelectric point of about 6.0. In some embodiments, the protein has an isoelectric point of about 6.1. In some embodiments, the protein has an isoelectric point of about 6.2. In some -28- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT embodiments, the protein has an isoelectric point of about 6.3. In some embodiments, the protein has an isoelectric point of about 6.4. In some embodiments, the protein has an isoelectric point of about 6.5. In some embodiments, the protein has an isoelectric point of about 6.6. In some embodiments, the protein has an isoelectric point of about 6.7. In some embodiments, the protein has an isoelectric point of about 6.8. In some embodiments, the protein has an isoelectric point of about 6.9. In some embodiments, the protein has an isoelectric point of about 7.0. In some embodiments, the protein has an isoelectric point of about 7.1. In some embodiments, the protein has an isoelectric point of about 7.2. In some embodiments, the protein has an isoelectric point of about 7.3. In some embodiments, the protein has an isoelectric point of about 7.4. In some embodiments, the protein has an isoelectric point of about 7.5. In some embodiments, the protein has an isoelectric point of about 7.6. In some embodiments, the protein has an isoelectric point of about 7.7. In some embodiments, the protein has an isoelectric point of about 7.8. In some embodiments, the protein has an isoelectric point of about 7.9. In some embodiments, the protein has an isoelectric point of about 8.0.

[0096] In some embodiments, the protein solution comprises a protein selected from, but not limited to, myoglobulin, serum albumin, bovine serum albumin, human serum albumin, immunoglobulin, immunoglobulin fragments, fibronectin, vitronectin, or a combination thereof. In some embodiments, the protein solution comprises serum albumin. In some embodiments, the protein solution comprises human serum albumin. In some embodiments, the human serum albumin is recombinant human serum albumin. In some embodiments, the human serum albumin is non-recombinant human serum albumin. In some embodiments, the non-recombinant human serum albumin is US Pharmacopeia (USP) grade non-recombinant human serum albumin. In some embodiments, the non-recombinant human serum albumin is FDA approved non-recombinant human serum albumin. The human serum albumin may be produced from any source. Methods of making human serum albumin are known in the art, and any such method is within the scope of the present application. In some embodiments, human serum albumin is produced from a bacteria source (e.g. bacteria cells). In some embodiments, human serum albumin is produced from an insect source (e.g. insect cells). In some embodiments, human serum albumin is produced from a mammalian source (e.g. mammalian cells). In some embodiments, human serum albumin is produced from a plant source. Likewise, the recombinant human serum albumin may be recombinantly produced from any source. Methods of making recombinant human serum albumin are known in the art, and any such method is within the scope of the present application. In some embodiments, -29- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT human serum albumin is recombinantly produced from a bacteria source (e.g. bacteria cells). In some embodiments, recombinant human serum albumin is recombinantly produced from an insect source (e.g. insect cells). In some embodiments, recombinant human serum albumin is recombinantly produced from a mammalian source (e.g. mammalian cells). In some embodiments, recombinant human serum albumin is recombinantly produced from a plant source.

[0097] In some embodiments, the protein solution comprises about 0.1% to about 3% w / v human serum albumin or any value or range therein. In some embodiments, the protein solution comprises about 0.2% to about 3% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.3% to about 3% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.4% to about 3% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.5% to about 3% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.6% to about 3% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.7% to about 3% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.8% to about 3% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.9% to about 3% w / v human serum albumin. In some embodiments, the protein solution comprises about 1% to about 3% w / v human serum albumin. In some embodiments, the protein solution comprises about 1.2% to about 3% w / v human serum albumin. In some embodiments, the protein solution comprises about 1.4% to about 3% w / v human serum albumin. In some embodiments, the protein solution comprises about 1.6% to about 3% w / v human serum albumin. In some embodiments, the protein solution comprises about 1.8% to about 3% w / v human serum albumin. In some embodiments, the protein solution comprises about 2% to about 3% w / v human serum albumin. In some embodiments, the protein solution comprises about 2.2% to about 3% w / v human serum albumin. In some embodiments, the protein solution comprises about 2.4% to about 3% w / v human serum albumin. In some embodiments, the protein solution comprises about 2.6% to about 3% w / v human serum albumin. In some embodiments, the protein solution comprises about 2.8% to about 3% w / v human serum albumin.

[0098] In some embodiments, the protein solution comprises about 0.1% to about 3% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 2.8% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 2.6% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 2.4% w / v human serum albumin. In some -30- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT embodiments, the protein solution comprises about 0.1% to about 2.2% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 2% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 1.8% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 1.6% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 1.4% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 1.2% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 1.0% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.9% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.8% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.7% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.6% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.5% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.4% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.3% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.2% w / v human serum albumin.

[0099] In some embodiments, the protein solution comprises about 0.1% w / v, about 0.2% w / v, about 0.3% w / v, about 0.4% w / v, about 0.5% w / v, about 0.6% w / v, about 0.7% w / v, about 0.8% w / v, about 0.9% w / v, about 1.0% w / v, about 1.2% w / v, about 1.4% w / v, about 1.6% w / v, about 1.8% w / v, about 2.0% w / v, about 2.2% w / v, about 2.4% w / v, about 2.6% w / v, about 2.8% w / v, or about 3.0% w / v human serum albumin, or any value therein. In some embodiments, the protein solution comprises about 0.1% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.2% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.3% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.4% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.5% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.6% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.7% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.8% w / v human serum albumin. In some embodiments, the protein solution comprises about 0.9% w / v human serum albumin. In some embodiments, the protein solution comprises about 1.0% w / v human serum albumin. In some embodiments, the protein solution comprises about 1.2% w / v human serum albumin. In some -31- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT embodiments, the protein solution comprises about 1.4% w / v human serum albumin. In some embodiments, the protein solution comprises about 1.6% w / v human serum albumin. In some embodiments, the protein solution comprises about 1.8% w / v human serum albumin. In some embodiments, the protein solution comprises about 2.0% w / v human serum albumin. In some embodiments, the protein solution comprises about 2.2% w / v human serum albumin. In some embodiments, the protein solution comprises about 2.4% w / v human serum albumin. In some embodiments, the protein solution comprises about 2.6% w / v human serum albumin. In some embodiments, the protein solution comprises about 2.8% w / v human serum albumin. In some embodiments, the protein solution comprises about 3.0% w / v human serum albumin.

[0100] In some embodiments, the protein solution comprises about 0.1% to about 3% w / v recombinant human serum albumin or any value or range therein. In some embodiments, the protein solution comprises about 0.2% to about 3% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.3% to about 3% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.4% to about 3% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.5% to about 3% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.6% to about 3% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.7% to about 3% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.8% to about 3% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.9% to about 3% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 1% to about 3% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 1.2% to about 3% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 1.4% to about 3% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 1.6% to about 3% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 1.8% to about 3% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 2% to about 3% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 2.2% to about 3% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 2.4% to about 3% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 2.6% to about 3% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 2.8% to about 3% w / v recombinant human serum albumin. -32- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT

[0101] In some embodiments, the protein solution comprises about 0.1% to about 3% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 2.8% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 2.6% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 2.4% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 2.2% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 2% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 1.8% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 1.6% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 1.4% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 1.2% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 1.0% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.9% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.8% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.7% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.6% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.5% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.4% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.3% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.2% w / v recombinant human serum albumin.

[0102] In some embodiments, the protein solution comprises about 0.1% w / v, about 0.2% w / v, about 0.3% w / v, about 0.4% w / v, about 0.5% w / v, about 0.6% w / v, about 0.7% w / v, about 0.8% w / v, about 0.9% w / v, about 1.0% w / v, about 1.2% w / v, about 1.4% w / v, about 1.6% w / v, about 1.8% w / v, about 2.0% w / v, about 2.2% w / v, about 2.4% w / v, about 2.6% w / v, about 2.8% w / v, or about 3.0% w / v recombinant human serum albumin, or any value therein. In some embodiments, the protein solution comprises about 0.1% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.2% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises -33- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT about 0.3% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.4% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.5% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.6% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.7% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.8% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.9% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 1.0% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 1.2% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 1.4% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 1.6% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 1.8% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 2.0% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 2.2% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 2.4% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 2.6% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 2.8% w / v recombinant human serum albumin. In some embodiments, the protein solution comprises about 3.0% w / v recombinant human serum albumin.

[0103] In some embodiments, the recombinant human serum albumin is any recombinant human serum albumin. In some embodiments, the recombinant human serum albumin is Exbumin, Cellastim S, Albagen, Recombumin, or Optibumin. In some embodiments, the recombinant human serum albumin is Exbumin. In some embodiments, the recombinant human serum albumin is Cellastim S. In some embodiments, the recombinant human serum albumin is Albagen. In some embodiments, the recombinant human serum albumin is Recombumin. In some embodiments, the recombinant human serum albumin is Optibumin.

[0104] In some embodiments, the protein solution comprises about 0.1% to about 3% w / v non-recombinant human serum albumin or any value or range therein. In some embodiments, the protein solution comprises about 0.2% to about 3% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.3% to about 3% w / v non-recombinant human serum albumin. In some embodiments, the protein -34- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT solution comprises about 0.4% to about 3% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.5% to about 3% w / v non- recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.6% to about 3% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.7% to about 3% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.8% to about 3% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.9% to about 3% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 1% to about 3% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 1.2% to about 3% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 1.4% to about 3% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 1.6% to about 3% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 1.8% to about 3% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 2% to about 3% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 2.2% to about 3% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 2.4% to about 3% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 2.6% to about 3% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 2.8% to about 3% w / v non-recombinant human serum albumin.

[0105] In some embodiments, the protein solution comprises about 0.1% to about 3% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 2.8% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 2.6% w / v non- recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 2.4% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 2.2% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 2% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 1.8% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 1.6% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 1.4% w / v -35- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 1.2% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 1.0% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.9% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.8% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.7% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.6% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.5% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.4% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.3% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.1% to about 0.2% w / v non-recombinant human serum albumin.

[0106] In some embodiments, the protein solution comprises about 0.1% w / v, about 0.2% w / v, about 0.3% w / v, about 0.4% w / v, about 0.5% w / v, about 0.6% w / v, about 0.7% w / v, about 0.8% w / v, about 0.9% w / v, about 1.0% w / v, about 1.2% w / v, about 1.4% w / v, about 1.6% w / v, about 1.8% w / v, about 2.0% w / v, about 2.2% w / v, about 2.4% w / v, about 2.6% w / v, about 2.8% w / v, or about 3.0% w / v non-recombinant human serum albumin, or any value therein. In some embodiments, the protein solution comprises about 0.1% w / v non- recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.2% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.3% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.4% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.5% w / v non- recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.6% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.7% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.8% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 0.9% w / v non- recombinant human serum albumin. In some embodiments, the protein solution comprises about 1.0% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 1.2% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 1.4% w / v non-recombinant human serum -36- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT albumin. In some embodiments, the protein solution comprises about 1.6% w / v non- recombinant human serum albumin. In some embodiments, the protein solution comprises about 1.8% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 2.0% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 2.2% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 2.4% w / v non- recombinant human serum albumin. In some embodiments, the protein solution comprises about 2.6% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 2.8% w / v non-recombinant human serum albumin. In some embodiments, the protein solution comprises about 3.0% w / v non-recombinant human serum albumin.

[0107] In some embodiments, the non-recombinant human serum albumin is any non-recombinant human serum albumin. In some embodiments, the non-recombinant human serum albumin is FDA approved, USP grade non-recombinant human serum albumin. The FDA approved, USP grade non-recombinant human serum albumin may be obtained via any appropriate commercial vendor, such as but not limited to, Nova, Octapharma, Grifols Bio Supplies, and the like.

[0108] In some embodiments, the method of purifying a viral vector further comprises utilizing a protein coated sterile membrane with a positive charge prior to being coated with a protein. In some embodiments, the membrane is a polyethersulfone (PES) membrane, a nylon membrane, or a polyvinylidene fluoride (PVDF) membrane. In some embodiments, the membrane is a polyethersulfone (PES) membrane. In some embodiments, the membrane is a nylon membrane. In some embodiments, the membrane is a polyvinylidene fluoride (PVDF) membrane.

[0109] In some embodiments, the methods of purifying a viral vector or producing a concentration sterile solution comprising the viral vector comprise digesting DNA in the solution comprising the cell culture media and the viral vector prior to clarifying the solution. In some embodiments, the method of purifying a viral vector comprises the step of digesting DNA in the solution comprising the viral vector after filtering the clarified solution. In some embodiments, the method of purifying a viral vector comprises a first DNA digestion step comprising digesting DNA in the solution comprising the cell culture media and the viral vector prior to clarifying the solution; and a second DNA digestion step comprising digesting DNA in the solution comprising the viral vector after filtering the clarified solution. -37- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT

[0110] In some embodiments, the methods comprise mixing the concentrated sterilized solution comprising the viral vector with one or more sterile, pyrogen-free buffers and / or excipients in a sterile environment to produce a sterile pharmaceutical composition comprising the viral vector. In some embodiments, the methods comprises mixing the concentrated sterilized solution comprising the viral vector with one or more sterile, pyrogen- free buffers and / or excipients to produce a sterile composition comprising the viral vector. In some embodiments, the sterile, pyrogen-free buffers and / or excipients include, for example and without limitation, TRIS, HEPES, histidine buffer, phosphate buffer, sucrose, trehalose, polyethylene glycol, or any combination thereof.

[0111] In some embodiments, the methods comprise digesting DNA in a solution comprising the viral vector to produce a first digested solution comprising the viral vector; clarifying the digested solution comprising the viral vector; filtering the clarified, digested solution comprising the viral vector through a first chromatographic filter to produce a filtered, clarified, digested solution comprising the viral vector; digesting DNA in the filtered, clarified, digested solution comprising the viral vector to produce a filtered, clarified, twice-digested solution comprising viral vector; filtering the filtered, clarified, twice-digested solution comprising the viral vector through a second chromatographic filter to produce a twice-filtered, clarified, twice-digested solution comprising viral vector; passing the twice- filtered, clarified, twice-digested solution comprising the viral vector through a protein-coated sterile membrane to produce a sterilized solution comprising the viral vector; and concentrating the sterilized solution comprising the viral vector to produce a concentrated sterilized solution comprising the viral vector.

[0112] In some embodiments, the methods comprise digesting DNA in a solution comprising the viral vector to produce a first digested solution comprising the viral vector; clarifying the digested solution comprising the viral vector; filtering the clarified, digested solution comprising the viral vector through a first chromatographic filter to produce a filtered, clarified, digested solution comprising the viral vector; digesting DNA in the filtered, clarified, digested solution comprising the viral vector to produce a filtered, clarified, twice-digested solution comprising viral vector; filtering the filtered, clarified, twice-digested solution comprising the viral vector through a second chromatographic filter to produce a twice-filtered, clarified, twice-digested solution comprising viral vector; concentrating the twice-filtered, clarified, twice-digested solution comprising the viral vector to produce a concentrated, twice-filtered, clarified, twice digested solution comprising the viral vector; and passing the concentrated, twice-filtered, clarified, twice digested solution comprising the viral -38- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT vector through a protein-coated sterile membrane to produce a concentrated sterilized solution comprising the viral vector.

[0113] In some embodiments, the DNA digestion is performed using an endonuclease. In some embodiments, the endonuclease is a DNAase. In some embodiments, the endonuclease has both DNAase and RNAase activity. In some embodiments, the endonuclease is selected from the group including, but not limited to, DNAse I, DENARASE, Cryonase, or combinations thereof. In some embodiments, the endonuclease is DNAse I. In some embodiments, the endonuclease is DENARASE. In some embodiments, the endonuclease is Cryonase.

[0114] In some embodiments, the filtering of the clarified, digested solution through the first chromatographic filter comprises filtration of the clarified, digested solution via capture chromatography. In some embodiments, the capture chromatography is ion exchange chromatography, affinity chromatography, hydrophobic chromatography, size exclusion chromatography, or multimodal chromatography. In some embodiments, the capture chromatography is ion exchange chromatography. In some embodiments, the capture chromatography is affinity chromatography. In some embodiments, the capture chromatography is hydrophobic chromatography. In some embodiments, the capture chromatography is size exclusion chromatography. In some embodiments, the capture chromatography is multimodal chromatography.

[0115] In some embodiments, the second chromatographic filter is a filter as defined herein. In some embodiments, the second chromatographic step is a membrane as provided for herein, a membrane filter unit as provided for herein, or a stationary phase as provided for herein. In some embodiments, the stationary phase is a membrane filter unit. In some embodiments, the stationary phase is a resin.

[0116] In some embodiments, the second chromatographic filter is a resin.

[0117] In some embodiments, the filtering of the filtered, clarified, twice digested solution through the second chromatographic filter comprises filtration of the filtered, clarified, twice digested solution via polishing chromatography. In some embodiments, the polishing chromatography is ion exchange chromatography, hydrophobic interaction chromatography, size exclusion chromatography, or multimodal chromatography. In some embodiments, the polishing chromatography is ion exchange chromatography. In some embodiments, the polishing chromatography is hydrophobic interaction chromatography. In some embodiments, the polishing chromatography is size exclusion chromatography. In some embodiments, the polishing chromatography is multimodal chromatography. -39- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT

[0118] In some embodiments, the method of purifying a viral vector further comprises the step of collecting media from a cell culture producing the viral vector. In some embodiments, the method of purifying a viral vector further comprises the step of collecting media from a cell culture producing the viral vector prior to digesting DNA. In some embodiments, the method of purifying a viral vector further comprises the step of collecting media from a cell culture producing the viral vector prior to clarifying the digested solution.

[0119] In some embodiments, the protein-coated sterile membrane comprises a sterile membrane pre-flushed with a protein solution. In some embodiments, a sterile membrane is pre-flushed with a protein solution to produce the protein coated sterile membrane. In some embodiments, the method further comprises the step of pre-flushing a sterile membrane with a protein solution to produce a protein coated sterile membrane. In some embodiments, the method further comprises the step of coating a sterile membrane with a protein solution to produce a protein coated sterile membrane.

[0120] In some embodiments, the membrane is a charged sterile membrane. In some embodiments, the protein coated sterile membrane has a charge prior to being coated with a protein solution. In some embodiments, the membrane has a positive charge. In some embodiments, the protein coated sterile membrane has a positive charge prior to being coated with a protein solution. In some embodiments, the membrane is a polyethersulfone (PES) membrane, a nylon membrane, or a polyvinylidene fluoride (PVDF) membrane. In some embodiments, the membrane is a polyethersulfone (PES) membrane. In some embodiments, the membrane is a nylon membrane. In some embodiments, the membrane is a polyvinylidene fluoride (PVDF) membrane.

[0121] In some embodiments, the protein coated sterile membrane has a pore size as provided for herein. In some embodiments, the protein coated sterile membrane has a pore size of about 0.01µm to about 0.45µm, or any value or range therein, as provided for herein. In some embodiments, the protein coated sterile membrane has a pore size of 0.01µm, 0.10µm, 0.11µm, 0.12µm, 0.13µm, 0.14µm, 0.15µm, 0.16µm, 0.17µm, 0.18µm, 0.19µm, 0.2µm, 0.21µm, 0.22µm, 0.23µm, 0.24µm, 0.25µm, 0.26µm, 0.27µm, 0.28µm, 0.29µm, 0.30µm, 0.31µm, 0.32µm, 0.33µm, 0.34µm, 0.35µm, 0.36µm, 0.37µm, 0.38µm, 0.39µm, 0.40µm, 0.41µm, 0.42µm, 0.43µm, 0.44µm, or 0.45µm as provided for herein.

[0122] In some embodiments, the protein coated sterile membrane does not significantly retain the viral vector. In some embodiments, the recovery of the solution comprising the viral vector is as provided for herein. In some embodiments, the recovery of the solution comprising the viral vector is at least about 60%, 61%, 62%, 63%, 64%, 65%, -40- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, as provided for herein.

[0123] In some embodiments, the method of purifying a viral vector further comprises freezing the concentrated sterilized solution comprising the viral vector. In some embodiments, the freezing is performed by a controlled rate freeze.

[0124] In some embodiments, the method of purifying a viral vector further comprises diluting the concentrated sterilized solution comprising the viral vector prior to one or more of the steps.

[0125] In some embodiments, the viral vector is at a concentration of about 1x103to about 1x107transduction units (TU) / mL before passing the solution through the sterile filter. In some embodiments, the viral vector is at a concentration of about 1x104to about 1x106transduction units (TU) / mL before passing the solution through the sterile filter. In some embodiments, the viral vector is at a concentration of 1x106to 1x1010viral particles (vp) / mL before filtering the solution. In some embodiments, the viral vector is at a concentration of 1x107to 1x109viral particles (vp) / mL before filtering the solution.

[0126] In some embodiments, the methods provided for herein may be used to purify any viral vector. In some embodiments, the viral vector is an adenovirus, an adeno- associated virus, a lentivirus, or a retrovirus. In some embodiments, the viral vector is an adenovirus. In some embodiments, the viral vector is an adeno-associated virus. In some embodiments, the viral vector is a lentivirus. In some embodiments, the viral vector is a retrovirus. In some embodiments, the viral vector is a pseudotyped viral vector. In some embodiments, the viral vector is a pseudotyped lentivirus. In some embodiments, the viral vector is a pseudotyped retrovirus. EXEMPLARY METHODS

[0127] FIG.1 illustrates a non-limiting exemplary method of purifying a viral vector and / or producing a concentrated sterilized solution comprising a viral vector as provided for herein. It is to be understood that the method illustrated in FIG. 1 is exemplary only, and is not meant to be limiting in any way.

[0128] As shown in FIG.1, step 110 involves thawing a vial of cells. The cells may be any appropriate cell line capable of producing a viral vector, such as a viral vector disclosed herein. In some embodiments, the cells are HEK293 cells. In some embodiments, -41- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT the cells are HEK293T cells. In some embodiments, the cells are adherent. In some embodiments, the cells are cultured in suspension.

[0129] Cells are passaged (step 115) to allow the cells to return to proper growth rate and then transfected (step 120) with a nucleic acid molecule or nucleic acid molecules encoding one or more viral vectors. In some embodiments, the nucleic acid molecule or nucleic acid molecules encode for a viral vector as provided for herein. In some embodiments, the nucleic acid molecule or nucleic acid molecules further encode for additional elements for viral production, such as but not limited to Gag-Pol and Rev accessory elements. The skilled artisan will readily recognize that additional elements for viral production will vary depending on the viral vector being generated, and thus any such elements are within the scope of the present disclosure.

[0130] At step 125, the transfection is allowed to proceed for a pre-determined amount of time. In various embodiments, the transfection proceeds for between 1 hour and 120 hours. In various embodiments, the transfection proceeds for between 2 hours and 90 hours, between 3 hours and 80 hours, between 5 hours and 70 hours, between 10 hours and 60 hours, between 20 hours and 55 hours, or between 30 hours and 50 hours. In various embodiments, the transfection proceeds for between 40 and 50 hours, such as for 41 hours, for 42 hours, for 43 hours, for 44 hours, for 45 hours, for 46 hours, for 47 hours, for 48 hours, for 49 hours, or for 50 hours. In one non-limiting aspect, the transfection proceeds for 48 hours. After transfection, the cell supernatant is collected. The cell supernatant is then treated with an endonuclease to digest the DNA in the supernatant (step 130). In some embodiments, the DNA digestion is performed using an endonuclease as provided for herein.

[0131] Post DNA digestion, the digested cell culture media is clarified (step 135). In some embodiments, the clarification step comprises passing the digested cell culture media through a filter. In some embodiments, the clarification step comprises passing the digested cell culture media through a series of filters. In some embodiments, the clarification step comprises passing the digested cell culture media through a first filter and then passing the digested cell culture media through a second filter, wherein the first filter has a larger pore size than the second filter.

[0132] The clarified solution is then filtered using capture chromatography (step 140). Further details of capture chromatography are provided for herein. Post chromatographic filtration, the filtered solution is subject to a second round of DNA digestion (step 145). In various embodiments, the DNA digestion is carried out using an endonuclease. -42- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT Further details of example endonucleases are as provided for herein. In some embodiments, the endonuclease used in step 145 is the same endonuclease as used in step 130.

[0133] The twice digested solution is then filtered using polishing chromatography (step 150). Further details of the polishing chromatography are provided for herein. Post polishing filtration, the solution is subject to sterile filtration (step 155). In some embodiments, the sterile filtration step comprises a first step of pre-flushing the sterile filter membrane with a protein solution prior to passing the digested solution through the sterile filter. Further details of example protein solutions are provided for herein. In some embodiments, the protein in the protein solution is a human serum albumin (HSA), such as a recombinant HSA or a non-recombinant HSA. Further details of example proteins in protein solutions are provided for herein.

[0134] The sterile filtered solution is then concentrated and a buffer exchange is performed to the product formulation (step 160). In some embodiments, the concentration step is performed via ultra-centrifugation. In some embodiments, the concentrating step is performed via dialysis. In some embodiments, the concentrating step is performed via centrifugation. In some embodiments, the concentrating step is performed via chromatographic methods. In some embodiment, the concentration step is performed via tangential flow filtration. In some embodiments, the tangential flow filtration is a two stage tangential flow filtration. Once the solution is appropriately concentrated, a buffer exchange is performed to provide the final, concentrated, viral particle formulation. After buffer exchange, the product may be subject to an additional concentration step via the methods provided for herein. Once the final product is prepared, the formulation is subject to a controlled rate freeze (step 165) for long term storage. VIRAL VECTORS

[0135] In some embodiments, the pseudotyped viral vector comprises a VSV- G polypeptide.

[0136] In some embodiments, a VSV-G protein is provided that comprises a mutation at position 198 as compared to SEQ ID NO: 1 or at position 182 as compared to SEQ ID NO: 2. SEQ ID NO: 1 is the full length protein and SEQ ID NO: 2 is the ectodomain of the VSV-G protein. The 16-mer signal peptide of MKCLLYLAFLFIGVNC (SEQ ID NO: 65) as shown at the N-terminus of SEQ ID NO: 1 is cleaved leaving a protein of SEQ ID NO: 2. Thus, although a mutation may be referred to in the context of SEQ ID NO: 2, it should be understood to also be made in the context of SEQ ID NO: 1, which contains the leader sequence, and thus -43- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT would be a position number that is 16 more than the position recited for SEQ ID NO: 2. In some embodiments, the mutation inhibits or decreases the binding of the VSV-G protein to the LDL receptor (LDL-R). In some embodiments, the mutation is a I182D mutation as compared to SEQ ID NO: 2. In some embodiments, the mutation is a I182E mutation as compared to SEQ ID NO: 2.

[0137] In some embodiments, a VSV-G protein is provided that comprises a mutation at position 198 as compared to SEQ ID NO: 10 or at position 182 as compared to SEQ ID NO: 11. SEQ ID NO: 10 is the full length protein and SEQ ID NO: 11 is the ectodomain of the VSV-G protein. The 16-mer signal peptide of MLSYLIFALVVSPILG (SEQ ID NO: 66) as shown at the N-terminus of SEQ ID NO: 10 is cleaved leaving a protein of SEQ ID NO: 11. Thus, although a mutation may be referred to in the context of SEQ ID NO: 11, it should be understood to also be made in the context of SEQ ID NO: 10, which contains the leader sequence, and thus would be a position number that is 16 more than the position recited for SEQ ID NO: 11. In some embodiments, the mutation inhibits or decreases the binding of the VSV-G protein to the LDL receptor (LDL-R). In some embodiments, the mutation is a T182D mutation as compared to SEQ ID NO: 11. In some embodiments, the mutation is a T182E mutation as compared to SEQ ID NO: 11.

[0138] In some embodiments, a VSV-G protein is provided that comprises a mutation at position 198 as compared to SEQ ID NO: 12 or at position 182 as compared to SEQ ID NO: 13. SEQ ID NO: 12 is the full length protein and SEQ ID NO: 13 is the ectodomain of the VSV-G protein. The 16-mer signal peptide of MLRLFLFCFLALGAHS (SEQ ID NO: 67) as shown at the N-terminus of SEQ ID NO: 12 is cleaved leaving a protein of SEQ ID NO: 13. Thus, although a mutation may be referred to in the context of SEQ ID NO: 13, it should be understood to also be made in the context of SEQ ID NO: 12, which contains the leader sequence, and thus would be a position number that is 16 more than the position recited for SEQ ID NO: 13. In some embodiments, the mutation inhibits or decreases the binding of the VSV-G protein to the LDL receptor (LDL-R). In some embodiments, the mutation is a A182D mutation as compared to SEQ ID NO: 13. In some embodiments, the mutation is a A182E mutation as compared to SEQ ID NO: 13.

[0139] In some embodiments, a VSV-G protein is provided that comprises a mutation at position 203 as compared to SEQ ID NO: 14 or at position 182 as compared to SEQ ID NO: 15. SEQ ID NO: 14 is the full length protein and SEQ ID NO: 15 is the ectodomain of the VSV-G protein. The 21-mer signal peptide of MKMKMVIAGLILCIGILPAIG (SEQ ID NO: 68) as shown at the N-terminus of SEQ ID NO: -44- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT 14 is cleaved leaving a protein of SEQ ID NO: 15. Thus, although a mutation may be referred to in the context of SEQ ID NO: 15, it should be understood to also be made in the context of SEQ ID NO: 14, which contains the leader sequence, and thus would be a position number that is 21 more than the position recited for SEQ ID NO: 15. In some embodiments, the mutation inhibits or decreases the binding of the VSV-G protein to the LDL receptor (LDL-R). In some embodiments, the mutation is a V182D mutation as compared to SEQ ID NO: 15. In some embodiments, the mutation is a V182E mutation as compared to SEQ ID NO: 15.

[0140] In some embodiments, a VSV-G protein is provided that comprises a mutation at position 199 as compared to SEQ ID NO: 16 or at position 182 as compared to SEQ ID NO: 17. SEQ ID NO: 16 is the full length protein and SEQ ID NO: 17 is the ectodomain of the VSV-G protein. The 17-mer signal peptide of MTPAFILCMLLAGSSWA (SEQ ID NO: 69) as shown at the N-terminus of SEQ ID NO: 16 is cleaved leaving a protein of SEQ ID NO: 17. Thus, although a mutation may be referred to in the context of SEQ ID NO: 17, it should be understood to also be made in the context of SEQ ID NO: 16, which contains the leader sequence, and thus would be a position number that is 17 more than the position recited for SEQ ID NO: 17. In some embodiments, the mutation inhibits or decreases the binding of the VSV-G protein to the LDL receptor (LDL-R). In some embodiments, the mutation is a V182D mutation as compared to SEQ ID NO: 17. In some embodiments, the mutation is a V182E mutation as compared to SEQ ID NO: 17.

[0141] In some embodiments, a VSV-G protein is provided that comprises a mutation at position 199 as compared to SEQ ID NO: 18 or at position 182 as compared to SEQ ID NO: 19. SEQ ID NO: 18 is the full length protein and SEQ ID NO: 19 is the ectodomain of the VSV-G protein. The 17-mer signal peptide of MNFLLLTFIVLPLCSHA (SEQ ID NO: 70) as shown at the N-terminus of SEQ ID NO: 18 is cleaved leaving a protein of SEQ ID NO: 19. Thus, although a mutation may be referred to in the context of SEQ ID NO: 19, it should be understood to also be made in the context of SEQ ID NO: 18, which contains the leader sequence, and thus would be a position number that is 17 more than the position recited for SEQ ID NO: 19. In some embodiments, the mutation inhibits or decreases the binding of the VSV-G protein to the LDL receptor (LDL-R). In some embodiments, the mutation is a V182D mutation as compared to SEQ ID NO: 19. In some embodiments, the mutation is a V182E mutation as compared to SEQ ID NO: 19.

[0142] In some embodiments, a VSV-G protein is provided that comprises a mutation at position 199 as compared to SEQ ID NO: 20 or at position 182 as compared to SEQ ID NO: 21. SEQ ID NO: 20 is the full length protein and SEQ ID NO: 21 is the -45- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT ectodomain of the VSV-G protein. The 17-mer signal peptide of MLVLYLLLSLLALGAQC (SEQ ID NO: 71) as shown at the N-terminus of SEQ ID NO: 20 is cleaved leaving a protein of SEQ ID NO: 21. Thus, although a mutation may be referred to in the context of SEQ ID NO: 21, it should be understood to also be made in the context of SEQ ID NO: 20, which contains the leader sequence, and thus would be a position number that is 17 more than the position recited for SEQ ID NO: 21. In some embodiments, the mutation inhibits or decreases the binding of the VSV-G protein to the LDL receptor (LDL-R). In some embodiments, the mutation is a I182D mutation as compared to SEQ ID NO: 21. In some embodiments, the mutation is a I182E mutation as compared to SEQ ID NO: 21.

[0143] As used herein, when a polypeptide is said to have a mutation as compared to a reference sequence, such comparison is based on an alignment such as using BlastP or ClustalW or ClutalOmega alignment software using default parameters. For example, position 182 can be found in SEQ ID NO: 2 and also as compared to the other strains as illustrated in FIG.3. FIG.3 illustrates a clustal alignment of the wild-type sequences of the ectodomains of the various strains of the VSV-G protein. The residue that is bolded and underlined are the residues that align to position 182 of SEQ ID NO: 2 of the various strains. SEQ ID NO: 2 refers to ectodomain of the VSV-G protein of the Indiana strain. SEQ ID NO: 11 refers to ectodomain of the VSV-G protein of the New Jersey strain. SEQ ID NO: 13 refers to ectodomain of the VSV-G protein of the Marraba strain. SEQ ID NO: 15 refers to ectodomain of the VSV-G protein of the Carajas strain. SEQ ID NO: 17 refers to ectodomain of the VSV-G protein of the Alagoa strain. SEQ ID NO: 19 refers to ectodomain of the VSV- G protein of the Cocal strain. SEQ ID NO: 21 refers to ectodomain of the VSV-G protein of the Morreton strain. Accordingly, the residue that aligns to residues 182 as compared to SEQ ID NO: 2 can also be mutated as provided for herein.

[0144] In some embodiments, the mutation at position 182 as compared to SEQ ID NO: 2 is not an alanine. In some embodiments, the mutation at position 182 as compared to SEQ ID NO: 2 is not a valine.

[0145] In some embodiments, the mutation at position 182 as compared to SEQ ID NO: 2 is I182S, I182H, I182T, I182Q, or I182N. In some embodiments, the mutation at position 182 as compared to SEQ ID NO: 11 is T182S, T182H, T182Q, or T182N. In some embodiments, the mutation at position 182 as compared to SEQ ID NO: 13 is A182S, A182H, A182T, A182Q, or A182N. In some embodiments, the mutation at position 182 as compared to SEQ ID NO: 15 is V182S, V182H, V182T, V182Q, or V182N. In some embodiments, the mutation at position 182 as compared to SEQ ID NO: 17 is V182S, V182H, V182T, V182Q, -46- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT or V182N. In some embodiments, the mutation at position 182 as compared to SEQ ID NO: 19 is V182S, V182H, V182T, V182Q, or V182N. In some embodiments, the mutation at position 182 as compared to SEQ ID NO: 21 is I182S, I182H, I182T, I182Q, or I182N. In some embodiments, the mutation at position 182 is not a hydrophobic residue. In some embodiments, the mutation at position 182 is a charged residue. In some embodiments, the mutation at position 182 is a negatively charged residue.

[0146] Although, the mutations may be described in reference to SEQ ID NO: 1 or SEQ ID NO: 2, which is the VSV-G protein from the Indiana strain, the mutation can also be used in other strains of the VSV-G protein. For example, the mutation can be made in the New Jersey Strain of VSV-G, the Marraba strain of VSV-G, the Carajas strain of VSV-G, the Alagoa strain of VSV-G, the Cocal strain of VSV-G, or the Morreton strain of VSV-G. In some embodiments, the sequences of each are as provided herein. Examples of these can be found, for example in U.S. Patent Application Publication No.20200216502, which is hereby incorporated by reference. For example, the wild-type full length or ectodomain of the New Jersey Strain of VSV-G are SEQ ID NO: 10 and SEQ ID NO: 11, respectively, the wild-type full length or ectodomain of Marraba strain of VSV-G are SEQ ID NO: 12 and SEQ ID NO: 13, respectively, the wild-type full length or ectodomain of Carajas strain of VSV-G are SEQ ID NO: 14 and SEQ ID NO: 15, respectively, the wild-type full length or ectodomain of Alagoa strain of VSV-G are SEQ ID NO: 16 and SEQ ID NO: 17, respectively, the wild-type full length or ectodomain of Cocal strain of VSV-G are SEQ ID NO: 18 and SEQ ID NO: 19, respectively, or the wild-type full length or ectodomain of Morreton strain of VSV-G are SEQ ID NO: 20 and SEQ ID NO: 21, respectively.

[0147] A VSV-G protein comprising a mutation at position 182 as compared to SEQ ID NO: 2 can also comprise other mutations, such as those described in U.S. Patent Application Publication No. 20200216502, which is hereby incorporated by reference in its entirety. For example, the VSV-G protein can comprise a mutation at a position that corresponds to positions of 8, 47, 209 and / or 354 of SEQ ID NO: 2.

[0148] In some embodiments, the substitution at position 8 is by any amino acid different from the amino acid indicated at that position in the sequence SEQ ID NO: 2, except Y. In some embodiments, the substitution at position 209 is by any amino acid different from the amino acid indicated at that position in the sequence SEQ ID NO: 2, except H. In some embodiments, the substitution at position 47 is by any amino acid different from the amino acid indicated at that position in the sequence SEQ ID NO: 2, except K or R. In some embodiments, -47- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT the substitution at position 354 is by any amino acid different from the amino acid indicated at that position in the sequence SEQ ID NO: 2, except K or R.

[0149] In some embodiments, the substitution is at position 47 or at position 354, or at both positions 47 and 354 are substituted by A, G, F or Q. In some embodiments, the substitution is A or Q.

[0150] In some embodiments, the substitution at position 8 is an alanine, i.e., H8A.

[0151] In some embodiments, the substitution at position 47 is Q or N, i.e., K47Q or K47N.

[0152] In some embodiments, the protein comprises a mutation (substitution) at position 10. In some embodiments, the substitution / mutation is Q10A, Q10R, or Q10K.

[0153] In some embodiments, a protein comprising a mutation at position 182 as compared to SEQ ID NO: 2 comprises a mutation at position 182 and at least, or about, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical as compared to SEQ ID NO: 2 (or SEQ ID NO: 1 if using the full length protein). In some embodiments, the polypeptide comprises a I182D or I182E mutation. In some embodiments, the VSV-G protein comprises a I182S, I182H, I182T, I182Q, or I182N mutation.

[0154] In some embodiments, a protein comprising a mutation at position 182 as compared to SEQ ID NO: 11 comprises a mutation at position 182 and at least, or about, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical as compared to SEQ ID NO: 11 (or SEQ ID NO: 10 if using the full length protein). In some embodiments, the polypeptide comprises a T182D or T182E mutation. In some embodiments, the VSV-G protein comprises a T182S, T182H, T182Q, or T182N mutation.

[0155] In some embodiments, a protein comprising a mutation at position 182 as compared to SEQ ID NO: 13 comprises a mutation at position 182 and at least, or about, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical as compared to SEQ ID NO: 13 (or SEQ ID NO: 12 if using the full length protein). In some embodiments, the polypeptide comprises a A182D or A182E mutation. In some embodiments, the VSV-G protein comprises a A182S, A182H, A182T, A182Q, or A182N mutation. -48- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT

[0156] In some embodiments, a protein comprising a mutation at position 182 as compared to SEQ ID NO: 15 comprises a mutation at position 182 and at least, or about, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical as compared to SEQ ID NO: 15 (or SEQ ID NO: 14 if using the full length protein). In some embodiments, the polypeptide comprises a V182D or V182E mutation. In some embodiments, the VSV-G protein comprises a V182S, V182H, V182T, V182Q, or V182N mutation.

[0157] In some embodiments, a protein comprising a mutation at position 182 as compared to SEQ ID NO: 17 comprises a mutation at position 182 and at least, or about, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical as compared to SEQ ID NO: 17 (or SEQ ID NO: 16 if using the full length protein). In some embodiments, the polypeptide comprises a V182D or V182E mutation. In some embodiments, the VSV-G protein comprises a V182S, V182H, V182T, V182Q, or V182N mutation.

[0158] In some embodiments, a protein comprising a mutation at position 182 as compared to SEQ ID NO: 19 comprises a mutation at position 182 and at least, or about, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical as compared to SEQ ID NO: 19 (or SEQ ID NO: 18 if using the full length protein). In some embodiments, the polypeptide comprises a V182D or V182E mutation. In some embodiments, the VSV-G protein comprises a V182S, V182H, V182T, V182Q, or V182N mutation.

[0159] In some embodiments, a protein comprising a mutation at position 182 as compared to SEQ ID NO: 21 comprises a mutation at position 182 and at least, or about, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical as compared to SEQ ID NO: 21 (or SEQ ID NO: 20 if using the full length protein). In some embodiments, the polypeptide comprises a I182D or I182E mutation. In some embodiments, the VSV-G protein comprises a I182S, I182H, I182T, I182Q, or I182N mutation. Viral Particles -49- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT

[0160] The mutant VSV-G proteins can be used, for example, to pseudotype a virus, such as, but not limited to a lentivirus. Accordingly, in some embodiments, a viral particle comprising a mutant VSV-G protein as provided herein are provided. In some embodiments, the viral particle comprises a VSV-G protein comprising a mutation at position 198 as compared to SEQ ID NO: 1. In some embodiments, a protein comprising a mutation at position 182 as compared to SEQ ID NO: 2 comprises a mutation at position 182 and at least, or about, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical as compared to SEQ ID NO: 2 (or SEQ ID NO: 1 if using the full length protein). In some embodiments, the polypeptide comprises a I182D or I182E mutation as compared to SEQ ID NO: 2. In some embodiments, the VSV-G protein comprises a I182S, I182H, I182T, I182Q, or I182N mutation.

[0161] In some embodiments, the viral particle comprises a VSV-G protein comprising a mutation at position 198 as compared to SEQ ID NO: 10. In some embodiments, a protein comprising a mutation at position 182 as compared to SEQ ID NO: 11 comprises a mutation at position 182 and at least, or about, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical as compared to SEQ ID NO: 11 (or SEQ ID NO: 10 if using the full length protein). In some embodiments, the polypeptide comprises a T182D or T182E mutation as compared to SEQ ID NO: 11. In some embodiments, the VSV- G protein comprises a T182S, T182H, T182Q, or T182N mutation.

[0162] In some embodiments, the viral particle comprises a VSV-G protein comprising a mutation at position 198 as compared to SEQ ID NO: 12. In some embodiments, a protein comprising a mutation at position 182 as compared to SEQ ID NO: 13 comprises a mutation at position 182 and at least, or about, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical as compared to SEQ ID NO: 13 (or SEQ ID NO: 12 if using the full length protein). In some embodiments, the polypeptide comprises a A182D or A182E mutation as compared to SEQ ID NO: 13. In some embodiments, the VSV- G protein comprises a A182S, A182H, A182T, A182Q, or A182N mutation.

[0163] In some embodiments, the viral particle comprises a VSV-G protein comprising a mutation at position 203 as compared to SEQ ID NO: 14. In some embodiments, a protein comprising a mutation at position 182 as compared to SEQ ID NO: 15 comprises a mutation at position 182 and at least, or about, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, -50- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical as compared to SEQ ID NO: 15 (or SEQ ID NO: 14 if using the full length protein). In some embodiments, the polypeptide comprises a V182D or V182E mutation as compared to SEQ ID NO: 15. In some embodiments, the VSV- G protein comprises a V182S, V182H, V182T, V182Q, or V182N mutation.

[0164] In some embodiments, the viral particle comprises a VSV-G protein comprising a mutation at position 199 as compared to SEQ ID NO: 16. In some embodiments, a protein comprising a mutation at position 182 as compared to SEQ ID NO: 17 comprises a mutation at position 182 and at least, or about, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical as compared to SEQ ID NO: 17 (or SEQ ID NO: 16 if using the full length protein). In some embodiments, the polypeptide comprises a V182D or V182E mutation as compared to SEQ ID NO: 17. In some embodiments, the VSV- G protein comprises a V182S, V182H, V182T, V182Q, or V182N mutation.

[0165] In some embodiments, the viral particle comprises a VSV-G protein comprising a mutation at position 199 as compared to SEQ ID NO: 18. In some embodiments, a protein comprising a mutation at position 182 as compared to SEQ ID NO: 19 comprises a mutation at position 182 and at least, or about, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical as compared to SEQ ID NO: 19 (or SEQ ID NO: 18 if using the full length protein). In some embodiments, the polypeptide comprises a V182D or V182E mutation as compared to SEQ ID NO: 19. In some embodiments, the VSV- G protein comprises a V182S, V182H, V182T, V182Q, or V182N mutation.

[0166] In some embodiments, the viral particle comprises a VSV-G protein comprising a mutation at position 199 as compared to SEQ ID NO: 20. In some embodiments, a protein comprising a mutation at position 182 as compared to SEQ ID NO: 21 comprises a mutation at position 182 and at least, or about, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical as compared to SEQ ID NO: 21 (or SEQ ID NO: 20 if using the full length protein). In some embodiments, the polypeptide comprises a I182D or I182E mutation as compared to SEQ ID NO: 21. In some embodiments, the VSV- G protein comprises a I182S, I182H, I182T, I182Q, or I182N mutation.

[0167] In some embodiments, the VSV-G protein further comprises a mutation at position that corresponds to positions 214 and / or 352 of SEQ ID NO: 2. In some -51- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT embodiments, the residue that corresponds to position 214 of SEQ ID NO: 2 is T214. In some embodiments, the residue that corresponds to position 352 of SEQ ID NO: is T352. In some embodiments, the VSV-G protein comprises mutation that corresponds to T214N mutation as compared to SEQ ID NO: 2. In some embodiments, the VSV-G protein comprises mutation that corresponds to T352A mutation as compared to SEQ ID NO: 2. In some embodiments, the VSV-G protein comprises a T214N and T352A mutations as compared to SEQ ID NO: 2. These mutations can be combined with any other mutations as provided for herein. In some embodiments, the T214N and / or T352A mutations are combined with the I182E or I182D mutations. In some embodiments, a VSV-G protein comprises an amino acid sequence of SEQ ID NO: 22 and SEQ ID NO: 23, which combines the I182D or I182E, respectively, with the T214N and T352A mutations. The sequences are also illustrated below with the leader sequences, which are removed during protein processing. VSV-G Protein_ I196D, T230N and T368A mutations (with leader sequence and adjusted numbering) MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPS SSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTC DFRWYGPKYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFP PQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQFINGK CSNYICPTVHNSTTWHSDYKVKGLCDSNLDSMDITFFSEDGE LSSLGKEGTGFRSNYFAYENGGKACKMQYCKHWGVRLPSGVW FEMADKDLFAAARFPECPEGSSISAPSQTSVDVSLIQDVERI LDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIIN GTLKYFETRYIRVDIAAPILSRMVGMISGTTAERELWDDWAP YEDVEIGPNGVLRTSSGYKFPLYMIGHGMLDSDLHLSSKAQV FEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWK SSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDI EMNRLGK (SEQ ID NO: 24) VSV-G Protein_I182D, T214N and T352A mutations (without leader sequence) KFTIVFPHNQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQ VKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRS FTPSVEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVI VQVTPHHVLVDEYTGEWVDSQFINGKCSNYICPTVHNSTTWH SDYKVKGLCDSNLDSMDITFFSEDGELSSLGKEGTGFRSNYF AYENGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPE CPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAG LPISPVDLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIA APILSRMVGMISGTTAERELWDDWAPYEDVEIGPNGVLRTSS GYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDD ESLFFGDTGLSKNPIELVEGWFSSWKSSIASFFFIIGLIIGL FLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK (SEQ ID NO: 22) -52- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT VSV-G Protein with I196E, T230N and T368A mutations (with leader sequence and adjusted numbering) MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPS SSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTC DFRWYGPKYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFP PQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQFINGK CSNYICPTVHNSTTWHSDYKVKGLCDSNLESMDITFFSEDGE LSSLGKEGTGFRSNYFAYENGGKACKMQYCKHWGVRLPSGVW FEMADKDLFAAARFPECPEGSSISAPSQTSVDVSLIQDVERI LDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIIN GTLKYFETRYIRVDIAAPILSRMVGMISGTTAERELWDDWAP YEDVEIGPNGVLRTSSGYKFPLYMIGHGMLDSDLHLSSKAQV FEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWK SSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDI EMNRLGK (SEQ ID NO: 25) VSV-G Protein with I182E, T214N and T352A mutations (without leader sequences) KFTIVFPHNQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQ VKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRS FTPSVEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVI VQVTPHHVLVDEYTGEWVDSQFINGKCSNYICPTVHNSTTWH SDYKVKGLCDSNLESMDITFFSEDGELSSLGKEGTGFRSNYF AYENGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPE CPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAG LPISPVDLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIA APILSRMVGMISGTTAERELWDDWAPYEDVEIGPNGVLRTSS GYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDD ESLFFGDTGLSKNPIELVEGWFSSWKSSIASFFFIIGLIIGL FLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK (SEQ ID NO: 23)

[0168] In some embodiments, the VSV-G protein comprising a mutation at position 182 as compared to SEQ ID NO: 2 further comprises a mutation at position that corresponds to positions 38 and / or 320 or SEQ ID NO: 2. In some embodiments, the residue that corresponds to position 38 of SEQ ID NO: 2 is T38. In some embodiments, the residue that corresponds to position 320 of SEQ ID NO: 2 is T320. In some embodiments, the VSV- G protein comprises mutation that corresponds to T38A mutation as compared to SEQ ID NO: 2. In some embodiments, the VSV-G protein comprises mutation that corresponds to T320A mutation as compared to SEQ ID NO: 2. In some embodiments, the VSV-G protein comprises a T38A and T320A mutations as compared to SEQ ID NO: 2. These mutations can be combined with any other mutations as provided for herein.

[0169] In some embodiments, the other strains of the VSV-G protein as described herein can further comprise one or more mutations corresponding to any of the other -53- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT mutations as compared to SEQ ID NO: 2 and as provided for herein. For example, the other strains of the VSV-G protein as described herein can also comprises the mutations that correspond to T38A, T214N, T320A, and / or T352A in SEQ ID NO: 2. In some embodiments, the other strains of the VSV-G protein as described herein can also comprises the mutations that correspond to T214N and / or T352A in SEQ ID NO: 2 and as illustrated in SEQ ID NO: 22 and SEQ ID NO: 23.

[0170] In some embodiments, the composition comprises a mutation as described in Hwang et al., Gene Ther 2013 Aug;20(8):807-15. (Epub 2013 Jan 31), which is hereby incorporated by reference in its entirety. For example, the mutations can be. at positions 230, 368, 66, and / or 162 that corresponds to SEQ ID NO: 1. The positions will be 16 positions less as compared to SEQ ID NO: 2, when the leader sequence is removed. In some embodiments, the mutations at those positions are, for example, T230N, T368A, K66T, S162T, or any combination thereof. In some embodiments, the VSV-G protein comprises a T230N and a T368A mutation. In some embodiments, the VSV-G polypeptide comprises a K66T, S162T, T230N, and a T368A. These positions are those that correspond to the positions in the full length protein (SEQ ID NO: 1). In some embodiments, the VSV-G protein comprises T230N mutation, a T368A mutation, a K66T mutation, a S162T mutation, or any combination thereof. In some embodiments, the VSV-G protein further comprises one or more mutations in addition to the mutation that corresponds to position 182 of SEQ ID NO: 2, such as those described in U.S. Patent Application Publication No. 20200216502, which is hereby incorporated by reference in its entirety. For example, the VSV-G protein can further comprise a mutation at a position that corresponds to positions of 8, 47, 209 and / or 354 of SEQ ID NO: 2.

[0171] In some embodiments, the substitution at position 8 is by any amino acid different from the amino acid indicated at that position in the sequence SEQ ID NO: 2, except Y. In some embodiments, the substitution at position 209 is by any amino acid different from the amino acid indicated at that position in the sequence SEQ ID NO: 2, except H. In some embodiments, the substitution at position 47 is by any amino acid different from the amino acid indicated at that position in the sequence SEQ ID NO: 2, except K or R. In some embodiments, the substitution at position 354 is by any amino acid different from the amino acid indicated at that position in the sequence SEQ ID NO: 2, except K or R. In some embodiments, the substitution is at position 47 or at position 354, or at both positions 47 and 354 are substituted by A, G, F or Q. In some embodiments, the substitution is A or Q. In some embodiments, the substitution at position 8 is an alanine, i.e., H8A. In some embodiments, the substitution at -54- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT position 47 is Q or N, i.e., K47Q or K47N. In some embodiments, the protein comprises a mutation (substitution) at position 10. In some embodiments, the substitution / mutation is Q10A, Q10R, or Q10K.

[0172] Additionally, in some embodiments, instead of the VSV-G protein or mutant thereof, the viruses can be pseudotyped with other viral structural proteins. Suitable examples of alternate viral structural proteins may be found at least in WO2023 / 064884, WO2023 / 114698, WO2023 / 114884, and WO2023 / 154858, each of which are incorporated by reference in their entirety. Targeting Moieties

[0173] In some embodiments, the viral particle comprises a targeting moiety. The targeting moiety can be used to target the viral particle comprising the mutant VSV-G protein to a cell that expresses the target to which the targeting moiety binds to. In some embodiments, the targeting moiety is an antibody, a scFv antibody, an antigen binding domain, an ankyrin repeat (e.g., DARPIN), a VHH domain antibody, a nanobody, single domain antibody, a FN3 domain, or any combination thereof. The targeting moiety can be attached to the viral surface through an IgG Fc stalk. In some embodiments, the stalk comprises a transmembrane domain. In some embodiments, the transmembrane domain comprises the CD28 transmembrane domain. In some embodiments, the targeting moiety is attached (fused or linked) an envelope glycoprotein G or H of a virus of the Paramyxoviridae family, such as a morbillivirus, such as Measles virus, or a henipavirus, such as Nipah virus, Cedar virus, or Hendra virus. In some embodiments, the targeting moiety can be attached (fused or linked) to a glycoprotein of a virus of the Rhabdoviridae family, such as a vesicular stomatitis New Jersey virus, a vesicular stomatitis Indiana virus, a vesicular stomatitis Alagoas virus, a vesicular stomatitis Maraba virus, a vesicular stomatitis Carajas virus, Parainfluenza virus, Spodoptera frugiperda rhabdovirus isolate Sf G, Drosophila obscura sigmavirus 10A, Wuhan insect virus 7, Perch virus, or Spring viremia of carp virus. In some embodiments, the VSV protein is the mutated proteins, such as those provided for herein. In some embodiments, the targeting moiety is attached to a glycoprotein of a virus of the Filoviridae family, such as Ebola virus or a glycoprotein of a virus of the Arenaviridae family, such as Machupo virus.

[0174] In some embodiments, the targeting moiety is a scFv. In some embodiments, the targeting moiety is a single domain antibody. In some embodiments, the targeting moiety is a VHH. -55- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT

[0175] In some embodiments, the targeting moiety binds to CD7, CD8, cKit (CD117), CD4, CD3, CD5, CD6, CD2, TCR alpha, TCR beta, TCR gamma, TCR delta, CD10, CD34, CD110, CD33, CD14, CD68, CCR7, CD62L, CD25, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, or CXCR3, A glycosylated CD43 epitope expressed on acute leukemia or lymphoma but not on hematopoietic progenitors; A glycosylated CD43 epitope expressed on non-hematopoietic cancers; A kinase anchor protein 4 (AKAP-4); Adrenoceptor beta 3 (ADRB3); AFP; Anaplastic lymphoma kinase (ALK); Androgen receptor; Angiopoietin- binding cell surface receptor 2 (Tie 2); Auto antibody to desmoglein 1 (Dsgl); Auto antibody to desmoglein 3 (Dsg3); B7H3 (CD276); Biotin; Bone marrow stromal cell antigen 2 (BST2); BST1 / CD157; Cancer / testis antigen 1 (NY-ESO-1); Cancer / testis antigen 2 (LAGE-la); Carbonic anhydrase IX (CA1X); Carcinoembryonic antigen (CEA); CCCTC-Binding Factor (Zinc Finger Protein)-Like (BORIS or Brother of the Regulator of lmprinted Sites); CCR4; CD5; CD19; CD20; CD22; CD24; CD30; CD32 (FCGR2A); CD33; CD34; CD38; CD44v6; CD72; CD79a; CD79b; CD97; CD99; CD123; CD171; CD179a; CD179b-IGLll; CD200R; CD276 / B7H3; CD300 molecule-like family member f (CD300LF); CDH1-CD324; CDH6; CDH17; CDH19; Chromosome X open reading frame 61 (CXORF61); Claudin 6 (CLDN6); Claudinl8.2 (CLD18A2 or CLDN18A.2); CMV pp65; C-MYC epitope Tag; Cripto; CS1 (also referred to as CD2 subset 1 or CRACC or SLAMF7 or CD319 or 19A24); CSF2RA (GM- CSFR-alpha); C-type lectin domain family 12 member A (CLEC12A); C-type lectin-like molecule-1 (CLL-1 or CLECL1); Cyclin Bl; Cytochrome P450 IB 1 (CYP1B 1); DLL3; EBV- EBNA3c; EGF-bke module- containing mucin-like hormone receptor-like 2 (EMR2); Elongation factor 2 mutated (ELF2M); Ephrin B2; Ephrin type-A receptor 2 (EphA2); Epidermal growth factor receptor (EGFR); Epidermal growth factor receptor variant III (EGFRviii); Epithelial cell adhesion molecule (EPCAM); ERG; ETS translocation-variant gene 6 located on chromosome 12p (ETV6-AML); Fc fragment of IgA receptor (FCAR or CD89); Fc receptor-like 5 (FCRL5); Fibroblast activation protein alpha (FAP); FITC; Fms Like Tyrosine Kinase 3 (FLT3); Folate receptor alpha (FRa or FR1); Folate receptor beta (FRb); Follicle stimulating hormone receptor (FSHR); Fos-related antigen 1; Fucosyl-GMl; G protein coupled receptor class C group 5 member D (GPRC5D); G protein-coupled receptor 20 (GPR20); GAD; Ganglioside G2 (GD2) ; Ganglioside GD3 (aNeu5Ac(2-8)aNeu5Ac(2- 3)bDGalp(l-4 )bDGlcp(l-l)Cer); Ganglioside GM3 (aNeu5Ac(2-3)bDClalp(l- 4)bDGlcp(l- l)Cer); GD3; GFRalpha4; Glycoprotein 100 (gplOO); Glypican-3 (GPC3); Gonadotropin Hormone receptor (CGHR or GR); GpA33; GpNMB; GPRC5D; Guanylyl cyclase C (GCC); Heat shock protein 70-2 mutated (mut hsp70-2); Hepatitis A virus cellular receptor 1 -56- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT (HAVCR1); Hexasaccharide portion of globoH glycoceramide (GloboH); High molecular weight-melanoma associated antigen (HMWMAA); HIV1 envelope glycoprotein; HLA; HLA- DOA; HLA-A; HLA-A2; HLA-B; HLA-C; HLA-DM; HLA-DOB; HLA-DP; HLA-DQ; HLA- DR; HLA-G; HTLVl-Tax; Human papilloma virus E6 (HPV E6); Human papilloma virus E7 (HPV E7); Human Telomerase reverse transcriptase (hTERT); IgE; IL13Ra2; ILl lRa; Immunoglobulin lambda-like polypeptide 1 (IGLL1); Influenza A hemagglutinin (HA); Insulin-like growth factor 1 receptor (IGF-I receptor); Interleukin 11 receptor alpha (IL-llRa); Interleukin- 13 receptor subunit alpha-2 (IL-13Ra2 or CD213A2); Intestinal carboxyl esterase; KIT (CD117); KSHV K8.1; KSHV-gH; LAMP1 ; Legumain; Leukocyte immunoglobulin-like receptor subfamily A member 2 (LILRA2); Leukocyte-associated immunoglobulin-like receptor 1 (LAIR1); Leutenizing hormone receptor (LHR); Lewis(Y) antigen; Lews Ag; Livl; Locus K 9 (LY6K); Low conductance chloride channel; Lymphocyte antigen 6 complex; Lymphocyte antigen 75 (LY75); Lymphocyte-specific protein tyrosine kinase (LCK); Mammary gland differentiation antigen (NY-BR-1); Melanoma antigen recognized by T cells 1 (MelanA or MARTI); Melanoma- associated antigen 1 (MAGE-A1); Melanoma cancer testis antigen-1 (MAD-CT-1); Melanoma cancer testis antigen-2 (MAD-CT-2); Melanoma inhibitor of apoptosis (ML-IAP); Mesothelin; MPL; Mucin 1 cell surface associated (MUC1); N-Acetyl glucosaminyl-transferase V (NA17); Nectin-4; Neural cell adhesion molecule (NCAM); NKG2D; NYBR1; O-acetyl-GD2 ganglioside (OAcGD2); Olfactory receptor 51E2 (OR51E2); Oncogene fusion protein consisting of breakpoint cluster region (BCR) and Abelson murine leukemia viral oncogene homolog 1 (Abl) (bcr-abl); P53 mutant; Paired box protein Pax-3 (PAX3); Paired box protein Pax-5 (PAX5); Pannexin 3 (PANX3); PDL1; P-glycoprotein; Placenta-specific 1 (PLAC1); Platelet-derived growth factor receptor beta (PDGFR-beta); Polysialic acid; Proacrosin binding protein sp32 (OY-TES1); Prostase; Prostate carcinoma tumor antigen-1 (PCT A-l or Galectin 8); Prostate stem cell antigen (PSCA); Prostate-specific membrane antigen (PSMA); Prostatic acid phosphatase (PAP); Prostein; Protease Serine 21 (Testisin or PRSS21); Proteasome (Prosome Macropain) Subunit Beta Type 9 (LMP2); PTK7; Ras G12V; Ras Homolog Family Member C (RhoC); Rat sarcoma (Ras) mutant; Receptor for Advanced Gly cation Endproducts (RAGE-1); Receptor tyrosine kinase-like orphan receptor 1 (ROR1); Receptor tyrosine-protein kinase ERBB2 or Her-22 / neu; Renal ubiquitous 1 (RU1); Renal ubiquitous 2 (RU2); Sarcoma translocation breakpoints; Serine 2 (TMPRSS2) ETS fusion gene; Sialyl Lewis adhesion molecule (sLe); SLAMF4; SLAMF6; Slea (CA19.9 or Sialyl Lewis Antigen); Sperm protein 17 (SPA17); Squamous Cell Carcinoma Antigen Recognized By T Cells 3 (SART3); Stage-specific embryonic antigen-4 (SSEA-4); STEAP1; -57- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT Survivin; Synovial sarcoma X breakpoint 2 (SSX2); TCR Gamma Alternate Reading Frame Protein (TARP); TCR-beta1 chain; TCR-beta2 chain; TCR-delta chain; TCR-gamma chain; TCRgamma-delta; Telomerase; TGFbetaR2; The antigen recognized by TNT antibody; Thyroid stimulating hormone receptor (TSHR); Timl- / HVCR1; Tissue Factor 1 (TF1); Tn ag; Tn antigen ((Tn Ag) or (GalNAca-Ser / Thr)); TNF receptor family member B cell maturation (BCMA); Transglutaminase 5 (TGS5); Transmembrane protease; TROP2; Tumor endothelial marker 1 (TEM1 / CD248); Tumor endothelial marker 7-related (TEM7R); Tumor protein p53 (p53); Tumor-associated glycoprotein 72 (TAG72); Tyrosinase; Tyrosinase-related protein 2 (TRP-2); Uroplakin 2 (UPK2); Vascular endothelial growth factor receptor 2 (VEGFR2); V- myc avian myelocytomatosis viral oncogene neuroblastoma derived homolog (MYCN); Wilms tumor protein (WT1); or X Antigen Family Member 1A (XAGE1). In some embodiments, the targeting moiety binds to CD7. In some embodiments, the targeting moiety binds to CD8.

[0176] In some embodiments, the targeting moiety binds to a target that is present on a cell, such as an immune cell. In some embodiments, the cell is an immune cell, such as, but not limited to, T cell, B cell; NK cell, dendritic cell, neutrophils, macrophages, a cancer cell; or, for example, CD3+ T cell; CD4+ T cell; CD7+ T cell, CD8+ T cell; CD19+ B cell; CD19+ cancer cell; CD20+ B cell; CD20+ cancer cell; CD30+ lung epithelial cell; CD34+ haematopoietic stem cell; CD105+ endothelial cell; CD105+ haematopoietic stem cell; CD117+ haematopoietic stem cell; CD133+ cancer cell; EpCAM+ cancer cell; GluA2+ neuron; GluA4+ neuron; Haematopoietic stem cell; Hepatocyte; Her2 / Neu+ cancer cell; NKG2D+ natural killer cell; SLC1A3+ astrocyte; SLC7A10+ adipocyte. In some embodiments, the cell is a T cell. In some embodiments, the cell is a B cell. In some embodiments, the cell is a CD7+ T cell and / or CD8+ T cell.

[0177] In some embodiments, the targeting moiety (a polypeptide) can bind to CD7.

[0178] In some embodiments, the polypeptide binds to CD7. In some embodiments, the polypeptide that binds to CD7 is an antibody which binds to non-human primate CD7. In some embodiments, the polypeptide that binds to CD7 is an antibody which binds to human CD7. The sequence of human CD7 (UniProtKB P09564) is as follows (SEQ ID NO: 29): MAGPPRLLLLPLLLALARGLPGALAAQEVQQSPHCTTVPVGASVNITCSTSGGLRGI YLRQLGPQPQDIIYYEDGVVPTTDRRFRGRIDFSGSQDNLTITMHRLQLSDTGTYTC QAITEVNVYGSGTLVLVTEEQSQGWHRCSDAPPRASALPAPPTGSALPDPQTASALP -58- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT DPPAASALPAALAVISFLLGLGLGVACVLARTQIKKLCSWRDKNSAACVVYEDMSHS RCNTLSSPNQYQ (SEQ ID NO: 29)

[0179] In some embodiments, the CD7 antibody comprises a Fc region. The Fc region can be linked to the heavy or light chain of the antibody. The Fc region may be fused directly to the heavy or light chain of the antibody or may be fused indirectly to the heavy or light chain of the antibody via, for example, a peptide linker as provided for herein. In some embodiments, the Fc region is an IgG Fc. In some embodiments, the IgG is selected from IgG1, IgG2, IgG3, or IgG4. In some embodiments, the IgG Fc is IgG1 Fc. In some embodiments, the antibody comprises an Fc constant region of SEQ ID NO: 26 as set forth below: ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPA PELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAK TKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDG SFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 26)

[0180] In some embodiments, the IgG fc is IgG2 Fc. In some embodiments, the antibody comprises an Fc constant region of SEQ ID NO: 27 as set forth below: STKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVA GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPRE EQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYT LPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 27)

[0181] In some embodiments, the IgG fc is IgG4 Fc. In some embodiments, the antibody comprises an Fc constant region of SEQ ID NO: 28 as set forth below: STKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPSCPAPEFL GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPR EEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVY -59- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT TLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFL YSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 28)

[0182] In some embodiments, the IgG Fc is a variant of an IgG1 Fc protein (SEQ ID NO: 26). In some embodiments, the variant IgG1 Fc protein comprises one or more of the mutations that corresponds to those selected from the group consisting of: L234A, L235A, N297A, P329G, I253A, H310A, and H435A of SEQ ID NO: 26. Any of the mutations L234A, L235A, N297A, P329G, I253A, H310A, and H435A of SEQ ID NO: 26 may be present or absent and the mutations may be combined in any combination. In some embodiments, the variant IgG1 Fc protein comprises a mutation that corresponds to L234A and L235A of SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein comprises a mutation that corresponds to N297A of SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein comprises a mutation that corresponds to P329G of SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein comprises a mutation that corresponds to L234A, L235A, N297A, and P329G of SEQ ID N: 83. In some embodiments, the variant IgG1 Fc protein comprises a mutation that corresponds to I253A of SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein comprises a mutation that corresponds to H310A of SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein comprises a mutation that corresponds to H435A of SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein comprises a mutation that corresponds to I253A, H310A, and H435A of SEQ ID NO: 26.

[0183] In some embodiments, the IgG Fc is a variant of an IgG2 Fc protein (SEQ ID NO: 27). In some embodiments, the variant IgG2 Fc protein comprises one or more mutations selected from the group consisting of: N297A, P329G, I253A, H310A, and H435A as those position correspond to SEQ ID NO: 27. Any of the mutations N297A, P329G, I253A, H310A, and H435A of SEQ ID NO: 27 may be present or absent and the mutations may be combined in any combination. In some embodiments, the variant IgG2 Fc protein comprises a mutation that corresponds to N297A of SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein comprises a mutation that corresponds to P329G of SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein comprises a mutation that corresponds to N297A and P329G of SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein comprises a mutation that corresponds to I253A of SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein comprises a mutation that corresponds to H310A of SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein comprises a mutation that corresponds to H435A of -60- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein comprises a mutation that corresponds to I253A, H310A, and H435A of SEQ ID NO: 27.

[0184] In some embodiments, the IgG Fc protein is a variant of an IgG4 Fc protein (SEQ ID NO: 28). In some embodiments, the variant IgG4 Fc protein comprises one or more mutations selected from the group consisting of: S228P, L235E, N297A, P329G, I253A, H310A, and H435A as those positions correspond to SEQ ID NO: 28. Any of the mutations S228P, L235E, N297A, P329G, I253A, H310A, and H435A of SEQ ID NO: 28 may be present or absent and the mutations may be combined in any combination. In some embodiments, the variant IgG4 Fc protein comprises a mutation that corresponds to S228P of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein comprises a mutation that corresponds to L235E of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein comprises a mutation that corresponds to N297A of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein comprises a mutation that corresponds to P329G of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein comprises a mutation that corresponds to S228P, L235E, N297A, and P329G of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein comprises a mutation that corresponds to I253A of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein comprises a mutation that corresponds to H310A of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein comprises a mutation that corresponds to H435A of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein comprises a mutation that corresponds to I253A, H310A, and H435A of SEQ ID NO: 28.

[0185] In some embodiments, the Fc region comprises a variant Fc polypeptide. In some embodiments, the variant Fc polypeptide is a variant Fc polypeptide as provided for in PCT Publication No. WO2024026284 which is hereby incorporated by reference in its entirety. In some embodiments, the variant Fc polypeptide comprises an amino acid sequence that is at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 82: EPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMASRTPEVTCVVVDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLAQDWLNGKEY KCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNAYTQKSLSLSPGK (SEQ ID NO: 82) In some embodiments, the variant Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 82. -61- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT

[0186] In some embodiments, the targeting moiety binds to CD7 and comprises an HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, LCDR3, VH, and VL as recited in Table 1 and Table 2 below: Table 1 - CD7 targeting moiety CD7AB1 CDR sequences Numbering HCDR1 HCDR2 HCDR3 LCDR1 LCDR2 LCDR3 System P Qa e - a ge g o e y a a e eg o s SEQ ID NO: AB ID NO: Region Sequence 42 CD7AB1 VHQVQLQQPGAELVKPGASVKLSCKASGYPFTSYWIHWVKQRPGRGLEWLC I T

[0187] The VH and the VL sequences can be in any format, including, but not limited to an scFv format where the VH and VL regions are linked with a peptide linker. Examples of peptide linkers that can be used to link various peptides provided for herein include, but are not limited to: (GGGGS)n(SEQ ID NO: 64), wherein each n is independently 1-4. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, the variable regions are not linked with a peptide linker. In some embodiments, the polypeptide comprises SEQ ID NO: 42 and SEQ ID NO: 43. In some embodiments, the targeting moiety comprises a linked peptide represented by a formula of VL-Z-VH, wherein Z is a peptide linker. In some embodiments, the targeting moiety comprises a heavy chain variable region as set forth in SEQ ID NO: 42 linked via a linker sequence GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 72) to a light chain variable region as set forth in SEQ ID NO: 43. In some embodiments, a targeting moiety comprising a VL linked via a peptide linker to a VH has the sequence as set forth below: -62- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT DILLTQSPAILSVSPGERVSFSCRASQSIGTSIHWYQQRTNDSPRLLIKYASESISG IPSRFSGSGSGTDFTLSINSVESEDIADYYCQQSNSWPTTFGGGTKLEIKRGGGGSG GGGSGGGGSGGGGSQVQLQQPGAELVKPGASVKLSCKASGYPFTSYWIHWVKQRPGR GLEWLGRIDPNSGDTKYNEKFKNKATLTVDKSSTTAYMQLSSLTSEDSAVYYCARSP YYSNDNSMDYWGQGTSVTVSS (SEQ ID NO: 44)

[0188] In some embodiments, the targeting moiety comprises a linked peptide represented by a formula of VH-Z-VL, wherein Z is a peptide linker. In some embodiments, the targeting moiety comprises a heavy chain variable region as set forth in SEQ ID NO: 42 linked via a linker sequence GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 72) to a light chain variable region as set forth in SEQ ID NO: 43. In some embodiments, a targeting moiety comprising a VH linked via a peptide linker to a VL has the sequence as set forth below: QVQLQQPGAELVKPGASVKLSCKASGYPFTSYWIHWVKQRPGRGLEWLGRIDPNSGD TKYNEKFKNKATLTVDKSSTTAYMQLSSLTSEDSAVYYCARSPYYSNDNSMDYWGQG TSVTVSSGGGGSGGGGSGGGGSGGGGSDILLTQSPAILSVSPGERVSFSCRASQSIG TSIHWYQQRTNDSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYY CQQSNSWPTTFGGGTKLEIKR (SEQ ID NO: 45)

[0189] In some embodiments, the targeting moiety (a polypeptide) can bind to CD8.

[0190] In some embodiments, the polypeptide binds to CD8. In some embodiments, the polypeptide binds to CD8-alpha. In some embodiments, the polypeptide binds to CD8-beta. In some embodiments, the polypeptide binds to CD8 heterodimer. In some embodiments, the CD8 heterodimer comprises CD8-alpha and CD8-beta subunits. In some embodiments, the polypeptide binds to CD8-alpha homodimer. In some embodiments, the polypeptide that binds to CD8 is an antibody which binds to non-human primate CD8. In some embodiments, the antibody that binds to non-human primate CD8 is an antibody which binds to non-human primate CD8-alpha. In some embodiments, the antibody that binds to non-human primate CD8 is an antibody which binds to non-human primate CD8-beta. In some embodiments, the antibody that binds to non-human primate CD8 is an antibody which binds to non-human primate CD8-alpha homodimer. In some embodiments, the antibody that binds to non-human primate CD8 is an antibody which binds to non-human primate CD8 heterodimer. In some embodiments, the polypeptide that binds to CD8 is an antibody which binds to human CD8. In some embodiments, the antibody that binds to human CD8 is an -63- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT antibody which binds to human CD8-alpha. In some embodiments, the antibody that binds to human CD8 is an antibody which binds to human CD8-beta. In some embodiments, the antibody that binds to human CD8 is an antibody which binds to human CD8-alpha homodimer. In some embodiments, the antibody that binds to human CD8 is an antibody which binds to human CD8 heterodimer. The sequence of human CD8-alpha (UniProtKB Q8TAW8) is as follows (SEQ ID NO: 46): MALPVTALLLPLALLLHAARPSQFRVSPLDRTWNLGETVELKCQVLLSNPTSGCSWL FQPRGAAASPTFLLYLSQNKPKAAEGLDTQRFSGKRLGDTFVLTLSDFRRENEGCYF CSALSNSIMYFSHFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGA VHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNHRNRRRVCKCPRPVVKSGDKP SLSARYV (SEQ ID NO: 46)

[0191] The sequence of human CD8-beta (UniProtKB Q8TD28) is as follows (SEQ ID NO: 47): MRPRLWLLLAAQLTVLHGNSVLQQTPAYIKVQTNKMVMLSCEAKISLSNMRIYWLRQ RQAPSSDSHHEFLALWDSAKGTIHGEEVEQEKIAVFRDASRFILNLTSVKPEDSGIY FCMIVGSPELTFGKGTQLSVVDFLPTTAQPTKKSTLKKRVCRLPRPETQKGPLCSPI TLGLLVAGVLVLLVSLGVAIHLCCRRRRARLRFMKQLYK (SEQ ID NO: 47)

[0192] In some embodiments, the CD8 antibody comprises a Fc region. The Fc region can be linked to the heavy or light chain of the antibody. The Fc region may be fused directly to the heavy or light chain of the antibody or may be fused indirectly to the heavy or light chain of the antibody via, for example, a peptide linker as provided for herein. In some embodiments, the Fc region is an IgG Fc as provided for herein. In some embodiments, the IgG is selected from IgG1, IgG2, IgG3, or IgG4. In some embodiments, the IgG fc is IgG1 Fc as provided for herein. In some embodiments, the antibody comprises an Fc constant region of SEQ ID NO: 26. In some embodiments, the IgG fc is IgG2 Fc as provided for herein. In some embodiments, the antibody comprises an Fc constant region of SEQ ID NO: 27. In some embodiments, the IgG fc is IgG4 Fc as provided for herein. In some embodiments, the antibody comprises an Fc constant region of SEQ ID NO: 28.

[0193] In some embodiments, the targeting moiety binds to CD8 and comprises an HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, LCDR3, VH, and VL as recited in Table 3 and Table 4 below: Table 3 - CD8 targeting moiety CD8AB1 CDR sequences -64- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT Numberin HCDR1 HCDR2 HCDR3 LCDR1 LCDR2 LCDR3 g System hhiYPYN DHRYNE V RA E VD F LA NLE NNEDP Qy SEQ ID NO: AB ID NO: Region Sequence 60 CD8AB1 VHEVQLQQSGPELVKPGASVKISCKASRYTFTDYNLHWVKLSHEKSLEWIC P N

[0194] The VH and the VL sequences can be in any format, including, but not limited to an scFv format where the VH and VL regions are linked with a peptide linker. Examples of peptide linkers that can be used to link various peptides provided for herein include, but are not limited to: (GGGGS)n (SEQ ID NO: 64), wherein each n is independently 1-4. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, the variable regions are not linked with a peptide linker. In some embodiments, the polypeptide comprises SEQ ID NO: 60 and SEQ ID NO: 61.

[0195] In some embodiments, the targeting moiety comprises a linked peptide represented by a formula of VL-Z-VH, wherein Z is a peptide linker. In some embodiments, the targeting moiety comprises a heavy chain variable region as set forth in SEQ ID NO: 60 linked via a linker sequence GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 72) to a light chain variable region as set forth in SEQ ID NO: 61. In some embodiments, a targeting moiety comprising a VL linked via a peptide linker to a VH has the sequence as set forth below: NIVLTQSPASLAVSLGQRATISCRASESVDGFGNSFMNWYQQKPGQSPKLLIYLASN LESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQNNEDPYTFGGGTKLEIKRGG GGSGGGGSGGGGSGGGGSEVQLQQSGPELVKPGASVKISCKASRYTFTDYNLHWVKL -65- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT SHEKSLEWIGFIYPYNGGTGYNQKFKNKAKLTVDYSSSTAYMELRSLTSVDAAVYYC ARDHRYNEGVSFDYWGQGTTLTVSS (SEQ ID NO: 62)

[0196] In some embodiments, the targeting moiety comprises a linked peptide represented by a formula of VH-Z-VL, wherein Z is a peptide linker. In some embodiments, the targeting moiety comprises a heavy chain variable region as set forth in SEQ ID NO: 60 linked via a linker sequence GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 72) to a light chain variable region as set forth in SEQ ID NO: 61. In some embodiments, a targeting moiety comprising a VH linked via a peptide linker to a VL has the sequence as set forth below: EVQLQQSGPELVKPGASVKISCKASRYTFTDYNLHWVKLSHEKSLEWIGFIYPYNGG TGYNQKFKNKAKLTVDYSSSTAYMELRSLTSVDAAVYYCARDHRYNEGVSFDYWGQG TTLTVSSGGGGSGGGGSGGGGSGGGGSNIVLTQSPASLAVSLGQRATISCRASESVD GFGNSFMNWYQQKPGQSPKLLIYLASNLESGVPARFSGSGSRTDFTLTIDPVEADDA ATYYCQQNNEDPYTFGGGTKLEIKR (SEQ ID NO: 63)

[0197] In some embodiments, the targeting moiety comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 44, or is substantially similar to SEQ ID NO: 44, or is an active fragment of SEQ ID NO: 44. In some embodiments, the targeting moiety comprises an amino acid sequence having least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NO: 44. In some embodiments, the targeting moiety comprises an amino acid sequence of SEQ ID NO: 44.

[0198] In some embodiments, the targeting moiety comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 45, or is substantially similar to SEQ ID NO: 45, or is an active fragment of SEQ ID NO: 45. In some embodiments, the targeting moiety comprises an amino acid sequence having least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NO: 45. In some embodiments, the targeting moiety comprises an amino acid sequence of SEQ ID NO: 45.

[0199] In some embodiments, the targeting moiety comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 62, or is substantially similar to SEQ ID NO: 62, or is an active fragment of SEQ ID NO: 62. In some embodiments, the targeting -66- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT moiety comprises an amino acid sequence having least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NO: 62. In some embodiments, the targeting moiety comprises an amino acid sequence of SEQ ID NO: 62.

[0200] In some embodiments, the targeting moiety comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 63, or is substantially similar to SEQ ID NO: 63, or is an active fragment of SEQ ID NO: 63. In some embodiments, the targeting moiety comprises an amino acid sequence having least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NO: 63. In some embodiments, the targeting moiety comprises an amino acid sequence of SEQ ID NO: 63.

[0201] In some embodiments, the targeting moiety as provided for herein is attached to the surface of the virus through a stalk portion, S1. In some embodiments, the targeting moiety is represented by the formula T-S1, wherein T is a targeting moiety as provided for herein and S1 is the stalk portion. In some embodiments, the stalk portion, S1, is as provided in PCT Publication No. WO2024026284, which is hereby incorporated by reference in its entirety. In some embodiments, the stalk portion, S1, comprises a variant Fc protein as provided for herein and is given by the formula L1-Fc-L2-X1, wherein L1 is a linker or absent; Fc is the variant Fc protein; L2 is a linker or absent; and X1 is a polypeptide comprising a transmembrane domain. Thus, in some embodiments, the formula representing the targeting moiety may also be written T-L1-Fc-L2-X1. In some embodiments, the stalk portion, S1, does not comprise a variant Fc region and is given by the formula L3-X1, wherein L3 is a flexible polypeptide linker and X1 is a polypeptide comprising a transmembrane domain. Thus, in some embodiments, the formula representing the targeting moiety may also be written T-L3-X1. In some embodiments, the polypeptide comprising a transmembrane domain, X1, comprises a polypeptide having a formula of ECD-TM-ICD, wherein ECD is an extracellular domain, or a fragment thereof, of a cell surface protein, or is absent, TM is a transmembrane domain of a transmembrane protein, and ICD is an intracellular domain of a protein or is a protein that facilitates the incorporation of the targeting moiety into the envelope of the viral particle, or is absent. Accordingly, the formulas representing the targeting moieties linked to the stalk portions may also be written as T-L1-Fc-L2-ECD-TM-ICD or T-L3-ECD-TM-ICD. Exemplary -67- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT identities for L1, L2, L3, Fc, ECD, TM, and ICD may be found in PCT Publication No. WO2024026284, which is hereby incorporated by reference in its entirety.

[0202] In some embodiments, the targeting moiety comprises a formula of T- L1-Fc-L2-ECD-TM-ICD. In some embodiments, T comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 62, or SEQ ID NO: 63. In some embodiments, L1 is present and comprises the amino acid sequence of SEQ ID NO: 72. In some embodiments, Fc comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 82. In some embodiments, L2 is absent. In some embodiments, ECD comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 83: KIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP (SEQ ID NO: 83) In some embodiments, TM comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 84: FWVLVVVGGVLACYSLLVTVAFIIFWV (SEQ ID NO: 84) In some embodiments, ICD comprises an amino acid sequence comprising an env incorporation motif, wherein the env incorporation motif comprises an amino acid sequence of SEQ ID NO: 85: NRVRQGYS (SEQ ID NO: 85)

[0203] In some embodiments, the targeting moiety comprises a formula of T- L1-Fc-L2-ECD-TM-ICD. In some embodiments, T comprises an amino acid sequence of SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 62, or SEQ ID NO: 63. In some embodiments, L1 is present and comprises the amino acid sequence of SEQ ID NO: 72. In some embodiments, Fc comprises an amino acid sequence of SEQ ID NO: 82. In some embodiments, L2 is absent. In some embodiments, ECD comprises an amino acid sequence of SEQ ID NO: 83. In some embodiments, TM comprises an amino acid sequence of SEQ ID NO: 84. In some embodiments, ICD comprises an amino acid sequence comprising an env incorporation motif, wherein the env incorporation motif comprises an amino acid sequence of SEQ ID NO: 85.

[0204] In some embodiments, the targeting moiety comprises a formula of T- L1-Fc-L2-ECD-TM-ICD. In some embodiments, T comprises an amino acid sequence of SEQ ID NO: 44 or SEQ ID NO: 45. In some embodiments, L1 is present and comprises the amino acid sequence of SEQ ID NO: 72. In some embodiments, Fc comprises an amino acid sequence of SEQ ID NO: 82. In some embodiments, L2 is absent. In some embodiments, ECD comprises an amino acid sequence of SEQ ID NO: 83. In some embodiments, TM comprises an amino acid sequence of SEQ ID NO: 84. In some embodiments, ICD comprises an amino acid -68- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT sequence comprising an env incorporation motif, wherein the env incorporation motif comprises an amino acid sequence of SEQ ID NO: 85.

[0205] In some embodiments, the targeting moiety comprises a formula of T- L1-Fc-L2-ECD-TM-ICD. In some embodiments, T comprises an amino acid sequence of SEQ ID NO: 45. In some embodiments, L1 is present and comprises the amino acid sequence of SEQ ID NO: 72. In some embodiments, Fc comprises an amino acid sequence of SEQ ID NO: 82. In some embodiments, L2 is absent. In some embodiments, ECD comprises an amino acid sequence of SEQ ID NO: 83. In some embodiments, TM comprises an amino acid sequence of SEQ ID NO: 84. In some embodiments, ICD comprises an amino acid sequence comprising an env incorporation motif, wherein the env incorporation motif comprises an amino acid sequence of SEQ ID NO: 85.

[0206] In some embodiments, the targeting moiety comprises a formula of T- L3-ECD-TM-ICD. In some embodiments, T comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 62, or SEQ ID NO: 63. In some embodiments, L3 comprises an amino acid sequence of SEQ ID NO: 64, wherein n is 1, 2, or 4. In some embodiments, ECD comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 86: FVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 86) In some embodiments, TM comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 87: IYIWAPLAGTCGVLLLSLVITLYCNHRN (SEQ ID NO: 87) In some embodiments, ICD comprises an amino acid sequence comprising an env incorporation motif, wherein the env incorporation motif comprises an amino acid sequence of SEQ ID NO: 85 or SEQ ID NO: 88: GGTETSQVAPA (SEQ ID NO: 88).

[0207] In some embodiments, the targeting moiety comprises a formula of T- L3-ECD-TM-ICD. In some embodiments, T comprises an amino acid sequence of SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 62, or SEQ ID NO: 63. In some embodiments, L3 comprises an amino acid sequence of SEQ ID NO: 64, wherein n is 1, 2, or 4. In some embodiments, ECD comprises an amino acid sequence of SEQ ID NO: 86. In some embodiments, TM comprises an amino acid sequence of SEQ ID NO: 87. In some embodiments, ICD comprises an amino acid sequence comprising an env incorporation motif, wherein the env incorporation motif comprises an amino acid sequence of SEQ ID NO: 85 or SEQ ID NO: 88. -69- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT

[0208] In some embodiments, the targeting moiety comprises a formula of T- L3-ECD-TM-ICD. In some embodiments, T comprises an amino acid sequence of SEQ ID NO: 44 or SEQ ID NO: 45. In some embodiments, L3 comprises an amino acid sequence of SEQ ID NO: 64, wherein n is 1, 2, or 4. In some embodiments, ECD comprises an amino acid sequence of SEQ ID NO: 86. In some embodiments, TM comprises an amino acid sequence of SEQ ID NO: 87. In some embodiments, ICD comprises an amino acid sequence comprising an env incorporation motif, wherein the env incorporation motif comprises an amino acid sequence of SEQ ID NO: 85 or SEQ ID NO: 88.

[0209] In some embodiments, the targeting moiety comprises a formula of T- L3-ECD-TM-ICD. In some embodiments, T comprises an amino acid sequence of SEQ ID NO: 45. In some embodiments, L3 comprises an amino acid sequence of SEQ ID NO: 64, wherein n is 1, 2, or 4. In some embodiments, ECD comprises an amino acid sequence of SEQ ID NO: 86. In some embodiments, TM comprises an amino acid sequence of SEQ ID NO: 87. In some embodiments, ICD comprises an amino acid sequence comprising an env incorporation motif, wherein the env incorporation motif comprises an amino acid sequence of SEQ ID NO: 85 or SEQ ID NO: 88.

[0210] In some embodiments, the targeting moiety comprising a formula of T- S1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 89: METDTLLLWVLLLWVPGSTGDSAQVQLQQPGAELVKPGASVKLSCKASGYPFTSYWIHWVKQ RPGRGLEWLGRIDPNSGDTKYNEKFKNKATLTVDKSSTTAYMQLSSLTSEDSAVYYCARSPYYS NDNSMDYWGQGTSVTVSSGGGGSGGGGSGGGGSGGGGSDILLTQSPAILSVSPGERVSFSCR ASQSIGTSIHWYQQRTNDSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQS NSWPTTFGGGTKLEIKRASGGGGSGGGGSGGGGSEPKSCDKTHTCPPCPAPEAAGGPSVFLFP PKPKDTLMASRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLT VLAQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHN AYTQKSLSLSPGKKIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLA CYSLLVTVAFIIFWVRSKRSRLLHSDYMNRVRQGYS (SEQ ID NO: 89) In some embodiments, the targeting moiety comprising a formula of T-S1 comprises an amino acid sequence having 90% identity to SEQ ID NO: 89. In some embodiments, the targeting moiety comprising a formula of T-S1 comprises an amino acid sequence having 95% identity to SEQ ID NO: 89. In some embodiments, the targeting moiety comprising a formula of T-S1 comprises an amino acid sequence having 98% identity to SEQ ID NO: 89. In some embodiments, the targeting moiety comprising a formula of T-S1 comprises an amino acid sequence of SEQ ID NO: 89. -70- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT

[0211] In some embodiments, the viral particle comprising a mutant VSV-G protein as provided for herein and comprising a targeting moiety as provided for herein further comprises a nucleic acid molecule encoding for a heterologous molecule of interest or “cargo.” For example, heterologous molecule of interest is meant to refer to any product that may be encoded by a nucleic acid molecule. As non-limiting examples, “cargo” or “heterologous molecule of interest” may refer to an siRNA, an shRNA, a peptide, a polypeptide, a protein, a viral payload, a viral genome, or a combination thereof. In some embodiments, the polypeptide is a chimeric antigen receptor (“CAR”).

[0212] A “chimeric antigen receptor” or “CAR” as used herein refers to an antigen-binding domain that is fused, directly, or indirectly (e.g. via a hinge or transmembrane domain to an intracellular signaling domain capable of activating or stimulating an immune cell. Most commonly, the CAR's extracellular binding domain is composed of a single chain variable fragment (scFv) derived from fusing the variable heavy and light regions of a murine or humanized monoclonal antibody. Alternatively, scFvs may be used that are derived from Fab's (instead of from an antibody, e.g., obtained from Fab libraries). In various embodiments, this scFv is fused to a transmembrane domain and then to an intracellular signaling domain. However, the antigen binding domain can be any molecule that can bind to the to target on the cell. For example, the antigen binding domain of a CAR can be an antibody, a scFv antibody, an antigen binding domain, an ankyrin repeat (e.g. DARPIN), a VHH domain antibody, a nanobody, single domain antibody, a FN3 domain, or any combination thereof. In some embodiments, a CAR includes those that solely provide CD3ζ signals upon antigen binding. In some embodiments, the CAR includes those that provide both costimulation (e.g. CD28 or CD137) and activation (CD3 ζ). In some embodiments, the CARs include those that provide multiple costimulation (e.g. CD28 and CD137) and activation (CD3ζ). In various embodiments, the CAR is selected to have high affinity or avidity for the antigen. In some embodiments, the CAR comprises the 4-1BB domain as well. These are merely illustrative in nature and are not limiting to the present embodiments and any chimeric antigen receptor can be delivered in conjunction with the viral particles and vectors provided for herein. These are non-limiting examples of CARs and any CAR construct could be encoded for by the nucleic acid molecule.

[0213] In some embodiments, the antigen-binding domain of the CAR comprises a VH domain, a VL domain, or a VH and a VL domain. In some embodiments, the VH domain comprises an amino acid sequence having at least 75%, 80%, 85%, 85%, 87%, -71- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 73, or any value or range in-between. EVQLVESGGGLVQPGRSLRLSCAASGFTFNDYAMHWVRQAPGKGLEWVSTISWNSGS IGYADSVKGRFTISRDNAKKSLYLQMNSLRAEDTALYYCAKDIQYGNYYYGMDVWGQ GTTVTVSS(SEQ ID NO: 73) In some embodiments, the VH domain comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 73. In some embodiments, the VH domain comprises an amino acid sequence having at least 95% identity to SEQ ID NO: 73. In some embodiments, the VH domain comprises an amino acid sequence having at least 98% identity to SEQ ID NO: 73. In some embodiments, the VH domain comprises an amino acid sequence having at least 99% identity to SEQ ID NO: 73. In some embodiments, the VH domain comprises an amino acid sequence having the sequence of SEQ ID NO: 73.

[0214] In some embodiments, the VLdomain comprises an amino acid sequence having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 74, or any value or range in- between. EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATG IPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPITFGQGTRLEIK (SEQ ID NO: 74) In some embodiments, the VL domain comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 74. In some embodiments, the VL domain comprises an amino acid sequence having at least 95% identity to SEQ ID NO: 74. In some embodiments, the VL domain comprises an amino acid sequence having at least 98% identity to SEQ ID NO: 74. In some embodiments, the VL domain comprises an amino acid sequence having at least 99% identity to SEQ ID NO: 74. In some embodiments, the VL domain comprises an amino acid sequence having the sequence of SEQ ID NO: 74.

[0215] In some embodiments, the antigen-binding domain of the CAR comprises a VH domain and a VL domain. In some embodiments, the VH and VL domain are not linked by a linker peptide. In some embodiments, the VH and VL domain are linked by a linker peptide, such as those as provided for herein, including but not limited to: (GGGGS)n (SEQ ID NO: 64), wherein each n is independently 1-5. In some embodiment n is 1. In some -72- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT embodiment n is 2. In some embodiment n is 3. In some embodiment n is 4. In some embodiment n is 5.

[0216] In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 73, and comprises a VL having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 74. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 73, and comprises a VL having at least 90% identity to SEQ ID NO: 74. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 73, and comprises a VL having at least 95% identity to SEQ ID NO: 74. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 73, and comprises a VL having at least 98% identity to SEQ ID NO: 74. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 73, and comprises a VL having at least 99% identity to SEQ ID NO: 74. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 73, and comprises a VL having the sequence of SEQ ID NO: 74. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 90% identity to SEQ ID NO: 73, and comprises a VL having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 74. In some embodiments, the antigen- binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 95% identity to SEQ ID NO: 73, and comprises a VL having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 74. In some embodiments, the antigen-binding domain of the CAR -73- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT comprising a VH domain and a VL domain comprises a VH domain having at least 98% identity to SEQ ID NO: 73, and comprises a VL having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 74. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 99% identity to SEQ ID NO: 73, and comprises a VL having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 74. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having an amino acid sequence of SEQ ID NO: 73, and comprises a VL having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 74. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 90% identity to SEQ ID NO: 73, and comprises a VL having at least 90% identity to SEQ ID NO: 74. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 95% identity to SEQ ID NO: 73, and comprises a VL having at least 90% identity to SEQ ID NO: 74. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 90% identity to SEQ ID NO: 73, and comprises a VL having at least 95% identity to SEQ ID NO: 74. In some embodiments, the antigen- binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 95% identity to SEQ ID NO: 73, and comprises a VL having at least 95% identity to SEQ ID NO: 74. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 98% identity to SEQ ID NO: 73, and comprises a VL having at least 98% identity to SEQ ID NO: 74. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 99% identity to SEQ ID NO: 73, and comprises a VL having at least 99% identity to SEQ ID NO: 74. In some embodiments, the antigen- binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having an amino acid sequence of SEQ ID NO: 73, and comprises a VL having an amino acid sequence of SEQ ID NO: 74.

[0217] In some embodiments, the antigen-binding domain of the CAR comprises a formula of VH-Z-VL, wherein VH is a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 73, Z is a linker comprising the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO: 77), and VL is a light chain variable region comprising -74- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT the amino acid sequence of SEQ ID NO: 74. In some embodiments, the antigen-binding domain of the CAR comprising a formula of VH-Z-VL has an amino acid sequence as set forth below: EVQLVESGGGLVQPGRSLRLSCAASGFTFNDYAMHWVRQAPGKGLEWVSTISWNSGS IGYADSVKGRFTISRDNAKKSLYLQMNSLRAEDTALYYCAKDIQYGNYYYGMDVWGQ GTTVTVSSGGGGSGGGGSGGGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLA WYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR SNWPITFGQGTRLEIK (SEQ ID NO: 75) In some embodiments, the antigen-binding domain of the CAR comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to a sequence of SEQ ID NO: 75. In some embodiments, the antigen-binding domain of the CAR comprises an amino acid sequence having at least 90% identity to a sequence of SEQ ID NO: 75. In some embodiments, the antigen-binding domain of the CAR comprises an amino acid sequence having at least 95% identity to a sequence of SEQ ID NO: 75. In some embodiments, the antigen-binding domain of the CAR comprises an amino acid sequence having at least 98% identity to a sequence of SEQ ID NO: 75. In some embodiments, the antigen-binding domain of the CAR comprises an amino acid sequence having at least 99% identity to a sequence of SEQ ID NO: 75. In some embodiments, the antigen-binding domain of the CAR comprises the amino acid sequence of SEQ ID NO: 75.

[0218] In some embodiments, the antigen-binding domain of the CAR comprises a formula of VL-Z-VH, wherein VL is a light chain variable region comprising the amino acid sequence of SEQ ID NO: 74, Z is a linker comprising the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO: 77), and VH is a light chain variable region comprising the amino acid sequence of SEQ ID NO: 73. In some embodiments, the antigen-binding domain of the CAR comprising a formula of VL-Z-VH has an amino acid sequence as set forth below: EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATG IPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPITFGQGTRLEIKGGGGSGG GGSGGGGSEVQLVESGGGLVQPGRSLRLSCAASGFTFNDYAMHWVRQAPGKGLEWVS TISWNSGSIGYADSVKGRFTISRDNAKKSLYLQMNSLRAEDTALYYCAKDIQYGNYY YGMDVWGQGTTVTVSS (SEQ ID NO: 76) -75- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT In some embodiments, the antigen-binding domain of the CAR comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to a sequence of SEQ ID NO: 76. In some embodiments, the antigen-binding domain of the CAR comprises an amino acid sequence having at least 90% identity to a sequence of SEQ ID NO: 76. In some embodiments, the antigen-binding domain of the CAR comprises an amino acid sequence having at least 95% identity to a sequence of SEQ ID NO: 76. In some embodiments, the antigen-binding domain of the CAR comprises an amino acid sequence having at least 98% identity to a sequence of SEQ ID NO: 76. In some embodiments, the antigen-binding domain of the CAR comprises an amino acid sequence having at least 99% identity to a sequence of SEQ ID NO: 76. In some embodiments, the antigen-binding domain of the CAR comprises the amino acid sequence of SEQ ID NO: 76.

[0219] In some embodiments, the antigen-binding domain of the CAR comprises a VH domain, a VL domain, or a VH and a VL domain. In some embodiments, the VH domain comprises an amino acid sequence having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 78, or any value or range in-between. DIVLTQSPAILSASPGEKVTMTCRASSSVNYMDWYQKKPGSSPKPWIYATSNLASGV PARFSGSGSGTSYSLTISRVEAEDAATYYCQQWSFNPPTFGGGTKLEIKGSTS (SEQ ID NO: 78) In some embodiments, the VH domain comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 78. In some embodiments, the VH domain comprises an amino acid sequence having at least 95% identity to SEQ ID NO: 78. In some embodiments, the VH domain comprises an amino acid sequence having at least 98% identity to SEQ ID NO: 78. In some embodiments, the VH domain comprises an amino acid sequence having at least 99% identity to SEQ ID NO: 78. In some embodiments, the VH domain comprises an amino acid sequence having the sequence of SEQ ID NO: 78.

[0220] In some embodiments, the VL domain comprises an amino acid sequence having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 79, or any value or range in- between. EVQLQQSGAELVKPGASVKMSCKASGYTFTSYNMHWVKQTPGQGLEWIGAIYPGNGD TSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSADYYCARSNYYGSSYWFFDVWGA GTTVTVSS (SEQ ID NO: 79) -76- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT In some embodiments, the VL domain comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 79. In some embodiments, the VL domain comprises an amino acid sequence having at least 95% identity to SEQ ID NO: 79. In some embodiments, the VL domain comprises an amino acid sequence having at least 98% identity to SEQ ID NO: 79. In some embodiments, the VL domain comprises an amino acid sequence having at least 99% identity to SEQ ID NO: 79. In some embodiments, the VL domain comprises an amino acid sequence having the sequence of SEQ ID NO: 79.

[0221] In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 78, and comprises a VL having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 79. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 78, and comprises a VL having at least 90% identity to SEQ ID NO: 79. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 78, and comprises a VL having at least 95% identity to SEQ ID NO: 79. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 78, and comprises a VL having at least 98% identity to SEQ ID NO: 79. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 78, and comprises a VL having at least 99% identity to SEQ ID NO: 79. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 78, and comprises a VL having the sequence of SEQ ID NO: 79. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 90% identity to SEQ ID NO: 78, and comprises -77- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT a VL having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 79. In some embodiments, the antigen- binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 95% identity to SEQ ID NO: 78, and comprises a VL having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 79. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 98% identity to SEQ ID NO: 78, and comprises a VL having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 79. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 99% identity to SEQ ID NO: 78, and comprises a VL having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 79. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having an amino acid sequence of SEQ ID NO: 78, and comprises a VL having at least 75%, 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 79. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 90% identity to SEQ ID NO: 78, and comprises a VL having at least 90% identity to SEQ ID NO: 79. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 95% identity to SEQ ID NO: 78, and comprises a VL having at least 90% identity to SEQ ID NO: 79. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 90% identity to SEQ ID NO: 78, and comprises a VL having at least 95% identity to SEQ ID NO: 79. In some embodiments, the antigen- binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 95% identity to SEQ ID NO: 78, and comprises a VL having at least 95% identity to SEQ ID NO: 79. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 98% identity to SEQ ID NO: 78, and comprises a VL having at least 98% identity to SEQ ID NO: 79. In some embodiments, the antigen-binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain having at least 99% identity to SEQ ID NO: 78, and comprises a VL having at least 99% identity to SEQ ID NO: 79. In some embodiments, the antigen- binding domain of the CAR comprising a VH domain and a VL domain comprises a VH domain -78- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT having an amino acid sequence of SEQ ID NO: 78, and comprises a VL having an amino acid sequence of SEQ ID NO: 79.

[0222] In some embodiments, the antigen-binding domain of the CAR comprises a formula of VH-Z-VL, wherein VH is a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 78, Z is a linker comprising the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO: 77), and VL is a light chain variable region comprising the amino acid sequence of SEQ ID NO: 79. In some embodiments, the antigen-binding domain of the CAR comprising a formula of VH-Z-VL has an amino acid sequence as set forth below: DIVLTQSPAILSASPGEKVTMTCRASSSVNYMDWYQKKPGSSPKPWIYATSNLASGV PARFSGSGSGTSYSLTISRVEAEDAATYYCQQWSFNPPTFGGGTKLEIKGSTSGGGG SGGGGSGGGGSSEVQLQQSGAELVKPGASVKMSCKASGYTFTSYNMHWVKQTPGQGL EWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSADYYCARSNYY GSSYWFFDVWGAGTTVTVSS (SEQ ID NO: 80) In some embodiments, the antigen-binding domain of the CAR comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to a sequence of SEQ ID NO: 80. In some embodiments, the antigen-binding domain of the CAR comprises an amino acid sequence having at least 90% identity to a sequence of SEQ ID NO: 80. In some embodiments, the antigen-binding domain of the CAR comprises an amino acid sequence having at least 95% identity to a sequence of SEQ ID NO: 80. In some embodiments, the antigen-binding domain of the CAR comprises an amino acid sequence having at least 98% identity to a sequence of SEQ ID NO: 80. In some embodiments, the antigen-binding domain of the CAR comprises an amino acid sequence having at least 99% identity to a sequence of SEQ ID NO: 80. In some embodiments, the antigen-binding domain of the CAR comprises the amino acid sequence of SEQ ID NO: 80.

[0223] In some embodiments, the antigen-binding domain of the CAR comprises a formula of VL-Z-VH, wherein VL is a light chain variable region comprising the amino acid sequence of SEQ ID NO: 79, Z is a linker comprising the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO: 77), and VHis a light chain variable region comprising the amino acid sequence of SEQ ID NO: 78. In some embodiments, the antigen-binding domain of the CAR comprising a formula of VL-Z-VH has an amino acid sequence as set forth below: -79- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT SEVQLQQSGAELVKPGASVKMSCKASGYTFTSYNMHWVKQTPGQGLEWIGAIYPGNG DTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSADYYCARSNYYGSSYWFFDVWG AGTTVTVSSGGGGSGGGGSGGGGSDIVLTQSPAILSASPGEKVTMTCRASSSVNYMD WYQKKPGSSPKPWIYATSNLASGVPARFSGSGSGTSYSLTISRVEAEDAATYYCQQW SFNPPTFGGGTKLEIKGSTS (SEQ ID NO: 81) In some embodiments, the antigen-binding domain of the CAR comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to a sequence of SEQ ID NO: 81. In some embodiments, the antigen-binding domain of the CAR comprises an amino acid sequence having at least 90% identity to a sequence of SEQ ID NO: 81. In some embodiments, the antigen-binding domain of the CAR comprises an amino acid sequence having at least 95% identity to a sequence of SEQ ID NO: 81. In some embodiments, the antigen-binding domain of the CAR comprises an amino acid sequence having at least 98% identity to a sequence of SEQ ID NO: 81. In some embodiments, the antigen-binding domain of the CAR comprises an amino acid sequence having at least 99% identity to a sequence of SEQ ID NO: 81. In some embodiments, the antigen-binding domain of the CAR comprises the amino acid sequence of SEQ ID NO: 81.

[0224] In some embodiments, the antigen-binding domain of the CAR comprises rituximab, ocrelizumab, obinutuzumab, ofatumumab, ibritumomab tiuxetan, tositumomab, or ublituximab. In some embodiment, the antigen-binding domain comprises rituximab. In some embodiment, the antigen-binding domain comprises ofatumumab. In some embodiments, the CAR comprises the 4-1BB domain as well. These are merely illustrative in nature and are not limiting to the present embodiments and any chimeric antigen receptor can be delivered in conjunction with the viral particles and vectors provided for herein. These are non-limiting examples of CARs and any CAR construct could be encoded for by the nucleic acid molecule.

[0225] In some embodiments, the CAR comprises an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a sequence of SEQ ID NO: 90: MALPVTALLLPLALLLHAARPGSEVQLVESGGGLVQPGRSLRLSCAASGFTF NDYAMHWVRQAPGKGLEWVSTISWNSGSIGYADSVKGRFTISRDNAKKSLYL QMNSLRAEDTALYYCAKDIQYGNYYYGMDVWGQGTTVTVSSGGGGSGGGGSG GGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPITFGQ GTRLEIKSGLDFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAA GGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNHRNKRGRKKLLYI -80- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT FKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQL YNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYS EIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR (SEQ ID NO: 90) or is substantially similar to SEQ ID NO: 90, or is an active fragment of SEQ ID NO: 90. In some embodiments, the CAR comprises an amino acid sequence that is at least 90% identical to a sequence of SEQ ID NO: 90. In some embodiments, the CAR comprises an amino acid sequence that is at least 95% identical to a sequence of SEQ ID NO: 90. In some embodiments, the CAR comprises an amino acid sequence that is at least 98% identical to a sequence of SEQ ID NO: 90. In some embodiments, the CAR comprises an amino acid sequence of SEQ ID NO: 90. In some embodiments, the sub domains of the CAR (e.g., the antigen binding domain, hinge domain, transmembrane domain, costimulatory domain, signaling domain) are as provided in PCT Publication No. WO2024026284, which is hereby incorporated by reference in its entirety.

[0226] In some embodiments, the pseudotyped viral particle further comprises a heterologous nucleic acid molecule encoding a cargo of interest. The nucleic acid molecule may be useful for modulating the expression of a target gene. In some embodiments, the cargo can be used to modulate the activity of a cell or express a protein that is trafficked to the surface of the target cell. Therefore, in some embodiments, the nucleic acid may comprise an siRNA or an shRNA. The nucleic acid may also encode for a cargo of interest. Therefore, in some embodiments, the cargo of interest may comprise a polypeptide or portion thereof, a protein or portion thereof, a chimeric antigen receptor or portion thereof, or a tumor antigen or a portion thereof. In some embodiments, the cargo of interest is an antibody that is produced by the virus, which can then be secreted by the cell that is infected with the virus. The term “protein” can refer to any polypeptide that carries a native function in a cellular environment. Therefore, in some embodiments, the protein encoded by the nucleic acid cargo of interest may comprise an enzyme, a nuclear receptor, a transporter, a ribosomal protein, a membrane bound protein, a cytoplasmic protein, a G-protein coupled receptor, a voltage gated ion channel, a secretory protein, a mitochondria protein, a cytokine, a chimeric antigen receptor, a tumor antigen, or a portion or chimeric species thereof.

[0227] Without being bound to any particular theory, the viral particle comprising the mutant VSV-G protein as provided for herein that comprises a targeting moiety can be used to express the heterologous molecule of interest in the target cell. Thus, for example, the CAR can be expressed in a T cell that is targeted by a viral particle pseudotyped -81- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT with a VSV-G protein as provided for herein. Where the T cell is the intended target the viral particle can comprise a targeting moiety that binds to a target on the surface of a T cell, such as, but not limited to CD2, CD3, CD4, CD5, CD7 or CD8. In some embodiments, the target is CD2. In some embodiments, the target is CD3. In some embodiments, the target is CD4. In some embodiments, the target is CD5. In some embodiments, the target is CD6. In some embodiments, the target is CD7. In some embodiments, the target is CD8.

[0228] In some embodiments, the pseudotyped viral particle is a recombinant lentivirus. In some embodiments, the recombinant pseudotyped viral particle is replication competent. In some embodiments, the recombinant pseudotyped viral particle is replication incompetent. Exemplary viral particles:

[0229] In some embodiments, a viral particle is provided, the the viral particle comprising a heterologous viral glycoprotein and a targeting moiety, wherein the heterologous viral glycoprotein comprises an amino acid sequence selected from SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, or SEQ ID NO: 25 and wherein the targeting moiety comprises an amino acid sequence of SEQ ID NO: 89. In some embodiments, the viral particle further comprises a nucleic acid molecule encoding a heterologous molecule of interest. In some embodiments, the heterologous molecule of interest is as provided for herein. In some embodiments, the heterologous molecule of interest is a CAR as provided for herein. In some embodiments, the CAR comprises an amino acid sequence having at least 85% identity to SEQ ID NO: 90, at least 90% identity to SEQ ID NO: 90, at least 95% identity to SEQ ID NO: 90. at least 99% identity to SEQ ID NO: 90, or at least 100% identity to SEQ ID NO: 90.

[0230] In some embodiments, a viral particle is provided, the the viral particle comprising a heterologous viral glycoprotein and a targeting moiety, wherein the heterologous viral glycoprotein comprises an amino acid sequence selected from SEQ ID NO: 22 or SEQ ID NO: 23, and wherein the targeting moiety comprises an amino acid sequence of SEQ ID NO: 89. In some embodiments, the viral particle further comprises a nucleic acid molecule encoding a heterologous molecule of interest. In some embodiments, the heterologous molecule of interest is as provided for herein. In some embodiments, the heterologous molecule of interest is a CAR as provided for herein. In some embodiments, the CAR comprises an amino acid sequence having at least 85% identity to SEQ ID NO: 90, at least 90% identity to SEQ ID NO: 90, at least 95% identity to SEQ ID NO: 90. at least 99% identity to SEQ ID NO: 90, or at least 100% identity to SEQ ID NO: 90. -82- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT

[0231] In some embodiments, a viral particle is provided, the the viral particle comprising a heterologous viral glycoprotein and a targeting moiety, wherein the heterologous viral glycoprotein comprises an amino acid sequence of SEQ ID NO: 22 and wherein the targeting moiety comprises an amino acid sequence of SEQ ID NO: 89. In some embodiments, the viral particle further comprises a nucleic acid molecule encoding a heterologous molecule of interest. In some embodiments, the heterologous molecule of interest is as provided for herein. In some embodiments, the heterologous molecule of interest is a CAR as provided for herein. In some embodiments, the CAR comprises an amino acid sequence having at least 85% identity to SEQ ID NO: 90, at least 90% identity to SEQ ID NO: 90, at least 95% identity to SEQ ID NO: 90. at least 99% identity to SEQ ID NO: 90, or at least 100% identity to SEQ ID NO: 90.

[0232] In some embodiments, a viral particle is provided, the the viral particle comprising a heterologous viral glycoprotein and a targeting moiety, wherein the heterologous viral glycoprotein comprises an amino acid sequence of SEQ ID NO: 23 and wherein the targeting moiety comprises an amino acid sequence of SEQ ID NO: 89. In some embodiments, the viral particle further comprises a nucleic acid molecule encoding a heterologous molecule of interest. In some embodiments, the heterologous molecule of interest is as provided for herein. In some embodiments, the heterologous molecule of interest is a CAR as provided for herein. In some embodiments, the CAR comprises an amino acid sequence having at least 85% identity to SEQ ID NO: 90, at least 90% identity to SEQ ID NO: 90, at least 95% identity to SEQ ID NO: 90. at least 99% identity to SEQ ID NO: 90, or at least 100% identity to SEQ ID NO: 90. Enumerated Embodiments

[0233] The following examples are illustrative, but not limiting, of the compounds, compositions and methods described herein. Other suitable modifications and adaptations known to those skilled in the art are within the scope of the following embodiments. 1. A method of producing a concentrated sterilized solution comprising a viral vector, the method comprising the steps of: clarifying a solution comprising cell culture media and the viral vector; filtering the clarified solution comprising the viral vector through a first chromatographic filter to produce a filtered clarified solution comprising the viral vector; -83- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT passing the filtered clarified solution comprising the viral vector through a protein coated sterile membrane to produce a sterilized solution comprising the viral vector; and concentrating the sterilized solution comprising the viral vector to produce the concentrated sterilized solution comprising the viral vector. 2. The method of embodiment 1, further comprising mixing the concentrated sterilized solution comprising the viral vector with one or more sterile, pyrogen-free buffers and / or excipients to produce a sterile pharmaceutical composition comprising the viral vector. 3. The method of embodiment 1, further comprising collecting the cell culture media from a cell culture producing the viral vector prior to clarifying the solution. 4. The method of any one of embodiments 1-3, further comprising filtering the filtered clarified solution comprising the viral vector through a second chromatographic filter to produce a twice-filtered clarified solution comprising the viral vector prior to passing filtered clarified solution through the protein coated sterile membrane. 5. The method of embodiment 1, wherein the protein coated sterile membrane comprises a membrane pre-flushed with a protein solution. 6. The method of embodiment 1, the method further comprising coating a sterile membrane with a protein solution to produce the protein coated sterile membrane. 7. The method of embodiment 5 or 6, wherein the protein solution comprises one or more component selected from a protein, 2-Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS), histidine, phosphate, HEPES ((4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), sodium chloride, or combinations thereof. 8. The method of embodiment 7, wherein the protein solution comprises about 1% w / v of a protein, about 20 mM 2-Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS), about 100 mM to about 400 mM sodium chloride, and a pH of about 6.0 to about 8.0. 9. The method of embodiment 7, wherein the protein comprises human serum albumin, -84- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT myoglobulin, bovine serum albumin, immunoglobulin, immunoglobulin fragments, fibronectin, vitronectin, or any combination thereof. 10. The method of embodiment 9, wherein the human serum albumin is recombinant human serum albumin, such as being produced from a plant. 11. The method of embodiment 9, wherein the human serum albumin is produced from a plant. 12. The method of embodiment 9, wherein the human serum albumin is a non- recombinant human serum albumin. 13. The method of embodiment 12, wherein the non-recombinant human serum albumin is USP grade non-recombinant human serum albumin. 14. The method of embodiment 6, wherein the protein coated sterile membrane has a positive charge prior to being coated with a protein. 15. The method of embodiment 6, wherein the membrane a polyethersulfone (PES) membrane, a nylon membrane, or a polyvinylidene fluoride (PVDF) membrane. 16. The method of any one of embodiments 1-15, wherein the clarified solution is filtered through the first chromatographic filter by capture chromatography. 17. The method of embodiment 16, wherein the capture chromatography is ion exchange chromatography, affinity chromatography, hydrophobic chromatography, size exclusion chromatography, or multimodal chromatography. 18. The method of embodiment 4, wherein the filtered solution is filtered through the second chromatographic filter by polishing chromatography. 19. The method of embodiment 18, wherein the polishing chromatography is ion exchange chromatography, hydrophobic interaction chromatography, size-exclusion, or multimodal chromatography. -85- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT 20. The method of any one of embodiments 1-19, further comprising digesting DNA in the solution comprising the cell culture media and the viral vector prior to the clarifying the solution. 21. The method of any one of embodiments 1-20, further comprising digesting DNA in the solution comprising the viral vector after filtering the clarified solution. 22. The method of any one of embodiments 1-21, wherein the concentrating step comprises concentrating the viral vector by tangential flow filtration. 23. The method of embodiment 22, wherein the tangential flow filtration comprises a two stage tangential flow filtration. 24. The method of embodiment 22 or 23, wherein the tangential flow filtration utilizes a 1.0 mm ID fiber and 750 kDa pore size. 25. The method of any one of embodiments 1-24, wherein the concentrated sterilized solution comprising the viral vector is mixed in a sterile environment with the one or more sterile, pyrogen-free buffers and / or excipients to produce the sterile pharmaceutical composition comprising the viral vector. 26. The method of embodiment 25, wherein the mixing with one or more sterile, pyrogenic free buffers and / or excipients comprises a buffer exchange step. 27. The method of embodiment 1, further comprising: prior to clarifying the solution: i) obtaining a solution comprising cell culture media and the viral vector; and ii) digesting DNA in the solution comprising cell culture media and the viral vector to produce a first digested solution comprising the viral vector; wherein clarifying the solution comprises clarifying the digested solution comprising the viral vector to produce a clarified, digested solution comprising the viral vector; -86- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT subsequent to filtering the clarified solution and prior to passing the filtered clarified solution through a protein coated sterile membrane: i) digesting DNA in the filtered, clarified, digested solution comprising the viral vector to produce a filtered, clarified, twice-digested solution comprising viral vector; and ii) filtering the filtered, clarified, twice-digested solution comprising the viral vector through a second chromatographic filter to produce a twice-filtered, clarified, twice-digested solution comprising viral vector; wherein the protein coated sterile membrane comprises a membrane pre-flushed with a protein solution, wherein the protein solution comprises about 0.1% w / v to 2.0% w / v of a protein, about 10 mM to about 30 mM 2-Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS), about 100 mM to about 400 mM sodium chloride, and a pH of about 6.0 to about 8.0. 28. A method of producing a concentrated sterilized solution comprising a viral vector, the method comprising the steps of: digesting DNA in a solution comprising the viral vector to produce a first digested solution comprising the viral vector; clarifying the digested solution comprising the viral vector to produce a clarified, digested solution comprising the viral vector; filtering the clarified, digested solution comprising the viral vector through a first chromatographic filter to produce a filtered, clarified, digested solution comprising the viral vector; digesting DNA in the filtered, clarified, digested solution comprising the viral vector to produce a filtered, clarified, twice-digested solution comprising viral vector; filtering the filtered, clarified, twice-digested solution comprising the viral vector through a second chromatographic filter to produce a twice-filtered, clarified, twice-digested solution comprising viral vector; passing the twice-filtered, clarified, twice-digested solution comprising the viral vector through a protein coated sterile membrane to produce a sterilized solution comprising the viral vector; and concentrating the sterilized solution comprising the viral vector to produce the concentrated sterilized solution comprising the viral vector. 29. The method of embodiment 28, wherein the digesting DNA is performed using an -87- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT endonuclease having DNAase activity, RNAase activity, or a combination thereof, such as DNAse I, DENARASE, Cryonase, and the like. 30. The method of embodiment 28, wherein the second chromatographic filter is a resin. 31. The method of embodiment 28, further comprising mixing the concentrated sterilized solution comprising the viral vector with one or more sterile, pyrogen-free buffers and / or excipients to produce a sterile pharmaceutical composition comprising the viral vector. 32. The method of embodiment 28, further comprising collecting media from a cell culture producing the viral vector. 33. The method of embodiment 28, further comprising collecting media from a cell culture producing the viral vector prior to digesting DNA. 34. The method of embodiment 28, further comprising collecting media from a cell culture producing the viral vector prior to clarifying the digested solution. 35. The method of embodiment 28, wherein the protein coated sterile membrane comprises a membrane pre-flushed with a protein solution. 36. The method of embodiment 35, the method further comprising coating a sterile membrane with a protein solution to produce the protein coated sterile membrane. 37. The method of embodiment 36, wherein the protein solution comprises one or more of a protein, 2-Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS), histidine, phosphate, HEPES ((4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), sodium chloride, or combinations thereof. 38. The method of embodiment 37, wherein the protein solution comprises about 1% w / v of a protein, about 20 mM 2-Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS), about 100 mM to about 400 mM sodium chloride, and a pH of about 6.0 to about 8.0. 39. The method of any one of embodiments 37-38, wherein the protein is a human serum -88- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT albumin. 40. The method of embodiment 39, wherein the human serum albumin is a recombinant human serum albumin. 41. The method of embodiment 39, wherein the human serum albumin is produced from a plant. 42. The method of embodiment 39, wherein the human serum albumin is a non- recombinant human serum albumin. 43. The method of embodiment 42, wherein the non-recombinant human serum albumin is USP grade non-recombinant human serum albumin. 44. The method of any one of embodiments 35-43, wherein the protein coated sterile membrane has a positive charge prior to being coated with a protein. 45. The method of any one of embodiments 35-44, wherein the membrane is a polyethersulfone (PES), a nylon membrane, or a polyvinylidene fluoride (PVDF) membrane. 46. The method of any one of embodiments 28-45, wherein the protein coated sterile membrane has a pore size of about 0.01µm, about 0.10µm, about 0.11µm, about 0.12µm, about 0.13µm, about 0.14µm, about 0.15µm, about 0.16µm, about 0.17µm, about 0.18µm, about 0.19µm, about 0.2µm, about 0.21µm, about 0.22µm, about 0.23µm, about 0.24µm, about 0.25µm, about 0.26µm, about 0.27µm, about 0.28µm, about 0.29µm, about 0.30µm, about 0.31µm, about 0.32µm, about 0.33µm, about 0.34µm, about 0.35µm, about 0.36µm, about 0.37µm, about 0.38µm, about 0.39µm, about 0.40µm, about 0.41µm, about 0.42µm, about 0.43µm, about 0.44µm, or about 0.45µm. 47. The method of any one of embodiments 28-46, wherein the protein coated sterile membrane does not significantly retain the viral vector. 48. The method of embodiment 47, wherein the recovery of the viral vector from the solution is at least about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, -89- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or about 100%. 49. The method of any one of embodiments 28-48, wherein the clarified solution is filtered through the first chromatographic filter by capture chromatography. 50. The method of embodiment 49, wherein the capture chromatography is ion exchange chromatography, affinity chromatography, hydrophobic chromatography, size exclusion chromatography, or multimodal chromatography. 51. The method of embodiment 28, wherein the first filtered solution is filtered by polishing chromatography through the second filter. 52. The method of embodiment 51, wherein the polishing chromatography is ion exchange chromatography, hydrophobic interaction chromatography, size-exclusion, or multimodal chromatography. 53. The method of any one of embodiments 28-52, wherein the concentrating step comprises concentrating the viral vector by tangential flow filtration. 54. The method of embodiments 53, wherein the tangential flow filtration comprises a two stage tangential flow filtration. 55. The method of embodiment 53 or embodiment 54, wherein the tangential flow filtration utilizes a 1.0 mm ID fiber and 750 kDa pore size. 56. The method of any one of embodiments 28-55, wherein the concentrated sterilized solution comprising the viral vector is mixed in a sterile environment with excipients to produce the sterile pharmaceutical composition comprising the viral vector. 57. The method of embodiment 56, wherein the mixing with excipients comprises a buffer exchange step. -90- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT 58. The method of any one of embodiments 1-57, further comprising freezing the concentrated sterilized solution comprising the viral vector or the sterile pharmaceutical composition comprising the viral vector. 59. The method of embodiment 58, wherein the freezing is performed by a controlled rate freeze. 60. The method of any one of embodiments 1-59, wherein the viral vector is a lentiviral vector, an adenoviral vector, an adeno-associated viral vector, or a retroviral vector. 61. The method of any one of embodiments 1-60, wherein the concentrated sterilized solution comprising the viral vector is diluted prior to one or more of the steps. 62. The method of any one of embodiments 1-61, wherein the viral vector is at a concentration of about 1x104to about 1x106transduction units (TU) / mL before passing the solution through the sterile filter. 63. The method of any one of embodiments 1-62, wherein the viral vector is at a concentration of 1x107to 1x109viral particles (vp) / mL before passing the solution through the sterile filter. 64. The method of any one of the preceding embodiments, wherein the viral vector comprises a VSV-G polypeptide. 65. The method of any one of the preceding embodiments, wherein the viral vector, such as a lentiviral vector, is pseudotyped with a VSV-G polypeptide comprising a mutation that corresponds to a mutation at position 182 of SEQ ID NO: 2. 66. The method of embodiments 64 or 65, wherein the VSV-G polypeptide comprises an amino acid sequence of SEQ ID NO: 2 with a mutation at position 182 and has at least 95% identity to SEQ ID NO: 2. 67. The method of embodiments 64-66, wherein the VSV-G polypeptide comprises a I182E or I182D mutation as compared to SEQ ID NO: 2. -91- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT 68. The method of any one of embodiments 64-67, wherein the VSV-G polypeptide comprises an amino acid sequence of SEQ ID NO: 1 with a mutation at position 198 and has at least 70% identity to SEQ ID NO: 2. 69. The method of any one of embodiments 64-68, wherein the VSV-G polypeptide comprises a mutation that corresponds to I182D or I182E as compared to a sequence of SEQ ID NO: 2. 70. The method of any one of embodiments 64-69, wherein the VSV-G polypeptide comprises an amino acid sequence at least 95% identical to a sequence of SEQ ID NO: 4. 71. The method of any one of embodiments 64-70, wherein the VSV-G polypeptide comprises an amino acid sequence of SEQ ID NO: 4. 72. The method of any one of embodiments 64-69, wherein the VSV-G polypeptide comprises an amino acid sequence at least 95% identical to a sequence of SEQ ID NO: 5. 73. The method of any one of embodiments 64-72, wherein the VSV-G polypeptide comprises an amino acid sequence of SEQ ID NO: 5. 74. The method of any one of embodiments 64-73, wherein the VSV-G polypeptide further comprises a mutation in the VSV-G protein that corresponds to a position of 8, 10, 47, 209 and / or 354 as compared to SEQ ID NO: 2. 75. The method of any one of embodiments 64-74, wherein the VSV-G polypeptide further comprises a mutation that corresponds to position 8 in SEQ ID NO: 2, wherein the mutation is any amino acid different from the amino acid indicated at that position in SEQ ID NO: 2, except Y. 76. The method of any one of embodiments 64-75, wherein the VSV-G polypeptide further comprises a mutation that corresponds to position 209 in SEQ ID NO: 2, wherein the mutation is any amino acid different from the amino acid indicated at that position in SEQ ID NO: 2, except H. -92- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT 77. The method of any one of embodiments 64-76, wherein the VSV-G polypeptide further comprises a mutation that corresponds to position 47 in SEQ ID NO: 2, wherein the mutation is any amino acid different from the amino acid indicated at that position in SEQ ID NO: 2, except K or R. 78. The method of any one of embodiments 64-77, wherein the VSV-G polypeptide further comprises a mutation that corresponds to position 354 in SEQ ID NO: 2, wherein the mutation is any amino acid different from the amino acid indicated at that position in SEQ ID NO: 2, except K or R. 79. The method of any one of embodiments 64-78, wherein the VSV-G polypeptide further comprises a mutation that corresponds to position 10 in SEQ ID NO: 2, wherein the mutation is any amino acid different from the amino acid indicated at that position in SEQ ID NO: 2, except Q or N. 80. The method of any one of embodiments 64-79, wherein the VSV-G polypeptide further comprises a substitution at position 47 or at position 354, or at both positions 47 and 354, wherein each position is, independently, substituted by A, G, F, Q, or N. 81. The method of any one of embodiments 64-80, wherein the VSV-G polypeptide comprises a substitution at position 8, wherein the substitution is H8A, H8I, H8V, H8L, and the like. 82. The method of any one of embodiments 64-81, wherein the VSV-G polypeptide comprises a substitution at position 47, wherein the substitution is K47Q or K47N. 83. The method of any one of embodiments 64-82, wherein the VSV-G polypeptide comprises a substitution H8A and / or K47Q mutation. 84. The method of any one of embodiments 94-83, wherein the VSV-G polypeptide comprises a Q10A, Q10R, or Q10K substitution. 85. The method of any one of embodiments 64-84, wherein the VSV-G polypeptide further -93- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT comprises a mutation that corresponds to a mutation at positions 214 and / or 352 of SEQ ID NO: 2. 86. The method of embodiment 85, wherein the VSV-G polypeptide comprises a T214N and / or T352A mutation. 87. The method of claim any one of embodiments 64-86, wherein the viral particle comprises a VSV-G polypeptide comprising a substitution at positions I182 and at least one of T214, and T352 of SEQ ID NO: 2. 88. The method of embodiment 87, wherein the VSV-G polypeptide comprises substitutions at positions I182, T214, and T352 of SEQ ID NO: 2. 89. The method of embodiments 87 or 88, wherein the substitution at position 182 is I182D or I182E, the substitution at position 214 is T214N, and the substitution at position 352 is T352A. 90. The method of any one of embodiments 87-89, wherein the VSV-G polypeptide comprises a sequence of SEQ ID NO: 23, SEQ ID NO: 22, SEQ ID NO: 24 or SEQ ID NO: 25. 91. The method of any one of embodiments 87-89, wherein the VSV-G polypeptide comprises a sequence of SEQ ID NO: 23. 92. The method of any one of embodiments 87-89, wherein the VSV-G polypeptide comprises a sequence of SEQ ID NO: 22. 93. The method of any one of embodiments 87-89, wherein the VSV-G polypeptide comprises a sequence of SEQ ID NO: 24. 94. The method of any one of embodiments 87-89, wherein the VSV-G polypeptide comprises a sequence of SEQ ID NO: 25. 95. The method of any one of embodiments 1-94, wherein the viral vector comprises a -94- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT targeting moiety. 96. The method of embodiment 95, wherein the targeting moiety binds to CD7. 97. The method of embodiment 96, wherein the targeting moiety comprises a polypeptide comprising: (i) a heavy chain variable region comprising heavy chain CDRl, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 30; the heavy chain CDR2 has the amino acid sequence of SEQ ID NO: 31; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 32, or variants of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 33; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 34; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 35; or variants of any of the foregoing. 98. The method of embodiment 96 or 97, wherein the targeting moiety comprises a heavy chain variable region (VH) comprising an amino acid sequence of SEQ ID NO: 42 and a light chain variable region (VL) comprising an amino acid sequence of SEQ ID NO: 43. 99. The method of any one of embodiments 96-98, wherein the targeting moiety comprises an antigen binding domain comprising an amino acid sequence of SEQ ID NO: 44 or SEQ ID NO: 45. 100. The method of embodiment 95, wherein the targeting moiety binds to CD8. 101. The method of embodiment 100, wherein the targeting moiety comprises a polypeptide comprising: (i) a heavy chain variable region comprising heavy chain CDRl, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 48; the heavy chain CDR2 has the amino acid sequence of SEQ ID NO: 49; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 50, or variants of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 51; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 52; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 53; or variants -95- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT of any of the foregoing. 102. The method of embodiment 100 or 101, wherein the targeting moiety comprises a heavy chain variable region (VH) comprising an amino acid sequence of SEQ ID NO: 60 and a light chain variable region (VL) comprising an amino acid sequence of SEQ ID NO: 61. 103. The method of any one of embodiments 100-102, wherein the targeting moiety comprises an antigen binding domain comprising an amino acid sequence of SEQ ID NO: 62 or SEQ ID NO: 63. 104. The method of any one of embodiments 95-103, wherein the targeting moiety comprises a formula of T-S1, wherein T is a target binding domain and S1 is a stalk portion, wherein the targeting moiety is attached to the surface of the viral vector through a stalk portion, S1,. 105. The method of embodiment 104, wherein the stalk portion, S1 comprises a variant Fc protein. 106. The method of embodiment 105, wherein the stalk portion, S1, comprises a formula of L1-Fc-L2-X1, wherein: L1 is a linker or absent; Fc is a variant Fc protein; L2 is a linker or absent; and X1 is a polypeptide comprising the transmembrane domain, wherein the targeting moiety having the formula T-S1 has a formula of T-L1-Fc-L2-X1. 107. The method of embodiment 106, wherein X1 comprises a polypeptide having a formula of ECD-TM-ICD, wherein: ECD is an extracellular domain, or a fragment thereof, of a cell surface protein, or absent; TM is a transmembrane domain of a transmembrane protein; and ICD is an intracellular domain or a protein that facilitates incorporation of the targeting moiety into the envelope of the viral particle, or absent, wherein the targeting moiety having the formula of T-L1-Fc-L2-X1 has a formula of T-L1-Fc- -96- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT L2-ECD-TM-ICD. 108. The method of embodiment 107, wherein: T comprises an amino acid sequence of SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 62, or SEQ ID NO: 63; L1 comprises the amino acid sequence of SEQ ID NO: 72; Fc comprises the amino acid sequence of SEQ ID NO: 82; L2 is absent; ECD comprises the amino acid sequence of SEQ ID NO: 83; TM comprises the amino acid sequence of SEQ ID NO: 84; and ICD comprises an amino acid sequence comprising an env incorporation motif, wherein the env incorporation motif comprises an amino acid sequence of SEQ ID NO: 85. 109. The method of embodiment 104, wherein the stalk portion, S1 does not comprise a variant Fc protein. 110. The method of embodiment 109, wherein the stalk portion S1 comprises a formula of L3-X1, wherein: L3 is a flexible peptide linker, and X1 is a polypeptide comprising a transmembrane domain, wherein the targeting moiety having the formula T-S1 has a formula of T-L3-X1. 111. The method of embodiment 110, wherein X1 comprises a polypeptide having a formula of ECD-TM-ICD, wherein: ECD is an extracellular domain, or a fragment thereof, of a cell surface protein, or absent; TM is a transmembrane domain of a transmembrane protein; and ICD is an intracellular domain or a protein that facilitates incorporation of the targeting moiety into the envelope of the viral particle, or absent, wherein the targeting moiety having the formula of T-L3-X1 has a formula of T-L3-ECD-TM- ICD. 112. The method of embodiment 111, wherein: T comprises an amino acid sequence of SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: -97- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT 62, or SEQ ID NO: 63; L3 comprises the amino acid sequence of SEQ ID NO: 64, wherein n is 1, 2, or 4; ECD comprises the amino acid sequence of SEQ ID NO: 86; TM comprises the amino acid sequence of SEQ ID NO: 87; and ICD comprises an amino acid sequence comprising an env incorporation motif, wherein the env incorporation motif comprises an amino acid sequence of SEQ ID NO: 85 or SEQ ID NO: 88. 113. The method of embodiment 104, wherein the targeting moiety comprising a formula of T-S1 comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 89. 114. The method of embodiment 104, wherein the targeting moiety comprising a formula of T-S1 comprises an amino acid sequence having at least 95% identity to SEQ ID NO: 89. 115. The method of embodiment 104, wherein the targeting moiety comprising a formula of T-S1 comprises an amino acid sequence of SEQ ID NO: 89. 116. The method of any one of embodiments 1-115, wherein the viral vector further comprises a nucleic acid molecule encoding for a heterologous molecule of interest. 117. The method of embodiment 116, wherein the heterologous molecule of interest is an siRNA, an shRNA, a non-coding RNA (e.g. a guide RNA for a CRISPR system), a peptide, a polypeptide, a protein, a viral payload, a viral genome, or a combination thereof. 118. The method of embodiments 116 or 117, wherein the heterologous molecule of interest is a chimeric antigen receptor (“CAR”). 119. The method of embodiment 118, wherein the CAR comprises an antigen binding domain comprising an antibody, or a fragment thereof. 120. The method of embodiment 119, wherein the antibody, or fragment thereof is an antibody, a scFv antibody, an antigen binding domain, an ankyrin repeat, a VHH domain antibody, a nanobody, a single domain antibody, or an FN3 antibody. -98- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT 121. The method of embodiment 119 or 120, wherein the antigen binding domain of the CAR binds to CD20. 122. The method of embodiment 121, wherein the antigen binding domain that binds to CD20 comprises a polypeptide comprising a light chain and a heavy chain comprising: a heavy chain variable region of the heavy chain having at least 90% identity to an amino acid sequence of SEQ ID NO: 73; and a light chain variable region of the light chain having at least 90% identity to an amino acid sequence of SEQ ID NO: 74. 123. The method of embodiment 121 or 122, wherein the antigen binding domain that binds to CD20 comprises an amino acid sequence of SEQ ID NO: 75 or SEQ ID NO: 76. 124. The method of embodiment 121, wherein the antigen binding domain that binds to CD20 comprises a polypeptide comprising a light chain and a heavy chain comprising: a heavy chain variable region of the heavy chain having at least 90% identity to an amino acid sequence of SEQ ID NO: 78; and a light chain variable region of the light chain having at least 90% identity to an amino acid sequence of SEQ ID NO: 79. 125. The method of embodiment 123 or 124, wherein the antigen binding domain that binds to CD20 comprises an amino acid sequence of SEQ ID NO: 80 or SEQ ID NO: 81. 126. The method of any one of embodiments 118-121, wherein the CAR comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 90. 127. The method of any one of embodiments 118-121, wherein the CAR comprises an amino acid sequence having at least 95% identity to SEQ ID NO: 90. 128. The method of any one of embodiments 118-121, wherein the CAR comprises an amino acid sequence of SEQ ID NO: 90. Examples

[0234] Various embodiments are further described in detail by reference to the following examples. These examples are provided for purposes of illustration only, and are not -99- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT intended to be limiting unless otherwise specified. Thus, embodiments should in no way be construed as being limited to the following examples, but rather, should be construed to encompass any and all variations which become evident as a result of the teaching provided herein.

[0235] Example 1: Viral Vector Manufacturing Process.

[0236] Viral Vector Production: Viral vectors were produced in 293T cells. In transfection, self-inactivating Lentivirus (LV) pseudotyped with a VSV-G protein comprising a mutation that corresponds to I182E, T214N and T352A , expressing GFP or other gene of interest such as a chimeric antigen receptor (CAR20, a chimeric antigen receptor with an antigen binding domain that binds to CD20) under the EF1A promoter, was produced using a five-plasmid system (pModified-VSVg , pGag-Pol, pRev, pBinder, and LV plasmid expressing GFP or CAR20). 300 ng / cm2of plasmids was used for transfection with PEIpro- mediated transfection with a 1 mcg:1mcL DNA:polyethylenimine (PEI) ratio in T175 and CellSTACK™ cell culture vessels. The cell culture medium used during transfection is DMEM supplemented with either 10% or 3% Fetal Bovine Serum (FBS).

[0237] DNAse Digestion: 48 hour post transfection, cell culture was spiked with 2 mM MgCl2 and 150 Unit of DNAse / mL for 2 hours. The cell culture was incubated at 37° Celsius in an incubator. After completion of incubation, the cell culture was cooled down to room temperature.

[0238] Clarification: The digested cell culture was clarified using a peristaltic pump with Sartopore® PP31.2 micron in series with Sartopore® PP30.45 micron at a flux of 250 L / m2h (LMH). The clarification filters were loaded with 80-200 L of digested cell culture per m2of each clarification filter area. The filters were preconditioned with water, with final equilibration with the DMEM + 3% FBS (10L / m2) followed by DMEM (5L / m2). Post filtration, the filters were flushed with minimum filter void volume of DMEM and emptied to product collection.

[0239] Ion Exchange Chromatography: Mustang ® Q ion exchange membrane was used as the LV capture and purification step after clarification. The membrane was conditioned using 20 mM Tris, 100 mM NaCl, pH 7.5 with or without 4% sucrose for 25 membrane volume (MV) at 5MV / min flow rate. Post conditioning, the clarified cell culture was pumped through the Mustang ® Q membrane using the ÄKTA Avant 25 or Avant 150 chromatography systems at 5MV / min flow rate. After loading, the membrane was washed with 25 MV of 20 mM Tris 100 mM NaCl pH 7.5 with or without 4% sucrose to remove residual -100- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT loads and weakly-bound molecules and eluted with 20 mM Tris 1000 mM NaCl pH 7.5 with or without 4% sucrose to recover LV. Eluted LV was collected 1 MV after start of elution step until the UV signal at 280 nm flattened. Immediately after elution was completed, the LV eluate was diluted with 20 mM Tris pH 7.5 with or without 4% sucrose at a ratio of 1:2 or 1:6 of LV eluate and dilution buffer to stabilize the LV functionality. The dilution ratio of 1:2 resulted in higher conductivity (~50 mS / cm) while the dilution ratio of 1:6 resulted in conductivity of ~12.5 mS / cm.

[0240] Post-Chromatography DNAse: DNase digestion of the diluted chromatography product was performed using the same type of DNase as the first digestion post-harvest. Diluted chromatography product was spiked with 2 mM MgCl2 and 100 Unit of DNAse / mL and stored inside a 4° Celsius cold storage for 16-24 hours. After the incubation is completed, the product was warmed up to room temperature.

[0241] Polishing Chromatography: Capto Core 700 resin was packed to resin bed height of 10-20 cm. Post packing, the packed Capto Core 700 column was conditioned with 10 column volume (CV) of 20 mM Tris, 100 mM NaCl, pH 7.5 with or without 4% sucrose. The Mustang® Q chromatography product was loaded onto the Capto Core 700 column at column loading of 10-20 mL per mL of resin volume. LV flows through Capto Core 700 and after load is completed, the column is washed to recover the remaining LV flow through. Product collection was based on the UV absorbance at 280 nm wavelength with collection starting when UV280 starts to rise and ended when the UV280 decreases after wash.

[0242] Sterile Filtration: Two types of sterile filtration filters were used to generate the data for positively charged sterile filters described in the present disclosure: Sartopore ® 2 XLG with dual layer of 0.8 micron / 0.2 micron and Pall Supor ® with dual layer of 0.8 micron / 0.2 micron membranes. The sterile filters were flushed with water and were run with either no pre-flush or with pre-flush of 1% w / v Exbumin® (20 mL flush / cm2filter area) prior to the introduction of Capto Core 700 product at a loading ratio of 50-200 L / m2and flow rate of 250 LMH. After the feed was loaded, the sterile filters were flushed with 20 mM Tris 100 mM NaCl pH 7.5 to recover the hold up volume in the filter and tubing.

[0243] Final Concentration, Buffer Exchange, and Controlled Rate Freeze: For the final concentration step and buffer exchange, 2-stage Tangential Flow Filtration (TFF) was utilized to allow for high concentration factor to be achieved. For both stages, modified PES (mPES) hollow fiber module with 1.0 mm ID fiber and 750 kDa pore size were used with the exception that the first stage filter area is ~20 times larger than the second stage filter area. The hollow fibers were flushed with water and 20 mM Tris, 100 mM NaCl, pH 7.5 prior to use. In -101- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT the first stage, the sterile filtered product was concentrated approximately 20x. The 20x concentrated product was then concentrated further using the second stage TFF to approximately 15x at which point the product is buffer exchanged against 20 mM Tris, 100 mM NaCl, 4% sucrose pH 7.5 for 6-8 diavolumes (DV). After buffer exchange was completed, the product was concentrated 2x further and human serum albumin was added to a final concentration of 0.5% v / v. The product was frozen at -80 ° Celsius.

[0244] Example 2: Viral Vector recovery with and without pre-flushing of a charged sterile filter.

[0245] In the initial sterile filtration step development, the recovery of the non- concentrated purified lentivirus vector (~2x108vp / mL) across 3 types of sterile filters (Pall Supor® 0.8 / 0.2 micron, Sartopore® 2 XLG 0.8 / 0.2 micron, and Millipore Multimedia Durapore® 1.2 / 0.5 / 0.22 micron) in a 20 mM Tris 100 mM NaCl pH 7.5 buffer system was less than 50%. The sterile filters tested were composed of modified Polyethersulfone (PES) or Polyvinylidene Fluoride (PVDF) which has been reported to have positive charge from about pH 6 to about 8 (Basic Aspects of Membrane Science and Engineering, N. Hilal, D. Johnson, in Comprehensive Membrane Science and Engineering, 2010)

[0246] Vector recovery was compared across PES sterile filters not pre-flushed and pre-flushed with 1% recombinant human albumin for 2 feed conductivities (12.5 mS / cm and 41.0 mS / cm). At the lower conductivity, pre-flushing with 1% recombinant human albumin solution prior to sterile filtration resulted in double the recovery compared to no pre- flushing of the PES filter. For the higher conductivity feed, pre-flushing the sterile filters resulted in marginal yield increase compared to no pre-flushing. These results are illustrated in FIG.2.

[0247] In contrast, when Sartobran P sterile filter was used, pre-flushing with a recombinant human albumin solution led to moderate increase in viral recoverey (FIG. 3). Sartobran P is made of cellulose acetate material, which has been reported to bear a negative charge (Li, Nancy et al. “Synthesis and Characterization of a High Flux Nanocellulose- Cellulose Acetate Nanocomposite Membrane.” Membranes vol. 9,6 70. 6 Jun. 2019, doi:10.3390 / membranes9060070). This demonstrates that the viral vector loss during sterile filtration is largely due to adsorption by charge onto the filter, which can be avoided by coating with a protein, such as recombinant human serum albumin.

[0248] Example 3: Initial Pre-Coating of a Sterile Filter.

[0249] The impact of pre-coating of the sterile filter with protein to reduce vector loss was investigated. Viral vector recovery for non-concentrated, purified viral vectors -102- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT was assessed at 12.5 mS / cm and 30 mS / cm conductivity using sterile filters flushed with water and 20 mM Tris 100 M NaCl pH 7.5, or water followed by 20 mM Tris 100 mM NaCl + 1% exbumin® pH 7.5, prior to lentivirus filtration. The sterile filter used in this study is Pall Supor® 0.8 / 0.2 micron, composed of modified polyethersulfone (PES). The protein for pre- coating was a recombinant human serum albumin, such as Exbumin® (Invitria). Results are shown in FIG. 4. The experimental filtration results are denoted with Exb (+) and the control filtration results, with no recombinant human serum albumin flush, are denoted with Exb (-) in FIG.4. Pre-coating of a sterile filter with recombinant human serum albumin solution improved lentivirus vector recovery across sterile filtration by up to 2 fold as evidenced by three orthogonal analytical methods (FIG. 4). Thus, this demonstrates that the viral vector loss during sterile filtration is largely due to adsorption onto the filter, which can be avoided by coating with a protein, such as recombinant human serum albumin.

[0250] Example 4: Pre-Coating of a Negatively Charged Sterile Filter .

[0251] The effect of protein pre-coating on a sterile filter with a negative charge was also investigated. Protein pre-coating was applied to a Sartobran® P sterile filter, composed of cellulose acetate material, which has been reported to bear a negative charge (Synthesis and Characterization of a High Flux Nanocellulose-Cellulose Acetate Nanocomposite Membrane; Nancy Li, Jackie Zheng, Pejman Hadi, Mengying Yang, Xiangyu Huang, Hongyang Ma, Harold W. Walker, & Bejamin S. Hsiao; Membranes volume 9(6), 70, (2019).. A smaller magnitude of improvement was observed (Figure 5), relative to the improvement seen with positively charged sterile filters. The control experiments without protein pre-coating produced approximately 80% vector recovery, and the experiments with protein pre-coating has yield of approximately 70%. This result indicated that the lentivirus vector is not prone to loss due to charge interaction with the Sartobran P filter (See, FIG. 5). Lentiviral vectors are themselves negatively charged at neutral pH (see Perry C, Rayat ACME. Lentiviral Vector Bioprocessing. Viruses. 2021 Feb 9;13(2):268. doi: 10.3390 / v13020268. PMID: 33572347; PMCID: PMC7916122; and / or Rodrigues, Teresa et al. “Removal of envelope protein-free retroviral vectors by anion-exchange chromatography to improve product quality.” Journal of separation science vol. 31,20 (2008): 3509-18. doi:10.1002 / jssc.200800195). Thus, it is likely that viral vector recovery when using a negatively charged filter is not impacted when pre-flushing the filters because the lentiviral vectors bear the same charge as the filter.

[0252] Example 5: Impact of pre-coating of the sterile filter with HSA vs recombinant HSA. -103- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT

[0253] The impact of pre-coating of the sterile filter with Human Serum Albumin (HSA) to reduce vector loss was compared to Recombinant Human Serum Albumin (Exbumin®). In the first condition (FIG. 6, left most data), viral vector recovery for non- concentrated, purified viral vectors was assessed at 12.5 mS / cm using sterile filters flushed with water and 20 mM Tris 100 M NaCl pH 7.5, or water followed by 20 mM Tris 100 mM NaCl + 1% Exbumin® pH 7.5, prior to lentivirus filtration. In the second condition (FIG. 6, middle data), viral vector recovery for non-concentrated, purified viral vectors was assessed at 12.5 mS / cm using sterile filters flushed with water and 20 mM Tris 100 M NaCl pH 7.5, or water followed by 20 mM Tris 100 mM NaCl + 1% HSA pH 7.5, prior to lentivirus filtration. In the third condition (FIG. 6, right most data), viral vector recovery for non-concentrated, purified viral vectors was assessed at 12.5 mS / cm using sterile filters flushed with water and 20 mM Tris 100 M NaCl pH 7.5, or water followed by 20 mM Tris 100 mM NaCl + 0.1% HSA pH 7.5, prior to lentivirus filtration. The sterile filter used in this study was Sartopore® 2 XLG 0.8 / 0.2 micron, composed of modified polyethersulfone (PES). Pre-coating of a sterile filter with HSA, either 1% or 0.1%, or Exbumin® solution resulted in comparable lentiviral vector recovery when assayed by Genome Copy or viral particle count by p24. This demonstrates that both HSA (e.g., non-recombinant HSA) and recombinant HSA could be used as a sterile filter precoating protein solution.

[0254] Example 6: Mutation at position 182 of VSV-G abrogates LDL-R interaction, but retains fusing properties as illustrated in FIG 7A-8B.

[0255] Plasmids / Sequences. All VSV-G plasmids were derived from pCMV- VSV-G Envelope Vector (Cell Bio Labs, catalog RV-110). Point mutations and combinations thereof were introduced using site directed mutagenesis (New England Biolabs). Individual mutations H8A and K47Q were previously shown to partially “blind” VSV-G, reducing its binding to LDL-R, the native cellular receptor for VSV (PMID: 29531262, DOI: 10.1038 / s41467-018-03432-4). In this experiment, a single binder molecule consisting of a CD7-targeting scFv (clone MT701) fused to an IgG “stalk” bearing a CD28 transmembrane domain was used.

[0256] Cells. HEK293T cells were grown in DMEM with 10% FBS. SupT1 cells were maintained in RPMI media with 10% FBS. Human PBMCs were purchased from AllCells and cultured in X-Vivo 10 (Lonza) supplemented with 20ng / mL IL-2 (Peprotech). PBMCs were activated 48 hours prior to transduction using anti-CD3 / CD28 Dynabeads (Cell Therapy Systems). -104- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT

[0257] Generation of lentiviral particles. The recombinant lentiviral particles co-expressing VSV-G glycoprotein and binder molecules were generated by plasmid transection into HEK293T cells using Lipofectamine 3000 (ThermoFisher Scientific). A total of 5 plasmids were transfected: (1) plasmid expressing the VSV-G glycoprotein, (2) plasmid expressing the binder protein (3) plasmid expressing the lentiviral transfer genome encoding for eGFP, (4) plasmid expressing gag-pol, and (5) plasmid expressing rev. Transfected cell supernatant was harvested 48 hours later. Virus in the cell supernatant was concentrated by centrifugation through a sucrose cushion and resuspended in PBS. Lentiviral particle titer was determined using the Lenti-X p24 Rapid Titer Kit (Takara Bio, San Jose, CA).

[0258] Lentivirus transduction assay. A series of 10-fold dilutions (in cell culture media) of the concentrated lentivirus was performed and used to infect SupT1 and activated human PBMCs. Media was replaced 6 hours later, and the transduced cells were analyzed by flow cytometry on days 4 and 7 after transduction. Cells were stained with a viability stain and an anti-CD7 antibody to detect CD7 positive cells (PeCy7 mouse-anti- human CD7, clone CD7-6B7, BD Biosciences). Expression of eGFP was measured to calculate transduction efficiency.

[0259] Structure-guided design of novel blinding mutations. Using a published crystal structures of VSV-G bound to CR2 and CR3 of the LDL-R (pdb 50YL and 50Y9, respectively), we identified two putative positions in VSV-G with side chains oriented toward the binding interface on LDL-R (FIG. 7A and 7B). Residue Q10 (SEQ ID NO: 2) appeared to form several interactions with residues in both CR2 and CR3. In CR3, this included interactions with a positively charged arginine residue. Thus, three substitutions were tested: Q10A to reduce side-chain interactions that potentially stabilize LDL-R binding and Q10R and Q10K to create electrostatic repulsion.

[0260] Residue I182 (SEQ ID NO: 2) appeared to contact several residues in both CR2 and CR3 as well. Three substitutions were tested: I182A to reduce side-chain interactions that potentially stabilize LDL-R binding and I182D and I182E to create electrostatic repulsion against the primary binding interfaces on LDL-R.

[0261] Addition of negative charges in the binding interface ablate native tropism without altering fusogenicity. Titration of viral supernatants on the CD7+ T cell line SupT1 validated the structural predictions for residue I182. In the absence of any compensatory binder molecule, WT VSV-G reached titers of 3.0e8 while both I182D and I182E were ~3 orders of magnitude lower (Fig. 8A. Substitutions at residue I182 preserved fusogenicity, as titers were restored to 1e8 in the presence of a binder redirecting the virions to CD7. -105- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT

[0262] The data is illustrated in FIG.8A and 8B which shows that the addition of negative charges in the binding interface ablate native tropism without altering fusogenicity. FIG.8A (Top) shows the titration of VSV-G constructs on SupT1 cells. Plotted is the percentage of SupT1 cells expressing GFP at each amount of viral input in terms of p24 antigen. Dashed lines / open circles indicate VSV-G constructs alone, solid lines / filled circles indicate the same construct with a CD7 targeting molecule expressed in trans. FIG. 8B illustrates Functional titer of each construct calculated from the titration in A, expressed as transducing units per mL of concentrated virus supernatant (TU / mL).

[0263] Thus these examples demonstrate that a mutation position 182 is sufficient to abrogate the LDL-R interaction, but retain fusogenic properties when combining with a targeting moiety that binds to a target on the target cell.

[0264] Thus, the examples provided for herein demonstrate that viral recovery can be significantly increased when pre-coating a sterile filter, such as a positively charged sterile filter, with a protein, such as recombination human serum albumin. This effect appears to be greater with a positively charged filter, which was surprising and could not have been predicted. Additionally, the methods provided for herein lead to increased recovery even while being able to produce a sterile composition even though steps are performed after passing a solution through a sterile filter. This is in contrast to other methods that have the final step being the sterile filtration step. Thus, the present methods and embodiments lead to increased recovery without sacrificing the sterility of a product, which is important for a pharmaceutical composition. These methods can be used to administer a higher dose of a pharmaceutical composition comprising a viral vector in a smaller volume, which is advantageous to patients being treated with viral vector based therapies.

[0265] This specification contains numerous citations to patents, patent applications, and publications. Each is hereby incorporated by reference for all purposes.

[0266] The specification also makes reference to various sequences, such as those provided herein and below. The alignment of the ectodomains of different VSV-G proteins from different strains is illustrated in FIG 9. VSV-G Indiana Full length WT: MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMC HASKWVTTCDFRWYGPKYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHV LVDEYTGEWVDSQFINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLISMDITFFSEDGELSSLGKEGTGFRSNY FAYETGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQTSVDVSLIQDVERILDYSLC QETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTTERELWDD WAPYEDVEIGPNGVLRTSSGYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNP IELVEGWFSSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK (SEQ ID NO: -106- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT 1) VSV-G Indiana Ectodomain WT: KFTIVFPHNQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGP KYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQFIN GKCSNYICPTVHNSTTWHSDYKVKGLCDSNLISMDITFFSEDGELSSLGKEGTGFRSNYFAYETGGKACKMQYCK HWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPV DLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTTERELWDDWAPYEDVEIGPNGVLR TSSGYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIA SFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK (SEQ ID NO: 2) VSV-G Indiana ectodomain I182A: KFTIVFPHNQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGP KYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQFIN GKCSNYICPTVHNSTTWHSDYKVKGLCDSNLASMDITFFSEDGELSSLGKEGTGFRSNYFAYETGGKACKMQYCK HWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPV DLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTTERELWDDWAPYEDVEIGPNGVLR TSSGYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIA SFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK (SEQ ID NO: 3) VSV-G Indiana ectodomain I182D: KFTIVFPHNQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGP KYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQFIN GKCSNYICPTVHNSTTWHSDYKVKGLCDSNLDSMDITFFSEDGELSSLGKEGTGFRSNYFAYETGGKACKMQYCK HWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPV DLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTTERELWDDWAPYEDVEIGPNGVLR TSSGYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIA SFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK (SEQ ID NO: 4) VSV-G Indiana ectodomain I182E: KFTIVFPHNQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGP KYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQFIN GKCSNYICPTVHNSTTWHSDYKVKGLCDSNLESMDITFFSEDGELSSLGKEGTGFRSNYFAYETGGKACKMQYCK HWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPV DLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTTERELWDDWAPYEDVEIGPNGVLR TSSGYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIA SFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK (SEQ ID NO: 5) VSV-G Indiana ectodomain H8A + K47Q: KFTIVFPANQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPQSHKAIQADGWMCHASKWVTTCDFRWYGP KYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQFIN GKCSNYICPTVHNSTTWHSDYKVKGLCDSNLISMDITFFSEDGELSSLGKEGTGFRSNYFAYETGGKACKMQYCK HWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPV DLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTTERELWDDWAPYEDVEIGPNGVLR TSSGYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIA SFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK (SEQ ID NO: 6) VSV-G Indiana ectodomain Q10A: KFTIVFPHNAKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGP KYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQFIN GKCSNYICPTVHNSTTWHSDYKVKGLCDSNLISMDITFFSEDGELSSLGKEGTGFRSNYFAYETGGKACKMQYCK HWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPV DLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTTERELWDDWAPYEDVEIGPNGVLR TSSGYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIA SFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK (SEQ ID NO: 7) VSV-G Indiana ectodomain Q10R: KFTIVFPHNRKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGP KYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQFIN GKCSNYICPTVHNSTTWHSDYKVKGLCDSNLISMDITFFSEDGELSSLGKEGTGFRSNYFAYETGGKACKMQYCK -107- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT HWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPV DLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTTERELWDDWAPYEDVEIGPNGVLR TSSGYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIA SFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK (SEQ ID NO: 8) VSV-G Indiana ectodomain Q10K: KFTIVFPHNKKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGP KYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQFIN GKCSNYICPTVHNSTTWHSDYKVKGLCDSNLISMDITFFSEDGELSSLGKEGTGFRSNYFAYETGGKACKMQYCK HWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPV DLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTTERELWDDWAPYEDVEIGPNGVLR TSSGYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIA SFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK (SEQ ID NO: 9) VSV-G New Jersey Full length WT: MLSYLIFALVVSPILGKIEIVFPQHTTGDWKRVPHEYNYCPTSADKNSHGTQTGIPVELTMPKGLTTHQVDGFMC HSALWMTTCDFRWYGPKYITHSIHNEEPTDYQCLEAIKAYKDGVSFNPGFPPQSCGYGTVTDAEAHIVTVTPHSV KVDEYTGEWIDPHFIGGRCKGQICETVHNSTKWFTSSDGESVCSQLFTLVGGTFFSDSEEITSMGLPETGIRSNY FPYVSTEGICKMPFCRKPGYKLKNDLWFQITDPDLDKTVRDLPHIKDCDLSSSIVTPGEHATDISLISDVERILD YALCQNTWSKIEAGEPITPVDLSYLGPKNPGAGPVFTIINGSLHYFMSKYLRVELESPVIPRMEGKVAGTRIVRQ LWDQWFPFGEVEIGPNGVLKTKQGYKFPLHIIGTGEVDNDIKMERIVKHWEHPHIEAAQTFLKKDDTEEVLYYGD TGVSKNPVELVEGWFSGWRSSIMGVLAVIIGFVILIFLIRLIGVLSSLFRQKRRPIYKSDVEMAHFR (SEQ ID NO: 10) VSV-G New Jersey ectodomain WT: KIEIVFPQHTTGDWKRVPHEYNYCPTSADKNSHGTQTGIPVELTMPKGLTTHQVDGFMCHSALWMTTCDFRWYGP KYITHSIHNEEPTDYQCLEAIKAYKDGVSFNPGFPPQSCGYGTVTDAEAHIVTVTPHSVKVDEYTGEWIDPHFIG GRCKGQICETVHNSTKWFTSSDGESVCSQLFTLVGGTFFSDSEEITSMGLPETGIRSNYFPYVSTEGICKMPFCR KPGYKLKNDLWFQITDPDLDKTVRDLPHIKDCDLSSSIVTPGEHATDISLISDVERILDYALCQNTWSKIEAGEP ITPVDLSYLGPKNPGAGPVFTIINGSLHYFMSKYLRVELESPVIPRMEGKVAGTRIVRQLWDQWFPFGEVEIGPN GVLKTKQGYKFPLHIIGTGEVDNDIKMERIVKHWEHPHIEAAQTFLKKDDTEEVLYYGDTGVSKNPVELVEGWFS GWRSSIMGVLAVIIGFVILIFLIRLIGVLSSLFRQKRRPIYKSDVEMAHFR (SEQ ID NO: 11) VSV-G Marraba Full length WT: MLRLFLFCFLALGAHSKFTIVFPHHQKGNWKNVPSTYHYCPSSSDQNWHNDLTGVSLHVKIPKSHKAIQADGWMC HAAKWVTTCDFRWYGPKYITHSIHSMSPTLEQCKTSIEQTKQGVWINPGFPPQSCGYATVTDAEVVVVQATPHHV LVDEYTGEWIDSQLVGGKCSKEVCQTVHNSTVWHADYKITGLCESNLASVDITFFSEDGQKTSLGKPNTGFRSNH FAYESGEKACRMQYCTQWGIRLPSGVWFELVDKDLFQAAKLPECPRGSSISAPSQTSVDVSLIQDVERILDYSLC QETWSKIRAKLPVSPVDLSYLAPKNPGSGPAFTIINGTLKYFETRYIRVDISNPIIPHMVGTMSGTTTERELWND WYPYEDVEIGPNGVLKTPTGFKFPLYMIGHGMLDSDLHKSSQAQVFEHPHAKDAASQLPDDETLFFGDTGLSKNP VELVEGWFSSWKSTLASFFLIIGLGVALIFIIRIIVAIRYKYKGRKTQKIYNDVEMSRLGNK (SEQ ID NO: 12) VSV-G Marraba ectodomain WT: KFTIVFPHHQKGNWKNVPSTYHYCPSSSDQNWHNDLTGVSLHVKIPKSHKAIQADGWMCHAAKWVTTCDFRWYGP KYITHSIHSMSPTLEQCKTSIEQTKQGVWINPGFPPQSCGYATVTDAEVVVVQATPHHVLVDEYTGEWIDSQLVG GKCSKEVCQTVHNSTVWHADYKITGLCESNLASVDITFFSEDGQKTSLGKPNTGFRSNHFAYESGEKACRMQYCT QWGIRLPSGVWFELVDKDLFQAAKLPECPRGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAKLPVSPV DLSYLAPKNPGSGPAFTIINGTLKYFETRYIRVDISNPIIPHMVGTMSGTTTERELWNDWYPYEDVEIGPNGVLK TPTGFKFPLYMIGHGMLDSDLHKSSQAQVFEHPHAKDAASQLPDDETLFFGDTGLSKNPVELVEGWFSSWKSTLA SFFLIIGLGVALIFIIRIIVAIRYKYKGRKTQKIYNDVEMSRLGNK (SEQ ID NO: 13) VSV-G Carajas Full length WT: MKMKMVIAGLILCIGILPAIGKITISFPQSLKGDWRPVPKGYNYCPTSADKNLHGDLIDIGLRLRAPKSFKGISA DGWMCHAARWITTCDFRWYGPKYITHSIHSFRPSNDQCKEAIRLTNEGNWINPGFPPQSCGYASVTDSESVVVTV -108- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT TKHQVLVDEYSGSWIDSQFPGGSCTSPICDTVHNSTLWHADHTLDSICDQEFVAMDAVLFTESGKFEEFGKPNSG IRSNYFPYESLKDVCQMDFCKRKGFKLPSGVWFEIEDAEKSHKAQVELKIKRCPHGAVISAPNQNAADINLIMDV ERILDYSLCQATWSKIQNKEALTPIDISYLGPKNPGPGPAFTIINGTLHYFNTRYIRVDIAGPVTKEITGFVSGT STSRVLWDQWFPYGENSIGPNGLLKTASGYKYPLFMVGTGVLDADIHKLGEATVIEHPHAKEAQKVVDDSEVIFF GDTGVSKNPVEVVEGWFSGWRSSLMSIFGIILLIVCLVLIVRILIALKYCCVRHKKRTIYKEDLEMGRIPRRA (SEQ ID NO: 14) VSV-G Carajas ectodomain WT: KITISFPQSLKGDWRPVPKGYNYCPTSADKNLHGDLIDIGLRLRAPKSFKGISADGWMCHAARWITTCDFRWYGP KYITHSIHSFRPSNDQCKEAIRLTNEGNWINPGFPPQSCGYASVTDSESVVVTVTKHQVLVDEYSGSWIDSQFPG GSCTSPICDTVHNSTLWHADHTLDSICDQEFVAMDAVLFTESGKFEEFGKPNSGIRSNYFPYESLKDVCQMDFCK RKGFKLPSGVWFEIEDAEKSHKAQVELKIKRCPHGAVISAPNQNAADINLIMDVERILDYSLCQATWSKIQNKEA LTPIDISYLGPKNPGPGPAFTIINGTLHYFNTRYIRVDIAGPVTKEITGFVSGTSTSRVLWDQWFPYGENSIGPN GLLKTASGYKYPLFMVGTGVLDADIHKLGEATVIEHPHAKEAQKVVDDSEVIFFGDTGVSKNPVEVVEGWFSGWR SSLMSIFGIILLIVCLVLIVRILIALKYCCVRHKKRTIYKEDLEMGRIPRRA (SEQ ID NO: 15) VSV-G Alagoa Full length WT: MTPAFILCMLLAGSSWAKFTIVFPQSQKGDWKDVPPNYRYCPSSADQNWHGDLLGVNIRAKMPKVHKAIKADGWM CHAAKWVTTCDYRWYGPQYITHSIHSFIPTKAQCEESIKQTKEGVWINPGFPPKNCGYASVSDAESIIVQATAHS VMIDEYSGDWLDSQFPTGRCTGSTCETIHNSTLWYADYQVTGLCDSALVSTEVTFYSEDGLMTSIGRQNTGYRSN YFPYEKGAAACRMKYCTHEGIRLPSGVWFEMVDKELLESVQMPECPAGLTISAPTQTSVDVSLILDVERMLDYSL CQETWSKVHSGLPISPVDLGYIAPKNPGAGPAFTIVNGTLKYFDTRYLRIDIEGPVLKKMTGKVSGTPTKRELWT EWFPYDDVEIGPNGVLKTPEGYKFPLYMIGHGLLDSDLQKTSQAEVFHHPQIAEAVQKLPDDETLFFGDTGISKN PVEVIEGWFSNWRSSVMAIVFAILLLVITVLMVRLCVAFRHFCCQKRHKIYNDLEMNQLRR (SEQ ID NO: 16) VSV-G Alagoa ectodomain WT: KFTIVFPQSQKGDWKDVPPNYRYCPSSADQNWHGDLLGVNIRAKMPKVHKAIKADGWMCHAAKWVTTCDYRWYGP QYITHSIHSFIPTKAQCEESIKQTKEGVWINPGFPPKNCGYASVSDAESIIVQATAHSVMIDEYSGDWLDSQFPT GRCTGSTCETIHNSTLWYADYQVTGLCDSALVSTEVTFYSEDGLMTSIGRQNTGYRSNYFPYEKGAAACRMKYCT HEGIRLPSGVWFEMVDKELLESVQMPECPAGLTISAPTQTSVDVSLILDVERMLDYSLCQETWSKVHSGLPISPV DLGYIAPKNPGAGPAFTIVNGTLKYFDTRYLRIDIEGPVLKKMTGKVSGTPTKRELWTEWFPYDDVEIGPNGVLK TPEGYKFPLYMIGHGLLDSDLQKTSQAEVFHHPQIAEAVQKLPDDETLFFGDTGISKNPVEVIEGWFSNWRSSVM AIVFAILLLVITVLMVRLCVAFRHFCCQKRHKIYNDLEMNQLRR (SEQ ID NO: 17) VSV-G Cocal Full length WT: MNFLLLTFIVLPLCSHAKFSIVFPQSQKGNWKNVPSSYHYCPSSSDQNWHNDLLGITMKVKMPKTHKAIQADGWM CHAAKWITTCDFRWYGPKYITHSIHSIQPTSEQCKESIKQTKQGTWMSPGFPPQNCGYATVTDSVAVVVQATPHH VLVDEYTGEWIDSQFPNGKCETEECETVHNSTVWYSDYKVTGLCDATLVDTEITFFSEDGKKESIGKPNTGYRSN YFAYEKGDKVCKMNYCKHAGVRLPSGVWFEFVDQDVYAAAKLPECPVGATISAPTQTSVDVSLILDVERILDYSL CQETWSKIRSKQPVSPVDLSYLAPKNPGTGPAFTIINGTLKYFETRYIRIDIDNPIISKMVGKISGSQTERELWT EWFPYEGVEIGPNGILKTPTGYKFPLFMIGHGMLDSDLHKTSQAEVFEHPHLAEAPKQLPEEETLFFGDTGISKN PVELIEGWFSSWKSTVVTFFFAIGVFILLYVVARIVIAVRYRYQGSNNKRIYNDIEMSRFRK (SEQ ID NO: 18) VSV-G Cocal ectodomain WT: KFSIVFPQSQKGNWKNVPSSYHYCPSSSDQNWHNDLLGITMKVKMPKTHKAIQADGWMCHAAKWITTCDFRWYGP KYITHSIHSIQPTSEQCKESIKQTKQGTWMSPGFPPQNCGYATVTDSVAVVVQATPHHVLVDEYTGEWIDSQFPN GKCETEECETVHNSTVWYSDYKVTGLCDATLVDTEITFFSEDGKKESIGKPNTGYRSNYFAYEKGDKVCKMNYCK HAGVRLPSGVWFEFVDQDVYAAAKLPECPVGATISAPTQTSVDVSLILDVERILDYSLCQETWSKIRSKQPVSPV DLSYLAPKNPGTGPAFTIINGTLKYFETRYIRIDIDNPIISKMVGKISGSQTERELWTEWFPYEGVEIGPNGILK TPTGYKFPLFMIGHGMLDSDLHKTSQAEVFEHPHLAEAPKQLPEEETLFFGDTGISKNPVELIEGWFSSWKSTVV TFFFAIGVFILLYVVARIVIAVRYRYQGSNNKRIYNDIEMSRFRK (SEQ ID NO: 19) VSV-G Morreton Full length WT: -109- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT MLVLYLLLSLLALGAQCKFTIVFPHNQKGNWKNVPANYQYCPSSSDLNWHNGLIGTSLQVKMPKSHKAIQADGWM CHAAKWVTTCDFRWYGPKYVTHSIKSMIPTVDQCKESIAQTKQGTWLNPGFPPQSCGYASVTDAEAVIVKATPHQ VLVDEYTGEWVDSQFPTGKCNKDICPTVHNSTTWHSDYKVTGLCDANLISMDITFFSEDGKLTSLGKEGTGFRSN YFAYENGDKACRMQYCKHWGVRLPSGVWFEMADKDIYNDAKFPDCPEGSSIAAPSQTSVDVSLIQDVERILDYSL CQETWSKIRAHLPISPVDLSYLSPKNPGTGPAFTIINGTLKYFETRYIRVDIAGPIIPQMRGVISGTTTERELWT DWYPYEDVEIGPNGVLKTATGYKFPLYMIGHGMLDSDLHISSKAQVFEHPHIQDAASQLPDDETLFFGDTGLSKN PIELVEGWFSGWKSTIASFFFIIGLVIGLYLVLRIGIALCIKCRVQEKRPKIYTDVEMNRLDR (SEQ ID NO: 20) VSV-G Morreton ectodomain WT: KFTIVFPHNQKGNWKNVPANYQYCPSSSDLNWHNGLIGTSLQVKMPKSHKAIQADGWMCHAAKWVTTCDFRWYGP KYVTHSIKSMIPTVDQCKESIAQTKQGTWLNPGFPPQSCGYASVTDAEAVIVKATPHQVLVDEYTGEWVDSQFPT GKCNKDICPTVHNSTTWHSDYKVTGLCDANLISMDITFFSEDGKLTSLGKEGTGFRSNYFAYENGDKACRMQYCK HWGVRLPSGVWFEMADKDIYNDAKFPDCPEGSSIAAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAHLPISPV DLSYLSPKNPGTGPAFTIINGTLKYFETRYIRVDIAGPIIPQMRGVISGTTTERELWTDWYPYEDVEIGPNGVLK TATGYKFPLYMIGHGMLDSDLHISSKAQVFEHPHIQDAASQLPDDETLFFGDTGLSKNPIELVEGWFSGWKSTIA SFFFIIGLVIGLYLVLRIGIALCIKCRVQEKRPKIYTDVEMNRLDR (SEQ ID NO: 21) -110- IPTS / 126938130.1

Claims

DOCKET NO: INH-024WO PATENT What is Claimed:

1. A method of producing a concentrated sterilized solution comprising a viral vector, the method comprising the steps of: clarifying a solution comprising cell culture media and the viral vector; filtering the clarified solution comprising the viral vector through a first chromatographic filter to produce a filtered clarified solution comprising the viral vector; passing the filtered clarified solution comprising the viral vector through a protein coated sterile membrane to produce a sterilized solution comprising the viral vector; and concentrating the sterilized solution comprising the viral vector to produce the concentrated sterilized solution comprising the viral vector.

2. The method of claim 1, further comprising mixing the concentrated sterilized solution comprising the viral vector with one or more sterile, pyrogen-free buffers and / or excipients to produce a sterile pharmaceutical composition comprising the viral vector.

3. The method of claim 1, further comprising collecting the cell culture media from a cell culture producing the viral vector prior to clarifying the solution.

4. The method of any one of claims 1-3, further comprising filtering the filtered clarified solution comprising the viral vector through a second chromatographic filter to produce a twice-filtered clarified solution comprising the viral vector prior to passing filtered clarified solution through the protein coated sterile membrane.

5. The method of claim 1, wherein the protein coated sterile membrane comprises a membrane pre-flushed with a protein solution.

6. The method of claim 1, the method further comprising coating a sterile membrane with a protein solution to produce the protein coated sterile membrane.

7. The method of claims 5 or 6, wherein the protein solution comprises one or more component selected from a protein, 2-Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS), -111- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT histidine, phosphate, HEPES ((4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), sodium chloride, or combinations thereof.

8. The method of claim 7, wherein the protein solution comprises about 1% w / v of a protein, about 20 mM 2-Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS), about 100 mM to about 400 mM sodium chloride, and a pH of about 6.0 to about 8.

0.

9. The method of claim 7, wherein the protein comprises human serum albumin, myoglobulin, bovine serum albumin, immunoglobulin, immunoglobulin fragments, fibronectin, vitronectin, or any combination thereof.

10. The method of claim 9, wherein the human serum albumin is recombinant human serum albumin, human serum albumin produced from a plant, or a non-recombinant human serum albumin.

11. The method of claim 10, wherein the human serum albumin is non-recombinant human serum albumin.

12. The method of claim 10 or 11, wherein the non-recombinant human serum albumin is USP grade non-recombinant human serum albumin.

13. The method of claim 6, wherein the protein coated sterile membrane has a positive charge prior to being coated with a protein.

14. The method of claim 6, wherein the membrane is a polyethersulfone (PES) membrane, a nylon membrane, or a polyvinylidene fluoride (PVDF) membrane.

15. The method of any one of claims 1-14, wherein the clarified solution is filtered through the first chromatographic filter by capture chromatography.

16. The method of claim 15, wherein the capture chromatography is ion exchange chromatography, affinity chromatography, hydrophobic chromatography, size exclusion chromatography, or multimodal chromatography. -112- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT 17. The method of claim 4, wherein the filtered solution is filtered through the second chromatographic filter by polishing chromatography.

18. The method of claim 17, wherein the polishing chromatography is ion exchange chromatography, hydrophobic interaction chromatography, size-exclusion, or multimodal chromatography.

19. The method of any one of claims 1-18, further comprising digesting DNA in the solution comprising the cell culture media and the viral vector prior to clarifying the solution.

20. The method of any one of claims 1-19, further comprising digesting DNA in the solution comprising the viral vector after filtering the clarified solution.

21. The method of claim 1, wherein the concentrating step comprises tangential flow filtration.

22. The method of claim 21, wherein the tangential flow filtration is a two stage tangential flow filtration.

23. The method of claim 21 or claim 22, wherein the tangential flow filtration utilizes a 1.0 mm ID fiber and 750 kDa pore size.

24. The method of claim 2, wherein the concentrated sterilized solution comprising the viral vector is mixed in a sterile environment with the one or more sterile, pyrogen-free buffers and / or excipients to produce the sterile pharmaceutical composition comprising the viral vector.

25. The method of claim 24, wherein the mixing with one or more sterile, pyrogenic free buffers and / or excipients comprises a buffer exchange step.

26. The method of claim 1, further comprising: prior to clarifying the solution: i) obtaining a solution comprising cell culture media and the viral vector; and -113- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT ii) digesting DNA in the solution comprising cell culture media and the viral vector to produce a first digested solution comprising the viral vector; wherein clarifying the solution comprises clarifying the first digested solution comprising the viral vector to produce a clarified, digested solution comprising the viral vector; subsequent to filtering the clarified solution and prior to passing the filtered clarified solution through a protein coated sterile membrane: i) digesting DNA in the filtered, clarified, digested solution comprising the viral vector to produce a filtered, clarified, twice-digested solution comprising the viral vector; and ii) filtering the filtered, clarified, twice-digested solution comprising the viral vector through a second chromatographic filter to produce a twice- filtered, clarified, twice-digested solution comprising the viral vector; wherein the protein coated sterile membrane comprises a membrane pre-flushed with a protein solution, wherein the protein solution comprises about 0.1% w / v to 2.0% w / v of a protein, about 10 mM to about 30 mM 2-Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS), about 100 mM to about 400 mM sodium chloride, and a pH of about 6.0 to about 8.

0.

27. A method of producing a concentrated sterilized solution comprising a viral vector, the method comprising the steps of: digesting DNA in a solution comprising the viral vector to produce a first digested solution comprising the viral vector; clarifying the digested solution comprising the viral vector to produce a clarified, digested solution comprising the viral vector; filtering the clarified, digested solution comprising the viral vector through a first chromatographic filter to produce a filtered, clarified, digested solution comprising the viral vector; digesting DNA in the filtered, clarified, digested solution comprising the viral vector to produce a filtered, clarified, twice-digested solution comprising viral vector; filtering the filtered, clarified, twice-digested solution comprising the viral vector through a second chromatographic filter to produce a twice-filtered, clarified, twice-digested solution comprising viral vector; -114- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT passing the twice-filtered, clarified, twice-digested solution comprising the viral vector through a protein coated sterile membrane to produce a sterilized solution comprising the viral vector; and concentrating the sterilized solution comprising the viral vector to produce the concentrated sterilized solution comprising the viral vector.

28. The method of claim 27, wherein the digesting DNA is performed using an endonuclease having DNAase activity, RNAase activity, or a combination thereof.

29. The method of claim 27, wherein the second chromatographic filter is a resin.

30. The method of claim 27, further comprising mixing the concentrated sterilized solution comprising the viral vector with one or more sterile, pyrogen-free buffers and / or excipients to produce a sterile pharmaceutical composition comprising the viral vector.

31. The method of claim 27, further comprising collecting media from a cell culture producing the viral vector prior to digesting DNA.

32. The method of claim 27, further comprising collecting media from a cell culture producing the viral vector prior to clarifying the digested solution.

33. The method of claim 27, wherein the protein coated sterile membrane comprises a membrane pre-flushed with a protein solution.

34. The method of claim 27, the method further comprising coating a sterile membrane with a protein solution to produce the protein coated sterile membrane.

35. The method of claim 34, wherein the protein solution comprises one or more of a protein, 2-Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS), histidine, phosphate, HEPES ((4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), sodium chloride, or combinations thereof. -115- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT 36. The method of claim 35, wherein the protein solution comprises about 1% w / v of a protein, about 20 mM 2-Amino-2-(hydroxymethyl)propane-1,3-diol (TRIS), about 100 mM to about 400 mM sodium chloride, and a pH of about 6.0 to about 8.

0.

37. The method of claims 35 or 36, wherein the protein is human serum albumin.

38. The method of claim 37, wherein the human serum albumin is recombinant human serum albumin, human serum albumin produced from a plant, or non-recombinant human serum albumin.

39. The method of claim 38, wherein the human serum albumin is non-recombinant human serum albumin.

40. The method of claims 38 or 39, wherein the non-recombinant human serum albumin is USP grade non-recombinant human serum albumin.

41. The method of any one of claims 27-40, wherein the protein coated sterile membrane has a positive charge prior to being coated with a protein.

42. The method of any one of claims 27-41, wherein the membrane is a polyethersulfone (PES) membrane, a nylon membrane, or a polyvinylidene fluoride (PVDF) membrane.

43. The method of any one of claims 27-42, wherein the protein coated sterile membrane has a pore size of about 0.01µm, about 0.10µm, about 0.11µm, about 0.12µm, about 0.13µm, about 0.14µm, about 0.15µm, about 0.16µm, about 0.17µm, about 0.18µm, about 0.19µm, about 0.2µm, about 0.21µm, about 0.22µm, about 0.23µm, about 0.24µm, about 0.25µm, about 0.26µm, about 0.27µm, about 0.28µm, about 0.29µm, about 0.30µm, about 0.31µm, about 0.32µm, about 0.33µm, about 0.34µm, about 0.35µm, about 0.36µm, about 0.37µm, about about 0.38µm, about 0.39µm, about 0.40µm, about 0.41µm, about 0.42µm, about 0.43µm, about 0.44µm, or about 0.45µm.

44. The method of any one of claims 27-43, wherein the protein coated sterile membrane does not significantly retain the viral vector. -116- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT 45. The method of claim 44, wherein the recovery of the viral vector from the solution is at least about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or about 100%.

46. The method of any one of claims 27-45, wherein the clarified solution is filtered through the first chromatographic filter by capture chromatography.

47. The method of claim 46, wherein the capture chromatography is ion exchange chromatography, affinity chromatography, hydrophobic chromatography, size exclusion chromatography, or multimodal chromatography.

48. The method of claim 27, wherein the first filtered solution is filtered by polishing chromatography through the second filter.

49. The method of claim 48, wherein the polishing chromatography is ion exchange chromatography, hydrophobic interaction chromatography, size-exclusion, or multimodal chromatography.

50. The method of any one of claims 27-49, wherein the concentrating step comprises concentrating the viral vector by tangential flow filtration.

51. The method of claim 50, wherein the tangential flow filtration is a two stage tangential flow filtration.

52. The method of claims 5051, wherein the tangential flow filtration utilizes a 1.0 mm ID fiber and 750 kDa pore size.

53. The method of claim 30, wherein the concentrated sterilized solution comprising the viral vector is mixed in a sterile environment with excipients to produce the sterile pharmaceutical composition comprising the viral vector.

54. The method of claim 53, wherein the mixing with excipients comprises a buffer exchange step. -117- IPTS / 126938130.1DOCKET NO: INH-024WO PATENT 55. The method of any one of claims 1-55, further comprising freezing the concentrated sterilized solution comprising the viral vector or the sterile pharmaceutical composition comprising the viral vector.

56. The method of claim 1, wherein the viral vector is a lentiviral vector, an adenoviral vector, an adeno-associated viral vector, or a retroviral vector.

57. The method of claim 27, wherein the viral vector is a lentiviral vector, an adenoviral vector, an adeno-associated viral vector, or a retroviral vector.

58. The method of any one of claims 1-57, wherein the concentrated sterilized solution comprising the viral vector is diluted prior to one or more of the steps.

59. The method of any one of claims 1-58, wherein the viral vector is at a concentration of about 1x104to about 1x106transduction units (TU) / mL before passing the solution through the sterile filter.

60. The method of any one of claims 1-59, wherein the viral vector is at a concentration of 1x107to 1x109viral particles (vp) / mL before passing the solution through the sterile filter.

61. The method of claim 1, wherein the viral vector is a lentiviral vector pseudotyped with a VSV-G polypeptide comprising a mutation that corresponds to a mutation at position 182 of SEQ ID NO:

2.

62. The method of claim 27, wherein the viral vector is a lentiviral vector pseudotyped with a VSV-G polypeptide comprising a mutation that corresponds to a mutation at position 182 of SEQ ID NO:

2.

63. The method of any one of claims 2-26 or 28-60, wherein the viral vector, such as a lentiviral vector, is pseudotyped with a VSV-G polypeptide comprising a mutation that corresponds to a mutation at position 182 of SEQ ID NO:

2. -118- IPTS / 126938130.1

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