Particle formation and morphology

Abstract

To provide particle formation and morphology.SOLUTION: A method of producing particles comprising a therapeutic biological agent comprises: forming droplets of an aqueous first liquid, wherein the first liquid comprises a therapeutic biological agent, and wherein the droplets are formed within a second liquid comprising an organic solvent; dehydrating the droplets in the second liquid to form particles comprising the therapeutic biological agent.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Related Applications This application claims the benefit of U.S. Provisional Patent Application No. 62 / 799,696, filed January 31, 2019. The entire teachings of the above application are incorporated herein by reference.

[0002] The present disclosure relates to compositions and methods that allow for the formation of pharmaceutically suitable particles that can be used for therapy. In particular, the methods disclosed herein allow for the formation of round particles with low internal void space that contain a bioactive therapeutic agent. [Background technology]

[0003] Applications of materials science and nanotechnology require more efficient, reproducible, and innovative techniques for synthesizing novel functional particles. Recent advances in the synthesis and controlled assembly of bioactive particles have enabled their application for therapeutic use. Current efforts have often been directed toward developing novel synthetic approaches for non-circular microparticles that exhibit physical properties that cannot be obtained by simply fine-tuning particle size and morphology. [Prior art documents] [Non-patent literature]

[0004] [Non-Patent Document 1] J.Zhang et al.Lab Chip.2016,16,10-34 Summary of the Invention [Problem to be solved by the invention]

[0005] However, the application of these techniques to round particles has been limited due to the lack of sufficient control over particle size uniformity, shape selectivity, surface functionality, and backbone density, which are often difficult to obtain. Therefore, there is a need for a highly robust and controlled method for the preparation of round particles. [Means for solving the problem]

[0006] Provided herein is a composition comprising a particle or a plurality of particles comprising an agent, wherein the particle comprises less than about 25% internal void space and the particle has a circularity of about 0.10 to about 1.00.

[0007] In one aspect, the present disclosure provides particles comprising an agent, wherein the particles comprise less than about 25% internal void space and the circularity of the particles is from about 0.10 to about 1.00.

[0008] In another aspect, the present disclosure provides a composition comprising a plurality of particles comprising an agent suspended in a liquid, wherein the particles comprise less than about 25% internal void space and the circularity of the particles is from about 0.10 to about 1.00.

[0009] The present disclosure also provides a method of forming a particle.

[0010] In one aspect, the present disclosure provides a method of forming a particle, the method comprising: a) providing a droplet comprising a first liquid and an agent; b) contacting the droplet with a second liquid; c) drying the droplets; d) removing the first and second liquids, thereby forming particles including the agent; the particles comprise less than about 25% internal void space, and the circularity of the particles is from about 0.10 to about 1.00 after the first and second liquids are removed.

[0011] Also provided herein are methods for controlling particle morphology.

[0012] In one aspect, the present disclosure provides a method for controlling particle morphology, the method comprising: a) providing a droplet comprising a first liquid and an agent; b) contacting the droplet with a second liquid under a specific Peclet number; c) drying the droplets; d) removing the first and second liquids; The specific Peclet number controls the morphology of the particles.

[0013] The present disclosure also provides methods for controlling the surface properties of particles herein.

[0014] In one aspect, the present disclosure provides a method for controlling surface properties of a particle, the method comprising: a) providing droplets comprising a first liquid, a first component, and a second component, wherein the first component is present in an amount closer to its solubility limit than the second component, the first component has a higher Peclet number than the second component, or a combination thereof; b) contacting the droplet with a second liquid; c) drying the droplets; d) removing the first and second liquids, thereby forming particles; wherein the first component is concentrated at the surface of the particle relative to the second component.

[0015] The present compositions and methods can be useful for the formation of pharmaceutically suitable particles that can be used for therapy. In preferred embodiments, the methods disclosed herein can enable the formation of round particles with low internal void space that contain a bioactive therapeutic agent.

[0016] The foregoing will become apparent from the following more particular description of example embodiments, as illustrated in the accompanying drawings, in which like reference characters refer to the same parts throughout the different views. The drawings are not necessarily to scale, emphasis instead being placed upon illustrating the embodiments.

[0017] That is, the gist of the present invention relates to the following. Item 1 1. A method of making particles comprising a therapeutic biological agent, comprising: forming droplets of an aqueous first liquid, wherein the first liquid comprises a therapeutic biological agent, the droplets being formed within a second liquid comprising an organic solvent; dehydrating the droplets in the second liquid to form particles comprising the therapeutic biological agent; A method comprising: Section 2 Item 10. The method of claim 1, wherein the droplets are formed in a microfluidic device or a homogenizer. Section 3 Item 3. The method of item 2, wherein the droplets formed in the microfluidic device are regularly spaced apart in the microfluidic device. Section 4 Item 10. The method of item 1, wherein dehydrating the droplets comprises contacting a second liquid with a dry substance. Section 5 Item 5. The method according to item 4, wherein the drying substance is a desiccant. Section 6 Item 10. The method of claim 1, further comprising drying the particles, wherein the plurality of particles have a content of the first or second liquid of less than 10% by weight after drying. Section 7 7. The method of claim 6, wherein drying the particles comprises removing the first and second liquids. Section 8 8. The method of claim 7, wherein the first and second liquids are removed by centrifugation, sieving, filtration, magnetic collection, solvent exchange, or decanting. Section 9 Item 8. The method according to Item 7, wherein the amount of the first and second liquids remaining in the particles after drying is about 0% by weight to about 10% by weight. Section 10 8. The method of claim 7, wherein drying the particles further comprises contacting the particles with a gas stream. Section 11 Item 11. The method according to Item 10, wherein the gas has a temperature of about -80°C to about 200°C. Section 12 Item 11. The method according to item 10, wherein the gas is helium, air, nitrogen or argon. Section 13 Item 8. The method of item 7, wherein drying the particles further comprises using freeze-drying or vacuum dehydration. Section 14 Item 10. The method of claim 1, further comprising removing particles from the second liquid. Section 15 15. The method of claim 14, wherein the particles are removed from the second liquid by centrifugation, sieving, filtration, magnetic collection, solvent exchange, inertial separation, hydrocyclone separation, or decanting. Section 16 Item 7. The method of item 6, further comprising washing the dried particles with a washing fluid. Section 17 Item 17. The method of item 16, wherein the cleaning fluid is an organic liquid, a supercritical fluid, a cryogenic liquid, or a combination thereof. Section 18 The method of paragraph 1, wherein the therapeutic biological agent is an antibody. Section 19 Item 19. The method of item 18, wherein the antibody is an IgG antibody. Section 20 Item 10. The method of claim 1, wherein the therapeutic biological agent in the particles has a unit activity of about 0.5 to about 1.0. Section 21 8. The method of claim 7, wherein the particles contain less than about 10% internal void space. Section 22 8. The method of claim 7, wherein the particles have less than about 10% aggregates of the therapeutic biological agent. Section 23 8. The method of paragraph 7, wherein the particles have greater than about 60% by weight of the therapeutic biological agent. Section 24 Item 8. The method according to Item 7, wherein the circularity of the particles is about 0.80 to about 1.00. Section 25 Item 10. The method of claim 1, wherein the first liquid is water, 0.9% saline, lactated Ringer's solution, buffer, dextrose 5%, or a combination thereof. Section 26 Item 1. The method according to Item 1, wherein the first liquid is water. Section 27 Item 10. The method according to Item 1, wherein the concentration of the therapeutic biological agent in the first liquid is about 10 mg / mL to about 500 mg / mL. Section 28 Item 1. The method according to item 1, wherein the first liquid further contains a surfactant. Section 29 Item 1. The method according to Item 1, wherein the organic solvent is ethyl acetate or butyl acetate. Item 30 Item 10. The method of claim 1, wherein the droplets are formed in a vessel comprising tubing. [Effects of the Invention]

[0018] The present invention can provide particle formation and morphology. [Brief explanation of the drawings]

[0019] [Figure 1A] 1 shows an image of human IgG particles produced using a second liquid that had a Peclet number substantially less than 1. [Figure 1B] 1 shows images of human IgG particles produced using a second liquid that had a Peclet number substantially greater than 1. [Figures 2A-2C] 1 shows images of human IgG particles formed by the method of the present disclosure using several second liquids with varying levels of pre-saturation with respect to the first liquid. [Figure 3A] 1 shows an image of the surface of human IgG particles formed by the method of the present disclosure. [Figure 3B] Shown is an image of a human IgG particle that has been cut to reveal an internal cross section. [Figure 4] 1 shows a graph of the dispersive surface energy profile for particles formed in a second liquid of varying polarity. [Figure 5]1 shows a graph of acid-base surface energy profiles for particles formed in a second liquid of varying polarity and acid-base properties. [Figure 6] 1 shows a graph of the X-ray diffraction profile for solid proteinaceous material from a common first liquid compared to a standard freeze-dried material. [Figure 7A-7C] 1 shows images of human IgG particles formed by the method of the present disclosure at different voltages. [Figure 8] 1 shows a graph of the volume-weighted size distribution for human IgG particles D10, D50, and D90 formed by the method of the present disclosure at different voltages. [Figure 9] 1 shows a graph of the relationship between particle mass and environmental humidity for particles formed by the method of the present disclosure at a fixed temperature. [Figure 10] 1 shows a graph of a differential scanning calorimetry used to measure glass transition temperatures for particles formed by the methods of the present disclosure. [Figure 11] FIG. 1 shows a graph of the percentage of dissolved particles after storage for several suspensions of human IgG particles suspended in aqueous solutions containing various crowding agents. [Figure 12] Photographs of the sedimentation and surface adhesion properties of human IgG particles suspended in non-aqueous suspension media are shown. [Figure 13] 1 shows a graph of the surface tension at the air-liquid interface for neat deionized water solution and for a solution of protein dissolved in deionized water at a concentration of 20 mg / mL. [Figure 14A] 1 shows an image of the surface of human IgG particles formed by the method of the present disclosure. [Figure 14B] 1 shows an image of a human IgG particle that has been cut to reveal an internal cross section. DETAILED DESCRIPTION OF THE INVENTION

[0020] Particles have been produced using a variety of techniques. For example, particle production can be achieved by generating droplets of a liquid containing an active agent dissolved in a solvent. The solvent can then be extracted from the droplets by depositing the droplets in a liquid in which the solvent, but not the active agent, is soluble, leaving behind solid particles. Particle isolation follows evaporation of the liquid. However, the application of these techniques to form functional round particles has been limited by the lack of sufficient control over particle size uniformity, shape selectivity, surface functionality, and skeletal density, which are often difficult to achieve. The present disclosure seeks to alleviate the control issues associated with forming functional particles by providing a robust and controlled method for particle preparation.

[0021] The present disclosure generally relates to compositions comprising a particle comprising an agent or a plurality of particles comprising an agent suspended in a liquid, wherein the particle or plurality of particles comprises less than about 25% internal void space and the particle has a circularity of about 0.10 to about 1.00.

[0022] The present disclosure also relates to a method of forming particles, the method comprising: a) providing droplets comprising a first liquid and an agent; b) contacting the droplets with a second liquid; c) drying the droplets; and d) removing the first and second liquids, thereby forming particles comprising the agent, wherein the particles comprise less than about 25% internal void space, and the circularity of the particles is from about 0.10 to about 1.00 after removing the first and second liquids.

[0023] In certain aspects, the present disclosure generally relates to a method of controlling particle morphology, the method comprising: a) providing a droplet comprising a first liquid and an agent; b) contacting the droplet with a second liquid under a specific Peclet number; c) drying the droplet; and d) removing the first and second liquids, wherein the specific Peclet number controls the particle morphology.

[0024] In certain other aspects, the present disclosure generally relates to a method for controlling surface properties of particles, the method including: a) providing droplets comprising a first liquid, a first component, and a second component, wherein the first component is present in an amount closer to its solubility limit than the second component, the first component has a higher Peclet number than the second component, or a combination thereof; b) contacting the droplets with the second liquid; c) drying the droplets; and d) removing the first and second liquids, thereby forming particles, wherein the first component is concentrated at the surface of the particles compared to the second component.

[0025] As described herein, the present disclosure provides methods for preparing particles containing one or more agents, such as therapeutic or diagnostic agents. Particles can be formed, for example, by forming droplets of a first liquid containing the agent, removing the first liquid, for example, by dispersing it in a second liquid and / or evaporating it, and solidifying the droplets. The process of forming particles as described herein can significantly alter the structure or morphology of the particles, enhancing the stability of the agent. For example, particles can be stored for extended periods without significant loss of activity or the need for refrigeration. These particles can be used to produce stabilized pharmaceutical compositions, pharmaceutical suspension formulations, pharmaceutical powder formulations (e.g., inhalable powders, injectable powders), creams or other topical pastes, dietary supplements, or cosmetics. The term "pharmaceutical composition," as used herein, refers to a composition in which a therapeutic or diagnostic agent retains or partially retains its intended biological activity or functional form and contains only pharmaceutically acceptable ingredients.

[0026] It is readily understood that the aspects and embodiments generally described herein are exemplary. The following more detailed description of various aspects and embodiments is not intended to limit the scope of the present disclosure, but is merely representative of various aspects and embodiments. Furthermore, the compositions and methods disclosed herein can be modified by those skilled in the art without departing from the scope of the present disclosure. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs. All publications and patents mentioned herein are incorporated by reference.

[0027] definition For purposes of this disclosure, the following definitions will be used unless expressly stated otherwise.

[0028] As used in the context of describing the present disclosure, the terms "a," "an," "the," and similar referents are intended to encompass both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. All methods described herein can be performed in any suitable order, unless otherwise indicated herein or clearly contradicted by context. The use of any examples or exemplary language (e.g., "etc.") provided herein is intended merely to better elucidate the disclosure and does not pose a limitation on the scope of the otherwise claimed disclosure. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the disclosure.

[0029] For example, with respect to given numerical values ​​for temperature and time period, the term "about" means including numerical values ​​within 10% of the particular value.

[0030] As used herein, an "alkyl" group or "alkane" is a straight-chain or branched, non-aromatic hydrocarbon that is completely saturated. Typically, a straight-chain or branched alkyl group has from 1 to about 20, preferably from 1 to about 10, carbon atoms, unless otherwise defined. Examples of straight-chain and branched alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, tert-pentyl, neo-pentyl, iso-pentyl, sec-pentyl, 3-pentyl, sec-iso-pentyl, activated pentyl, hexyl, heptyl, octyl, ethylhexyl, and the like. 1~8A straight-chain or branched alkyl group is also referred to as a "lower alkyl" group. An alkyl group with two open valences may be referred to as an alkylene group, e.g., methylene, ethylene, propylene, etc. Furthermore, the term "alkyl" (or "lower alkyl"), as used throughout the specification, examples, and claims, is intended to include both "unsubstituted alkyl" and "substituted alkyl," the latter referring to alkyl moieties having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone. Such substituents, unless otherwise specified, can include, for example, alkyl, halogen, hydroxyl, carbonyl (e.g., carboxyl and alkoxycarbonyl, formyl, or acyl), thiocarbonyl (e.g., thioester, thioacetate, or thioformate), alkoxyl, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic moieties. Those skilled in the art will understand that the moieties substituted on the hydrocarbon chain can themselves be substituted, if appropriate. For example, substituents of substituted alkyls can include amino, azido, imino, amido, phosphoryl (including phosphonates and phosphinates), sulfonyl (including sulfates, sulfonamides, sulfamoyl, and sulfonates), and silyl groups, as well as substituted and unsubstituted forms of ethers, alkylthio, carbonyl (including ketones, aldehydes, carboxylates, and esters), -CF3, -CN, and the like. Exemplary substituted alkyls are described below. Cycloalkyls can be further substituted with alkyls, alkenyls, alkoxys, alkylthios, aminoalkyls, carbonyl-substituted alkyls, -CF3, -CN, and the like. In other embodiments, the term "alkyl" can refer to "cycloalkyl," which refers to a non-aromatic carbocyclic ring having 3 to 10 carbon ring atoms, the carbon atoms being bonded together to form the ring. The ring can be saturated or can have one or more carbon-carbon double bonds.Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, and cycloheptyl, as well as bridged and caged saturated ring groups, such as norbornyl and adamantyl. As described herein, organic solvents include, but are not limited to, aliphatic hydrocarbon solvents, aromatic hydrocarbon solvents, alcohols or alkyl alcohols, alkyl ethers, sulfoxides, alkyl ketones, alkyl acetates, trialkylamines, alkyl formates, trialkylamines, or combinations thereof. Aliphatic hydrocarbon solvents can be pentane, hexane, heptane, octane, cyclohexane, etc., or combinations thereof. Aromatic hydrocarbon solvents can be benzene, toluene, etc., or combinations thereof. Alcohols or alkyl alcohols include, for example, methanol, ethanol, propanol, butanol, pentanol, hexanol, heptanol, octanol, decanol, amyl alcohol, or combinations thereof. Alkyl ethers include methyl, ethyl, propyl, butyl, etc., such as diethyl ether, diisopropyl ether, or combinations thereof. Sulfoxides include dimethyl sulfoxide (DMSO), decyl methyl sulfoxide, tetradecyl methyl sulfoxide, etc., or combinations thereof. The term "alkyl ketone" refers to a ketone substituted with an alkyl group, such as acetone, ethyl methyl ketone, etc., or combinations thereof. The term "alkyl acetate" refers to an alkyl acetate such as ethyl acetate, propyl acetate (n-propyl acetate, iso-propyl acetate), butyl acetate (n-butyl acetate, iso-butyl acetate, sec-butyl acetate, tert-butyl acetate), amyl acetate (n-pentyl acetate, tert-pentyl acetate, neo-pentyl acetate, iso-pentyl acetate, sec-pentyl acetate, 3-pentyl acetate, sec-iso-pentyl acetate, activated pentyl acetate), 2-ethylhexyl acetate, and the like, or combinations thereof.The term "alkyl formate" refers to a formic acid ester substituted with an alkyl group, such as methyl formate, ethyl formate, propyl formate, butyl formate, etc., or combinations thereof. The term "trialkylamine" refers to an amino group substituted with three alkyl groups, such as triethylamine.

[0031] As used herein, "amino acid" or "residue" refers to any natural or unnatural amino acid, any amino acid derivative, or any amino acid mimetic known in the art. Included are L- and D-forms of each amino acid, although the L-form is generally preferred. In some embodiments, the term refers to any one of the 20 naturally occurring amino acids in their L-form: glycine (Gly), alanine (Ala), valine (Val), leucine (Leu), isoleucine (Ile), proline (Pro), cysteine ​​(Cys), methionine (Met), serine (Ser), threonine (Thr), glutamine (Gin), asparagine (Asn), glutamic acid (Glu), aspartic acid (Asp), lysine (Lys), histidine (His), arginine (Arg), phenylalanine (Phe), tryptophan (Trp), and tyrosine (Tyr). In certain embodiments, the amino acid side chain can be the side chain of Gly, Ala, Val, Leu, Ile, Met, Cys, Ser, Thr, Trp, Phe, Lys, Arg, His, Tyr, Asn, Gln, Asp, Glu, or Pro.

[0032] As used herein, unless the context requires otherwise, the term "comprise" and variations of this term, such as "comprises," "including," and "comprised," are not intended to exclude additional additives, components, integers, or steps. The terms "comprises" and "comprising" may be used interchangeably. As used herein, the phrases "selected from the group consisting of," "selected from," and the like, include mixtures of specified materials. When a numerical limit or range is stated herein, the endpoints are included. Also, all values ​​and subranges within the numerical limit or range are specifically included as if expressly written out. Reference to an element in the singular is not intended to mean "only one" unless specifically stated, but rather "one or more." Unless specifically stated otherwise, terms such as "some" refer to one or more, and singular terms such as "a," "an," and "the" refer to one or more.

[0033] The term "oligopeptide" refers to a peptide with a smaller number of amino acid residues than a polypeptide or protein. Oligopeptides described herein typically consist of about 2 to about 40 amino acid residues. Oligopeptides include dipeptides (2 amino acids), tripeptides (3 amino acids), tetrapeptides (4 amino acids), pentapeptides (5 amino acids), hexapeptides (6 amino acids), heptapeptides (7 amino acids), octapeptides (8 amino acids), nonapeptides (9 amino acids), decapeptides (10 amino acids), undecapeptides (11 amino acids), dodecapeptides (12 amino acids), icosapeptides (20 amino acids), tricontapeptides (30 amino acids), and tetracontapeptides (40 amino acids). Oligopeptides can also be classified by molecular structure, including aeruginosins, cyanopeptolins, microcystins, microviridins, microginins, anabaenopeptins, and cyclamides. Homo-oligopeptides are oligopeptides that contain the same amino acids. In a preferred embodiment, the homo-oligopeptides contain polyvaline, polyalanine, and polyglycine hexamers of 10 amino acids.

[0034] The term "peptide" is defined as a small protein of two or more amino acids linked by the carboxyl group of one amino acid to the amino group of another. Thus, at its basic level, any type of peptide synthesis involves repeated steps of adding amino acids or peptide molecules to each other or to an existing peptide chain. The term "peptide" generally contains from about 2 to about 100 amino acids, while a polypeptide or protein contains more than about 100 amino acids, up to the full-length sequence that can be translated from a gene. Furthermore, as used herein, a peptide can be a subsequence or portion of a polypeptide or protein. In certain embodiments, the peptide is 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, The peptide may consist of 53, 54, 55, 56, 57, 58, 59, 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, 99, or 100 amino acid residues. In preferred embodiments, the peptide is about 30 to about 100 amino acids in length. In some embodiments, the peptide is about 40 to about 100 amino acids in length.

[0035] As used herein, the term "pharmaceutically acceptable" refers to a composition that is physiologically tolerable and does not typically produce an allergic or similar adverse reaction when administered to a subject, preferably a human subject. Preferably, as used herein, the term "pharmaceutically acceptable" means approved by a federal or state regulatory agency or listed in the United States Pharmacopoeia or other generally recognized pharmacopoeias for use in animals, more particularly humans.

[0036] As used herein, the term "prodrug" is intended to encompass therapeutic biological agents that are converted under physiological conditions to therapeutically active biological agents of the present disclosure. Common methods for making prodrugs include one or more selected moieties that are hydrolyzed under physiological conditions to reveal the desired molecule. In other embodiments, the prodrug is converted by enzymatic activity in the host animal. For example, esters or carbonates (e.g., esters or carbonates of alcohols or carboxylic acids) are preferred prodrugs of the present disclosure. In certain embodiments, some or all of the molecules in the compositions depicted above can be replaced with the corresponding appropriate prodrug, e.g., where a hydroxyl in the parent molecule is presented as an ester, or a carbonate or carboxylic acid present in the parent therapeutic biological agent is presented as an ester.

[0037] The term "protein" is defined as a linear polymer constructed from approximately 20 different amino acids. The types and sequence of amino acids in a protein are specified by the DNA from which the protein is produced. In certain embodiments, the sequence can be natural or non-natural. The sequence of amino acids determines the overall structure and function of the protein. In some embodiments, a protein can contain 50 or more residues. In preferred embodiments, a protein can contain more than about 101 residues in length. The net charge of a protein can be determined by two factors: 1) the total number of acidic amino acids relative to basic amino acids; and 2) the particular solvent pH environment that exposes positive or negative residues. As used herein, a "net positively or net negatively charged protein" is a protein that has a net positive or net negative charge in a non-denaturing pH environment. Those skilled in the art will understand that, in general, all proteins can be considered "net negatively charged proteins" depending on their pH and / or solvent environment, regardless of their amino acid composition. For example, different solvents can expose negative or positive side chains depending on the solvent pH. The protein or peptide is preferably selected from any type of enzyme or antibody or fragment thereof that exhibits substantially the same activity as the corresponding enzyme or antibody. Proteins or peptides may serve as structural materials (e.g., keratin), enzymes, hormones, transporters (e.g., hemoglobin), antibodies, or regulators of gene expression. Proteins or peptides are required for the structure, function, and regulation of cells, tissues, and organs.

[0038] The term "substantially," as used herein, refers to a majority or majority, such as in at least about 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, 99.99%, or at least about 99.999% or more.

[0039] It is understood that the specific order or hierarchy of steps in a disclosed method or process is an illustration of a sample approach. Based on design preferences, it is understood that the specific order or hierarchy of steps in a method or process can be rearranged. Some of the steps may occur simultaneously. The accompanying methods claim elements of the various steps in the illustrated order and are not meant to be limited to the specific hierarchy or order presented. The use of a phrase such as "an embodiment" does not imply that such an embodiment applies to all configurations of the subject technology. Disclosure regarding an embodiment may apply to all embodiments or to one or more embodiments. A phrase such as "one embodiment" may refer to one or more embodiments, and vice versa.

[0040] particle Unless otherwise defined, all technical terms, notations, and other scientific terminology used herein are intended to have the meaning commonly understood by one of ordinary skill in the art to which this disclosure pertains. In some cases, terms having commonly understood meanings are defined herein for clarity and / or ready reference, and the inclusion of such definitions herein should not necessarily be construed as representing a substantial deviation from what is commonly understood in the art. The techniques and procedures described or referenced herein are generally well understood and commonly used by those skilled in the art using conventional methodology. Where appropriate, procedures involving the use of commercially available kits and reagents are generally carried out according to manufacturer-specified protocols and / or parameters unless otherwise noted.

[0041] In some embodiments, the present disclosure relates to particles comprising an agent, wherein the particles comprise less than about 25% internal void space and the circularity of the particles is from about 0.10 to about 1.00.

[0042] The terms "particle" or "microparticles" are used interchangeably herein in the broadest sense to refer to a discrete body. The particles described herein are round, spheroidal, and of controlled dispersity, with characteristic sizes ranging from submicrometers to tens of micrometers, in contrast to the porous, monolithic "cakes" typically produced during conventional freeze-drying, for example. This morphology allows for a flowable powder (as described by a low Hausner ratio) without further processing. In some embodiments, the term "particle" refers to a quantity of an agent in a substantially solid state of matter compared to a liquid droplet or in gel form. In other embodiments, a particle may comprise a core and a shell, where the shell may be considered as encapsulating material. In still other embodiments, a particle does not comprise a shell, in which case the particle is composed entirely of the core. The term "primitive particle" refers to a stage of particle formation in which one or more of the components that make up the particle are at least partially dehydrated. The total liquid content of the primary particle is less than that of the droplet and more than that of the formed particle. Similarly, the average concentration of solute is higher than that of the droplets, but typically lower than that of the formed particles. The term "encapsulant" refers to a substance that can be dried or gelled around a particle core to form a shell.

[0043] As disclosed herein, the agent can be a therapeutic or diagnostic agent. In some embodiments, the agent is not a diagnostic or therapeutic agent. In other embodiments, the agent can be a metal or other element, silica, titania, a metal salt, a metal oxide, a metal nitride, a metal sulfide, a metal alkoxide, a polymer, or a combination thereof. Exemplary therapeutic or diagnostic agents include, but are not limited to, nucleic acids, oligonucleotides, antibodies or fragments thereof, amino acids, peptides, proteins, cells, bacteria, gene therapy agents, genome engineering therapeutic agents, epigenetic engineering therapeutic agents, carbohydrates, chemical drugs, contrast agents, magnetic particles, polymer beads, metal nanoparticles, metal microparticles, quantum dots, antioxidants, antibiotic agents, hormones, nucleoproteins, polysaccharides, glycoproteins, lipoproteins, steroids, analgesics, local anesthetics, anti-inflammatory agents, antimicrobial agents, chemotherapeutic agents, exosomes, outer membrane vesicles, vaccines, viruses, bacteriophages, adjuvants, vitamins, minerals, organelles, or a combination thereof. In a preferred embodiment, the therapeutic agent is a therapeutic biological agent. Therapeutic and diagnostic agents can have a molecular weight of about 20 to about 200 kDa, e.g., about 40 to about 150 kDa or about 50 to about 100 kDa. Table 1 provides a list of therapeutic and diagnostic agents and typical concentration ranges for general classes of compounds in pharmaceutical compositions.

[0044] [Table 1]

[0045] In other embodiments, the particles may include agents such as, but not limited to, silica, titania, metals or other elements, metal salts, metal oxides, metal nitrides, metal sulfides, metal alkoxides, and / or polymers. Methods as described herein may offer an alternative to sol-gel synthesis and can provide particles for a diverse array of applications, including semiconductor particles (e.g., lead sulfide), surface plasmon resonance (e.g., gold), magnetic (e.g., iron oxide), UV blocking (e.g., zinc oxide), contrast agents (e.g., silicon), or laser applications (e.g., poly(methyl methacrylate) and silicon dioxide mixtures).

[0046] Therapeutic biological agents, also known as biomedical products or biopharmaceuticals, are any pharmaceutical products manufactured in, extracted from, or semi-synthesized from biological sources. Therapeutic biological agents can include a wide range of products, such as vaccines, blood and blood components, allergens, somatic cells, gene therapy, tissues, and recombinant therapeutic proteins. In some embodiments, biological agents can be composed of sugars, proteins, or nucleic acids, or complex combinations of these substances, or can be living entities, such as cells and tissues. Biological agents can be isolated from a variety of natural sources, such as humans, animals, or microorganisms, or can be produced by biotechnology methods or other techniques. Gene-based and cellular biological agents are often used, for example, to treat various medical conditions for which no other treatments are available. In a preferred embodiment of the present disclosure, the therapeutic biological agent is an antibody.

[0047] The terms "antibody" and "immunoglobulin" are used interchangeably in the broadest sense and include monoclonal, polyclonal, polyvalent, and multispecific antibodies, regardless of how they are produced (i.e., using immunization, recombinant, or synthetic methodologies). Antibodies may be gamma globulin proteins found in the blood or other bodily fluids of vertebrates that function in the immune system to bind antigens, thus identifying and / or neutralizing foreign substances. Antibodies can be assigned to different classes or isotypes. There are five classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, with heavy chains designated alpha, delta, epsilon, gamma, and mu, respectively. The gamma class is further divided into subclasses based on differences in sequence and function; for example, humans express the following subclasses: IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. In certain embodiments of the present disclosure, the IgG antibody is an IgG1 antibody. In a preferred embodiment of the present disclosure, the IgG1 antibody is a monoclonal IgG1 antibody. Light chains from any vertebrate species can be assigned to one of two clearly distinct types, e.g., kappa and lambda, based on the amino acid sequences of their constant domains.

[0048] Recognized immunoglobulin genes include the kappa, lambda, alpha, gamma, delta, epsilon, and mu constant region genes, as well as the myriad immunoglobulin variable region genes. In some embodiments, light chains are classified as kappa or lambda. In other embodiments, heavy chains are classified as gamma, mu, alpha, delta, or epsilon, which define the immunoglobulin classes IgG, IgM, IgA, IgD, and IgE, respectively. In preferred embodiments of the present disclosure, the antibody is an IgG antibody.

[0049] An exemplary antibody (immunoglobulin) structural unit comprises a tetramer. Each tetramer is composed of two identical pairs of polypeptide chains, each pair having one "light" chain (about 25 kD) and one "heavy" chain (about 50-70 kD). The N-terminus of each chain defines a variable region of about 100-110 or more amino acids primarily responsible for antigen recognition. The terms "variable light" chain, domain, region, and component are used interchangeably and are referred to as "VL" or "V L " and refers to the light chain of an antibody or antibody fragment. Similarly, the terms "variable heavy" chain, domain, region, and component are used interchangeably and are abbreviated as "VH" or "V H " and refers to the heavy chain of an antibody or antibody fragment. Antibodies are generally heterotetrameric glycoproteins composed of two identical light (L) chains and two identical heavy (H) chains. Each L chain is linked to an H chain by one covalent disulfide bond. The two H chains are linked to each other by one or more disulfide bonds depending on the H chain isotype. Each H and L chain also has regularly spaced intrachain disulfide bridges. H and L chains define specific Ig domains. In particular, each H chain has a variable domain (VH) at the N-terminus, followed by three constant domains (CH) for each of the alpha and gamma chains and four CH domains for the p and c isotypes. Each L chain has a variable domain (VL) at the N-terminus, followed by a constant domain (CL) at the other end. The VL is aligned with the VH, and the CL is aligned with the first constant domain of the heavy chain (CHL). The constant domain comprises the Fc portion, which comprises the carboxy-terminal portions of both H chains held together by disulfides. Antibody effector functions, such as ADCC, are determined by sequences in the Fc region, which is the portion recognized by Fc receptors (FcRs), which are also found on certain types of cells.

[0050] As disclosed herein, the pairing of a VH and a VL together forms a "variable region" or "variable domain" comprising the amino-terminal domain of either the heavy or light chain of an antibody. The variable domain of a heavy chain may be referred to as "VH." The variable domain of a light chain may be referred to as "VL." The V domain contains the "antigen-binding site," which influences antigen binding and defines the specificity of a particular antibody for its particular antigen. The V region spans approximately 110 amino acid residues and consists of relatively invariant stretches of 15-30 amino acids called framework regions (FRs) (generally about four) separated by shorter, highly variable regions called "hypervariable regions" (generally about three), each generally 9-12 amino acids long. The FRs largely adopt a p-sheet configuration, and the hypervariable regions form loops that connect and occasionally form part of the p-sheet structure. In certain embodiments, "hypervariable region" refers to the region of an antibody variable domain that is hypervariable in sequence and / or forms structurally defined loops. Generally, antibodies contain six hypervariable regions: three in the VH (H1, H2, H3) and three in the VL (L1, L2, L3). "Framework" or "FR" residues are those variable domain residues other than the hypervariable region residues as herein defined.

[0051] The terms "full length antibody," "intact antibody," and "whole antibody" are used interchangeably herein to refer to an antibody in its substantially intact form, and not as an antibody fragment as defined above. These terms particularly refer to an antibody having a heavy chain containing an Fc region. A full length antibody may be a native sequence antibody or an antibody variant. In certain embodiments, an "intact" or "whole" antibody is one that comprises an antigen-binding site and a CL and at least heavy chain constant domains, CH1, CH2, and CH3. The constant domains may be native sequence constant domains (e.g., human native sequence constant domains) or amino acid sequence variants thereof.

[0052] As disclosed herein, "whole antibody fragments containing the variable domains" include Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies; single-chain antibody molecules; and multispecific antibodies formed from antibody fragments. A "Fab fragment" consists of an entire L chain together with the variable region domain of the H chain (VH) and the first constant domain of one heavy chain (CHI). Each Fab fragment is monovalent with respect to antigen binding, i.e., it has a single antigen-binding site. A "Fab' fragment" differs from a Fab fragment by having a few additional residues at the carboxy terminus of the CHI domain, including one or more cysteines from the antibody hinge region. Fab'-SH is the designation for Fab' herein, in which the cysteine ​​residues of the constant domains bear a free thiol group. A "F(ab')2 fragment" roughly corresponds to two disulfide-linked Fab fragments with bivalent antigen-binding activity and is still capable of cross-linking antigen. An "Fv" is the minimum antibody fragment containing a complete antigen-recognition and binding site. This fragment consists of a dimer of one heavy- and one light-chain variable region domain in tight, non-covalent association. In single-chain Fv (scFv) species, one heavy- and one light-chain variable domain can be covalently linked by a flexible peptide linker, allowing the light and heavy chains to associate in a "dimeric" structure similar to that in two-chain Fv species. Folding of these two domains results in six hypervariable loops (three loops from each H and L chain) that provide amino acid residues for antigen binding and confer antigen-binding specificity to the antibody. A "single-chain Fv," also abbreviated as "sFv" or "scFv," is an antibody fragment comprising VH and VL antibody domains connected to form a single-chain polypeptide. In a preferred embodiment, the scFv polypeptide further comprises a polypeptide linker between the VH and VL domains, which enables the scFv to form the desired structure for antigen binding.In some embodiments, a "single variable domain" is half of an Fv (comprising only three CDRs specific for an antigen) that has the ability to recognize and bind antigen, although at a lower affinity than the entire binding site.

[0053] In some embodiments, "diabody" refers to an antibody fragment having two antigen-binding sites, which fragments comprise a heavy-chain variable domain (VH) connected to a light-chain variable domain (VL) on the same polypeptide chain (VH-VL). Small antibody fragments are prepared by constructing sFv fragments with a short linker (approximately 5-10 residues) between the VH and VL domains such that interchain pairing rather than intrachain pairing of the V domains is achieved, resulting in bivalent fragments, i.e., fragments with two antigen-binding sites. In other embodiments, diabodies can be bivalent or bispecific. In certain embodiments, bispecific diabodies are heterodimers of two "crossover" sFv fragments, in which the VH and VL domains of the two antibodies are present on different polypeptide chains. Trispecific and tetraspecific antibodies are also generally known in the art.

[0054] An "antigen-binding fragment" of an antibody as described herein includes only a portion of the intact antibody that generally comprises the antigen-binding site of the intact antibody and thus retains the ability to bind antigen. Illustrative examples of antibody fragments encompassed by this definition include, but are not limited to, (i) a Fab fragment having the VL, CL, VH, and CH1 domains; (ii) a Fab' fragment, which is a Fab fragment with one or more cysteine ​​residues at the C-terminus of the CH1 domain; (iii) an Fd fragment having the VH and CH1 domains; (iv) an Fd' fragment having the VH and CH1 domains and one or more cysteine ​​residues at the C-terminus of the CH1 domain; (v) an Fv fragment having the VL and VH domains of a single arm of an antibody; and (vi) a dAb consisting of a VH domain. (vii) isolated CDR regions; (viii) F(ab')2 fragments (bivalent fragments comprising two Fab' fragments linked by a disulfide bridge at the hinge region); (ix) single-chain antibody molecules (e.g., single-chain Fv; scFv); (x) "diabodies" having two antigen-binding sites comprising a heavy-chain variable domain (VH) connected to a light-chain variable domain (VL) in the same polypeptide chain; and (xi) "linear antibodies" comprising a pair of tandem Fd segments (VH-CH1-VH-CH1) that, together with complementary light-chain polypeptides, form a pair of antigen-binding regions. In some embodiments, "antigen-binding site" generally refers to a molecule comprising at least the hypervariable and framework regions required to confer antigen-binding function to the V domain. The antigen-binding site may be in the form of an antibody or antibody fragment (e.g., dAb, Fab, Fd, Fv, F(ab')2, or scFv) in the methods described herein.

[0055] In some embodiments, the term "single-chain Fv" or "scFv" or "single-chain" antibody can refer to an antibody fragment comprising the VH and VL domains of an antibody, wherein these domains are present in a single polypeptide chain. Generally, the Fv polypeptide further comprises a polypeptide linker between the VH and VL domains that enables the sFv to form the desired structure for antigen binding. For a review of sFvs, see Pluckthun, THE PHARMACOLOGY OF MONOCLONAL ANTIBODIES, vol. 113, Rosenburg and Moore eds. Springer-Verlag, New York, pp. 269-315 (1994).

[0056] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible natural mutations that may be present in minor amounts. Monoclonal antibodies (mAbs) are highly specific, targeting a single antigenic site or determinant on an antigen. In addition to their specificity, monoclonal antibodies are advantageous in that they may be synthesized uncontaminated by other antibodies. Monoclonal antibodies may be prepared by hybridoma methodology. Monoclonal antibodies may also be isolated from phage antibody libraries using molecular engineering techniques. The monoclonal antibodies of the present disclosure may be generated by recombinant DNA methods and may be referred to as "recombinant antibodies" or "recombinant monoclonal antibodies" as described herein. In some embodiments, monoclonal antibodies are a single species of antibody, in which all antibody molecules recognize the same epitope because all antibody-producing cells are derived from a single B lymphocyte cell lineage. Methods for generating monoclonal antibodies (mAbs) generally begin along the same lines as those for preparing polyclonal antibodies. In other embodiments, rodents, such as mice and rats, are used in raising monoclonal antibodies. In certain embodiments, rabbit, sheep, or frog cells are used in raising monoclonal antibodies. The use of rats is well known and may offer certain advantages. Mice (e.g., BALB / c mice) are routinely used and generally provide a high percentage of stable fusions. In yet other embodiments of the present disclosure, the antibody is a monoclonal antibody. In a preferred embodiment of the present disclosure, the IgG antibody is monoclonal.

[0057] In other embodiments, recombinant antibody fragments can be isolated from phage antibody libraries using techniques well known in the art. See, e.g., Clackson et al., 1991, Nature 352:624-628; Marks et al., 1991, J. Mol. Biol. 222:581-597. Recombinant antibody fragments can be derived from large phage antibody libraries generated by recombination in bacteria (Sblattero and Bradbury, 2000, Nature Biotechnology 18:75-80; and as described herein). Polynucleotides encoding the VH and VL components of antibody fragments (i.e., scFv) can be used to generate recombinant full-length immunoglobulins using methods known in the art (see, e.g., Persic et al., 1997, Gene 187:9-18).

[0058] An "isolated antibody" is one that has been identified and separated and / or recovered from components of its existing environment. Contaminant components are materials that would interfere with therapeutic uses for the antibody, and may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes.

[0059] As used herein, "human antibody" refers to an antibody having an amino acid sequence corresponding to that of an antibody produced by a human. Human antibodies can be produced using a variety of techniques known in the art, including phage display libraries. Human antibodies can be prepared by administering antigen to transgenic animals that have been engineered to produce such antibodies in response to antigen challenge, but whose endogenous gene loci are impaired. "Humanized" forms of non-human (e.g., rodent) antibodies are chimeric antibodies that contain minimal sequence derived from the non-human antibody. For the most part, humanized antibodies are human immunoglobulins (recipient antibody) in which residues from the recipient's hypervariable region are replaced by residues from a hypervariable region of a non-human species (donor antibody), such as mouse, rat, rabbit, or non-human primate, possessing the desired antibody specificity, affinity, and performance. In some cases, framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies may comprise residues not found in the recipient antibody or the donor antibody. These modifications are made to further refine antibody performance. In general, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the FRs are those of a human immunoglobulin sequence. The humanized antibody optionally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin.

[0060] An "affinity matured" antibody is one that has one or more alterations in one or more hypervariable regions thereof that result in an improvement in the affinity of the antibody for an antigen, compared to a parent antibody that does not possess those alterations. In some embodiments, an affinity matured antibody may have micromolar affinity for the target antigen. In other embodiments, an affinity matured antibody may have nanomolar or even picomolar affinity for the target antigen. Affinity matured antibodies are produced by procedures known in the art.

[0061] A "blocking" or "antagonist" antibody is one that inhibits or reduces the biological activity of the antigen to which it binds. In some embodiments, a blocking or antagonist antibody substantially or completely inhibits the biological activity of the antigen. An "agonist antibody," as used herein, is an antibody that mimics at least one functional activity of a polypeptide of interest.

[0062] "Binding affinity" generally refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule, e.g., an antibody, and its binding partner, e.g., an antigen. Unless otherwise indicated, as used herein, "binding affinity" refers to the intrinsic binding affinity, reflecting a 1:1 interaction between members of a binding pair, e.g., an antibody and an antigen. The affinity of molecule X for its partner Y can generally be expressed by the dissociation constant (Kd). Affinity can be measured by common methods known in the art, including those described herein. Low-affinity antibodies generally bind antigen slowly and tend to dissociate easily, while high-affinity antibodies generally bind antigen more quickly and remain bound longer. Various methods for measuring binding affinity are known in the art, any of which can be used for purposes of the present disclosure. "Epitope" generally refers to that portion of an antigen bound by the antigen-binding site of an antibody. In some embodiments, an epitope can be "linear" in the sense that the hypervariable loops of the antibody CDRs that form the antigen-binding site bind to a sequence of amino acids as found in the primary protein structure. In another embodiment, the epitope is a "conformational epitope," ie, a hypervariable loop of a CDR binds to residues that are displayed in the tertiary or quaternary structure of a protein.

[0063] In some embodiments of any of the above compositions and methods, the therapeutic biological agent is an antibody. In other embodiments, the antibody is selected from the group consisting of, but not limited to, 3F8, abagovomab, abatacept, abciximab, abirulumab, abrezekimab, acritumomab, actoxumab, abituzumab, adalimumab-adbm, adalimumab-atto, adalimumab-bwwb, adecatumumab, Ado-trastuzumab emtansine, aducanumab, afacevicumab, afelimomab, aflibercept, afutuzumab, alacizumab pegol, ALD518, alefacept, alemtuzumab, alirocumab, Altumomab pentetate, amatuximab, anatumomab mafenatox, andecaliximab, anetumab ravtansine, anifrolumab, anrukinzumab, apolizumab, aprtumab ixadotin, arcitumomab, asclinbacumab, acelizumab, atezolizumab, atidortoxumab, atinumab, atlizumab, atolimu-mab, avelumab, azintuxizumab vedotin, bapineuzumab, basiliximab, bavituximab, BCD-100, brosozumab, begelomab, belantamab vedotin, belatase Puto, belimumab, bemarituzumab, benralizumab, bermekimab, bersanlimab, bertilimumab, besilesomab, bevacizumab, bevacizumab-awwb, bezlotoxumab, biciromab, bimagrumab, bimekizumab, viltamimab, bivatuzumab mertansine, bleselumab, blinatumomab, brontuximab, brosozumab, bococizumab, brazikumab, brentuximab vedotin, briakinumab, brodalumab, brolucizumab, brontiximab, burosumab, cabilalizumab, camidalumab tesirin , camrelizumab, canakinumab, cantuzumab mertansine, cantuzumab lavtansine, caplacizumab, capromab pendetide, carlumab, carotuximab, catumaxomab, cBR96-doxorubicin immunoconjugate, cedelizumab, cemiplimab, sergituzumab amnalleukin, sergituzumab amnalleukin, certolizumab pegol, cetrelimab, cetuximab, civisatamab, cimutuzumab, sitatuzumab bogatox, cixutumumab, clazakizumab, clenoliximab, clivatuzumab tetraxetan,Codrituzumab, cofetuzumab peridotin, coltuximab ravtansine, conatumumab, concizumab, cosafrobiximab, crenezumab, CR6261, crizanlizumab, clotezumab, cusatuzumab, dacetuzumab, daclizumab, darotuzumab, dapirolizumab pegol, daratumumab, dectrecumab, demcizumab, denileukin diftitox, denituzumab mafodotin, denosumab, depatuximab mafodotin, delrotuximab biotin, detumomab, dezamizumab, dinutuximab, zirconiumab, domagrozumab, dol Rimomab alitox, dostallimab, drozitumab, DS-8201, zurigotumab, dupilumab, zulvalumab, zusigitumab, zuvortuxizumab, ecromeximab, eculizumab, edovacomab, edrecolomab, efalizumab, efungumab, eldelumab, elezanumab, elgemtumab, elotuzumab, ersilimomab, emactuzumab, emibetuzumab, emicizumab, enapotamab vedotin, enavatuzumab, enfortumab vedotin, enlimomab pegol, enoblitzumab, enokizumab, enoticumab, ensituximab , epitumomab-situxetan, epoetin-alfa, epoetin-alfa-epbx, epratuzumab, eptinezumab, erenumab, erlizumab, ertumaxomab, etanercept, etanercept-szzs, etaracizumab, etigilimab, etrolizumab, evinacumab, evolocumab, exibirumab, factor VIII Fc fusion protein, factor IX Fc fusion protein, fanolesomab, faralimomab, faricimab, farletuzumab, fasinumab, felvizumab, fezakinumab, fivatuzumab, ficlatuzumab , figitumumab, filgrastim, filgrastim-sndz, filibumab, framvotumab, fretikumab, flotetuzumab, hontolizumab, foramumab, foravilumab, fremanezumab, fresolimumab, flubocimab, furunvetomab, furanumab, futuximab, galcanezumab, galiximab, ganitumab, gantenerumab, gatipotuzumab, gavirimomab, gezivumab, gemtuzumab ozogamicin, gevokizumab, gilvetomab, gimsilumab, girentuximab, glenbatumumab vedotin, golimumab, gomiliximab,Goslanemab, guselkumab, ibalizumab, IBI308, ibritumomab tiuxetan, icrucumab, idarucizumab, ifavotuzumab, igovomab, iradatuzumab vedotin, IMAB362, imalumab, imaprelimab, imciromab, imgatuzumab, inlacumab, indatuximab ravtansine, indusatuzumab vedotin, inebilizumab, infliximab, infliximab-abda, infliximab-dyyb, infliximab-qbtx, intetumumab, inolimomab, inotuzumab ozogamicin, ipilimumab , Iomab-B, Iratumumab, Isatuximab, Iscalimab, Istiratumab, Itolizumab, Ixekizumab, Keliximab, Labetuzumab, Lacnotuzumab, Radilatuzumab vedotin, Lambrolizumab, Lampalizumab, Lanadelumab, Landgrozumab, Laprituximab emtansine, Ralcabiximab, Lebrikizumab, Remaresomab, Lendarizumab, Lembervimab, Lenzilumab, Lerdelimumab, Leronlimab, Lesofabrumab, Letolizumab, Lexatumumab, Ribivirumab, Rifastuzumab vedotin, Ligelizumab, Ronca Stuximab tesirin, rosatuxizumab vedotin, rilotomab satetraxetan, lintuzumab, lirilumab, roderucizumab, loxivetumab, lorvotuzumab mertansine, lucatumumab, lurizumab pegol, rumiliximab, lumletuzumab, rupartumab amadotin, rutikizumab, mapatumumab, margetuximab, marstacimab, maslimomab, mavrilimumab, matuzumab, mepolizumab, metelimumab, milatuzumab, minletumomab, mirikizumab, mirvetuximab soravtansine, mitumomab, modutuximab, mogamulizumab , monalizumab, morolimumab, mosunetuzumab, motavizumab, moxetumomab pasudotox, muromonab-CD3, nacolomab butafenatox, namilumab, naptumomab estafenatox, naratuximab emtansine, narutumab, natalizumab, nabicixizumab, nabivumab, naxitamab, nebacumab, necitumumab, nemolizumab, NEOD001, nerelimomab, nesbacumab, netakimab, nimotuzumab, nirsevimab, nivolumab, nofetumomab merpentane, obilutoxaximab, obinutuzumab, ocaratuzumab,Ocrelizumab, ozlimomab, ofatumumab, olaratumab, oleculab, orendalizumab, olokizumab, omalizumab, omburtamab, OMS721, onartuzumab, ontuxizumab, onvatilimab, opicinumab, oportuzumab monatox, oregovomab, olticumab, otelixizumab, otilimab, otreltuzumab, oxelumab, ozanezumab, ozoralizumab, pagibaximab, palivizumab, pamrevlumab, panitumumab, pancomab, panobacumab, palsatuzumab, pascolizumab, pasotuxizumab, pasotuximab ... Teclizumab, patritumab, PDR001, pegfilgrastim-jmdb, pembrolizumab, pemtumomab, perakizumab, pertuzumab, pexelizumab, pidilizumab, pinatuzumab vedotin, pintumomab, placuramab, prosalizumab, pogalizumab, polatuzumab vedotin, ponezumab, polgabiximab, prasinezumab, prezalizumab, priliximab, pritoxaximab, pritumumab, PRO140, kilimumab, racotumomab, radletumab, rafivirumab, ralpancizumab, ramucirumab, ranevtomab, ranibizumab Ibuprofen, raxibacumab, ravagalimab, ravutolizumab, refanezumab, regavirumab, relatolimab, lemtolumab, reslizumab, rilonacept, rilotumumab, rinukumab, risankizumab, rituximab, rituximab-abbs, rituximab-pvvr, rivabazumab pegol, rivabazumab pegol, lobatumumab, Rmab, lorezumab, romilkimab, romiplostim, romosozumab, rontalizumab, rosmantuzumab, rovalpituzumab tesirin, rovalpituzumab tesirin, rovelizumab, rozanolixizumab, ruplizumab, sasi Tuzumab govitecan, samalizumab, samrotamab vedotin, sapelizumab, sarilumab, satralizumab (SA237), satumomab pendetide, secukinumab, selicrelumab, seribantumab, cetoxaximab, ceturusumab, cevirumab, sibrotuzumab, SGN-CD19A, SGN-CD33A, SHP647, sifalimumab, siltuximab, simtuzumab, siplizumab, siltratumab vedotin, sirukumab, sofituzumab vedotin, solanezumab, solitomab, sonepcizumab, sontuzumab, spartalizumab, stamulumab,Sulesomab, sputabumab, stimulimab, subizumab, sublatoxumab, tabalumab, tacatuzumab tetraxetan, tadocizumab, talaxuzumab, talizumab, tamtubetomab, tanezumab, taplitumomab paptokis, tarexuzumab, tabolimab, tefibazumab, terimomab alitokis, telisotuzumab vedotin, tenatumomab, teneliximab, teplizumab, tepositamab, teprotumumab, tesid lumab, tetulomab, tezepelumab, TGN1412, tiburizumab, ticilimumab, tildrakizumab, tigatuzumab, timigituzumab, timolumab, tiragotumab, tislelizumab, tisotumab vedotin, TNX-650, tocilizumab, tomzotuximab, toralizumab, tosatoxumab, tositumomab, tobetumab, tralokinumab, trastuzumab, trastuzumab-anns, trastuzumab-dkst, Trastuzumab emtansine, tregalizumab, tremelimumab, trevoglumab, tucotuzumab celmoleukin, tuvilumab, ublituximab, urocupulumab, urelumab, urtoxazumab, ustekinumab, utomilumab, vadastuximab butarilin, banalimab, bundletuzumab vedotin, vanticutuzumab, vanucizumab, bapaliximab, valisacumab, varlilumab, batelizumab, vedolizumab, These include veltuzumab, bepalimomab, besencumab, visilizumab, bovalilizumab, volociximab, bonlerolizumab, bopratelimab, borsetuzumab mafodotin, votumumab, xentuzumab, XMAB-5574, zalutumumab, zanolimumab, zatuximab, xenoctuzumab, ziralimumab, zolbetuximab (IMAB362, claudiximab), Ziv-aflibercept, or zolimomab alitoxin.

[0064] In other embodiments of any of the above compositions and methods, the antibody is monoclonal. In certain embodiments, the monoclonal antibody is selected from the group consisting of, but not limited to, 3F8, 8H9, abatacept, abagovomab, abciximab, abituzumab, adalimumab-adbm, adalimumab-atto, adalimumab-bwwb, abrilumab, actoxumab, abituzumab, abrezekimab, abrilumab, actoxumab, adalimumab, adecatumumab, adotrastuzumab emtansine, aducanumab, afacevicumab, afelimomab, aflibercept, afutuzumab, alacizumab pegol, ALD518 , alefacept, alemtuzumab, alirocumab, altumomab pentetate, amatuximab, anatumomab, mafenatox, andecaliximab, anetumab ravtansine, anifrolumab, anrukinzumab (IMA-638), apolizumab, arcitumomab, asclinbacumab, acelizumab, atezolizumab, atidortoxumab, atinumab, atlizumab (tocilizumab), atolimumab, avelumab, bapineuzumab, basiliximab, bevacizumab, bevacizumab-awwb, BCD-100, brosozumab , begelomab, belatacept, belimumab, bemarituzumab, benralizumab, bermekimab, bersanlimab, bertilimumab, besilesomab, bezlotoxumab, biciromab, bimagrumab, bimekizumab, vilutamimab, bivatuzumab mertansine, bleselumab, blinatumomab, brontuzumab, brosozumab, bococizumab, brazikumab, brentuximab vedotin, briakinumab, brodalumab, brolucizumab, brontixotuzumab, burosumab, cabilalizumab, camrelizumab, canakinumab, cantuzumab, Ibuprofen mertansine, cantuzumab lavtansine, caplacizumab, capromab pendetide, carlumab, carotuximab, catumaxomab, cedelizumab, cemiplimab, certolizumab pegol, cetrelimab, cetuximab, civisatamab, cirumtuzumab, Ch.14.18, sitatuzumab bogatox, cixutumumab, clazakizumab, clenoliximab, clivatuzumab tetraxetan, codrituzumab, cofetuzumab peridotin, coltuximab lavtansine, conatumumab, concizumab, cosfrobiximab,crenezumab, CR6261, crizanlizumab, clotezumab, cusatuzumab, dacetuzumab, daclizumab, darotuzumab, dapirolizumab pegol, daratumumab, dectrecumab, demcizumab, denileukin diftitox, denintuzumab mafodotin, denosumab, delrotuximab biotin, detumomab, dezamizumab, dinutuximab, zilidabumab, domaglotuzumab, dostallimab, drozitumab, zurigotumab, dupilumab, zulvalumab, zusigitumab, zuvortuxizumab, eclomeximab, exclomeximab lizumab, edovacomab, edrecolomab, efalizumab, efungumab, eldelumab, elezanumab, elgemtuzumab, elotuzumab, ersilimomab, emactuzumab, emibetuzumab, emicizumab, enavatuzumab, enfortumab vedotin, enlimomab pegol, enoblitzumab, enokizumab, enoticumab, ensituximab, epitumomab cituxetan, epoetin-alpha, epoetin-alpha-epbx, epratuzumab, eptinezumab, erenumab, erlizumab, ertumaxomab, etanercept, etanercept To-szzs, etaracizumab, etigilimab, etorolizumab, evinacumab, evolocumab, exibirumab, factor VIII Fc fusion protein, factor IX Fc fusion protein, fanolesomab, faralimomab, faricimab, farletuzumab, fasinumab, FBTA05, felvizumab, fezakinumab, fibatuzumab, ficlatuzumab, figitumumab, filgrastim, filgrastim-sndz, filibumab, framvotumab, fretikumab, flotetuzumab, hontolizumab, foralumab, foralumab, floravirumab, frema Nezumab, fresolimumab, fluvocimab, furunevetumab, futuximab, galcanezumab, galiximab, ganitumab, gantenerumab, gatipotuzumab, gavilimomab, gedivumab, gemtuzumab ozogamicin, gevokizumab, gilvetomab, gimsilumab, girentuximab, glenbatumumab vedotin, golimumab, gomiliximab, goslanemab, guselkumab, ibalizumab, IBI308, ibritumomab tiuxetan, icrucumab, idarucizumab, ifavotuzumab, igovomab, IMAB362, imalumab,Imaprelimab, imciromab, imgatuzumab, inlacumab, indatuximab ravtansine, indusatumab vedotin, inebilizumab, infliximab, infliximab-abda, infliximab-dyyb, infliximab-qbtx, intetumumab, inolimomab, inotuzumab ozogamicin, ipilimumab, iratumumab, isatuximab, iscalimab, istiratumab, itolizumab, ixekizumab, keliximab, labetuzumab, lacunotuzumab, lambrolizumab, lampalizumab, lanadelumab, landrozma rab, ralcabiximab, lebrikizumab, remaresomab, lendalizumab, lembervimab, lenzilumab, lerdelimumab, leronlimab, lesofabumab, letolizumab, lexatumumab, ribivirumab, rifastuzumab vedotin, ligelizumab, rilotumab satetraxetan, lintuzumab, lirilumab, roderucizumab, lokivetmab, lorvotuzumab mertansine, lucatumumab, lurizumab pegol, lumiliximab, lumletuzumab, rutikizumab, mapatumumab, margetuximab, marstacimab, maslimomab, mavrilimumab, matsu Ibuprofen, mepolizumab, metelimuab, milatuzumab, minletumomab, mirikizumab, mirvetuximab soravtansine, mitumomab, modotuximab, mogamulizumab, monalizumab, morolimuab, mosunetuzumab, motavizumab, moxetumomab pasudotoxin, muromonab-CD3, nacolomab butafenatox, namilumab, naptumomab estafenatox, narunatumab, natalizumab, navicixizumab, naxitamab, nebacumab, necitumumab, nemolizumab, NEOD001, nerelimomab, nesbacumab, netakima , nimotuzumab, nirsevimab, nivolumab, nofetumomab merpentan, obilutoxaximab, obinutuzumab, occaratuzumab, ocrelizumab, ozlimomab, ofatumumab, olaratumab, oleculumab, orendalizumab, olokizumab, omalizumab, omburtamab, OMS721, onartuzumab, ontuxizumab, ombatilimab, opicinumab, oportuzumab monatox, oregovomab, olticumab, otelixizumab, otilimab, otlertuzumab, oxelumab, ozanezumab, pagibaximab, palivizumab,Pamrevlumab, panitumumab, pancomab, panobacumab, palsatuzumab, pascolizumab, pasotuximab, pateclizumab, patritumab, PDR001, pegfilgrastim-jmdb, pembrolizumab, pemtumomab, perakizumab, pertuzumab, pexelizumab, pidilizumab, pinatuzumab vedotin, pintumomab, placumab, prosalizumab, pogalizumab, polatuzumab vedotin, ponezumab, polgabiximab, prasinezumab, prezalizumab, priliximab, pritoxaximab, pritumumab, PRO140, Kirizumab, tetulomab, racotumomab, radletumab, rafivirumab, ralpancizumab, ramucirumab, ranevetomab, ranibizumab, raxibacumab, ravagalimab, ravtolimab, refanezumab, regavirumab, relatolimab, lemtolumab, reslizumab, rilonacept, rilotumumab, linukumab, risankizumab-rzaa, rituximab, rituximab-abbs, rituximab-pvvr, lobatumumab, Rmab, lorezumab, romilukimab, romiplostim, romosozumab, rontalizumab, rosmantuzumab, rovelizumab, Rozanolixizumab, ruplizumab, sacituzumab govitecan, samalizumab, sarilumab, satralizumab (SA237), satumomab pendetide, secukinumab, selicrelumab, seribantumab, cetoxaximab, setrusumab, sevirumab, sibrotuzumab, SGN-CD19A, SGN-CD33A, SHP647, sifalimumab, siltuximab, simtuzumab, siplizumab, sirukumab, sofituzumab vedotin, solanezumab, solitomab, sonepcizumab, sontuzumab, spartalizumab, stamulumab, suresomab, sputab Mab, stimulimab, subizumab, subratoxumab, tabalumab, tacatuzumab tetraxetan, tadocizumab, talaxuzumab, talizumab, tamtubetomab, tanezumab, taplitumomab paptokis, tarexuzumab, tabolimab, tefibazumab, terimomab alitox, tenatumomab, teneliximab, teplizumab, tepositamab, teprotumumab, tesidolumab, tetulomab (rilotomab), tezepelumab, TGN1412, tiburizumab, ticilimumab (tremelimumab), tildrakizumab, tigatuzumab, timigtuzumab, timolumab,Chiragotumab, tislelizumab, TNX-650, tocilizumab (atlizumab), tomzotuximab, toralizumab, tosatoxumab, tositumomab, tobetumab, tralokinumab, trastuzumab, trastuzumab-anns, trastuzumab-dkst, trastuzumab emtansine, TRBS07, tregalizumab, tremelimumab, trevoglumab, tucotuzumab-celmoleukin, tuvilumab, ublituximab, urocuplumab, urelumab, urtoxazumab, ustekinumab, utomilumab, banalimab, bundeltuzumab vedotin, vantixumab The monoclonal antibody may be rituximab, vanucizumab, bapaliximab, valisacumab, varlilumab, batelizumab, vedolizumab, veltuzumab, bepalimomab, besencumab, visilizumab, bovalilizumab, volociximab, bonlerolizumab, vopratelimab, borsetuzumab mafodotin, votumumab, xentuzumab, XMAB-5574, zalutumumab, zanolimumab, zatuximab, xenoctuzumab, diralimumab, zolbetuximab (IMAB362, claudiximab), Ziv-aflibercept, zolimomab alitox, or a corresponding anti-drug antibody in a sample from a human patient. In a preferred embodiment, the monoclonal antibody is rituximab, rituximab-abbs, or rituximab-pvvr.

[0065] In some embodiments, the monoclonal antibody is a biosimilar. In other embodiments, biosimilars include, but are not limited to, adalimumab-adbm, adalimumab-atto, adalimumab-bwwb, bevacizumab-awwb, epoetin alfa-epbx, etanercept-szzs, infliximab-abda, infliximab-dyyb, infliximab-qbtx, filgrastim-sndz, pegfilgrastim-jmdb, pegfilgrastim-bmez, risankizumab-rzaa, rituximab-abbs, rituximab-pvvr, trastuzumab-anns, or trastuzumab-dkst. In certain embodiments, the active biosimilar substance is adalimumab, bevacizumab, enoxaparin sodium, epoetin alfa, epoetin zeta, etanercept, filgrastim, follitropin alfa, infliximab, insulin glargine, insulin lispro, pegfilgrastim, risankizumab, rituximab, rituximab-abbs, rituximab-pvvr, somatropin, teriparatide, or trastuzumab. In preferred embodiments, the biosimilar is rituximab, rituximab-abbs, or rituximab-pvvr.

[0066] In other embodiments, the targeting moiety is an antibody from an intact polyclonal antibody, an intact monoclonal antibody, an antibody fragment, a single-chain Fv (scFv) variant, a multispecific antibody, a bispecific antibody, a chimeric antibody, a humanized antibody, a human antibody, a fusion protein containing an antigenic determinant portion of an antibody, or other modified immunoglobulin molecule containing an antigen recognition site.

[0067] In some embodiments, the therapeutic biological agent is an immunotherapy. In other embodiments, the immunotherapy is an anti-CD20 antibody. In certain embodiments, the anti-CD20 antibody is rituximab. In certain other embodiments, the therapeutic biological agent is an anti-CD20 antibody. As described herein, any antibody capable of binding the CD20 antigen may be used in the methods of the present disclosure. Antibodies that bind the CD20 antigen include, for example, C2B8 (rituximab; RITUXAN.RTM.) (U.S. Pat. No. 5,736,137, expressly incorporated herein by reference); yttrium-

[90] -labeled 2138 murine antibody designated Y2B8 (U.S. Pat. No. 5,736,137, expressly incorporated herein by reference); murine IgG2a131 (BEXXAR.RTM.) optionally labeled with 1311 to generate the 1311-B1 antibody (U.S. Pat. No. 5,595,721, expressly incorporated herein by reference); murine monoclonal antibody 1F5 (Press et al. Blood 69(2):584-591 (1987)); chimeric 2H7 antibody (U.S. Pat. No. 5,677,180, expressly incorporated herein by reference); and International Leukocyte Typing Monoclonal antibodies include monoclonal antibodies L27, G28-2, 93-1133, B--Cl, or NU--B2 available from the Workshop (Valentine et al., In: Leukocyte Typing III (McMichael, Ed., p. 440, Oxford University Press (1987)).

[0068] In a specific embodiment of the present disclosure, the anti-CD20 antibody is rituximab. Rituximab is a genetically engineered chimeric mouse / human monoclonal antibody. Rituximab is an IgG, kappa immunoglobulin containing mouse light and heavy chain variable region sequences and human constant region sequences. Rituximab has a binding affinity for the CD20 antigen of approximately 8.0 nM and is commercially available, for example, from Genentech (South San Francisco, Calif.).

[0069] In some embodiments, the therapeutic biological agent is an immunotherapeutic agent. In other embodiments, the immunotherapeutic agent is a PD-1 inhibitor, e.g., a PD-1 antibody, a PD-L1 inhibitor, e.g., a PD-L1 antibody, a CTLA-4 inhibitor, e.g., a CTLA-4 antibody, a CSF-1R inhibitor, an IDO inhibitor, an A1 adenosine inhibitor, an A2A adenosine inhibitor, an A2B adenosine inhibitor, an A3A adenosine inhibitor, an arginase inhibitor, or an HDAC inhibitor. In still other embodiments, the immunotherapeutic agent is a PD-1 inhibitor (e.g., nivolumab, pembrolizumab, pidilizumab, BMS936559, and MPDL328OA). In some embodiments, the immunotherapy is a PD-L1 inhibitor (e.g., atezolizumab and MEDI4736). In some embodiments, the immunotherapeutic agent is a CTLA-4 inhibitor (e.g., ipilimumab). In certain other embodiments, the immunotherapeutic agent is a CSF-1R inhibitor (e.g., pexidartinib and AZD6495). In certain embodiments, the immunotherapeutic agent is an IDO inhibitor (e.g., norharmane, rosmarinic acid, and alpha-methyl-tryptophan). In some embodiments, the immunotherapeutic agent is an A1 adenosine inhibitor (e.g., 8-cyclopentyl-1,3-dimethylxanthine, 8-cyclopentyl-1,3-dipropylxanthine, 8-phenyl-1,3-dipropylxanthine, bamifylline, BG-9719, BG-9928, FK-453, FK-838, rolofylline, or N-0861). In other embodiments, the immunotherapeutic agent is an A2A adenosine inhibitor (e.g., ATL-4444, istradefylline, MSX-3, preladenant, SCH-58261, SCH-412,348, SCH-442,416, ST-1535, VER-6623, VER-6947, VER-7835, viadenant, or ZM-241,385). In still other embodiments, the immunotherapeutic agent is an A2B adenosine inhibitor (e.g., ATL-801, CVT-6883, MRS-1706, MRS-1754, OSIP-339,391, PSB-603, PSB-0788, or PSB-1115).In certain other embodiments, the immunotherapeutic agent is an A3A adenosine inhibitor (e.g., KF-26777, MRS-545, MRS-1191, MRS-1220, MRS-1334, MRS-1523, MRS-3777, MRE-3005-F20, MRE-3008-F20, PSB-11, OT-7999, VUF-5574, and SSR161421). In certain embodiments, the immunotherapeutic agent is an arginase inhibitor (e.g., an arginase antibody, (2s)-(+)-amino-5-iodoacetamidopentanoic acid, NG-hydroxy-L-arginine, (2S)-(+)-amino-6-iodoacetamidohexanoic acid, or (R)-2-amino-6-borono-2-(2-(piperidin-1-yl)ethyl)hexanoic acid. In some embodiments, the immunotherapeutic agent is an HDAC inhibitor (e.g., valproic acid, SAHA, or romidepsin). In other embodiments, the immunotherapeutic agent is a Toll-like receptor activator. In yet other embodiments, the immunotherapy is an RIG-I-like receptor activator. In certain other embodiments, the immunotherapeutic agent is a stimulator of interferon genes (STING) pathway activator. In certain embodiments, the immunotherapeutic agent is an interleukin-1 receptor agonist, e.g., an IL-R1 antagonist. In some embodiments, the immunotherapeutic agent is a PTEN inhibitor, e.g., a bisperoxovanadium compound. In other embodiments, the immunotherapeutic agent is a tumor necrosis factor receptor (TNFR), e.g., a TNFR-1 or TNFR-2 inhibitor. In certain embodiments, the immunotherapeutic agent is a lymphocyte-activation gene 3 (LAG-3) inhibitor, e.g., GSK2831781.

[0070] In other embodiments, the therapeutic biologic agent is ledipasvir / sofosbuvir, insulin glargine, lenalidomide, pneumococcal 13-valent conjugate vaccine, fluticasone / salmeterol, elvitegravir / cobicistat / emtricitabine / tenofovir alafenamide, emtricitabine, rilpivirine and tenofovir alafenamide, emtricitabine / tenofovir alafenamide, grazoprevir / elbasvir, coagulation factor VIIa recombinant, epoetin alfa, aflibercept, or etanercept.

[0071] In some embodiments, the therapeutic or diagnostic agent is abatacept, abobotulinumtoxinA, agalsidase beta, albiglutide, aldesleukin, alglucosidase alfa, alteplase (cathflo activase), anakinra, asfotase alfa, asparaginase, asparaginase erwinia chrysanthemi, becaplermin, belatacept, collagenase, collagenase clostridium histolyticum histolyticum), darbepoetin alfa, denileukin diftitox, dornase alfa, dulaglutide, ecallantide, elosulfase alfa, etanercept-szzs, filgrastim, filgrastim-sndz, galsulfase, glucarpidase, idursulfase, incobotulinum toxin A, interferon alfa-2b, interferon alfa-n3, interferon beta-1a, interferon beta-1b, interferon gamma-1b, laronidase, methoxypolyethylene glycol-epoetin beta, methicone The active ingredient is replenisher, ocriplasmin, onabotulinumtoxinA, oprelvekin, palifermin, parathyroid hormone, pegaspargase, pegfilgrastim, peginterferon alfa-2a, peginterferon alfa-2a packaged with ribavirin, peginterferon alfa-2b, peginterferon beta-1a, pegloticase, rasburicase, reteplase, rilonacept, rimabotulinumtoxinB, romiplostim, sargramostim, sebelipase alfa, Tbo-filgrastim, tenecteplase, or Ziv-aflibercept.

[0072] In other embodiments, the diagnostic agent is tuberculin purified protein derivative, thyrotropin alpha, secretin, soluble transferrin receptor, troponin, B-type natriuretic peptide, iobenguane I123, florbetapir F18, perflutren, gadoterate meglumine, florbetaben F18, flutemetamol F18, gadoterate meglumine, isosulfan blue, regadenoson, technetium Tc99m tilmanocept, florbetaben F18, perflutren, regadenoson, or flutemetamol F18.

[0073] The therapeutic or diagnostic agent in the particles can have a unit activity of about 0.5 to about 1.0, about 0.75 to about 1.0, or about 0.9 to about 1.0. The activity is measured relative to the same therapeutic or diagnostic agent prior to particle formation. In certain embodiments, the therapeutic agent has a unit activity of about 0.5 to about 1.0. In preferred embodiments, the therapeutic biological agent has a unit activity of about 0.5 to about 1.0. The term "activity" refers to the ratio of a functional or structural aspect of an agent, e.g., a therapeutic or diagnostic agent, at two points in time. The denominator of the ratio corresponds to a measure of the functional or structural aspect of the agent in the feed solution immediately prior to droplet formation. The numerator of the ratio corresponds to the same measure of the functional or structural aspect of the agent at a later point in time, e.g., immediately after particle formation.

[0074] In certain embodiments, the particles comprise a loading of about 1 to about 100% by weight, e.g., about 50 to about 100% by weight, about 75 to about 100% by weight, about 90 to about 100% by weight, about 95 to about 100% by weight, about 99 to about 100% by weight, or about 99.9 to about 100% by weight of a therapeutic or diagnostic agent. With these loadings, the therapeutic or diagnostic agent retains about 0.5 to about 1.0 activity, e.g., about 0.75 to about 1.0 activity, about 0.9 to about 1.0 activity, about 0.95 to about 1.0 activity, about 0.99 to about 1.0 activity, or about 0.999 to about 1.0 activity during particle formation. This includes the activity retained through primary dehydration (i.e., dehydration using a second liquid) and, optionally, secondary dehydration.

[0075] In some embodiments, the particles have less than about 25% internal void space, such as less than about 24%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1%. In certain embodiments, the particles may contain less than 10% internal void space, less than 5%, less than 1%, less than 0.1%, or less than 0.01% internal void space. In preferred embodiments, the particles are substantially free of any internal void space. A suitable method for determining internal void space can be achieved by using focused ion beam scanning electron microscopy (FIB-SEM). For example, the internal void space can be calculated using the following formula: internal void space = A v / A p , where A v is the total area of ​​void space, and A p is the total area of ​​the particle.

[0076] In other embodiments, the particles may exhibit a porosity of about 0 to about 50%, e.g., about 0 to about 10%, about 0 to about 5%, about 0 to about 1%, about 0 to about 0.5%, about 0 to about 0.1%, or about 0 to about 0.01%. Exemplary pore size measurements include scanning electron microscopy (SEM), transmission electron microscopy (TEM), and confocal laser scanning microscopy analysis. The specific surface area of ​​porous microspheres and nanospheres can also be investigated by nitrogen adsorption / desorption analysis and the Brunauer-Emmett-Teller adsorption model. In certain embodiments where the pore size is sufficiently large, mercury intrusion porosimetry can be used.

[0077] Particles according to the present disclosure are circular. Circularity can serve as an indicator of particle shape. The particles described herein can have a characteristic circularity, e.g., a relative shape that is substantially circular. This characteristic describes and defines the particle's morphology based on its circularity. When a particle has a perfectly circular structure, the circularity is 1.0. Particles as described herein can have a circularity of about 0.8, 0.9, 0.95, 0.96, 0.97, 0.98, or 0.99; greater than about 0.80, greater than about 0.90, or greater than about 0.95. In some embodiments, the circularity of the particles is greater than about 0.88. In other embodiments, the circularity of the particles is greater than about 0.90. In certain embodiments, the circularity of the particles is greater than about 0.93. In preferred embodiments, the circularity of the particles is greater than about 0.97. The diameter and circularity of the particles can be determined by image processing of images observed under an electron microscope, for example, or by a flow particle image analyzer. The circularity can also be determined by subjecting particles to circularity measurement and averaging the values ​​thus obtained. For example, the circularity (circ) can be calculated using the following formula:

number

[0078] In other embodiments, the circularity of the particles is about 0.10 to about 1.00, e.g., about 0.20, 0.30, 0.40, 0.50, 0.60, 0.70, 0.75, 0.80, 0.81, 0.82, 0.83, 0.84, 0.85, 0.86, 0.87, 0.88, 0.89, 0.90, 0.91, 0.92, 0.93, 0.94, 0.95, 0.96, 0.97, 0.98, or 0.99 to about 1.00. In certain embodiments, the circularity of the particles is about 0.88 to about 1.00. In still other embodiments, the circularity of the particles is about 0.90 to about 1.00. In certain other embodiments, the circularity of the particles is about 0.93 to about 1.00. In preferred embodiments, the particle circularity is about 0.97 to about 1.00. In some embodiments, a method for measuring particle circularity involves image analysis of a scanning electron micrograph of the particle, where the average roundness is calculated based on the cross-sectional shape of the particle projected onto the image plane. Such a roundness factor can be expanded to identify the corresponding circularity.

[0079] The particles according to the present disclosure are spherical. The sphericity coefficient of a particle is the ratio of the particle's smallest diameter to its largest diameter. For a perfect sphere, this ratio is 1. The sphericity coefficient can be calculated by measuring the aspect ratio using any software adapted to handle images, such as images obtained by microscopy, particularly scanning electron microscopy (SEM) or transmission electron microscopy (TEM). In some embodiments, the sphericity of the particle is at least about 50%, for example, at least about 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or about 100%. In other embodiments, the sphericity of the particles is from about 0.10 to about 1.00, e.g., from about 0.20, 0.30, 0.40, 0.50, 0.60, 0.70, 0.75, 0.80, 0.81, 0.82, 0.83, 0.84, 0.85, 0.86, 0.87, 0.88, 0.89, 0.90, 0.91, 0.92, 0.93, 0.94, 0.95, 0.96, 0.97, 0.98, or 0.99 to about 1.00. In a preferred embodiment, the sphericity of the particles is about 1.00.

[0080] In certain embodiments, the sphericity of a particle can range from about 0.10 to about 1.00, such as at least about 0.20, about 0.40, about 0.60, or about 0.80 to about 1.00. In some embodiments, a method for measuring the sphericity of a particle involves image analysis of a scanning electron micrograph of the particle, where the average roundness is calculated based on the cross-sectional shape of the particle projected onto the image plane. Sphericity (ψ) is a measure of the roundness of an object. Sphericity is the ratio of the surface area of ​​a sphere (having the same volume as the particle being compared) to the surface of the particle being tested. Sphericity can be calculated by the following formula:

number

[0081] In other embodiments, the sphericity (minor axis / major axis) can be determined by using an image analysis instrument or an electron micrograph taken with a scanning electron microscope (SEM). For example, the average sphericity can be calculated as the average of the sphericity values ​​calculated for randomly selected particles in an electron micrograph by determining their minor and major axes based on visual observation.

[0082] In some embodiments of the present disclosure, the drying operation can be controlled to provide particles with particular characteristics, such as particles with a substantially smooth surface. "Surface roughness," as used herein, refers to a particle having numerous wrinkles or creases, e.g., ribbed or wrinkled. The term "pit," as used herein, refers to an indentation or crevice in a particle, which is an indentation or crevice in a two-dimensional image or an indentation or crevice in an object. The term "spike," as used herein, refers to a protrusion directed outward from the center of mass of a particle, where the protrusion is directed outward from the center of mass of a two-dimensional image, or a sharp protrusion is directed outward from an object.

[0083] In preferred embodiments of the present disclosure, the particles as described herein have a surface morphology that is smooth rather than ridged or wrinkled. The surface roughness of the particles can be reduced by controlling the formulation and / or process for forming the particles as described herein. In certain embodiments, drying conditions can be selected to control the particle morphology to promote the smoothness of the particle's surface. In particular, drying conditions can be selected to provide particles with a substantially smooth surface. In certain preferred embodiments, the particles have a substantially smooth surface. Those skilled in the art of the present disclosure can easily evaluate the surface morphology of the disclosed particles using routine and standard techniques.

[0084] In other embodiments, the particles have a diameter of about 0.1 to about 1000 μm, e.g., about 0.1 to about 900, 800, 700, 600, 500, 400, 300, 200, 100, 90, 80, 70, 60, 50, 45, 40, 35, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3 or about 0.2 μm. In certain embodiments, the particles have a diameter of about 1 to about 100 μm, e.g., about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, or 50 to about 100 μm. In still other embodiments, the particles have a diameter of about 4 to about 100 μm. In certain other embodiments, the particles have a diameter of about 10 to about 100 μm. In preferred embodiments, the particles have a diameter of about 20 to about 50 μm. In certain preferred embodiments, the diameter of the particles is intentionally controlled. In some embodiments, the particles have a diameter of about 0.1 to about 1000 μm, e.g., about 1 to about 400 μm, about 1 to about 200 μm, about 1 to about 100 μm, about 1 to about 50 μm, about 1 to about 25 μm, about 1 to about 10 μm, about 10 to about 100 μm, about 50 to about 100 μm, about 50 to about 75 μm, or about 75 to about 100 μm. In other embodiments, the particles have a diameter of about 1 to about 100 μm, e.g., about 4 to about 100 μm, about 10 to about 100 μm, or about 20 to about 50 μm.

[0085] In certain embodiments, the particles have a diameter of about 0.1 to about 100 μm. In certain other embodiments, the particles have a diameter of about 0.5 to about 50 μm. In still other embodiments, the particles have a diameter of about 20 to about 50 μm. In certain preferred embodiments, the particles have a diameter of about 1 to about 40 μm. In preferred embodiments, the particles have a diameter of about 2 to about 15 μm.

[0086] In some embodiments, the particles are less than about 10% by weight, e.g., less than about 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.9%, less than 0.8%, less than 0.7%, less than 0.6%, less than 0.5%, less than 0.4%, less than 0.3%, less than 0.2%, less than 0.1%, less than 0.09%, less than 0.08%, The surfactant content may be less than 0.07%, less than 0.06%, less than 0.05%, less than 0.04%, less than 0.03%, less than 0.02%, less than 0.01%, less than 0.009%, less than 0.008%, less than 0.007%, less than 0.006%, less than 0.005%, less than 0.004%, less than 0.003%, less than 0.002%, or less than 0.001% by weight. In other embodiments, the particles have a surfactant content of less than about 5% by weight. In certain embodiments, the particles have a surfactant content of less than about 3% by weight. In still other embodiments, the particles have a surfactant content of less than about 0.1% by weight. In certain other embodiments, the particles have a surfactant content of less than about 0.01% by weight. In some embodiments, the particles have a surfactant content of less than about 0.001% by weight. In preferred embodiments, the particles have a surfactant content of less than about 1% by weight. In certain preferred embodiments, the particles are substantially free of any surfactant content.

[0087] In other embodiments, the surfactant content of the particles is 0-10 wt%, e.g., 0-5 wt%, 0-3 wt%, 0-2 wt%, 0-1 wt%, 0-0.5 wt%, 0-0.2 wt%, 0-0.1 wt%, 0-0.01 wt%, or 0-0.001 wt%. An exemplary method for measuring surfactant content involves reconstitution of the particles in a suitable medium, such as deionized water, and subsequent analysis of the reconstituted solution by liquid chromatography. Chromatographic techniques may include the use of a charged aerosol detector (CAD) or an evaporative light scattering detector (ELSD).

[0088] In some embodiments, the surfactant is polysorbate, magnesium stearate, sodium dodecyl sulfate, TRITON™ N-101, glycerin, polyoxyethylated castor oil, docusate, sodium stearate, decyl glucoside, nonoxynol-9, cetyltrimethylammonium bromide, sodium bis(2-ethylhexyl) sulfosuccinate, sodium laureth sulfate, lecithin, or a combination thereof. In some embodiments, surfactants include, but are not limited to, (i) cationic surfactants, such as cetyltrimethylammonium chloride, hexadecyltrimethylammonium chloride, benzalkonium chloride, benzethonium chloride, dioctadecyldimethylammonium bromide; (ii) anionic surfactants, such as magnesium stearate, sodium dodecyl sulfate, dioctyl sodium sulfosuccinate, sodium myreth sulfate, perfluorooctane sulfonate, alkyl ether phosphates; (iii) nonionic surfactants, such as alkylphenol ethoxylates (Triton X-100), fatty alcohol ethoxylates (octaethylene glycol monododecyl ether, cocamide diethanolamine, poloxamer, glycerol monostearate, fatty acid esters of sorbitol (sorbitan monolaurate, Tween 80), etc. , Tween 20; and (iv) zwitterionic surfactants, such as cocamidopropyl hydroxysultaine and 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS). In other embodiments, the surfactant is polysorbate, magnesium stearate, sodium dodecyl sulfate, TRITON™ N-101, glycerin, polyoxyethylated castor oil, docusate, sodium stearate, decyl glucoside, nonoxynol-9, cetyltrimethylammonium bromide, sodium bis(2-ethylhexyl)sulfosuccinate, lecithin, sorbitan ester, or a combination thereof. In certain embodiments, the surfactant is polysorbate, docusate, or lecithin. In preferred embodiments, the surfactant is polysorbate 20, polysorbate 60, or polysorbate 80.In certain preferred embodiments, the surfactant is polysorbate 20 or polysorbate 80. In certain other embodiments, the fatty acid ester of sorbitol is a sorbitan ester, such as span 20, span 40, span 60, or span 80. In still other embodiments, the surfactant is an ionic surfactant.

[0089] In other embodiments, the particles have a density of about 1.00 to about 6.00 g / cm 3 , for example, about 1.00 to about 5.00 g / cm 3 , about 1.00~3.00g / cm 3 , about 1.00~2.00g / cm 3 , about 1.00~1.50g / cm 3 , about 1.30~1.50g / cm 3 , about 1.32~about 1.50g / cm 3 or about 1.10 to about 1.40 g / cm 3 In some embodiments, the particles exhibit a skeletal density of about 0.10 to about 5.00 g / cm 3 , for example, about 0.10 to about 2.50 g / cm 3 , about 0.10~about 1.40g / cm 3 , about 0.50~1.40g / cm 3 or about 1.00 to about 1.40 g / cm 3 In certain embodiments, the particles exhibit a skeletal density of from about 0.09 to about 1.60 g / cm 3 In yet another embodiment, the particles have a skeletal density of about 1.30 to about 1.58 g / cm 3 In a preferred embodiment, the particles have a skeletal density of about 1.32 to about 1.50 g / cm 3 An exemplary method for measuring skeletal density includes gas displacement pycnometry.

[0090] In certain embodiments, the particles have a concentration of about 1000 mg / mL to about 1500 mg / mL, about 1050 mg / mL to about 1500 mg / mL, about 1100 mg / mL to about 1500 mg / mL, about 1150 mg / mL to about 1500 mg / mL, about 1200 mg / mL to about 1500 mg / mL, about 1250 mg / mL to about 1500 mg / mL, about 1300 mg / mL to about 1500 mg / mL, about 1310 mg / mL to about 1500 mg / mL, about 1320 mg / mL to about 1500 mg / mL, about 1330 mg / mL to about 1500 mg / mL, about 1340 mg / mL to about 1500 mg / mL, about 1350 mg / mL to about 1500 mg / mL, or about 1360 mg / mL to about 1500 mg / mL L, about 1370mg / mL to about 1500mg / mL, about 1380mg / mL to about 1500mg / mL, about 1390mg / mL to about 1500mg / mL, about 1400mg / mL to about 1500mg / mL, about 1410mg / mL to about 1500mg / mL, about 1420mg / mL to about 1500mg / mL, about 1430mg / mL to about 1500mg / mL, about 1440mg / mL to about 1500mg / mL, about 1450mg / mL to about 1500mg / mL, about 1460mg / mL to about 1500mg / mL, about 1470mg / mL to about 1500mg / mL, about 1480mg / mL to about 1500mg / mL, or about 1490mg / mL to about 1500mg / mL.

[0091] In some embodiments, the particles can be characterized by a glass transition temperature of about 0°C to about 250°C, e.g., about 34°C to about 200°C, about 50°C to about 200°C, about 60°C to about 200°C, about 40°C to about 160°C, about 50°C to about 110°C, about 60°C to about 100°C, or about 75°C to about 80°C. The term "glass transition," as used herein, refers to a thermodynamic transition of an amorphous material characterized by a step change in specific heat capacity and elasticity. At temperatures above the glass transition temperature, molecular mobility increases, as do the rates of physical and chemical changes. Exemplary analytical methods for determining the glass transition temperature include differential scanning calorimetry and dynamic kinetic analysis. In other embodiments, the particles have a glass transition temperature of about 40°C to about 160°C. In yet other embodiments, the particles have a glass transition temperature of about 50°C to about 110°C. In certain embodiments, the particles have a glass transition temperature of about 60°C to about 100°C. In a preferred embodiment, the particles have a glass transition temperature of about 75 to about 80°C.

[0092] In certain embodiments, the particles have a glass transition temperature greater than about 160° C. In certain other embodiments, the particles have a glass transition temperature greater than about 90° C. In certain preferred embodiments, the particles have a glass transition temperature greater than about 50° C.

[0093] In other embodiments, the particles are heated to about ±30° C., such as about ±20, ±10, ±5, ±1° C., of the glass transition temperature of the particles during drying.

[0094] In some embodiments, the particles further comprise a carbohydrate, a pH adjuster, a salt, a chelating agent, a mineral, a polymer, a surfactant, a protein stabilizer, an emulsifier, a preservative, an amino acid, an antioxidant, a protein, an organic solvent, a paraben, a bactericide, a fungicide, a vitamin, a preservative, a nutrient medium, an oligopeptide, a biological additive, a chemical additive, or a combination thereof. In certain embodiments, the particles further comprise a carbohydrate, a pH adjuster, a salt, a surfactant, a protein stabilizer, an emulsifier, an amino acid, or a combination thereof.

[0095] In other embodiments, the carbohydrate can be from the monosaccharide, disaccharide, oligosaccharide, or polysaccharide family. In some embodiments, the carbohydrate is dextran, trehalose, sucrose, agarose, mannitol, lactose, sorbitol, maltose, starch, alginate, xanthan, galactomannan, agar, agarose, or a combination thereof. In certain embodiments, the carbohydrate is dextran, trehalose, sucrose, agarose, mannitol, lactose, sorbitol, maltose, hydroxypropyl beta-cyclodextrin, or a combination thereof. In preferred embodiments, the carbohydrate is trehalose, cyclodextrin, hydroxypropyl beta-cyclodextrin, or a combination thereof. Cyclodextrins are available in three different forms, α, β, and γ, based on the number of glucose monomers. The number of glucose monomers in α, β, and γ cyclodextrin can be 6, 7, or 8, respectively.

[0096] In some embodiments, the pH adjuster is acetate, citrate, glutamate, glycinate, histidine, lactate, maleate, phosphate, succinate, tartrate, bicarbonate, aluminum hydroxide, phosphoric acid, hydrochloric acid, DL-lactic acid / glycolic acid, phosphorylethanolamine, tromethamine, imidazole, glycylglycine, monosodium glutamate, sodium hydroxide, potassium hydroxide, or a combination thereof. In other embodiments, the pH adjuster is citrate, histidine, phosphate, succinate, sodium hydroxide, potassium hydroxide, or a combination thereof. In certain embodiments, the pH adjuster is hydrochloric acid or citric acid.

[0097] In other embodiments, the salt is sodium chloride, calcium chloride, potassium chloride, sodium hydroxide, stannous chloride, magnesium sulfate, sodium glucoheptonate, sodium pertechnetate, guanidine hydrochloride, potassium hydroxide, or a combination thereof. In a preferred embodiment, the salt is sodium chloride.

[0098] In some embodiments, the chelating agent is edetate disodium, ethylenediaminetetraacetic acid, pentetic acid, or a combination thereof. In other embodiments, the mineral is calcium, zinc, titanium dioxide, or a combination thereof. In certain embodiments, the polymer is propylene glycol, glucose star polymer, silicone polymer, polydimethylsiloxane, polyethylene glycol, carboxymethylcellulose, poly(glycolic acid), poly(lactic-co-glycolic acid), polylactic acid, polycaprolactone (PCL), polyvinylpyrrolidone (PVP), ficoll, dextran, or a combination thereof.

[0099] In other embodiments, the surfactant is polysorbate, magnesium stearate, sodium dodecyl sulfate, TRITON™ N-101, glycerin, polyoxyethylated castor oil, docusate, sodium stearate, decyl glucoside, nonoxynol-9, cetyltrimethylammonium bromide, sodium bis(2-ethylhexyl) sulfosuccinate, sodium laureth sulfate, lecithin, or a combination thereof. In some embodiments, surfactants include, but are not limited to, (i) cationic surfactants, such as cetyltrimethylammonium chloride, hexadecyltrimethylammonium chloride, benzalkonium chloride, benzethonium chloride, dioctadecyldimethylammonium bromide; (ii) anionic surfactants, such as magnesium stearate, sodium dodecyl sulfate, dioctyl sodium sulfosuccinate, sodium myreth sulfate, perfluorooctane sulfonate, alkyl ether phosphates; (iii) nonionic surfactants, such as alkylphenol ethoxylates (Triton X-100), fatty alcohol ethoxylates (octaethylene glycol monododecyl ether, cocamide diethanolamine, poloxamer, glycerol monostearate), glycerol monostearate, glycerol monoisopropyl ether ... and (iv) zwitterionic surfactants, such as cocamidopropyl hydroxysultaine and 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS). In certain embodiments, the surfactant is polysorbate, docusate, or lecithin. In preferred embodiments, the surfactant is polysorbate 20, polysorbate 60, or polysorbate 80. In certain preferred embodiments, the surfactant is polysorbate 20 or polysorbate 80. In certain other embodiments, the fatty acid ester of sorbitol is a sorbitan ester, such as span 20, span 40, span 60, or span 80.

[0100] In some embodiments, the protein stabilizer is acetyltryptophanate, caprylate, N-acetyltryptophan, trehalose, PEG200, PEG300, PEG3350, PEG8000, PEG10000, PEG20000, poloxamer, polyvinylpyrrolidone, polyacrylic acid, poly(vinyl) polymer, polyester, polyaldehyde, tertiary polymer, polyamino acid, hydroxyethyl starch, N-methyl-2-pyrrolidone, sorbitol, sucrose, mannitol, or a combination thereof. In certain embodiments, the protein stabilizer is trehalose, PEG200, PEG300, PEG3350, PEG8000, PEG10000, PEG20000, poloxamer, polyvinylpyrrolidone, polyacrylic acid, poly(vinyl) polymer, polyester, polyaldehyde, tertiary polymer, polyamino acid, hydroxyethyl starch, N-methyl-2-pyrrolidone, sorbitol, sucrose, mannitol, cyclodextrin, sugar, hydroxypropyl beta-cyclodextrin, or a combination thereof. In preferred embodiments, the protein stabilizer is trehalose, cyclodextrin, hydroxypropyl beta-cyclodextrin, or a combination thereof. Stabilizers, used synonymously with the term "stabilizer," can be salts, carbohydrates, sugars, or amino acids, preferably carbohydrates or sugars, recognized by authorities as suitable additives or additives in pharmaceutical compositions, as described herein. The term "additive" refers to an additive to a preparation or formulation that may be useful in achieving desired modifications to the characteristics of the preparation or formulation. Such modifications include, but are not limited to, physical stability, chemical stability, and therapeutic efficacy. Exemplary additives include, but are not limited to, carbohydrates, pH adjusters, salts, chelating agents, minerals, polymers, surfactants, amino acids, oligopeptides, biological additives, chemical additives, preservatives, antioxidants, parabens, bactericides, fungicides, vitamins, preservatives, analgesics, and / or nutrient media.

[0101] Examples of emulsifiers suitable for use in the particles include, but are not limited to, lipophilic drugs having an HLB of less than 7, such as mixed fatty acid monoglycerides; mixed fatty acid diglycerides; mixtures of fatty acid mono- and diglycerides; lipophilic polyglycerol esters; glycerol esters including glyceryl monooleate, glyceryl dioleate, glycerol monostearate, glyceryl distearate, glyceryl monopalmitate, and glyceryl dipalmitate; glyceryl-lactoesters of fatty acids; propylene glycol esters including propylene glycol monopalmitate, propylene glycol monostearate, and propylene glycol monooleate; sorbitan monostearate, sorbitan sesquioleate, and the like. sorbitan esters; fatty acids including stearic acid, palmitic acid, and oleic acid and soaps thereof; and mixtures thereof such as glyceryl monooleate, glyceryl dioleate, glycerin monostearate, glyceryl distearate, glyceryl monopalmitate, and glyceryl dipalmitate; glyceryl-lactoesters of fatty acids; propylene glycol esters including propylene glycol monopalmitate, propylene glycol monostearate, and propylene glycol monooleate; sorbitan esters including sorbitan monostearate and sorbitan sesquioleate; fatty acids including stearic acid, palmitic acid, and oleic acid and soaps thereof; or combinations thereof. In some embodiments, the emulsifier is polysorbate 80, polysorbate 60, polysorbate 20, sorbitan monooleate, ethanolamine, polyoxyl 35 castor oil, polyoxyl 40 hydrogenated castor oil, carbomer 1342, corn oil-mono-di-triglyceride, polyoxyethylated oleic acid glyceride, poloxamer, or a combination thereof. In preferred embodiments, the fatty acid ester of sorbitol is a sorbitan ester, such as span 20, span 40, span 60, or span 80. In certain preferred embodiments, the emulsifier is polysorbate 80, sorbitan monooleate, or a combination thereof.

[0102] In other embodiments, the preservative is phenol, m-cresol, benzyl alcohol, 2-phenyloxyethanol, chlorobutanol, neomycin, benzethonium chloride, glutaraldehyde, beta-propiolactone, or a combination thereof.

[0103] In certain embodiments, the amino acid is alanine, aspartic acid, cysteine, isoleucine, glutamic acid, leucine, methionine, phenylalanine, pyrrolidine, serine, selenocysteine, threonine, tryptophan, tyrosine, valine, asparagine, L-arginine, histidine, glycine, glutamine, proline, or various salts thereof (such as arginine hydrochloride, arginine glutamate), or combinations thereof. In preferred embodiments, the amino acid is L-arginine, histidine, proline, or combinations thereof.

[0104] In some embodiments, the antioxidant is glutathione, ascorbic acid, cysteine, N-acetyl-L-tryptophanate, tocopherol, histidine, methionine, or a combination thereof. In other embodiments, the protein is protamine, protamine sulfate, gelatin, or a combination thereof. In certain embodiments, the organic solvent is dimethyl sulfoxide, N-methyl-2-pyrrolidone, or a combination thereof. In still other embodiments, the preservative is methyl hydroxybenzoate, thimerosal, paraben, formaldehyde, castor oil, or a combination thereof. The paraben may be parahydroxybenzoate. In some embodiments, the disinfectant is benzalkonium chloride (a cationic surfactant), hypochlorite, peroxide, alcohol, a phenolic compound (e.g., carbolic acid), benzyl benzoate, or a combination thereof. In a preferred embodiment, the disinfectant is benzyl benzoate.

[0105] In other embodiments, the fungicide is acibenzolar, 2-phenylphenol, allilazine, carvone, natamycin, potassium azide, or a combination thereof. In a preferred embodiment, the fungicide is benzyl benzoate. In certain embodiments, the vitamin is thiamine, riboflavin, niacin, pantothenic acid, biotin, vitamin B6, vitamin B12, folic acid, niacin, ascorbic acid, calciferol, retinol, quinone, or a combination thereof. In yet other embodiments, the preservative is sodium nitrate, sulfur dioxide, potassium sorbate, sodium sorbate, sodium benzoate, benzoic acid, methyl hydroxybenzoate, thimerosal, parabens, formaldehyde, castor oil, or a combination thereof. In a preferred embodiment, the preservative is methyl hydroxybenzoate, thimerosal, parabens, formaldehyde, castor oil, or a combination thereof.

[0106] Several nutrient media, preferably serum-free, alone or in combination, may be used in the present disclosure, including commercially available media or other media known in the art. Examples of such media (all serum-free or serum-depleted) include ADC-1, LPM (without bovine serum albumin), F10 (HAM), F12 (HAM), DCCM1, DCCM2, RPMI 1640, BGJ medium (Fitton-Jackson modified), Basal Medium Eagle (BME supplemented with Earl's salt base), Dulbecco's Modified Eagle Medium (DMEM, serum-free), Glasgow's Modified Eagle Medium (GMEM), Leibovitz's L-15 medium, McCoy's 5A medium, Medium M199 (M199E - with Earl's salt base), Medium M199 (M199H - with Hank's salt base), Minimum Essential Medium Eagle (MEM-E - with Earl's salt base), Minimum Essential Medium Eagle (MEM-H - with Hank's salt base), and Minimum Essential Medium Eagle (MEM-NAA - with non-essential amino acids), among many others. Additionally, although serum-containing nutrient media may be used in compositions according to the present disclosure, the use of serum-containing media is less preferred due to the possibility that serum may be contaminated with microbial agents and because patients may develop immune responses to certain antigenic components contained in the serum.

[0107] In some embodiments, the oligopeptide is trileucine. In other embodiments, the biological additive is a nucleic acid, an oligonucleotide, an antibody or fragment thereof, an amino acid, a polyamino acid, a peptide, a protein, a cell, a bacterium, a gene therapy agent, a genome engineering therapy agent, an epigenome engineering therapy agent, a hormone, a nucleoprotein, a glycoprotein, a lipoprotein, an exosome, an outer membrane vesicle, a vaccine, a virus, a bacteriophage, an organelle, a nutrient medium, or a combination thereof. In certain embodiments, the chemical additive is a chemical drug, a contrast agent, a dye, a magnetic particle, a polymer bead, a metal nanoparticle, a metal microparticle, a quantum dot, an antioxidant, an antibiotic agent, a steroid, an analgesic, a local anesthetic, an anti-inflammatory agent, a paraben, an antimicrobial agent, a chemotherapeutic agent, a vitamin, a mineral, a disinfectant, a preservative, or a combination thereof.

[0108] In other embodiments, the particles have less than 20% aggregates or less than 20% fragmentation of the therapeutic biological agent, e.g., less than about 19%, less than 18%, less than 17%, less than 16%, less than 15%, less than 14%, less than 13%, less than 12%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1.9%, less than 1.8%, less than 1.7%, less than 1.6%, less than 1.5%, less than 1.4%, less than 1.3%, less than 1.2%, less than 1.1%, less than 1%, less than 0.9%, less than 0.8%, less than 0.7%, less than 0.6%, less than 0.5%, less than 0.4%, less than 0.3%, less than 0.2%, or less than 0.1%. In some embodiments, the particles have less than 10% aggregates or less than 10% fragments of the therapeutic biological agent, e.g., less than about 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1.9%, 1.8%, 1.7%, 1.6%, 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%, or 0.1%. In certain embodiments, the particles have between about 3% and about 1% aggregates of the therapeutic biological agent. In certain other embodiments, the particles have between about 1% and about 0.5% aggregates of the therapeutic biological agent. In preferred embodiments, the particles are substantially free of any aggregates of the therapeutic biological agent. In yet other embodiments, the particles have less than about 1% fragmentation of the therapeutic biological agent. In certain preferred embodiments, the particles are substantially free of any fragmentation of the therapeutic biological agent. A suitable method for measuring aggregates and fragmentation of biological agents can be achieved by using size exclusion chromatography (SEC).

[0109] In some embodiments, the particle formation process results in less than a 50% change in charge variants in a population of diagnostic or therapeutic agents, e.g., antibodies or antibody fragments, compared to the therapeutic or diagnostic agents prior to particle formation (e.g., less than 40%, 30%, 20%, 10%, 8%, 5%, 4%, 3%, or 1%). Charge variants can be acidic, basic, or neutral, and variations can be caused by post-translational modifications at terminal amino acids, such as asparagine deamidation or lysine glycosylation. For example, charge variants include loss of a positive charge due to loss of a C-terminal lysine residue, covalent attachment of the amine moieties of two lysine residues with a reducing sugar, or conversion of an N-terminal amine to a neutral amide via cyclization of an N-terminal glutamine. Negative charges on proteins, e.g., antibodies, can appear due to conversion of an asparagine residue to an aspartic acid and / or isoaspartic acid residue via a deamidation reaction. An exemplary method for measuring charge variants includes cation exchange chromatography (CIEX), in which variants are quantified by dividing the area under the peak corresponding to a variant, e.g., an acidic, basic, or neutral population, by the cumulative area contained under all peaks in the sample spectrum. The change in percentage of a charge variant population between two samples, e.g., sample A and sample B, is calculated as the numerical difference in the percentage of each population variant, i.e., by subtracting the percentage of a particular variant, e.g., acidic in sample B, from the percentage of a particular variant, e.g., acidic in sample A, or vice versa. In certain embodiments, the analysis may be extended to all variants in the population in a similar manner.

[0110] In certain embodiments, the particles have less than about 50%, e.g., less than about 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% change in charge variants of the therapeutic biological agent compared to the starting biological agent prior to particle formation. In preferred embodiments, the particles are substantially free of any change in charge variants of the therapeutic biological agent compared to the starting biological agent prior to particle formation. A suitable method for measuring changes in charge variants of a biological agent can be achieved by using cation exchange chromatography (CIEX).

[0111] In other embodiments, the residual moisture or solvent content of the dry ingredients is less than about 7% by weight, e.g., less than about 6%, 5%, 4%, 3%, 2%, 1.9%, 1.8%, 1.7%, 1.6%, 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%, or 0.1% by weight. In some embodiments, the particles have less than about 7% by weight residual moisture. In still other embodiments, the particles have less than about 5% by weight residual moisture. In certain embodiments, the particles have less than about 3% by weight residual moisture. In preferred embodiments, the particles have less than about 1% by weight residual moisture.

[0112] In some embodiments, the particles have a residual moisture content of about 1% to about 7% by weight. In still other embodiments, the particles have a residual moisture content of about 1% to about 5% by weight. In certain embodiments, the particles have a residual moisture content of about 1% to about 3% by weight. In preferred embodiments, the particles are substantially free of any residual moisture by weight.

[0113] Exemplary methods for measuring water content include chemical titration, such as Karl Fischer titration, which involves a vacuum oven. Various solvents, including water, can also be measured using weight loss methods, which involve thermal excitation. Exemplary methods include thermogravimetric analysis with infrared spectroscopy (TGA-IR).

[0114] In some embodiments, the particles comprise greater than about 60% by weight of the therapeutic biological agent, e.g., greater than about 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9% by weight of the therapeutic biological agent. In other embodiments, the particles comprise greater than about 90% by weight of the therapeutic biological agent. In certain embodiments, the particles have greater than about 95% by weight of the therapeutic biological agent. In still other embodiments, the particles have greater than about 98% by weight of the therapeutic biological agent. In preferred embodiments, the particles have greater than about 98% by weight of the therapeutic biological agent. In certain preferred embodiments, the particles have greater than about 99% by weight of the therapeutic biological agent.

[0115] Particles comprising at least one therapeutic biological agent as described herein can be prepared by many methods and any of the methods that form particles disclosed in, for example, International Application No. PCT / US2017 / 063150, International Application No. PCT / US2018 / 043774, International Application No. PCT / US2019 / 033875, and US Ser. No. 62 / 799,696, each of which is incorporated herein by reference in its entirety.

[0116] As used herein, the term "dispersity index" (DI) is a parameter that characterizes the degree of non-uniformity of particle size distribution. The term "polydispersity index" (PDI) is a parameter that characterizes the width of particle size distribution within a given sample. PDI values ​​range from 0.0 (a sample that is completely uniform in terms of particle size) to 1.0 or greater (a highly polydisperse sample with multiple particle size populations). As the value decreases, the particles have a more narrowly distributed particle size and greater uniformity of the particles. Particle diameters can be collected using microscopy (FlowCAM, SEM) and laser diffraction.

[0117] In some embodiments, the polydispersity index (PDI) calculation used for dynamic light scattering (DLS) measurements is the polydispersity index from DLS = (squared standard deviation) / (squared mean diameter). In other embodiments, the PDI calculation can be a statistical characteristic of the particles, namely, number average diameter (Dn), weight average diameter (Dw), and polydispersity index (PDI), where the calculation can be accomplished using the following equation, where di represents the diameter of the microsphere and n is the number of particles:

number

[0118] In certain embodiments, the particles can include one or more agents, such as therapeutic biological agents. In other embodiments, the particles can have a diameter of about 0.1 to about 1000 μm, e.g., about 0.1 to about 90 μm, about 90 to about 230 μm, or about 0.1 to about 1 μm. In still other embodiments, the particles can have a size dispersity of about 0 to about 0.9, e.g., about 0 to about 0.7, about 0 to about 0.5, or about 0 to about 0.2. Methods for measuring particle size and particle distribution include imaging flow cytometry and image analysis of scanning electron micrographs of particles, where the mean spherical radius or diameter can be calculated based on the cross-sectional area of ​​the particle projected onto the image plane. In certain other embodiments of the present disclosure, the particles can have a diameter of about 0.1 to about 1000 μm, a skeletal density of about 1.00 to about 6.00 g / cm3, and a glass transition temperature of about 0 to about 250°C.

[0119] While each of the elements of the present disclosure are described herein as containing multiple embodiments, it should be understood that, unless otherwise indicated, each of the embodiments of a given element of the present disclosure can be used with each of the embodiments of the other elements of the present disclosure, and each such use is intended to form a separate embodiment of the present disclosure.

[0120] Those skilled in the relevant art will appreciate that other suitable modifications and adaptations to the compositions and methods described herein will be readily apparent from the description of the disclosure contained herein, in view of the information known to those skilled in the art, and may be made without departing from the scope of the disclosure or any embodiment thereof.

[0121] Pharmaceutical Composition In certain embodiments, the present disclosure relates to a composition comprising a plurality of particles comprising any one of the above-mentioned agents suspended in a low-viscosity liquid. In certain preferred embodiments, the present disclosure relates to a pharmaceutical composition comprising a plurality of particles comprising any one of the above-mentioned therapeutic biological agents suspended in a low-viscosity pharmaceutically acceptable liquid.

[0122] In preferred embodiments according to the present disclosure as described herein, a composition comprising a plurality of particles has improved stability of a therapeutic biological agent compared to an aqueous composition comprising a therapeutic biological agent in monomeric form.

[0123] In another aspect, the present disclosure relates to a composition comprising a plurality of particles comprising an agent suspended in a liquid, wherein the particles comprise less than about 25% internal void space and the particles have a circularity of about 0.10 to about 1.00. As disclosed herein, the agent can be a therapeutic agent or a diagnostic agent. In certain embodiments, the therapeutic agent has a unit activity of about 0.5 to about 1.0. In a preferred embodiment, the therapeutic biological agent has a unit activity of about 0.5 to about 1.0.

[0124] In some embodiments, the present disclosure provides compositions containing a plurality of particles comprising an agent, e.g., a therapeutic or diagnostic agent, wherein the storage stability of the agent in the particles is improved relative to the storage stability of the agent in the first liquid. In other embodiments, the storage conditions are defined by time (e.g., greater than about 2 years, greater than about 1 year, greater than about 6 months, greater than about 3 months, greater than about 1 month, or greater than about 1 week) and temperature (e.g., about -80°C to about 100°C, about -80°C to about 60°C, about -20°C to about 60°C, or about 4 to about 60°C), potentially among other variables. In still other embodiments, the storage time is about 3 days, about 7 days, about 30 days, about 90 days, about 180 days, about 1 year, or about 2 years. In certain other embodiments, the temperature is about -80°C, about -40°C, about -20°C, about 4°C, about 25°C, about 40°C, or about 40 to about 60°C. In certain embodiments, the storage stability of the therapeutic or diagnostic agent in the particles is improved relative to the storage stability of the first liquid of the therapeutic or diagnostic agent.

[0125] In other embodiments, the particles have less than about 25% internal void space, e.g., less than about 24%, less than 20%, less than 15%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, or less than 0.1%. In certain embodiments, the particles may contain less than 10% internal void space, less than 5%, less than 1%, less than 0.1%, or less than 0.01% internal void space. In preferred embodiments, the particles are substantially free of any internal void space. In other embodiments, the particles may exhibit a porosity of about 0 to about 50%, e.g., about 0 to about 10%, about 0 to about 5%, about 0 to about 1%, about 0 to about 0.5%, about 0 to about 0.1%, or about 0 to about 0.01%. Exemplary pore size measurements include scanning electron microscopy (SEM), transmission electron microscopy (TEM), and confocal laser scanning microscopy analysis. A gallium focused ion beam (FIB) was used to cut one of the particles in half to reveal a cross-section of the particle's interior. The specific surface area of ​​porous microspheres and nanospheres can also be investigated by nitrogen adsorption / desorption analysis and the Brunauer-Emmett-Teller adsorption model. In certain embodiments where the pore size is sufficiently large, mercury intrusion porosimetry can be used.

[0126] In some embodiments, the circularity of the particles is at least about 10%, e.g., at least about 20, 30, 40, 50, 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or about 100%. In other embodiments, the circularity of the particles is at least about 88%. In certain embodiments, the circularity of the particles is at least about 90%. In still other embodiments, the circularity of the particles is at least about 93%. In preferred embodiments, the circularity of the particles is at least about 97%.

[0127] In other embodiments, the circularity of the particles is about 0.10 to about 1.00, e.g., about 0.20, 0.30, 0.40, 0.50, 0.60, 0.70, 0.75, 0.80, 0.81, 0.82, 0.83, 0.84, 0.85, 0.86, 0.87, 0.88, 0.89, 0.90, 0.91, 0.92, 0.93, 0.94, 0.95, 0.96, 0.97, 0.98, or 0.99 to about 1.00. In certain embodiments, the circularity of the particles is about 0.88 to about 1.00. In still other embodiments, the circularity of the particles is about 0.90 to about 1.00. In certain other embodiments, the circularity of the particles is about 0.93 to about 1.00. In a preferred embodiment, the circularity of the particles is from about 0.97 to about 1.00.

[0128] In certain embodiments, the circularity of the particles can be at least about 0.10 to about 1.00, such as at least about 0.88, about 0.90, about 0.93, or in the range of about 0.97 to about 1.00.

[0129] In some embodiments, the sphericity of the particles is at least about 50%, e.g., at least about 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or about 100%. In other embodiments, the sphericity of the particles is from about 0.10 to about 1.00, e.g., from about 0.20, 0.30, 0.40, 0.50, 0.60, 0.70, 0.75, 0.80, 0.81, 0.82, 0.83, 0.84, 0.85, 0.86, 0.87, 0.88, 0.89, 0.90, 0.91, 0.92, 0.93, 0.94, 0.95, 0.96, 0.97, 0.98, or 0.99 to about 1.00. In a preferred embodiment, the particles have a sphericity of about 1.00.

[0130] In certain embodiments, the sphericity of the particles can range from about 0.10 to about 1.00, such as at least about 0.20, about 0.40, about 0.60, or from about 0.80 to about 1.00.

[0131] In a preferred embodiment, the particles have a substantially smooth surface.

[0132] In some embodiments, the particles have a diameter of about 0.1 to about 1000 μm, e.g., about 0.1 to about 900, 800, 700, 600, 500, 400, 300, 200, 100, 90, 80, 70, 60, 50, 45, 40, 35, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, or about 0.2 μm. In certain embodiments, the particles have a diameter of about 1 to about 100 μm, e.g., about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, or 50 to about 100 μm. In still other embodiments, the particles have a diameter of about 4 to about 100 μm. In certain other embodiments, the particles have a diameter of about 10 to about 100 μm. In preferred embodiments, the particles have a diameter of about 20 to about 50 μm. In certain preferred embodiments, the diameter of the particles is intentionally controlled. In some embodiments, the particles have a diameter of about 0.1 to about 1000 μm, e.g., about 1 to about 400 μm, about 1 to about 200 μm, about 1 to about 100 μm, about 1 to about 50 μm, about 1 to about 25 μm, about 1 to about 10 μm, about 10 to about 100 μm, about 50 to about 100 μm, about 50 to about 75 μm, or about 75 to about 100 μm. In other embodiments, the particles have a diameter of about 1 to about 100 μm, e.g., about 4 to about 100 μm, about 10 to about 100 μm, or about 20 to about 50 μm.

[0133] In certain embodiments, the particles have a diameter of about 0.1 to about 100 μm. In certain other embodiments, the particles have a diameter of about 0.5 to about 50 μm. In still other embodiments, the particles have a diameter of about 20 to about 50 μm. In certain preferred embodiments, the particles have a diameter of about 1 to about 40 μm. In preferred embodiments, the particles have a diameter of about 2 to about 15 μm.

[0134] In some embodiments, the particles are less than about 10% by weight, e.g., less than about 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.9%, less than 0.8%, less than 0.7%, less than 0.6%, less than 0.5%, less than 0.4%, less than 0.3%, less than 0.2%, less than 0.1%, less than 0.09%, less than 0.08%, The surfactant content may be less than 0.07%, less than 0.06%, less than 0.05%, less than 0.04%, less than 0.03%, less than 0.02%, less than 0.01%, less than 0.009%, less than 0.008%, less than 0.007%, less than 0.006%, less than 0.005%, less than 0.004%, less than 0.003%, less than 0.002%, or less than 0.001% by weight. In other embodiments, the particles have a surfactant content of less than about 5% by weight. In certain embodiments, the particles have a surfactant content of less than about 3% by weight. In still other embodiments, the particles have a surfactant content of less than about 0.1% by weight. In certain other embodiments, the particles have a surfactant content of less than about 0.01% by weight. In some embodiments, the particles have a surfactant content of less than about 0.001% by weight. In preferred embodiments, the particles have a surfactant content of less than about 1% by weight. In certain preferred embodiments, the particles are substantially free of any surfactant content.

[0135] In other embodiments, the surfactant content of the particles is 0-10% by weight, e.g., 0-5% by weight, 0-3% by weight, 0-2% by weight, 0-1% by weight, 0-0.5% by weight, 0-0.2% by weight, 0-0.1% by weight, 0-0.01% by weight, or 0-0.001% by weight.

[0136] In some embodiments, the surfactant is polysorbate, magnesium stearate, sodium dodecyl sulfate, TRITON™ N-101, glycerin, polyoxyethylated castor oil, docusate, sodium stearate, decyl glucoside, nonoxynol-9, cetyltrimethylammonium bromide, sodium bis(2-ethylhexyl)sulfosuccinate, lecithin, sorbitan ester, or a combination thereof. In certain embodiments, the surfactant is polysorbate, docusate, or lecithin. In preferred embodiments, the surfactant is polysorbate 20, polysorbate 60, or polysorbate 80. In certain preferred embodiments, the surfactant is polysorbate 20 or polysorbate 80. In certain other embodiments, the fatty acid ester of sorbitol is a sorbitan ester, such as span 20, span 40, span 60, or span 80. In still other embodiments, the surfactant is an ionic surfactant.

[0137] In other embodiments, the particles have a density of about 1.00 to about 6.00 g / cm 3 , for example, about 1.00 to about 5.00 g / cm 3 , about 1.00~3.00g / cm 3 , about 1.00~2.00g / cm 3 , about 1.00~1.50g / cm 3 , about 1.30~1.50g / cm 3 , about 1.32~about 1.50g / cm 3 or about 1.10 to about 1.40 g / cm 3 In some embodiments, the particles exhibit a skeletal density of about 0.10 to about 5.00 g / cm 3 , for example, about 0.10 to about 2.50 g / cm 3 , about 0.10~about 1.40g / cm 3 , about 0.50~1.40g / cm 3 or about 1.00 to about 1.40 g / cm 3 In certain embodiments, the particles exhibit a skeletal density of from about 0.09 to about 1.60 g / cm 3 In yet another embodiment, the particles have a skeletal density of about 1.30 to about 1.58 g / cm 3In a preferred embodiment, the particles have a skeletal density of about 1.32 to about 1.50 g / cm 3 It has a skeletal density of

[0138] In certain embodiments, the particles have a concentration of about 1000 mg / mL to about 1500 mg / mL, about 1050 mg / mL to about 1500 mg / mL, about 1100 mg / mL to about 1500 mg / mL, about 1150 mg / mL to about 1500 mg / mL, about 1200 mg / mL to about 1500 mg / mL, about 1250 mg / mL to about 1500 mg / mL, about 1300 mg / mL to about 1500 mg / mL, about 1310 mg / mL to about 1500 mg / mL, about 1320 mg / mL to about 1500 mg / mL, about 1330 mg / mL to about 1500 mg / mL, about 1340 mg / mL to about 1500 mg / mL, about 1350 mg / mL to about 1500 mg / mL, or about 1360 mg / mL to about 1500 mg / mL L, about 1370mg / mL to about 1500mg / mL, about 1380mg / mL to about 1500mg / mL, about 1390mg / mL to about 1500mg / mL, about 1400mg / mL to about 1500mg / mL, about 1410mg / mL to about 1500mg / mL, about 1420mg / mL to about 1500mg / mL, about 1430mg / mL to about 1500mg / mL, about 1440mg / mL to about 1500mg / mL, about 1450mg / mL to about 1500mg / mL, about 1460mg / mL to about 1500mg / mL, about 1470mg / mL to about 1500mg / mL, about 1480mg / mL to about 1500mg / mL, or about 1490mg / mL to about 1500mg / mL.

[0139] In other embodiments, the particles can be characterized by a glass transition temperature of about 0°C to about 250°C, e.g., about 34°C to about 200°C, about 50°C to about 200°C, about 60°C to about 200°C, about 40°C to about 160°C, about 50°C to about 110°C, about 60°C to about 100°C, or about 75°C to about 80°C. In other embodiments, the particles have a glass transition temperature of about 40°C to about 160°C. In still other embodiments, the particles have a glass transition temperature of about 50°C to about 110°C. In certain embodiments, the particles have a glass transition temperature of about 60°C to about 100°C. In preferred embodiments, the particles have a glass transition temperature of about 75°C to about 80°C. In still other embodiments, the particles are heated during drying to about ±30°C, e.g., about ±20, ±10, ±5, or ±1°C, of ​​the glass transition temperature of the particles.

[0140] In certain embodiments, the particles have a glass transition temperature greater than about 160° C. In certain other embodiments, the particles have a glass transition temperature greater than about 90° C. In certain preferred embodiments, the particles have a glass transition temperature greater than about 50° C.

[0141] In some embodiments, the particles further comprise a carbohydrate, a pH adjuster, a salt, a chelating agent, a mineral, a polymer, a surfactant, a protein stabilizer, an emulsifier, a preservative, an amino acid, an antioxidant, a protein, an organic solvent, a paraben, a bactericide, a fungicide, a vitamin, a preservative, a nutrient medium, an oligopeptide, a biological additive, a chemical additive, or a combination thereof. In certain embodiments, the particles further comprise a carbohydrate, a pH adjuster, a salt, a surfactant, a protein stabilizer, an emulsifier, an amino acid, or a combination thereof.

[0142] In certain embodiments, the liquid is non-aqueous or aqueous. In other embodiments, the liquid is non-aqueous. In yet other embodiments, the liquid is aqueous.

[0143] In other embodiments, the non-aqueous liquid is an organic solvent or an ionic liquid. In some embodiments, the organic solvent is selected from the group consisting of benzyl benzoate, coconut oil, cottonseed oil, fish oil, grapeseed oil, hazelnut oil, hydrogenated vegetable oil, olive oil, palm kernel oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, sunflower oil, walnut oil, acetone, ethyl acetate, ethyl lactate, dimethylacetamide, dimethyl isosorbide, dimethyl sulfoxide, glycofurol, diglyme, methyl tert-butyl ether, N-methylpyrrolidone, perfluorodecalin, polyethylene glycol, ethanol, 2-pyrrolidone, tetrahydrofurfuryl alcohol, triglycerides, fractionated vegetable fatty acid C8 and C10 triglycerides, propylene glycol diesters of saturated vegetable fatty acids C8 and C10, ethyl oleate, ethyl caprate, dibutyl adipate, fatty acid esters, hexanoic acid, octanoic acid, triacetin, diethyl glycol monoether, gamma-butyrolactone, eugenol, clove bud oil, citral, limonene, polyoxyl 40 hydrogenated castor oil, polyoxyl 35 castor oil , simple alcohols such as ethanol, octanol, hexanol, decanol, propanol and butanol, gamma-butyrolactone, tocopherol, octa-fluoropropane, (perfluorohexyl)octane, n-acetyltryptophan, ethyl laurate, methyl caprylate, methyl caprate, methyl myristate, methyl oleate, methyl linoleate, dimethyl adipate, dibutyl suberate, diethyl sebacate, ethyl macadamiate, trimethylolpropane triisostearate, isopropyl laurate, isopropyl myristate, diethyl succinate, polysorbate esters, ethanolamine, propanoic acid, citral, anisole, anethole, benzaldehyde, linalool, caprolactone, phenol, thioglycerol, dimethylacetamide, diethylene glycol monoethyl ether, propylene carbonate, solketal, isosorbide dimethyl ether, ethyl formate and ethylhexyl acetate or combinations thereof.In preferred embodiments, the organic solvent is ethyl oleate, ethyl laurate, ethyl macadamiate, ethyl caprate, diethyl succinate, diethylene glycol monoethyl ether, propylene carbonate, or a combination thereof. In a particularly preferred embodiment, the organic solvent is ethyl oleate. Exemplary ionic liquids of the present disclosure contain (i) a cation, such as pyridinium, pyridazinium, pyrimidinium, pyrazinium, imidazolium, pyrazolium, thiazolium, oxazolium, triazolium, ammonium, or sulfonium; and (ii) an anion, such as a halide, sulfate, sulfonate, carbonate, phosphate, bicarbonate, nitrate, acetate, PF6-, BF4-, triflate, nonaflate, bis(triflyl)amide, trifluoroacetate, heptafluorobutanoate, haloaluminate, or a combination thereof. In certain embodiments, the ionic liquid comprises pyridinium, pyridazinium, pyrimidinium, pyrazinium, imidazolium, pyrazolium, thiazolium, oxazolium, triazolium, ammonium, sulfonium, halide, sulfate, sulfonate, carbonate, phosphate, bicarbonate, nitrate, acetate, PF6-, BF4-, triflate, nonaflate, bis(triflyl)amide, trifluoroacetate, heptafluorobutanoate, haloaluminate, or combinations thereof.

[0144] In certain embodiments, the organic solvent is acetonitrile, chlorobenzene, chloroform, cyclohexane, cumene, 1,2-dichloroethene, dichloromethane, 1,2-dimethoxyethane, N,N-dimethylacetamide, N,N-dimethylformamide, 1,4-dioxane, 2-ethoxyethanol, ethylene glycol, formamide, hexane, methanol, 2-methoxyethanol, methyl butyl ketone, methyl cyclohexane, methyl isobutyl ketone, N-methylpyrrolidone, nitromethane, pyridine, sulfolane, tetrahydrofuran, tetralin, toluene, 1,1,2-trichloroethene, xylene, acetic acid, acetone, anisole, 1-butanol, 2-butanol, butyl acetate, tetramethylpropane ... rt-butyl methyl ether, dimethyl sulfoxide, ethanol, ethyl acetate, ethyl ether, ethyl formate, formic acid, heptane, isobutyl acetate, isopropyl acetate, methyl acetate, 3-methyl-1-butanol, methyl ethyl ketone, 2-methyl-1-propanol, pentane, 1-pentanol, 1-propanol, 2-propanol, propyl acetate, triethylamine, 1,1-diethoxypropane, 1,1-dimethoxymethane, 2,2-dimethoxypropane, isooctane, isopropyl ether, methyl isopropyl ketone, methyl tetrahydrofuran, petroleum ether, trichloroacetic acid, trifluoroacetic acid, decanol, 2-ethylhexyl acetate, amyl acetate, or a combination thereof.

[0145] In some embodiments, the aqueous liquid is water, 0.9% saline, lactated Ringer's solution, buffer, dextrose 5%, or a combination thereof. In a preferred embodiment, the aqueous liquid is water. Exemplary buffers of the present disclosure may include acetate buffer, histidine buffer, succinate buffer, HEPES buffer, Tris buffer, carbonate buffer, citrate buffer, phosphate buffer, phosphate buffered saline, glycine buffer, barbital buffer, cacodylate buffer, ammonium formate buffer, urea solution, or a combination thereof.

[0146] The phrase "pharmaceutically acceptable" is used herein to refer to those therapeutic biological agents, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit / risk ratio. The term "pharmaceutically acceptable" can refer to particles and compositions, including a plurality of particles, that do not produce adverse, allergic, or other untoward reactions when administered to a mammal, e.g., a human, as needed. The preparation of pharmaceutical compositions containing an antibody or additional active ingredient will be known to those of skill in the art in light of the present disclosure. Furthermore, it is understood that for mammalian (e.g., human) administration, preparations should meet sterility, pyrogenicity, general safety, and purity standards as required by the FDA Office of Biological Standards.

[0147] The phrase "pharmaceutically acceptable liquid" includes any aqueous solvent (e.g., water, alcoholic / aqueous solutions, saline, parenteral vehicles such as sodium chloride, Ringer's dextrose, etc.), non-aqueous solvent (e.g., propylene glycol, polyethylene glycol, vegetable oils, and injectable organic esters such as ethyl oleate), dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antibacterial or antifungal agents, antioxidants, chelating agents, and inert gases), isotonic agents, absorption delaying agents, salts, drugs, drug stabilizers, gels, binders, additives, disintegrants, lubricants, sweeteners, flavoring agents, dyes, fluid and nutrient replenishers, such materials, and combinations thereof, as known to those skilled in the art. The pH and precise concentration of the various components in the pharmaceutical composition are controlled by well-known parameters. In certain preferred embodiments, the plurality of particles is suspended in a pharmaceutically acceptable liquid. In preferred embodiments, the liquid is a pharmaceutically acceptable liquid.

[0148] Pharmaceutical compositions (formulations) as disclosed herein can be administered to a subject by any of several routes of administration, including, for example, parenterally (including, for example, intramuscularly, intravenously, subcutaneously, or intrathecally, as a sterile solution or suspension); intraperitoneally; or subcutaneously. In certain embodiments, the compositions may simply be suspended in a non-aqueous liquid carrier. Details of suitable administration routes and compositions suitable for these can be found, for example, in U.S. Pat. Nos. 6,110,973; 5,763,493; 5,731,000; 5,541,231; 5,427,798; 5,358,970; and 4,172,896 and the patents cited therein. The term "suspension formulation" refers to a liquid formulation comprising solid particles disposed in a carrier liquid in which the solid particles are not soluble on a suitable time scale. The particles may settle over time, ie, the physical stability of the suspension is not indefinite, but they can be resuspended using some form of agitation or excitation.

[0149] A "therapeutic amount" refers to the amount of a therapeutic or diagnostic agent needed to produce a desired effect. As used herein, the terms "treat," "treated," and "treating" refer to both therapeutic treatment and preventative or prophylactic measures, where the objective is to prevent or slow (alleviate) an undesirable physiological condition, disorder, or disease, or to obtain a beneficial or desired clinical outcome. Beneficial or desired clinical outcomes include, but are not limited to, alleviation of symptoms; a reduction in the extent of a condition, disorder, or disease; a stabilized (i.e., non-worsening) state of a condition, disorder, or disease; a delay in the onset or progression of a condition, disorder, or disease; an improvement or remission (partial or total) of a condition, disorder, or pathology (detectable or undetectable); an improvement in at least one measurable physical parameter, not necessarily discernible by the patient; or an enhancement or amelioration of a condition, disorder, or disease. Treatment includes eliciting a clinically meaningful response without excessive levels of side effects. Treatment also includes prolonging survival compared to expected survival if not receiving treatment.

[0150] In certain embodiments, the liquid further comprises a carbohydrate, a pH adjuster, a salt, a chelating agent, a mineral, a polymer, a surfactant, a protein stabilizer, an emulsifier, a preservative, an amino acid, an antioxidant, a protein, an organic solvent, a paraben, a bactericide, a fungicide, a vitamin, a preservative, a nutrient medium, an analgesic, or a combination thereof. In preferred embodiments, the liquid further comprises a carbohydrate, a pH adjuster, a salt, a surfactant, a protein stabilizer, an emulsifier, an amino acid, or a combination thereof. In certain preferred embodiments, the aqueous liquid further comprises a carbohydrate, a pH adjuster, a salt, a surfactant, a protein stabilizer, an emulsifier, an amino acid, or a combination thereof.

[0151] In other embodiments, the carbohydrate can be from the monosaccharide, disaccharide, oligosaccharide, or polysaccharide family. In some embodiments, the carbohydrate is dextran, trehalose, sucrose, agarose, mannitol, lactose, sorbitol, maltose, starch, alginate, xanthan, galactomannan, agar, agarose, or a combination thereof. In certain embodiments, the carbohydrate is dextran, trehalose, sucrose, agarose, mannitol, lactose, sorbitol, maltose, hydroxypropyl beta-cyclodextrin, or a combination thereof. In preferred embodiments, the carbohydrate is trehalose, cyclodextrin, hydroxypropyl beta-cyclodextrin, or a combination thereof. Cyclodextrins are available in three different forms, α, β, and γ, based on the number of glucose monomers. The number of glucose monomers in α, β, and γ cyclodextrin can be 6, 7, or 8, respectively.

[0152] In some embodiments, the pH adjuster is acetate, citrate, glutamate, glycinate, histidine, lactate, maleate, phosphate, succinate, tartrate, bicarbonate, aluminum hydroxide, phosphoric acid, hydrochloric acid, DL-lactic acid / glycolic acid, phosphorylethanolamine, tromethamine, imidazole, glycylglycine, monosodium glutamate, sodium hydroxide, potassium hydroxide, or a combination thereof. In other embodiments, the pH adjuster is citrate, histidine, phosphate, succinate, sodium hydroxide, potassium hydroxide, or a combination thereof. In certain embodiments, the pH adjuster is hydrochloric acid or citric acid.

[0153] In other embodiments, the salt is sodium chloride, calcium chloride, potassium chloride, sodium hydroxide, stannous chloride, magnesium sulfate, sodium glucoheptonate, sodium pertechnetate, guanidine hydrochloride, potassium hydroxide, or a combination thereof. In a preferred embodiment, the salt is sodium chloride.

[0154] In some embodiments, the chelating agent is edetate disodium, ethylenediaminetetraacetic acid, pentetic acid, or a combination thereof. In other embodiments, the mineral is calcium, zinc, titanium dioxide, or a combination thereof. In certain embodiments, the polymer is propylene glycol, glucose star polymer, silicone polymer, polydimethylsiloxane, polyethylene glycol, carboxymethylcellulose, poly(glycolic acid), poly(lactic-co-glycolic acid), polylactic acid, polycaprolactone (PCL), polyvinylpyrrolidone (PVP), ficoll, dextran, or a combination thereof.

[0155] In other embodiments, the surfactant is polysorbate, magnesium stearate, sodium dodecyl sulfate, TRITON™ N-101, glycerin, polyoxyethylated castor oil, docusate, sodium stearate, decyl glucoside, nonoxynol-9, cetyltrimethylammonium bromide, sodium bis(2-ethylhexyl)sulfosuccinate, sodium laureth sulfate, lecithin, or a combination thereof. In some embodiments, the surfactant includes, but is not limited to, (i) cationic surfactants, such as cetyltrimethylammonium chloride, hexadecyltrimethylammonium chloride, benzalkonium chloride, benzethonium chloride, and dioctadecyldimethylammonium bromide; (ii) anionic surfactants, such as magnesium stearate, sodium dodecyl sulfate, dioctyl sodium sulfosuccinate, sodium myreth sulfate, perfluorooctane sulfonate, and alkyl ether phosphates; and (iii) nonionic surfactants, such as alkylphenol ethoxylates (Triton X-100), fatty alcohols, and the like. Ethoxylates (octaethylene glycol monododecyl ether, cocamide diethanolamine, poloxamer, glycerol monostearate, fatty acid esters of sorbitol (sorbitan monolaurate, Tween 80, Tween 20); and (iv) zwitterionic surfactants, such as cocamidopropyl hydroxysultaine and 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS). In certain embodiments, the surfactant is polysorbate, docusate, or lecithin. In preferred embodiments, the surfactant is polysorbate 20 or polysorbate 80.

[0156] In some embodiments, the protein stabilizer is acetyltryptophanate, caprylate, N-acetyltryptophan, trehalose, PEG200, PEG300, PEG3350, PEG8000, PEG10000, PEG20000, poloxamer, polyvinylpyrrolidone, polyacrylic acid, poly(vinyl) polymer, polyester, polyaldehyde, tertiary polymer, polyamino acid, hydroxyethyl starch, N-methyl-2-pyrrolidone, sorbitol, sucrose, mannitol, or a combination thereof. In certain embodiments, the protein stabilizer is trehalose, PEG200, PEG300, PEG3350, PEG8000, PEG10000, PEG20000, poloxamer, polyvinylpyrrolidone, polyacrylic acid, poly(vinyl) polymer, polyester, polyaldehyde, tertiary polymer, polyamino acid, hydroxyethyl starch, N-methyl-2-pyrrolidone, sorbitol, sucrose, mannitol, cyclodextrin, sugar, or combinations thereof. In preferred embodiments, the protein stabilizer is trehalose, PEG200, PEG300, PEG3350, PEG8000, PEG10000, PEG20000, cyclodextrin, hydroxypropyl beta-cyclodextrin, or combinations thereof. Stabilizers, used synonymously with the term "stabilizer," can be salts, carbohydrates, sugars, or amino acids, preferably carbohydrates or sugars, recognized by authorities as suitable additives or excipients in pharmaceutical compositions, as described herein. The term "stabilizer" refers to an additive or mixture of additives that stabilizes the physical and / or chemical properties of an agent, e.g., a therapeutic or diagnostic agent. In some embodiments, the stabilizer prevents degradation of the therapeutic or diagnostic agent, e.g., during droplet formation, dehydration, and / or storage of particulate matter. Exemplary stabilizers include, but are not limited to, sugars, salts, hydrophobic salts, detergents, reducing agents, cyclodextrins, polyols, carboxylic acids, and amino acids. A "stable" formulation, as described herein, refers to one in which the therapeutic or diagnostic agent retains an acceptable portion of its essential physical, chemical, or biological properties for an acceptable period of time.For proteins and peptides, for example, exemplary methods for assessing stability are reviewed in (i) Peptide and Protein Drug Delivery, 247-301, Vincent Lee Ed., Marcel Dekker, Inc., New York, NY, 1991, and (ii) Jones, A., Adv. Drug Delivery Rev. 10:29-90 (1993). In certain embodiments, the chemical stability of a protein is assessed by measuring the size distribution of samples at several stages. These include, for example, before particle formation (evaluation of the feed solution), immediately after particle formation, and again after a period of storage, where storage occurs in or without a suspension formulation carrier medium. In certain other embodiments, the size distribution is assessed by size exclusion chromatography (SEC-HPLC).

[0157] Examples of emulsifiers suitable for use in liquids include, but are not limited to, lipophilic agents having an HLB of less than 7, such as mixed fatty acid monoglycerides; mixed fatty acid diglycerides; mixtures of fatty acid mono- and diglycerides; lipophilic polyglycerol esters; glycerol esters including glyceryl monooleate, glyceryl dioleate, glycerol monostearate, glyceryl distearate, glyceryl monopalmitate, and glyceryl dipalmitate; glyceryl-lactoesters of fatty acids; propylene glycol esters including propylene glycol monopalmitate, propylene glycol monostearate, and propylene glycol monooleate; sorbitan monostearate, sorbitan sesquioleate. sorbitan esters; fatty acids including stearic acid, palmitic acid, and oleic acid and soaps thereof; and mixtures thereof such as glyceryl monooleate, glyceryl dioleate, glyceryl monostearate, glyceryl distearate, glyceryl monopalmitate, and glyceryl dipalmitate; glyceryl-lactoesters of fatty acids; propylene glycol esters including propylene glycol monopalmitate, propylene glycol monostearate, and propylene glycol monooleate; sorbitan esters including sorbitan monostearate, sorbitan sesquioleate; fatty acids including stearic acid, palmitic acid, and oleic acid and soaps thereof; or combinations thereof. In some embodiments, the emulsifier is polysorbate 80, polysorbate 20, sorbitan monooleate, ethanolamine, polyoxyl 35 castor oil, polyoxyl 40 hydrogenated castor oil, carbomer 1342, corn oil-mono-di-triglyceride, polyoxyethylated oleic acid glyceride, poloxamer, or a combination thereof. In a preferred embodiment, the emulsifier is polysorbate 80, sorbitan monooleate, or a combination thereof.

[0158] In other embodiments, the preservative is phenol, m-cresol, benzyl alcohol, 2-phenyloxyethanol, chlorobutanol, neomycin, benzethonium chloride, glutaraldehyde, beta-propiolactone, or a combination thereof.

[0159] In certain embodiments, the amino acid is alanine, aspartic acid, cysteine, isoleucine, glutamic acid, leucine, methionine, phenylalanine, pyrrolysine, serine, selenocysteine, threonine, tryptophan, tyrosine, valine, asparagine, L-arginine, histidine, glycine, glutamine, proline, or a combination thereof. In preferred embodiments, the amino acid is L-arginine, histidine, proline, or a combination thereof.

[0160] In some embodiments, the antioxidant is glutathione, ascorbic acid, cysteine, N-acetyl-L-tryptophanate, tocopherol, histidine, methionine, or tocopherol, or a combination thereof. In other embodiments, the protein is protamine, protamine sulfate, gelatin, or a combination thereof. In certain embodiments, the organic solvent is dimethyl sulfoxide, N-methyl-2-pyrrolidone, or a combination thereof. In still other embodiments, the preservative is methyl hydroxybenzoate, thimerosal, a paraben, formaldehyde, castor oil, or a combination thereof. In certain other embodiments, the preservative is sodium nitrate, sulfur dioxide, potassium sorbate, sodium sorbate, sodium benzoate, benzoic acid, methyl hydroxybenzoate, thimerosal, a paraben, formaldehyde, castor oil, or a combination thereof. The paraben can be parahydroxybenzoate. In some embodiments, the disinfectant is benzalkonium chloride (a cationic surfactant), hypochlorite, peroxide, alcohol, a phenolic compound (e.g., carbolic acid), or a combination thereof.

[0161] In other embodiments, the fungicide is acibenzolar, 2-phenylphenol, allilazine, carvone, natamycin, potassium azide, or a combination thereof. In particular embodiments, the vitamin is thiamine, riboflavin, niacin, pantothenic acid, biotin, vitamin B6, vitamin B12, folate, niacin, ascorbic acid, calciferol, retinol, quinone, or a combination thereof.

[0162] Several nutrient media, preferably serum-free, alone or in combination, may be used in the present disclosure, including commercially available media or other media known in the art. Examples of such media (all serum-free or serum-depleted) include ADC-1, LPM (without bovine serum albumin), F10 (HAM), F12 (HAM), DCCM1, DCCM2, RPMI 1640, BGJ medium (Fitton-Jackson modified), Basal Medium Eagle (BME supplemented with Earl's salt base), Dulbecco's Modified Eagle Medium (DMEM, serum-free), Glasgow's Modified Eagle Medium (GMEM), Leibovitz's L-15 medium, McCoy's 5A medium, Medium M199 (M199E - with Earl's salt base), Medium M199 (M199H - with Hank's salt base), Minimum Essential Medium Eagle (MEM-E - with Earl's salt base), Minimum Essential Medium Eagle (MEM-H - with Hank's salt base), and Minimum Essential Medium Eagle (MEM-NAA - with non-essential amino acids), among many others. Additionally, although serum-containing nutrient media may be used in compositions according to the present disclosure, the use of serum-containing media is less preferred due to the possibility that serum may be contaminated with microbial agents and because patients may develop immune responses to certain antigenic components contained in the serum.

[0163] In some embodiments, the analgesic agent is paracetamol, a histamine receptor antagonist (e.g., an H1 or H2 blocker), an NSAID, a COX-2 inhibitor, celecoxib, rofecoxib, valdecoxib, parecoxib, lumiracoxib, etoricoxib, firocoxib, acetaminophen, an opiate, dextropropoxyphene, codeine, tramadol, anileridine, pethidine, hydrocodone, morphine, oxycodone, methadone, diacetylmorphine, hydromorphone, oxymorphone, levorphanol, buprenorphine, fentanyl, sufentanil, etorphine, carfentanil, dihydromorphine, dihydrocodeine, thebaine, papaverine, diproqualone, flupirtine, a tricyclic antidepressant, acetaminophen, or lidocaine, or a combination thereof. In certain embodiments, the analgesic agent is acetaminophen or lidocaine.

[0164] In certain embodiments, the liquid further comprises at least one pharmaceutically acceptable additive, excipient, additive, carrier, or combination thereof. In certain other embodiments, the liquid further comprises a second agent. In other embodiments, the liquid further comprises a second diagnostic or therapeutic agent.

[0165] In some embodiments, the particles have less than 20% aggregates or less than 20% fragmentation of the therapeutic biological agent, e.g., less than about 19%, less than 18%, less than 17%, less than 16%, less than 15%, less than 14%, less than 13%, less than 12%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1.9%, less than 1.8%, less than 1.7%, less than 1.6%, less than 1.5%, less than 1.4%, less than 1.3%, less than 1.2%, less than 1.1%, less than 1%, less than 0.9%, less than 0.8%, less than 0.7%, less than 0.6%, less than 0.5%, less than 0.4%, less than 0.3%, less than 0.2%, or less than 0.1%. In other embodiments, the particles have less than 10% aggregates or less than 10% fragments of the therapeutic biological agent, e.g., less than about 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1.9%, 1.8%, 1.7%, 1.6%, 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%, or 0.1%. In certain embodiments, the particles have between about 3% and about 1% aggregates of the therapeutic biological agent. In certain other embodiments, the particles have between about 1% and about 0.5% aggregates of the therapeutic biological agent. In preferred embodiments, the particles are substantially free of any aggregates of the therapeutic biological agent. In yet other embodiments, the particles have less than about 1% fragmentation of the therapeutic biological agent. In certain preferred embodiments, the particles are substantially free of any fragmentation of the therapeutic biological agent.

[0166] In certain embodiments, the methods described herein may further include suspending the particles in a pharmaceutically acceptable medium, e.g., reconstituting dried particles. In some embodiments, dissolving or reconstituting the particles results in less than about a 10% increase in aggregates of the diagnostic or therapeutic agent, e.g., protein, compared to the therapeutic or diagnostic agent in the first liquid before processing (e.g., less than about 8%, less than about 5%, less than about 4%, less than about 3%, less than about 1%, less than about 0.5%, or less than about 0.1%). An exemplary method for measuring aggregates includes size-exclusion high-performance liquid chromatography (SEC-HPLC), in which the aggregate population is quantified by dividing the area under the peak corresponding to the aggregate population by the cumulative area contained under all peaks in the sample spectrum. The change in aggregate percentage between two samples, e.g., sample A and sample B, is calculated as the numerical difference in the respective aggregate percentages, i.e., by subtracting the aggregate percentage of sample B from the aggregate percentage of sample B, or vice versa. In certain other embodiments, dissolution or reconstitution of the particles results in less than about a 10% increase in fragments of the diagnostic or therapeutic agent, e.g., protein, compared to the therapeutic or diagnostic agent in the first liquid before processing (e.g., less than about 8%, less than about 5%, less than about 4%, less than about 3%, less than about 1%, less than about 0.5%, or less than about 0.1%). An exemplary method for measuring fragments includes size-exclusion high-performance liquid chromatography (SEC-HPLC), in which the fragment population is quantified by dividing the area under the peak corresponding to the fragment population by the cumulative area contained under all peaks in the sample spectrum. The change in fragment percentage between two samples, e.g., sample A and sample B, is calculated as the numerical difference in the respective fragment percentages, i.e., by subtracting the fragment percentage of sample B from the fragment percentage of sample A, or vice versa.

[0167] In other embodiments, the particle formation process achieves less than a 50% change in charge variants (e.g., less than 40%, less than 30%, less than 20%, less than 10%, less than 8%, less than 5%, less than 4%, less than 3%, or less than 1%) in a population of diagnostic or therapeutic agents, e.g., antibodies or antibody fragments, compared to the therapeutic or diagnostic agents prior to particle formation. In certain embodiments, the particles have less than about a 50% change in charge variants of the therapeutic biological agent, e.g., less than about 45%, less than 40%, less than 35%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, or less than 0.1%, compared to the starting biological agent prior to particle formation. In preferred embodiments, the particles are substantially free of any change in charge variants of the therapeutic biological agent, compared to the starting biological agent prior to particle formation.

[0168] In some embodiments, the residual moisture or solvent content of the dry ingredients is less than about 7% by weight, e.g., less than about 6%, 5%, 4%, 3%, 2%, 1.9%, 1.8%, 1.7%, 1.6%, 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%, or 0.1% by weight. In other embodiments, the particles have less than about 7% by weight residual moisture. In still other embodiments, the particles have less than about 5% by weight residual moisture. In certain embodiments, the particles have less than about 3% by weight residual moisture. In preferred embodiments, the particles have less than about 1% by weight residual moisture.

[0169] In other embodiments, the particles have a residual moisture content of about 1% to about 7% by weight. In still other embodiments, the particles have a residual moisture content of about 1% to about 5% by weight. In certain embodiments, the particles have a residual moisture content of about 1% to about 3% by weight. In preferred embodiments, the particles are substantially free of any residual moisture by weight.

[0170] In some embodiments, the particles comprise greater than about 60% by weight of the therapeutic biological agent, e.g., greater than about 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9% by weight of the therapeutic biological agent. In other embodiments, the particles comprise greater than about 90% by weight of the therapeutic biological agent. In certain embodiments, the particles have greater than about 95% by weight of the therapeutic biological agent. In still other embodiments, the particles have greater than about 98% by weight of the therapeutic biological agent. In preferred embodiments, the particles have greater than about 98% by weight of the therapeutic biological agent. In certain preferred embodiments, the particles have greater than about 99% by weight of the therapeutic biological agent.

[0171] The concentration of the therapeutic biological agent in the composition is typically about 20 mg / mL to about 650 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625 mg / mL to about 650 mg / mL. The therapeutic biological agent in the composition may have about 0.5 to about 1.0 unit activity, about 0.75 to about 1.0 unit activity, or about 0.9 to about 1.0 unit activity. Activity is measured relative to the same therapeutic biological agent prior to particle formation. In a preferred embodiment, the therapeutic biological agent has about 0.5 to about 1.0 unit activity.

[0172] In some embodiments, the compositions described herein contain a concentration of about 20 mg / mL to about 650 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625 mg / mL to about 650 mg / mL; , 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575, 600 mg / mL to about 625 mg / mL; about 20 mg / mL to about 600 mg / mL, for example, about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575 mg / mL to about 600 mg / mL; about 20 mg / mL to about 575 mg / mL, for example, about 20 , 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550 mg / mL to about 575 mg / mL; about 20 mg / mL to about 550 mg / mL, for example about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525 mg / mL to about 550 mg / mL; about 20 mg g / mL to about 525 mg / mL, for example, about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500 mg / mL to about 525 mg / mL; about 20 mg / mL to about 500 mg / mL, for example, about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475 mg / mL to about 500 mg / mL;about 20 mg / mL to about 475 mg / mL, for example, about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450 mg / mL to about 475 mg / mL; about 20 mg / mL to about 450 mg / mL, for example, about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 25 0, 275, 300, 325, 350, 375, 400, 425 mg / mL to about 450 mg / mL; about 20 mg / mL to about 425 mg / mL, for example, about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400 mg / mL to about 425 mg / mL; about 20 mg / mL to about 400 mg / mL, for example, about 20 , 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375 mg / mL to about 400 mg / mL; about 20 mg / mL to about 375 mg / mL, for example about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350 mg / mL to about 375 mg / mL; about 20 mg g / mL to about 350 mg / mL, for example, about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325 mg / mL to about 350 mg / mL; about 20 mg / mL to about 325 mg / mL, for example, about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300 mg / mL to about 325 mg / mL;Alternatively, concentrations of the therapeutic biological agent in the composition of about 20 mg / mL to about 300 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275 mg / mL to about 300 mg / mL, are used. In other embodiments, the concentration of the therapeutic biological agent in the composition is about 30 mg / mL to about 500 mg / mL. In certain embodiments, the concentration of the therapeutic biological agent in the composition is about 100 mg / mL to about 500 mg / mL. In still other embodiments, the concentration of the therapeutic biological agent in the composition is about 200 mg / mL to about 400 mg / mL. In preferred embodiments, the concentration of the therapeutic biological agent in the composition is about 300 mg / mL to about 400 mg / mL. In certain preferred embodiments, the concentration of the therapeutic biological agent in the composition is about 350 mg / mL to about 400 mg / mL. ;

[0173] In other embodiments, the composition has a molecular weight of less than about 200 mPa·s, less than about 150 mPa·s, less than about 125 mPa·s, less than about 100 mPa·s, less than about 75 mPa·s, less than about 75 mPa·s, less than about 70 mPa·s, less than about 65 mPa·s, less than about 60 mPa·s, less than about 55 mPa·s, less than about 50 mPa·s, less than about 45 mPa·s, less than about 40 mPa·s, less than about 35 mPa·s, less than about 30 mPa·s, less than about 25 mPa·s, less than about 20 mPa·s, less than about 19 mPa·s, less than about 18 mPa·s, less than about 17 mPa·s, less than about 16 mPa·s, less than about 15 mPa·s, less than about 14 mPa·s, less than about 13 mPa·s, less than about 12 mPa·s The viscosity of the composition is less than about 11 mPa·s, less than about 10 mPa·s, less than about 9.5 mPa·s, less than about 9 mPa·s, less than about 8.5 mPa·s, less than about 8 mPa·s, less than about 7.5 mPa·s, less than about 7 mPa·s, less than about 6.5 mPa·s, less than about 6 mPa·s, less than about 5.5 mPa·s, less than about 5 mPa·s, less than about 4.5 mPa·s, less than about 4 mPa·s, less than about 3.5 mPa·s, less than about 3 mPa·s, less than about 2.5 mPa·s, less than about 2 mPa·s, less than about 1.5 mPa·s, less than about 1 mPa·s, less than about 0.5 mPa·s, less than about 0.1 mPa·s, less than about 0.05 mPa·s, or less than about 0.01 mPa·s (1 milliPascal second). In other embodiments, the composition has a viscosity of about 0.01 mPa·s to about 10,000 mPa·s, e.g., about 0.01 mPa·s to about 1,000 mPa·s, about 0.01 mPa·s to about 100 mPa·s, about 0.01 mPa·s to about 50 mPa·s, about 0.01 mPa·s to about 25 mPa·s, about 0.01 mPa·s to about 10 mPa·s, about 0.01 mPa·s to about 5 mPa·s, or about 0.01 mPa·s to about 1 mPa·s. In certain embodiments, the viscosity of the composition can be in the range of about 0.27 mPa·s to about 200 mPa·s, e.g., about 0.27 mPa·s to about 50 mPa·s, about 1 mPa·s to about 30 mPa·s, or about 20 mPa·s to about 50 mPa·s.In yet other embodiments, the viscosity of the composition ranges from about 0.27 mPa·s to about 200 mPa·s, e.g., from about 0.27 mPa·s to about 100 mPa·s, from about 0.27 mPa·s to about 50 mPa·s, from about 0.27 mPa·s to about 30 mPa·s, from about 1 mPa·s to about 20 mPa·s, or from about 1 mPa·s to about 15 mPa·s. The term "viscosity" is used to describe the property of a fluid that acts to resist shear flow. For purposes of this disclosure, viscosity can be determined at 25°C at a specific shear rate using a rheometer fitted with a cone and plate (2° / 40 mm), such as an AR-G2 rheometer (TA Instruments, USA). In certain embodiments, viscosity is measured at shear rates in the Newtonian region. The term "Newtonian region" refers to a range of shear rates that are linearly proportional or nearly linearly proportional to the local strain rate at every point. In some embodiments, the viscosity is about 100 s. -1 More than, for example, about 1000s -1 or about 1000s -1 The viscosity is measured at a shear rate of greater than 1000 s. The composition may contain about 5 to about 90% by volume of particles, e.g., about 20 to about 90% by volume, about 40 to about 80% by volume, about 50 to about 60% by volume, or about 70 to about 90% by volume. The composition may have a concentration of therapeutic biological agent of about 0.0001 to about 1000 mg / mL, e.g., about 100 to about 900 mg / mL, about 150 to about 800 mg / mL, or about 200 to about 700 mg / mL. Methods for controlling viscosity include temperature regulation and viscosity-modifying additives. Liquid mixtures may also be used to control viscosity. The units "mPa·s" and "cP" are used interchangeably herein in their broadest sense.

[0174] In some embodiments, the composition has a viscosity of less than about 50 mPa·s. In other embodiments, the composition has a viscosity of less than about 30 mPa·s. In still other embodiments, the composition has a viscosity of less than about 20 mPa·s. In certain other embodiments, the composition has a viscosity of less than about 10 mPa·s. In certain embodiments, the composition has a viscosity of less than about 5 mPa·s. In preferred embodiments, the composition has a viscosity of less than about 3 mPa·s. In certain preferred embodiments, the composition has a viscosity of less than about 2.5 mPa·s.

[0175] In other embodiments of the compositions described herein, the plurality of particles has a polydispersity index of about 0.002 to about 1.000, e.g., about 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.010, 0.020, 0.030, 0.040, 0.050, 0.060, 0.070, 0.080, 0.090, 0.100, 0.200, 0.300, 0.400, 0.500, 0.600, 0.700, 0.800, 0.900 to about 1.000. In certain embodiments, the plurality of particles has a polydispersity index of about 0.002 to about 0.900.

[0176] Certain embodiments of the present disclosure described herein allow for high concentrations of therapeutic biological agents in particles and high concentrations of particles in liquids, which in some embodiments may be achieved by mixing particles of various sizes.

[0177] In preferred embodiments according to the present disclosure as described herein, a composition comprising a plurality of particles has improved stability of a therapeutic biological agent compared to an aqueous composition comprising a therapeutic biological agent in monomeric form.

[0178] In other embodiments, particles of the present disclosure can be suspended in an aqueous liquid carrier, a non-aqueous liquid carrier, such as an organic liquid, an ionic liquid carrier, a gel carrier, or a combination thereof, to form a suspension composition. The vehicle for the suspension can further include, for example, carbohydrates, pH adjusters, salts, chelating agents, minerals, polymers, surfactants, amino acids, oligopeptides, biological additives, chemical additives, preservatives, antioxidants, parabens, bactericides, fungicides, vitamins, preservatives, analgesics, and / or nutrient media. In some embodiments, each of the other components can independently comprise about 0.0001 to about 99% (w / v) of the vehicle, e.g., about 0.0001 to about 90% (w / v), about 0.0001 to about 50% (w / v), about 0.0001 to about 10% (w / v), about 0.0001 to about 1% (w / v), or about 0.0001 to about 0.1% (w / v). In certain embodiments, the present disclosure provides a plurality of particles described herein suspended in a liquid. The liquid can be an organic solvent, an ionic liquid, an aqueous liquid, or a combination thereof. The liquid can further comprise a second diagnostic or therapeutic agent.

[0179] In some embodiments, insoluble particulate matter having a characteristic size of about 100 μm or greater that persists upon dissolution in an aqueous liquid is referred to as a visible particle (VP). In preferred embodiments of the present disclosure described herein, the composition is substantially free of visible particles (VP). In certain preferred embodiments, the aqueous liquid is water, an aqueous buffer, or a physiologically suitable aqueous liquid. In other embodiments, insoluble particulate matter that is visible to the naked eye under certain lighting conditions persists upon reconstitution of the particles of the present disclosure into a liquid pharmaceutical composition. This type of insoluble particulate matter may be referred to as a visible particle (VP) and is typically greater than about 100 μm in size. The VP is present in an amount of about 0 to about 1 per mL, e.g., about 0 to about 0.01 per mL, about 0 to about 0.001 per mL, or about 0 to about 0.0001 per mL. An exemplary method for measuring VP is to measure VP using the USP 1000 mg / mL (1000 mg / mL) of a therapeutic or diagnostic agent after reconstitution and dilution to a standard concentration, e.g., about 100 mg / mL or about 1 mg / mL. <790> In some embodiments, fewer than 65 samples out of 10,000 (0.65%) meet the U.S.P. <790> Alternative inspection strategies include light obscuration, automated optical imaging systems, or USP <1790> This is X-ray imaging.

[0180] In another embodiment, insoluble particulate matter having a characteristic size of about 1 μm to about 100 μm that persists upon dissolution in an aqueous liquid is referred to as subvisible particles (SvP). SvP is present in an amount of about 0 to 100,000,000 per about 1 mL, for example, about 0 to about 10,000,000 per about 1 mL, about 0 to about 1,000,000 per about 1 mL, about 0 to about 500,000 per about 1 mL, about 0 to about 100,000 per about 1 mL, about 0 to about 50,000 per about 1 mL, about 0 to about 10,000 per about 1 mL, about 0 to about 6,000 per about 1 mL, about 0 to about 1,000 per about 1 mL, about 0 to about 600 per about 1 mL, about 0 to about 250 per about 1 mL, about 0 to about 100 per about 1 mL, about 0 to about 60 per about 1 mL, or about 0 to about 10 per about 1 mL. In other embodiments, the number of particles having a characteristic size of 10 μm or greater is about 0 to about 6,000 per mL, e.g., about 0 to about 1,000 per mL, about 0 to about 100 per mL, about 0 to about 10 per mL, about 0 to about 5 per mL, about 0 to about 3 per mL, or about 0 to about 1 per mL. In certain embodiments, the number of particles having a characteristic size of 25 μm or greater is about 0 to about 600 per mL, e.g., about 0 to about 100 per mL, about 0 to about 10 per mL, about 0 to about 3 per mL, about 0 to about 1 per mL, about 0 to about 0.5 per mL, or about 0 to about 0.1 per mL. Exemplary methods for measuring SvP include analysis of the therapeutic biological agent by a Coulter counter, HIAC Royco, or microfluidic imaging system after reconstitution and dilution of the therapeutic biological agent to a standard concentration, e.g., about 100 mg / mL or about 1 mg / mL. In still other embodiments, the composition, when dissolved in an aqueous liquid, has a concentration of about 0 to about 100,000,000 insoluble sub-visible particles per mL of particles greater than about 10 μm. In certain embodiments, the composition, when dissolved in an aqueous liquid, has a concentration of about 0 to about 6,000 insoluble sub-visible particles per mL of particles greater than about 10 μm. In preferred embodiments, the composition, when dissolved in an aqueous liquid, has a concentration of about 0 to about 600 insoluble sub-visible particles per mL of particles greater than about 25 μm.In certain preferred embodiments, the composition is substantially free of insoluble sub-visible particles when dissolved in an aqueous liquid, hi preferred embodiments, the aqueous liquid is water, an aqueous buffer, or a physiologically suitable aqueous liquid.

[0181] In some embodiments, insoluble particulate matter having a characteristic size of about 100 nm to about 1 μm that persists upon dissolution in an aqueous liquid is referred to as a submicron particle (SMP), sometimes known as a nanoparticle. Quantitatively, SMPs have a particle size of about 0-5×10 per mL. 12 , for example, about 0 to about 0.5 × 10 per about 1 mL 12 , approximately 0 to approximately 50 × 10 per 1 mL 9 , approximately 0 to approximately 10 × 10 per 1 mL 9 , approximately 0 to approximately 5 × 10 per 1 mL 9 , approximately 0 to approximately 0.5 × 10 per 1 mL 9 , approximately 0 to approximately 50 × 10 per 1 mL 6 , approximately 0 to approximately 1 × 10 per 1 mL 6 , about 0 to about 500,000 per mL, about 0 to about 200,000 per mL, about 0 to about 100,000 per mL, about 0 to about 10,000 per mL, about 0 to about 5000 per mL, or about 0 to about 1000 per mL. Exemplary methods for quantitatively measuring SMP include analysis of the therapeutic biological agent by NanoSight, a microfluidic imaging system, asymmetric field-flow fractionation coupled to multi-angle laser light scattering (AF4MALS), or dynamic light scattering (DLS) after reconstitution and dilution of the therapeutic biological agent to a standard concentration, e.g., about 100 mg / mL, about 1 mg / mL, or about 1 μg / mL. Qualitatively, the SMP is in the range corresponding to the starting monomer therapeutic biological agent solution. In a preferred embodiment, the composition is substantially free of submicron particles (SMP) upon dissolution in an aqueous liquid. In certain preferred embodiments, the aqueous liquid is water, an aqueous buffer, or a physiologically suitable aqueous liquid. Qualitatively, as described herein, the SMP is in a range that corresponds to the feed solution.

[0182] In certain embodiments, the suspension contains insoluble particulate matter of 1 μm or less. The suspension contains about 1-5×10 particles per mL of suspension. 12 or in a suspension having insoluble particles with a characteristic size of about 100 nm or greater at a concentration of about 1-5 x 10 per mL. 12 In certain other embodiments, the particle suspension may have a concentration of insoluble particles having a characteristic size of about 1 μm or less. In yet other embodiments, the particle suspension may include insoluble particulate matter having a size of about 1 μm or greater. In certain other embodiments, the number of insoluble particles is from about 0 to about 100,000,000 per mL, e.g., less than about 10,000,000, less than 1,000,000, less than 100,000, less than 10,000, less than 1000, less than 10, or less than about 1 per mL. For example, the number of insoluble particles greater than about 10 μm is about 0 to about 6,000 per mL, e.g., less than about 5,000, less than about 4,000, less than about 3,000, less than about 2,000, less than about 1,000, less than about 500, less than about 100, less than about 10, or less than about 1 per mL, and / or the number of insoluble particles greater than about 25 μm is about 0 to about 600 per mL, e.g., less than about 500, less than about 400, less than about 300, less than about 200, less than about 100, less than about 50, less than about 10, or less than about 1 per mL.

[0183] In some embodiments, the present disclosure provides compositions, e.g., suspensions or dried forms, containing a plurality of particles comprising an agent, e.g., a therapeutic or diagnostic agent. The compositions preferably have a concentration of insoluble particles, e.g., SvP, per mL, either in suspension or upon reconstitution. In other embodiments, the concentration of insoluble particles is from about 0 to about 1,000,000 per mL, either in suspension or upon reconstitution. In still other embodiments, the concentration of insoluble particles is from about 0 to about 10,000 per mL, either in suspension or upon reconstitution. In certain other embodiments, the concentration of insoluble particles having a characteristic size of about 10 μm or greater is from about 0 to about 6,000 per mL, either in suspension or upon reconstitution. In certain embodiments, the concentration of insoluble particles having a characteristic size of about 25 μm or greater is from about 0 to about 600 per mL, either in suspension or upon reconstitution.

[0184] In other embodiments, after dissolution or reconstitution of the particles following storage, SvP is present in an amount of about 0 to about 100,000,000 per mL, e.g., about 0 to about 10,000,000 per mL, about 0 to about 1,000,000 per mL, about 0 to about 500,000 per mL, about 0 to about 100,000 per mL, about 0 to about 50,000 per mL, about 0 to about 10,000 per mL, about 0 to about 6,000 per mL, about 0 to about 1,000 per mL, about 0 to about 600 per mL, about 0 to about 250 per mL, about 0 to about 100 per mL, about 0 to about 60 per mL, or about 0 to about 10 per mL. In some embodiments, the number of particles having a characteristic size of about 10 μm or greater is about 0 to about 6,000 per mL, e.g., about 0 to about 1,000 per mL, about 0 to about 100 per mL, about 0 to about 10 per mL, about 0 to about 5 per mL, about 0 to about 3 per mL, or about 0 to about 1 per mL. In certain embodiments, the number of particles having a characteristic size of about 25 μm or greater is about 0 to about 600 per mL, e.g., about 0 to about 100 per mL, about 0 to about 10 per mL, about 0 to about 3 per mL, about 0 to about 1 per mL, about 0 to about 0.5 per mL, or about 0 to about 0.1 per mL. In still other embodiments, after dissolution or reconstitution of the particles following storage, the therapeutic or diagnostic agent retains about 0.5 to about 1.0 activity, e.g., about 0.75 to about 1.0 activity, about 0.9 to about 1.0 activity, about 0.95 to about 1.0 activity, about 0.99 to about 1.0 activity, or about 0.999 to about 1.0 activity. In certain other embodiments, dissolution or reconstitution of the particles following storage achieves less than about a 10% increase in agent, e.g., protein, aggregates (e.g., less than about 8%, less than about 5%, less than about 4%, less than about 3%, less than about 1%, less than about 0.5%, or less than about 0.1%) compared to the agent in the first liquid before processing. In certain embodiments, dissolution or reconstitution of the particles after storage achieves less than about a 10% increase in fragments of the agent, e.g., protein, compared to the therapeutic or diagnostic agent in the first liquid before processing (e.g., less than about 8%, less than about 5%, less than about 4%, less than about 3%, less than about 1%, less than about 0.5%, or less than about 0.1%).In some embodiments, dissolution or reconstitution of the particles following storage achieves less than about a 50% (e.g., less than about 40%, less than 30%, less than 20%, less than 10%, less than 8%, less than 5%, less than 4%, less than 3%, or less than about 1%) change in charge variants in the population of agents, e.g., antibodies or antibody fragments, compared to the therapeutic or diagnostic agent prior to particle formation.

[0185] In still other embodiments, after dissolution or reconstitution of the particles following storage, SvP is present in an amount of about 0 to about 100,000,000 per mL, e.g., about 0 to about 10,000,000 per mL, about 0 to about 1,000,000 per mL, about 0 to about 500,000 per mL, about 0 to about 100,000 per mL, about 0 to about 50,000 per mL, about 0 to about 10,000 per mL, about 0 to about 6,000 per mL, about 0 to about 1,000 per mL, about 0 to about 600 per mL, about 0 to about 250 per mL, about 0 to about 100 per mL, about 0 to about 60 per mL, or about 0 to about 10 per mL. In certain embodiments, the number of particles having a characteristic size of about 10 μm or more is about 0 to about 6,000 per mL, e.g., about 0 to about 1,000 per mL, about 0 to about 100 per mL, about 0 to about 10 per mL, about 0 to about 5 per mL, about 0 to about 3 per mL, or about 0 to about 1 per mL. In certain other embodiments, the number of particles having a characteristic size of about 25 μm or more is about 0 to about 600 per mL, e.g., about 0 to about 100 per mL, about 0 to about 10 per mL, about 0 to about 3 per mL, about 0 to about 1 per mL, about 0 to about 0.5 per mL, or about 0 to about 0.1 per mL. In some embodiments, dissolution or reconstitution of the particles following storage provides less than about a 10% (e.g., less than about 8%, less than about 5%, less than about 4%, less than about 3%, less than about 1%, less than about 0.5%, or less than about 0.1%) increase in diagnostic or therapeutic agent, e.g., protein aggregates, compared to the therapeutic or diagnostic agent in the first liquid before processing. In other embodiments, dissolution or reconstitution of the particles after storage provides less than about a 10% (e.g., less than about 8%, less than about 5%, less than about 4%, less than about 3%, less than about 1%, less than about 0.5%, or less than about 0.1%) increase in diagnostic or therapeutic agent, e.g., protein fragments, compared to the therapeutic or diagnostic agent in the first liquid before processing.In certain other embodiments, dissolution or reconstitution of the particles following storage achieves less than a 50% (e.g., less than about 40%, less than about 30%, less than about 20%, less than about 10%, less than about 8%, less than about 5%, less than about 4%, less than about 3%, or less than about 1%) change in charge variants in the population of diagnostic or therapeutic agents, e.g., antibodies or antibody fragments, compared to the therapeutic or diagnostic agents prior to particle formation.

[0186] In certain embodiments, particles of the present disclosure can be suspended in an aqueous liquid, an organic liquid, an ionic liquid, a gel, or a combination thereof to form a suspension formulation. The vehicle for the suspension can further include, for example, carbohydrates, pH adjusters, salts, chelating agents, minerals, polymers, surfactants, amino acids, oligopeptides, biological additives, chemical additives, preservatives, antioxidants, parabens, bactericides, fungicides, vitamins, preservatives, analgesics, and / or nutrient media. In some embodiments, each of the other components independently accounts for about 0.0001 to about 99% (w / v) of the vehicle, e.g., about 0.0001 to about 90% (w / v), about 0.0001 to about 50% (w / v), about 0.0001 to about 10% (w / v), about 0.0001 to about 1% (w / v), or about 0.0001 to about 0.1% (w / v).

[0187] For aqueous suspension formulations, highly concentrated trehalose solutions can stabilize particles in suspension and prevent premature dissolution. The sugar acts as a steric stabilizer when adsorbed onto the particle surface, but when non-adsorbed, it can also act as a "crowder" molecule. Crowder molecules may function by enhancing depletion repulsion. This stabilizing effect has also been described for other crowding agents in water, such as (i) polymers, such as PEG200, PEG300, PEG3350, PEG8000, PEG10000, PEG20000, poloxamers, polyvinylpyrrolidone, polyacrylic acid, poly(vinyl) polymers, polyesters, polyaldehydes, tertiary polymers, polyamino acids, and hydroxyethyl starch (note that these can be used alone or in combination); (ii) organic molecules, such as N-methyl-2-pyrrolidone (Miller et al. J. Pharm. Sci., 2012, 101, 3763-3778); and (iii) sugars and sugar alcohols, such as sorbitol, sucrose, and mannitol, among others. Other "crowding agents" include salts, such as ammonium sulfate, which can compete for water of hydration, and water-soluble organic liquids, such as N-methylpyrrolidone (NMP), which lowers the solvent's dielectric constant and creates an excluded volume effect. In a preferred embodiment, the crowding agent is PEG3350, dextran 40k or dextran 6k.

[0188] In some embodiments, surfactants in liquid suspensions (either aqueous or non-aqueous) act as charge stabilizers. The surfactants adsorb onto the surface of the particles and control the electrostatic interactions between them. The repulsive electrostatic forces generated by the addition of surfactants to the suspension are, in some embodiments, sufficient to prevent significant aggregation of the particles. The surfactants can also prevent adhesion to containers. In other embodiments, polymers can be added to the liquid suspension to act as steric stabilizers.

[0189] In certain embodiments, the therapeutic or diagnostic agent has about 0.5 to about 1.0 unit activity, for example, about 0.75 to about 1.0 unit activity or about 0.9 to about 1.0 unit activity (eg, about 0.99 unit activity).

[0190] In certain preferred embodiments, the present disclosure as described herein relates to highly concentrated compositions comprising a plurality of particles comprising at least one therapeutic biological agent suspended in a low-viscosity pharmaceutically acceptable liquid carrier, wherein, upon dissolution in water, buffer, or other physiologically suitable aqueous liquid, e.g., a biological fluid within a patient's body, the composition has substantially similar turbidity compared to a similar aqueous composition comprising a monomeric therapeutic biological agent. The term "turbidity" refers to the cloudiness or haze of a fluid caused by individual particles that remain insoluble after dissolution in water, buffer, or other physiologically suitable aqueous liquid, e.g., a biological fluid within a patient's body, at a desired concentration. As used herein, "physiologically suitable" conditions such as those that may be encountered in a mammal or human can be applied. One of skill in the art can determine the most appropriate set of conditions to test according to the ultimate use of the compositions described herein. In some embodiments, upon dissolution in an aqueous liquid, the composition has substantially similar turbidity compared to an aqueous composition comprising a monomeric therapeutic biological agent. In preferred embodiments, upon dissolution in an aqueous liquid, the composition is substantially free of turbidity. In certain preferred embodiments, the aqueous liquid is water, an aqueous buffer solution, or a physiologically suitable aqueous liquid.

[0191] In some embodiments, particles of the present disclosure can be reconstituted into a liquid pharmaceutical composition and assessed for turbidity or cloudiness (USP <855> Turbidity may be measured in units of FTU (Formazin Turbidity Units). This is accomplished by comparing the turbidity of a sample to the turbidity of a formazin suspension. Turbidity may also be measured in Nephelometric Turbidity Units (NTU), where 1 NTU = 1 FTU. In other embodiments, when 10 mg of particles are dissolved in 1 mL of liquid, the turbidity may be about 0 to about 4000 FTU, about 0 to about 1000 FTU, about 0 to about 500 FTU, about 0 to about 50 FTU, about 0 to about 20 FTU, about 0 to about 10 FTU, about 0 to about 5 FTU, about 0 to about 1 FTU, about 0 to about 0.1 FTU, or about 0 to about 0.01 FTU. In certain embodiments, the composition has a turbidity of about 0 to about 4000 Formazin Turbidity Units (FTU). In certain other embodiments, upon dissolution in an aqueous liquid, the composition has substantially similar turbidity as compared to an aqueous composition comprising a therapeutic biological agent in monomeric form. In preferred embodiments, upon dissolution in an aqueous liquid, the composition is substantially free of turbidity. In certain preferred embodiments, the aqueous liquid is water, an aqueous buffer, or a physiologically suitable aqueous liquid.

[0192] In other embodiments, the present disclosure relates to a highly concentrated composition with low turbidity comprising a plurality of particles comprising a carbohydrate, a pH adjuster, a salt, a surfactant, a protein stabilizer, an emulsifier, an amino acid, and a therapeutic biological agent in a non-aqueous liquid carrier. In a preferred embodiment, the present disclosure relates to a highly concentrated composition with low turbidity comprising a plurality of particles comprising trehalose, arginine hydrochloride, sodium succinate, succinic acid, citric acid, sodium citrate, histidine, histidine hydrochloride, sodium chloride, hydroxypropyl beta-cyclodextrin, polysorbate, polysorbate 80, or sorbitan monooleate, and an antibody in ethyl oleate. In certain preferred embodiments, the composition is substantially free of turbidity upon dissolution in water, an aqueous buffer, or any physiologically suitable aqueous liquid.

[0193] Compositions comprising a plurality of particles comprising at least one therapeutic biological agent as described herein can be prepared in many ways and by any method of forming particles as disclosed in, for example, International Application No. PCT / US2017 / 063150, International Application No. PCT / US2018 / 043774, International Application No. PCT / US2019 / 033875, and U.S. Patent Application No. 62 / 799,696, each of which is incorporated herein by reference in its entirety.

[0194] Methods of the present disclosure Methods described herein generally provide for forming particles, the methods including: a) providing a droplet comprising a first liquid and an agent; b) contacting the droplet with a second liquid; c) drying the droplet; and d) removing the first and second liquids, thereby forming particles comprising the agent, wherein the particles comprise less than about 25% internal void space and have a circularity of about 0.10 to about 1.00 after removing the first and second liquids. As disclosed herein, the agent can be a therapeutic or diagnostic agent. In certain embodiments, the therapeutic agent has a unit activity of about 0.5 to about 1.0. In certain preferred embodiments, the therapeutic agent is a therapeutic biological agent. In preferred embodiments, the therapeutic biological agent has a unit activity of about 0.5 to about 1.0. In other embodiments, the first liquid contains an agent that generates particles for non-therapeutic or non-diagnostic uses.

[0195] In some embodiments, the particles have less than about 25% internal void space after removal of the first and second liquids, e.g., less than about 24%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or less than 0.1% internal void space after removal of the first and second liquids. In certain embodiments, the particles may contain less than 10% internal void space after removal of the first and second liquids, less than 5% internal void space after removal of the first and second liquids, less than 1% internal void space after removal of the first and second liquids, less than 0.1% internal void space after removal of the first and second liquids, or less than 0.01% internal void space after removal of the first and second liquids. In preferred embodiments, the particles are substantially free of any internal void space after removal of the first and second liquids.

[0196] In other embodiments, the circularity of the particles is at least about 10% after the first and second liquids are removed, e.g., at least about 20, 30, 40, 50, 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or about 100% after the first and second liquids are removed. In some embodiments, the circularity of the particles is at least about 88% after the first and second liquids are removed. In certain embodiments, the circularity of the particles is at least about 90% after the first and second liquids are removed. In still other embodiments, the circularity of the particles is at least about 93% after the first and second liquids are removed. In preferred embodiments, the circularity of the particles is at least about 97% after the first and second liquids are removed.

[0197] In some embodiments, the circularity of the particles after the first and second liquids are removed is about 0.10 to about 1.00, e.g., about 0.20, 0.30, 0.40, 0.50, 0.60, 0.70, 0.75, 0.80, 0.81, 0.82, 0.83, 0.84, 0.85, 0.86, 0.87, 0.88, 0.89, 0.90, 0.91, 0.92, 0.93, 0.94, 0.95, 0.96, 0.97, 0.98, or 0.99 to about 1.00 after the first and second liquids are removed. In certain embodiments, the circularity of the particles after the first and second liquids are removed is about 0.88 to about 1.00. In still other embodiments, the circularity of the particles is from about 0.90 to about 1.00 after the first and second liquids have been removed. In certain other embodiments, the circularity of the particles is from about 0.93 to about 1.00 after the first and second liquids have been removed. In preferred embodiments, the circularity of the particles is from about 0.97 to about 1.00 after the first and second liquids have been removed.

[0198] In other embodiments, the sphericity of the particles is at least about 50% after the first and second liquids are removed, e.g., at least about 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or about 100% after the first and second liquids are removed. In some embodiments, the sphericity of the particles after removal of the first and second liquids is about 0.10 to about 1.00, e.g., about 0.20, 0.30, 0.40, 0.50, 0.60, 0.70, 0.75, 0.80, 0.81, 0.82, 0.83, 0.84, 0.85, 0.86, 0.87, 0.88, 0.89, 0.90, 0.91, 0.92, 0.93, 0.94, 0.95, 0.96, 0.97, 0.98, or 0.99 to about 1.00 after removal of the first and second liquids. In a preferred embodiment, the sphericity of the particles after removal of the first and second liquids is about 1.00.

[0199] In certain embodiments, the sphericity of the particles can range from about 0.10 to about 1.00 after removing the first and second liquids, e.g., at least about 0.20, about 0.40, about 0.60, or about 0.80 to about 1.00 after removing the first and second liquids.

[0200] In a preferred embodiment, the particles have a substantially smooth surface after the first and second liquids have been removed.

[0201] In some embodiments, the particles have a diameter of about 0.1 to about 1000 μm after the first and second liquids are removed, for example, about 0.1 to about 900, 800, 700, 600, 500, 400, 300, 200, 100, 90, 80, 70, 60, 50, 45, 40, 35, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3 or about 0.2 μm after the first and second liquids are removed. In certain embodiments, the particles have a diameter of about 1 to about 100 μm after the first and second liquids are removed, e.g., about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, or 50 to about 100 μm after the first and second liquids are removed. In still other embodiments, the particles have a diameter of about 4 to about 100 μm after the first and second liquids are removed. In certain other embodiments, the particles have a diameter of about 10 to about 100 μm after the first and second liquids are removed. In preferred embodiments, the particles have a diameter of about 20 to about 50 μm after the first and second liquids are removed. In certain preferred embodiments, the diameter of the particles is intentionally controlled. In some embodiments, the particles have a diameter of about 0.1 to about 1000 μm after the first and second liquids are removed, e.g., about 1 to about 400 μm, about 1 to about 200 μm, about 1 to about 100 μm, about 1 to about 50 μm, about 1 to about 25 μm, about 1 to about 10 μm, about 10 to about 100 μm, about 50 to about 100 μm, about 50 to about 75 μm, or about 75 to about 100 μm after the first and second liquids are removed. In other embodiments, the particles have a diameter of about 1 to about 100 μm after the first and second liquids are removed, e.g., about 4 to about 100 μm, about 10 to about 100 μm, or about 20 to about 50 μm after the first and second liquids are removed.

[0202] In other embodiments, the particles may comprise less than about 10% by weight after the first and second liquids have been removed, e.g., less than about 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%, or less than 0.1% by weight after the first and second liquids have been removed. , less than 0.09%, less than 0.08%, less than 0.07%, less than 0.06%, less than 0.05%, less than 0.04%, less than 0.03%, less than 0.02%, less than 0.01%, less than 0.009%, less than 0.008%, less than 0.007%, less than 0.006%, less than 0.005%, less than 0.004%, less than 0.003%, less than 0.002%, or less than 0.001% by weight. In some embodiments, the particles have a surfactant content of less than about 5% by weight after the first and second liquids have been removed. In certain embodiments, the particles have a surfactant content of less than about 3% by weight after the first and second liquids have been removed. In still other embodiments, the particles have a surfactant content of less than about 0.1% by weight after the first and second liquids have been removed. In certain other embodiments, the particles have a surfactant content of less than about 0.01% by weight after the first and second liquids have been removed. In some embodiments, the particles have a surfactant content of less than about 0.001% by weight after the first and second liquids have been removed. In preferred embodiments, the particles have a surfactant content of less than about 1% by weight after the first and second liquids have been removed. In certain preferred embodiments, the particles are substantially free of any surfactant content after the first and second liquids have been removed.

[0203] In some embodiments, the surfactant content of the particles is 0 to 10% by weight after the first and second liquids are removed, e.g., 0 to 5%, 0 to 3%, 0 to 2%, 0 to 1%, 0 to 0.5%, 0 to 0.2%, 0 to 0.1%, 0 to 0.01% or 0 to 0.001% by weight after the first and second liquids are removed.

[0204] In other embodiments, the particles have a density of about 1.00 to about 6.00 g / cm after the first and second liquids are removed. 3 After removing the first and second liquids, the concentration is, for example, about 1.00 to about 5.00 g / cm 3 , about 1.00~3.00g / cm 3 , about 1.00~2.00g / cm 3 , about 1.00~1.50g / cm 3 , about 1.30~1.50g / cm 3 , about 1.32~about 1.50g / cm 3 or about 1.10 to about 1.40 g / cm 3 In some embodiments, the particles have a skeletal density of about 0.10 to about 5.00 g / cm after the first and second liquids are removed. 3 After removing the first and second liquids, the concentration is, for example, about 0.10 to about 2.50 g / cm 3 , about 0.10~about 1.40g / cm 3 , about 0.50~1.40g / cm 3 or about 1.00 to about 1.40 g / cm 3 In certain embodiments, the particles exhibit a skeletal density of about 0.09 to about 1.60 g / cm after the first and second liquids are removed. 3 In yet another embodiment, the particles have a skeletal density of about 1.30 to about 1.58 g / cm after the first and second liquids are removed. 3 In a preferred embodiment, the particles have a skeletal density of about 1.32 to about 1.50 g / cm after the first and second liquids are removed. 3 It has a skeletal density of

[0205] In certain embodiments, the particles have a concentration of about 1000 mg / mL to about 1500 mg / mL after the first and second liquids are removed, e.g., about 1050 mg / mL to about 1500 mg / mL, about 1100 mg / mL to about 1500 mg / mL, about 1150 mg / mL to about 1500 mg / mL, about 1200 mg / mL to about 1500 mg / mL, about 1250 mg / mL to about 1500 mg / mL, about 1300 mg / mL to about 1500 mg / mL, about 1310 mg / mL to about 1500 mg / mL, about 1320 mg / mL to about 1500 mg / mL, about 1330 mg / mL to about 1500 mg / mL, about 1340 mg / mL to about 1500 mg / mL, about 1350 mg / mL to about 1500 mg / mL, about 1 360mg / mL ~ approx. 1500mg / mL, approx. 1370mg / mL ~ approx. 1500mg / mL, approx. 1380mg / mL ~ approx. 1500mg / mL, approx. 1390mg / mL ~ approx. 150 0mg / mL, about 1400mg / mL to about 1500mg / mL, about 1410mg / mL to about 1500mg / mL, about 1420mg / mL to about 1500mg / mL, about 1430m The compound has a skeletal density of about 1440 mg / mL to about 1500 mg / mL, about 1450 mg / mL to about 1500 mg / mL, about 1460 mg / mL to about 1500 mg / mL, about 1470 mg / mL to about 1500 mg / mL, about 1480 mg / mL to about 1500 mg / mL, or about 1490 mg / mL to about 1500 mg / mL.

[0206] In some embodiments, the particles can be characterized by a glass transition temperature of about 0°C to 250°C after removal of the first and second liquids, e.g., about 34°C to 200°C, about 50°C to 200°C, about 60°C to 200°C, about 40°C to about 160°C, about 50°C to about 110°C, about 60°C to about 100°C, or about 75°C to about 80°C after removal of the first and second liquids. In other embodiments, the particles have a glass transition temperature of about 40°C to about 160°C after removal of the first and second liquids. In still other embodiments, the particles have a glass transition temperature of about 50°C to about 110°C after removal of the first and second liquids. In certain embodiments, the particles have a glass transition temperature of about 60°C to about 100°C after removal of the first and second liquids. In preferred embodiments, the particles have a glass transition temperature of about 75°C to about 80°C after removal of the first and second liquids. In still other embodiments, the particles are heated to about ±30° C., such as about ±20, ±10, ±5, ±1° C., of the glass transition temperature of the particles during drying.

[0207] In certain embodiments, the particles have a glass transition temperature of greater than about 160° C. after the first and second liquids are removed. In certain other embodiments, the particles have a glass transition temperature of greater than about 90° C. after the first and second liquids are removed. In certain preferred embodiments, the particles have a glass transition temperature of greater than about 50° C. after the first and second liquids are removed.

[0208] In other embodiments, the particles further comprise a carbohydrate, a pH adjuster, a salt, a chelating agent, a mineral, a polymer, a surfactant, a protein stabilizer, an emulsifier, a preservative, an amino acid, an antioxidant, a protein, an organic solvent, a paraben, a bactericide, a fungicide, a vitamin, a preservative, a nutrient medium, an oligopeptide, a biological additive, a chemical additive, or a combination thereof. In certain embodiments, the particles further comprise a carbohydrate, a pH adjuster, a salt, a surfactant, a protein stabilizer, an emulsifier, an amino acid, or a combination thereof.

[0209] In some embodiments, the particles have less than 20% aggregates or less than 20% fragments of the therapeutic biological agent after the first and second liquids are removed, e.g., less than about 19%, less than 18%, less than 17%, less than 16%, less than 15%, less than 14%, less than 13%, less than 12%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1.9%, less than 1.8%, less than 1.7%, less than 1.6%, less than 1.5%, less than 1.4%, less than 1.3%, less than 1.2%, less than 1.1%, less than 1%, less than 0.9%, less than 0.8%, less than 0.7%, less than 0.6%, less than 0.5%, less than 0.4%, less than 0.3%, less than 0.2%, or less than 0.1% after the first and second liquids are removed. In other embodiments, the particles have less than 10% aggregates or less than 10% fragments of the therapeutic biological agent after the first and second liquids are removed, e.g., less than about 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1.9%, 1.8%, 1.7%, 1.6%, 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%, or 0.1% after the first and second liquids are removed. In certain embodiments, the particles have between about 3% and about 1% aggregates of the therapeutic biological agent after the first and second liquids are removed. In certain other embodiments, the particles have about 1% to about 0.5% aggregates of the therapeutic biological agent after the first and second liquids are removed. In preferred embodiments, the particles are substantially free of any aggregates of the therapeutic biological agent after the first and second liquids are removed. In still other embodiments, the particles have less than about 1% fragments of the therapeutic biological agent after the first and second liquids are removed. In certain preferred embodiments, the particles are substantially free of any fragments of the therapeutic biological agent after the first and second liquids are removed.

[0210] In other embodiments, the particle formation process achieves less than a 50% change in charge variants in the population of diagnostic or therapeutic agents, e.g., antibodies or antibody fragments, after removing the first and second liquids, compared to the therapeutic or diagnostic agent before particle formation (e.g., less than 40%, less than 30%, less than 20%, less than 10%, less than 8%, less than 5%, less than 4%, less than 3%, or less than 1% after removing the first and second liquids). In certain embodiments, the particles have less than about 50% change in charge variant of the therapeutic biological agent after removal of the first and second liquids compared to the starting biological agent before particle formation, e.g., less than about 45%, less than 40%, less than 35%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1% change in charge variant of the therapeutic biological agent after removal of the first and second liquids compared to the starting biological agent before particle formation. In preferred embodiments, the particles have substantially no change in charge variant of the therapeutic biological agent after removal of the first and second liquids compared to the starting biological agent before particle formation.

[0211] In some embodiments, the particles comprise less than about 3% by weight of the remaining first and second liquids remaining after removing the first and second liquids, e.g., less than about 2%, less than 1.9%, less than 1.8%, less than 1.7%, less than 1.6%, less than 1.5%, less than 1.4%, less than 1.3%, less than 1.2%, less than 1.1%, less than 1%, less than 0.9%, less than 0.8%, less than 0.7%, less than 0.6%, less than 0.5%, less than 0.4%, %, less than 0.3 wt.%, less than 0.2 wt.%, less than 0.1 wt.%, less than 0.09 wt.%, less than 0.08 wt.%, less than 0.07 wt.%, less than 0.06 wt.%, less than 0.05 wt.%, less than 0.04 wt.%, less than 0.03 wt.%, less than 0.02 wt.%, less than 0.01 wt.%, less than 0.009 wt.%, less than 0.008 wt.%, less than 0.007 wt.%, less than 0.006 wt.%, less than 0.005 wt.%, less than 0.004 wt.%, less than 0.003 wt.%, less than 0.002 wt.%, or less than 0.001 wt.%. In other embodiments, the particles have less than about 3 wt.% residual moisture remaining after the first and second liquids have been removed. In yet other embodiments, the particles have less than about 2 wt.% residual moisture remaining after the first and second liquids have been removed. In certain other embodiments, the particles have less than about 1% by weight residual moisture remaining after removal of the first and second liquids. In certain other embodiments, the particles have less than about 0.1% by weight residual first and second liquids remaining after removal of the first and second liquids. In some preferred embodiments, the particles have less than about 0.01% by weight residual first and second liquids remaining after removal of the first and second liquids. In certain preferred embodiments, the particles have less than about 0.001% by weight residual first and second liquids remaining after removal of the first and second liquids. In preferred embodiments, the particles are substantially free of any residual first and second liquids by weight after removal of the first and second liquids.

[0212] In other embodiments, the particles have greater than about 60% by weight of the therapeutic biological agent after the first and second liquids have been removed, e.g., greater than about 65%, greater than 70%, greater than 75%, greater than 80%, greater than 85%, greater than 86%, greater than 87%, greater than 88%, greater than 89%, greater than 90%, greater than 91%, greater than 92%, greater than 93%, greater than 94%, greater than 95%, greater than 96%, greater than 97%, greater than 98%, greater than 99%, greater than 99.1%, greater than 99.2%, greater than 99.3%, greater than 99.4%, greater than 99.5%, greater than 99.6%, greater than 99.7%, greater than 99.8%, greater than 99.9% by weight of the therapeutic biological agent after the first and second liquids have been removed. In some embodiments, the particles have greater than about 90% by weight of the therapeutic biological agent after the first and second liquids have been removed. In certain embodiments, the particles have greater than about 95% by weight of the therapeutic biological agent after the first and second liquids have been removed. In still other embodiments, the particles have greater than about 98% by weight of the therapeutic biological agent after the first and second liquids have been removed. In preferred embodiments, the particles have greater than about 98% by weight of the therapeutic biological agent after the first and second liquids have been removed. In certain preferred embodiments, the particles have greater than about 99% by weight of the therapeutic biological agent after the first and second liquids have been removed.

[0213] As described herein, particles can include both a core and a shell. In some embodiments, particles do not include a shell. In some embodiments, the core is a gel core or a dry solid core when no shell is present, but can exist in a liquid state when the particle includes a gel shell or a dry solid shell. In other embodiments, the particle morphology is generally spherical, mushroom-like, or raisin-like, potentially among other morphologies, depending on the conditions of particle formation. In certain embodiments, the particle surface can be wrinkled or jagged. When particles with a core-shell structure are used, the individual layers can contain the same or different agents, such as therapeutic or diagnostic agents, or no agents at all. Furthermore, layers having the same agent, such as a therapeutic or diagnostic agent, may or may not contain the same concentration of the agent.

[0214] In some embodiments, the residual amount of the first liquid in the particles after dehydration is about 0 to about 10% by weight, e.g., about 0 to about 5% by weight, about 0 to about 3% by weight, about 0 to about 1% by weight, about 0.01 to about 5% by weight, about 0.01 to about 3% by weight, or about 0.01 to about 1% by weight. Exemplary methods for measuring the residual solvent content include Karl Fischer titration, headspace gas chromatography mass spectrometry, and various weight loss methods. In other embodiments, the residual amount of the second liquid in the particles after dehydration is about 0 to about 10% by weight, e.g., about 0 to about 5% by weight, about 0 to about 3% by weight, about 0 to about 1% by weight, about 0.01 to about 5% by weight, about 0.01 to about 3% by weight, or about 0.01 to about 1% by weight. Exemplary methods for measuring the residual solvent content include Karl Fischer titration, headspace gas chromatography mass spectrometry, and various weight loss methods. In certain embodiments, the residual amount of one or more shell liquids in the particles after dehydration is about 0 to about 10 wt%, e.g., about 0 to about 5 wt%, about 0 to about 3 wt%, or about 0 to about 1 wt%. Exemplary methods for measuring residual solvent content include Karl Fischer titration, headspace gas chromatography-mass spectrometry, and various weight loss methods.

[0215] In other embodiments, the particles have a residual net charge of either polarity, i.e., a net positive charge or a net negative charge. In terms of size, the particles can have from about 0 to about 10 billion charges, e.g., from about 0 to about 100 million charges, from about 0 to about 1 million charges, from about 0 to about 10,000 charges, or from about 0 to about 100 charges. The magnitude of the charge is the magnitude of the charge carried by an electron, i.e., the elementary charge, 1.6×10 -19The charge is defined as a coulomb. Exemplary methods for measuring particle charge include those involving analysis of particle motion, e.g., electrical mobility, in response to an externally applied electric field. In some embodiments, measurements can be performed while the particles are suspended in an insulating liquid, e.g., oil. In certain embodiments, the therapeutic or diagnostic agent has a zeta potential of about −90 to about 90 mV; e.g., about −60 to about 60 mV, about −40 to about 40 mV, about −20 to about 20 mV, or about −5 to about 5 mV. An exemplary method for measuring zeta potential involves reconstituting the therapeutic or diagnostic agent by dissolving the particles in water and analyzing the solution by electrophoretic light scattering, similar to dynamic light scattering (DLS) measurements performed in the presence of a positive or negative electric field.

[0216] In certain embodiments, the major component of the particle, e.g., the agent, is characterized by a Peclet of from about 0 to about 10, e.g., from about 0 to about 9, from about 0 to about 8, from about 0 to about 7, from about 0 to about 6, from about 0 to about 5, from about 0 to about 4, from about 0 to about 3, from about 0 to about 2, from about 0 to about 1, from about 0 to about 0.5, from about 0 to about 0.25, or from about 0 to about 0.1 during the particle formation process. In certain other embodiments, the major component of the particle, e.g., the agent, is characterized by a Peclet of from about 0 to about 10,000 μm during the particle formation process. 2 / s, for example, about 0 to about 1,000 μm 2 / s, about 0 to about 100μm 2 / s, about 0 to about 50μm 2 / s, about 0 to about 25μm 2 / s, about 0 to about 10μm 2 / s, about 0 to about 5μm 2 / s, about 0 to about 2.5μm 2 / s or about 0 to about 1 μm 2 The diffusion coefficient can be characterized by the average diffusivity of the agent in s / s.

[0217] In some embodiments, the particles may be flowable. The Hausner ratio may be about 1.0 to greater than about 3.0, such as about 1.0 to about 3.0, about 1.0 to about 2.0, about 1.0 to about 1.70 (e.g., very poor), about 1.0 to about 1.59, about 1.0 to about 1.35, about 1.0 to about 1.25, or about 1.0 to about 1.11 (e.g., excellent). An exemplary method for measuring powder flowability includes the tapped density method (Carr RLChem.Eng., 1965;72:163-168). Bulk density can be first obtained by adding a known mass of powder to a graduated cylinder. The density can be calculated as mass / volume. The same sample can then be mechanically tapped until no further volume change is observed. The tapped density can then be calculated as mass divided by the final volume of the powder. A comparison of tapped density and bulk density can be used to indicate the ability of a powder to flow. In other embodiments, the Hausner ratio (unsettled apparent volume or bulk volume, V) is used to calculate the final tap volume, V f The density (divided by ) is a measure of the ability of a product to settle and allows for an assessment of the relative importance of particle-particle interactions. These interactions are less significant in free-flowing powders. The bulk density and tapped density for such free-flowing powders are close in value, and the Hausner ratio is close to about 1.0.

[0218] In other embodiments, the particles have one or more of the following characteristics: a size of about 1 to about 50 μm; a solid core; a gel or solid shell; and a density of about 1 to about 1.5 g / cm. 3 a density of about 0 to about 5 wt% residual solvent content; a porosity of about 0 to about 10%; a net charge of either polarity, i.e., positive or negative, from about 0 to about 1 million charges; a therapeutic or diagnostic component having a zeta potential of about -60 to about 60 mV; an SvP of about 0 to about 1,000,000 per mL upon reconstitution; a therapeutic or diagnostic agent loading of about 50 to about 100 wt% where the activity of the therapeutic or diagnostic agent is about 0.9 to about 1.0 upon reconstitution; a surfactant loading of about 0 to about 3 wt%; a major component, e.g., an active agent (the Peclet number was about 1 or less during the particle formation process); a major component, e.g., an active agent (the diffusivity was about 500 μm during the particle formation process). 2 / s or less); less than about 10% aggregates upon reconstitution; less than about 10% fragments upon reconstitution; and / or a Hausner ratio of about 1.0 to about 1.35 or about 1.0 to about 1.11.

[0219] The term "core-shell morphology" refers to a morphology having multiple layers containing different components and / or concentrations of components. The "dry" particle component containing the agent, i.e., the dry core or dry shell, is subjected to a dehydration step or series of dehydration steps such that its moisture content or solvent content is substantially reduced relative to the moisture content or solvent content prior to dehydration. As described herein, particles can have a core-shell morphology, where the shell can comprise multiple layers. In certain embodiments, the core is a solid, gel, or liquid. In some embodiments, the shell is a gel, particularly a hydrogel, ionogel, or organogel. In other embodiments, the shell is crystalline or semi-crystalline. In preferred embodiments, the particles have a morphology comprising a core and a shell.

[0220] In some embodiments, the particles have a morphology comprising a core and a shell, wherein the shell may comprise multiple layers. In certain embodiments, the core is a solid, gel, or liquid. In some embodiments, the shell is a gel, particularly a hydrogel, ionogel, or organogel. Exemplary hydrogels, ionogels, and organogels include collagen hydrogels, chitosan hydrogels, methylcellulose hydrogels, dextran hydrogels, alginate hydrogels, agarose hydrogels, poly(methyl methacrylate) hydrogels, poly(amidoamine) hydrogels, poly(ethyleneimine) hydrogels, polyethylene oxide hydrogels, gelatin hydrogels, hyaluronic acid hydrogels, 4-tert-butyl-1-arylcyclohexanol organogels, L-lysine derivative organogels, poly(ethylene glycol) organogels, polycarbonate organogels, polyester organogels, polyalkene organogels, oxalylamide derivative organogels, or combinations thereof.

[0221] Particle core: The core of each particle typically contains one or more therapeutic or diagnostic agents. When no shell is present, the core is a solid dry core or gel, but when the particle contains a gel shell or a solid dry shell, it can exist in a liquid state. When a shell is present, the shell may contain a therapeutic or diagnostic agent, while the core may not.

[0222] Particle Shell: Generally, any additive is suitable as a shell material. Exemplary additives include, but are not limited to, sugars, salts, and amino acids. Therapeutic agents, diagnostic agents, and biocompatible polymers can also be used to form the shell. This includes small molecule drugs. Non-limiting examples of hydrophilic biocompatible polymers include poly(vinyl alcohol), poly(acrylic acid), poly(acrylamide), poly(ethylene oxide), or copolymers or combinations of any two or more thereof. Hydrophilic polymers can be modified to adjust their characteristics. The shell component can alternatively or additionally include one or more biocompatible hydrophobic polymers. Hydrophobic polymers can be modified to adjust their characteristics. Non-limiting examples of hydrophobic polymers include polycaprolactam, poly(lactic acid), poly(glycolic acid), polycaprolactone, PLGA, or copolymers, or combinations of any two or more thereof. In some embodiments, PLGA (50:50) polymer is used as a shell to encapsulate a therapeutic agent, such as an antibody or antibody fragment, in an amount just below its solubility limit. The polymer can also be prepared with PLGA in various lactic acid-glycolic acid ratios, and can be copolymers with other polymers, such as chitosan, cellulose, etc.

[0223] In some embodiments, the thickness of the particle shell can range from about 0 to about 90% of the particle diameter. The shell does not need to be uniform or perfectly formed for encapsulation. In other embodiments, the interface between the shell and the core is partially blended such that there is no clear line of demarcation. One or more therapeutic or diagnostic agents can be included in the particle shell, as described herein. In still other embodiments, the therapeutic or diagnostic agent can be the same or different as in the core. In certain other embodiments, the concentration of the therapeutic or diagnostic agent in the shell can range from about 0.0001 to about 2000 mg / mL (or the crystalline density of the therapeutic or diagnostic agent, if higher).

[0224] Core-shell ratio: For those embodiments in which the particles contain a shell, a core-shell volume ratio of about 1:99% to about 99:1% by volume, e.g., about 10:90% by volume, about 90:10% by volume, or about 95:5% by volume, is expected to be most useful. Complete coverage is not necessary for sufficient encapsulation. In certain embodiments, a thick shell may be advantageous, for example, for a highly concentrated core. In some embodiments, the core-shell ratio may be useful in modulating the release kinetics of a therapeutic or diagnostic agent. In other embodiments, it may be advantageous to have a polydisperse system, for example, to reduce the viscosity of a pharmaceutical suspension formulation containing the particles. In this case, various core-shell ratios may be important.

[0225] droplet Droplets as described herein can be formed by any of several techniques known in the art. These include rotary atomization, pneumatic atomization, ultrasonic atomization, acoustic atomization, vibrating mesh atomization, jet atomization, microfluidic droplet generation, flow focusing, membrane emulsification, electrospray, or homogenization. The term "droplets" or "droplets" refers to a material having a liquid outer surface. In certain embodiments, the droplets of step a) are formed by electrospray, an ultrasonic atomizer, or a microfluidic device. In preferred embodiments, the droplets of step a) are formed in a microfluidic device. In certain preferred embodiments, the droplets formed in the microfluidic device are regularly spaced in the microfluidic device.

[0226] The term "feed solution" refers to a preparation of a therapeutic or diagnostic agent in a first liquid as a solution, slurry, or some other liquid form. In some embodiments, the preparation contains an additive. In other embodiments, the preparation further contains a buffer.

[0227] In some embodiments, the first liquid is aqueous, an organic solvent, an ionic liquid, a hydrogel, an ionogel, or a combination thereof. In other embodiments, the first liquid is aqueous. In certain embodiments, the first liquid is water, 0.9% saline, lactated Ringer's solution, buffer, dextrose 5%, or a combination thereof. In certain other embodiments, the buffer is acetate buffer, histidine buffer, succinate buffer, HEPES buffer, Tris buffer, carbonate buffer, citrate buffer, phosphate buffer, phosphate buffered saline, glycine buffer, barbital buffer, cacodylate buffer, ammonium formate buffer, urea solution, or a combination thereof. In a preferred embodiment, the first liquid is water.

[0228] In other embodiments, the organic liquid is acetone, acetonitrile, acyclic alkanes (e.g., hexane, heptane, pentane), amyl acetate, butanol, butyl acetate, chlorobenzene, chloroform, cumene, cyclohexane, 1,2-dichloroethene, dichloromethane, diethyl ether, dimethoxyethane, dimethylacetamide, dimethylformamide, dimethylsulfoxide, 1,4-dioxane, ethanol, 2-ethoxyethanol, ethyl acetate, ethyl nitrate, ethylene glycol, hydrazine, isopropanol, methanol, methyl acetate, 2 -Methyl-1-butanol, 2-methyl-1-propanol, methyl butyl ketone, methyl cyclohexane, methyl ethyl ketone, methylpyrrolidone, methyl tert-butyl ether, nitromethane, propanol, propyl acetate, sulfolane, propylene glycol, tetrahydrofuran, tetralin, toluene, 1,1,2-trichloroethane, triethylamine, xylene, benzyl benzoate, ethyl lactate, dimethyl isosorbide, dimethyl sulfoxide, glycofurol, diglyme, methyl tert-butyl ether, polyethylene glycol, 2-Pyrrolidone, tetrahydrofurfuryl alcohol, triglycerides, octyl acetate, ethanol, butanol, octanol, decanol, diglyme, tocopherol, octafluoropropane, (perfluorohexyl)octane, n-acetyltryptophan, triglycerides, fractionated vegetable fatty acid C8 and C10 triglycerides, propylene glycol diesters of saturated vegetable fatty acids C8 and C10, ethyl laurate, methyl caprylate, methyl caprate, methyl myristate, methyl oleate, methyl linoleate, dimethyl adipate, Dibutyl benzoate, diethyl sebacate, macadamia nut fatty acid ethyl ester, trimethylolpropane triisostearate, isopropyl laurate, isopropyl myristate, diethyl succinate, polysorbate esters, ethanolamine, propanoic acid, triacetin, citral, anisole, anethole, benzaldehyde, linalool, caprolactone, phenol, thioglycerol, dimethylacetamide, ethyl formate, ethylhexyl acetate, eugenol, clove bud oil, diethyl glycol monoether, dimethyl isosorbide,In another embodiment, the first liquid is an oil, such as isopropyl acetate, methyl isobutyl ketone, methyl tert-butyl ether, N-methylpyrrolidone, perfluorodecalin, 2-pyrrolidone, ethyl oleate, ethyl caprate, dibutyl adipate, hexanoic acid, octanoic acid, diethyl glycol monoether, gamma-butyrolactone, polyoxyl 40 hydrogenated castor oil, polyoxyl 35 castor oil, propylene carbonate, octanol, hexanol, sorbitan monooleate, n-acetyltryptophan, solketal, alkyl acetate, aryl acetate, aryl alkyl acetate, tolyl acetate, benzyl acetate, polysorbate 80, phenethyl acetate, phenyl acetate, glycerol, or a combination thereof. In certain embodiments, the oil is palm oil, cottonseed oil, fish oil, grapeseed oil, hazelnut oil, hydrogenated vegetable oil, lime oil, olive oil, palm kernel oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, sunflower oil, walnut oil, silica oil, mineral oil, or a combination thereof. In yet other embodiments, the first liquid is an ionic liquid. In certain other embodiments, the ionic liquid contains (i) a cation, such as pyridinium, pyridazinium, pyrimidinium, pyrazinium, imidazolium, pyrazolium, thiazolium, oxazolium, triazolium, ammonium, sulfonium; and (ii) anion, such as a halide, sulfate, sulfonate, carbonate, phosphate, bicarbonate, nitrate, acetate, PF6-, BF4-, triflate, nonaflate, bis(triflyl)amide, trifluoroacetate, heptafluorobutanoate, haloaluminate, or a combination thereof.

[0229] In certain embodiments, the first liquid is a hydrogel, an ionogel, or a combination thereof. Exemplary hydrogels are prepared from polymers such as collagen, chitosan, methylcellulose, dextran, alginate, agarose, poly(methyl methacrylate), poly(amidoamine), poly(ethyleneimine), polyethylene oxide, gelatin, hyaluronic acid, or a combination thereof, and may contain water, aqueous solutions, and other polar solvents. Exemplary organogels are prepared from organic gelators such as 4-tert-butyl-1-arylcyclohexanol, L-lysine derivatives, poly(ethylene glycol), polycarbonates, polyesters, polyalkenes, oxalylamide derivatives containing alkyl ester groups, or low-molecular-weight compounds such as fatty acids and n-alkanes, and contain a nonpolar solvent phase. Ionogels are similar to organogels, except that the solvent phase is an ionic liquid.

[0230] In some embodiments, the concentration of the therapeutic agent in the first liquid as described herein is from about 10 mg / mL to about 650 mg / mL, e.g., from about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625 mg / mL to about 650 mg / mL; from about 20 mg / mL to about 625 mg / mL, e.g., from about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625 mg / mL to about 650 mg / mL; 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575, 600 mg / mL to about 625 mg / mL; about 20 mg / mL to about 600 mg / mL, for example, about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575 mg / mL to about 600 mg / mL; about 20 mg / mL to about 575 mg / mL, for example about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550 mg / mL to about 575 mg / mL; about 20 mg / mL to about 550 mg / mL, for example, about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525 mg / mL to about 550 mg / mL; about 2 0 mg / mL to about 525 mg / mL, for example, about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500 mg / mL to about 525 mg / mL; about 20 mg / mL to about 500 mg / mL, for example, about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475 mg / mL to about 500 mg / mL;about 20 mg / mL to about 475 mg / mL, for example, about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450 mg / mL to about 475 mg / mL; about 20 mg / mL to about 450 mg / mL, for example, about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 25 0, 275, 300, 325, 350, 375, 400, 425 mg / mL to about 450 mg / mL; about 20 mg / mL to about 425 mg / mL, for example, about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400 mg / mL to about 425 mg / mL; about 20 mg / mL to about 400 mg / mL, for example, about 20 , 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375 mg / mL to about 400 mg / mL; about 20 mg / mL to about 375 mg / mL, for example about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350 mg / mL to about 375 mg / mL; about 20 mg g / mL to about 350 mg / mL, for example, about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325 mg / mL to about 350 mg / mL; about 20 mg / mL to about 325 mg / mL, for example, about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300 mg / mL to about 325 mg / mL;Alternatively, the concentration of the therapeutic agent in the first liquid may be about 20 mg / mL to about 300 mg / mL, for example, about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275 mg / mL to about 300 mg / mL. In other embodiments, the concentration of the therapeutic agent in the first liquid is about 10 mg / mL to about 500 mg / mL. In certain embodiments, the concentration of the therapeutic agent in the first liquid is about 10 mg / mL to about 100 mg / mL. In a preferred embodiment, the concentration of the therapeutic agent in the first liquid is about 20 mg / mL to about 100 mg / mL. In other embodiments of the present disclosure, the concentration of the therapeutic or diagnostic agent in the first liquid is about 0.0001 to about 1000 mg / mL, e.g., about 100 to about 800, about 200 to about 700, about 200 to about 600, or about 300 to about 700 mg / mL. In still other embodiments, the particles have a mass loading of the therapeutic or diagnostic agent of about 1% to about 100%.

[0231] In other embodiments, the first liquid has a viscosity of less than about 200 mPa·s, less than about 150 mPa·s, less than about 125 mPa·s, less than about 100 mPa·s, less than about 75 mPa·s, less than about 75 mPa·s, less than about 70 mPa·s, less than about 65 mPa·s, less than about 60 mPa·s, less than about 55 mPa·s, less than about 50 mPa·s, less than about 45 mPa·s, less than about 40 mPa·s, less than about 35 mPa·s, less than about 30 mPa·s, less than about 25 mPa·s, less than about 20 mPa·s, less than about 19 mPa·s, less than about 18 mPa·s, less than about 17 mPa·s, less than about 16 mPa·s, less than about 15 mPa·s, less than about 14 mPa·s, less than about 13 mPa·s, less than about 12 mPa·s s, less than about 11 mPa·s, less than about 10 mPa·s, less than about 9.5 mPa·s, less than about 9 mPa·s, less than about 8.5 mPa·s, less than about 8 mPa·s, less than about 7.5 mPa·s, less than about 7 mPa·s, less than about 6.5 mPa·s, less than about 6 mPa·s, less than about 5.5 mPa·s, less than about 5 mPa·s, less than about 4.5 mPa·s, less than about 4 mPa·s, less than about 3.5 mPa·s, less than about 3 mPa·s, less than about 2.5 mPa·s, less than about 2 mPa·s, less than about 1.5 mPa·s, less than about 1 mPa·s, less than about 0.5 mPa·s, less than about 0.1 mPa·s, less than about 0.05 mPa·s or less than about 0.01 mPa·s (1 milliPascal second). In other embodiments, the first liquid has a viscosity of about 0.01 mPa·s to about 10,000 mPa·s, e.g., about 0.01 mPa·s to about 1,000 mPa·s, about 0.01 mPa·s to about 100 mPa·s, about 0.01 mPa·s to about 50 mPa·s, about 0.01 mPa·s to about 25 mPa·s, about 0.01 mPa·s to about 10 mPa·s, about 0.01 mPa·s to about 5 mPa·s, or about 0.01 mPa·s to about 1 mPa·s. In certain embodiments, the first liquid has a viscosity that can range from about 0.27 mPa·s to about 200 mPa·s, such as from about 0.27 mPa·s to about 50 mPa·s, from about 1 mPa·s to about 30 mPa·s, or from about 20 mPa·s to about 50 mPa·s.In yet other embodiments, the first liquid has a viscosity ranging from about 0.27 mPa·s to about 200 mPa·s, e.g., from about 0.27 mPa·s to about 100 mPa·s, from about 0.27 mPa·s to about 50 mPa·s, from about 0.27 mPa·s to about 30 mPa·s, from about 1 mPa·s to about 20 mPa·s, or from about 1 mPa·s to about 15 mPa·s. Methods for controlling viscosity include temperature regulation and viscosity-modifying additives. Mixtures of liquids may also be used to control viscosity.

[0232] In some embodiments, the first liquid has a viscosity of about 0.01 to about 10,000 mPa·s. In other embodiments, the first liquid has a viscosity of less than about 100 mPa·s. In still other embodiments, the first liquid has a viscosity of less than about 10 mPa·s. In certain other embodiments, the first liquid has a viscosity of less than about 3 mPa·s. In certain embodiments, the first liquid has a viscosity of less than about 0.9 mPa·s. In preferred embodiments, the first liquid has a viscosity of less than about 0.5 mPa·s.

[0233] In certain embodiments, the first liquid further comprises a surfactant.

[0234] In some embodiments, the surfactant is polysorbate, magnesium stearate, sodium dodecyl sulfate, TRITON™ N-101, glycerin, polyoxyethylated castor oil, docusate, sodium stearate, decyl glucoside, nonoxynol-9, cetyltrimethylammonium bromide, sodium bis(2-ethylhexyl)sulfosuccinate, lecithin, sorbitan ester, or a combination thereof. In certain embodiments, the surfactant is polysorbate, docusate, or lecithin. In preferred embodiments, the surfactant is polysorbate 20, polysorbate 60, or polysorbate 80. In certain preferred embodiments, the surfactant is polysorbate 20 or polysorbate 80. In certain other embodiments, the fatty acid ester of sorbitol is a sorbitan ester, such as span 20, span 40, span 60, or span 80.

[0235] In other embodiments, the second liquid is aqueous, an organic solvent, an ionic liquid, a hydrogel, an ionogel, a protein stabilizer, or a combination thereof. In some embodiments, the second liquid is aqueous. In preferred embodiments, the second liquid is an organic solvent.

[0236] In some embodiments, the organic solvent is benzyl alcohol, benzyl benzoate, castor oil, coconut oil, corn oil, cottonseed oil, fish oil, grapeseed oil, hazelnut oil, hydrogenated palm kernel oil, olive oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, sunflower oil, vegetable oil, walnut oil, polyethylene glycol, glycofurol, acetone, diglyme, dimethylacetamide, dimethyl isosorbide, dimethyl sulfoxide, ethanol, ethyl acetate, butyl acetate, ethyl ether, ethyl lactate, isopropyl acetate, methyl acetate, methyl isobutyl ketone, methyl tert-butyl ether, N-methylpyrrolidone In certain embodiments, the organic solvent is ethyl acetate, perfluorodecalin, 2-pyrrolidone, triglycerides, tetrahydrofurfuryl alcohol, triglycerides of fractionated C8 and C10 vegetable fatty acids (e.g., MIGLYOL® 810 and MIGLOYL® 812N), propylene glycol diesters of saturated C8 and C10 vegetable fatty acids (e.g., MIGLYOL® 840), ethyl oleate, ethyl caprate, dibutyl adipate, fatty acid esters, hexanoic acid, octanoic acid, triacetin, diethyl glycol monoether, gamma-butyrolactone, eugenol, clove bud oil, citral, limonene, or a combination thereof. In certain embodiments, the organic solvent is ethyl acetate or butyl acetate.

[0237] In still other embodiments, the organic solvent is acetone, acetonitrile, acyclic alkanes (e.g., hexane, heptane, pentane), amyl acetate, butanol, butyl acetate, chlorobenzene, chloroform, cumene, cyclohexane, 1,2-dichloroethene, dichloromethane, diethyl ether, dimethoxyethane, dimethylacetamide, dimethylformamide, dimethyl sulfoxide, 1,4-dioxane, ethanol, 2-ethoxyethanol, ethyl acetate, ethyl nitrate, ethylene glycol, hydrazine, isopropanol, methanol, methyl acetate, 2-methyl-1-butanol, 2-methyl-1-propanol, methyl butyl ketone, methylcyclohexane, methyl ethyl ketone, methylpyrrolidone, methyl tert-butyl ether, nitromethane, propanol, propyl acetate, sulfolane, propylene glycol, tetrahydrofuran, tetralin, toluene, 1,1,2-Trichloroethane, triethylamine, xylene, benzyl benzoate, ethyl lactate, dimethyl isosorbide, dimethyl sulfoxide, glycofurol, diglyme, methyl tert-butyl ether, polyethylene glycol, 2-pyrrolidone, tetrahydrofurfuryl alcohol, triglyceride, octyl acetate, ethanol, butanol, octanol, decanol, diglyme, tocopherol, octa-fluoropropane, (perfluorohexyl)octane, n-acetyltryptophan, triglyceride Cerides, fractionated vegetable fatty acid C8 and C10 triglycerides, saturated vegetable fatty acid C8 and C10 propylene glycol diesters, ethyl laurate, methyl caprylate, methyl caprate, methyl myristate, methyl oleate, methyl linoleate, dimethyl adipate, dibutyl suberate, diethyl sebacate, macadamia nut fatty acid ethyl ester, trimethylolpropane triisostearate, isopropyl laurate, isopropyl myristate, diethyl succinate, polysorbate esters, ethanol Alcoholamine, Propanoic Acid, Triacetin, Citral, Anisole, Anethole, Benzaldehyde, Linalool, Caprolactone, Phenol, Thioglycerol, Dimethylacetamide, Ethyl Formate, Ethylhexyl Acetate, Eugenol, Clove Bud Oil, Diethyl Glycol Monoether, Dimethyl Isosorbide, Isopropyl Acetate, Methyl Isobutyl Ketone, Methyl Tert-Butyl Ether, N-Methylpyrrolidone, Perfluorodecalin, 2-Pyrrolidone, Ethyl Oleate, Ethyl Caprate ethanol, dibutyl adipate, hexanoic acid, octanoic acid, diethyl glycol monoether, gamma-butyrolactone, polyoxyl 40 hydrogenated castor oil, polyoxyl 35 castor oil, propylene carbonate, octanol, hexanol, sorbitan monooleate, n-acetyltryptophan, solketal, alkyl acetate, aryl acetate, aryl alkyl acetate, tolyl acetate, benzyl acetate, polysorbate 80, phenethyl acetate, phenyl acetate, glycerol, or a combination thereof.

[0238] In certain embodiments, the organic solvent is acetonitrile, chlorobenzene, chloroform, cyclohexane, cumene, 1,2-dichloroethene, dichloromethane, 1,2-dimethoxyethane, N,N-dimethylacetamide, N,N-dimethylformamide, 1,4-dioxane, 2-ethoxyethanol, ethylene glycol, formamide, hexane, methanol, 2-methoxyethanol, methyl butyl ketone, methyl cyclohexane, methyl isobutyl ketone, N-methylpyrrolidone, nitromethane, pyridine, sulfolane, tetrahydrofuran, tetralin, toluene, 1,1,2-trichloroethene, xylene, acetic acid, acetone, anisole, 1-butanol, 2-butanol, butyl acetate, tetramethylpropane ... rt-butyl methyl ether, dimethyl sulfoxide, ethanol, ethyl acetate, ethyl ether, ethyl formate, formic acid, heptane, isobutyl acetate, isopropyl acetate, methyl acetate, 3-methyl-1-butanol, methyl ethyl ketone, 2-methyl-1-propanol, pentane, 1-pentanol, 1-propanol, 2-propanol, propyl acetate, triethylamine, 1,1-diethoxypropane, 1,1-dimethoxymethane, 2,2-dimethoxypropane, isooctane, isopropyl ether, methyl isopropyl ketone, methyl tetrahydrofuran, petroleum ether, trichloroacetic acid, trifluoroacetic acid, decanol, 2-ethylhexyl acetate, amyl acetate, or a combination thereof.

[0239] In some embodiments, the second liquid is an ionic liquid. In certain embodiments, the second liquid is a protein stabilizer.

[0240] In other embodiments, the second liquid has a viscosity of less than about 200 mPa·s, less than about 150 mPa·s, less than about 125 mPa·s, less than about 100 mPa·s, less than about 75 mPa·s, less than about 75 mPa·s, less than about 70 mPa·s, less than about 65 mPa·s, less than about 60 mPa·s, less than about 55 mPa·s, less than about 50 mPa·s, less than about 45 mPa·s, less than about 40 mPa·s, less than about 35 mPa·s, less than about 30 mPa·s, less than about 25 mPa·s, less than about 20 mPa·s, less than about 19 mPa·s, less than about 18 mPa·s, less than about 17 mPa·s, less than about 16 mPa·s, less than about 15 mPa·s, less than about 14 mPa·s, less than about 13 mPa·s, less than about 12 mPa·s s, less than about 11 mPa·s, less than about 10 mPa·s, less than about 9.5 mPa·s, less than about 9 mPa·s, less than about 8.5 mPa·s, less than about 8 mPa·s, less than about 7.5 mPa·s, less than about 7 mPa·s, less than about 6.5 mPa·s, less than about 6 mPa·s, less than about 5.5 mPa·s, less than about 5 mPa·s, less than about 4.5 mPa·s, less than about 4 mPa·s, less than about 3.5 mPa·s, less than about 3 mPa·s, less than about 2.5 mPa·s, less than about 2 mPa·s, less than about 1.5 mPa·s, less than about 1 mPa·s, less than about 0.5 mPa·s, less than about 0.1 mPa·s, less than about 0.05 mPa·s or less than about 0.01 mPa·s (1 milliPascal second). In other embodiments, the second liquid has a viscosity of about 0.01 mPa·s to about 10,000 mPa·s, e.g., about 0.01 mPa·s to about 1,000 mPa·s, about 0.01 mPa·s to about 100 mPa·s, about 0.01 mPa·s to about 50 mPa·s, about 0.01 mPa·s to about 25 mPa·s, about 0.01 mPa·s to about 10 mPa·s, about 0.01 mPa·s to about 5 mPa·s, or about 0.01 mPa·s to about 1 mPa·s. In certain embodiments, the second liquid has a viscosity that can range from about 0.27 mPa·s to about 200 mPa·s, such as from about 0.27 mPa·s to about 50 mPa·s, from about 1 mPa·s to about 30 mPa·s, or from about 20 mPa·s to about 50 mPa·s.In yet other embodiments, the second liquid has a viscosity ranging from about 0.27 mPa·s to about 200 mPa·s, e.g., from about 0.27 mPa·s to about 100 mPa·s, from about 0.27 mPa·s to about 50 mPa·s, from about 0.27 mPa·s to about 30 mPa·s, from about 1 mPa·s to about 20 mPa·s, or from about 1 mPa·s to about 15 mPa·s. Methods for controlling viscosity include temperature regulation and viscosity-modifying additives. Mixtures of liquids may also be used to control viscosity.

[0241] In certain embodiments, the second liquid further comprises a surfactant, hi yet other embodiments, the surfactant is polysorbate, magnesium stearate, sodium dodecyl sulfate, TRITON™ N-101, glycerin, polyoxyethylated castor oil, docusate, sodium stearate, decyl glucoside, nonoxynol-9, cetyltrimethylammonium bromide, sodium bis(2-ethylhexyl)sulfosuccinate, lecithin, sorbitan ester, or a combination thereof.

[0242] In some embodiments, the second liquid has a viscosity of about 0.01 to about 10,000 mPa·s. In other embodiments, the second liquid has a viscosity of less than about 10 mPa·s. In still other embodiments, the second liquid has a viscosity of less than about 5 mPa·s. In certain other embodiments, the second liquid has a viscosity of less than about 2 mPa·s. In certain embodiments, the second liquid has a viscosity of less than about 0.70 mPa·s. In preferred embodiments, the second liquid has a viscosity of less than about 0.40 mPa·s.

[0243] Droplets as described herein can include a first liquid and one or more agents, such as therapeutic and / or diagnostic agents. In certain embodiments, the therapeutic agent is a therapeutic biological agent. In yet other embodiments, the therapeutic biological agent has a unit activity of about 0.5 to about 1.0. In certain other embodiments, the concentration of the agent, e.g., a therapeutic or diagnostic agent, in the first liquid can range from about 0.0001 to about 1000 mg / mL, e.g., from about 100 to about 900 mg / mL, from about 200 to about 800 mg / mL, from about 200 to about 700 mg / mL, from about 200 to about 600 mg / mL, or from about 300 to about 500 mg / mL.

[0244] In some embodiments, the first liquid is aqueous or an organic solvent, and the second liquid is oil, aqueous, or an ionic liquid. In other embodiments, the first liquid and / or the second liquid have a viscosity of about 0.01 mPa·s to about 10,000 mPa·s. In certain embodiments, the second liquid is a mixture of two or more liquids of different polarity, where the mixture includes a liquid having a different solubility from the first liquid. In still other embodiments, the first liquid or the second liquid further comprises a carbohydrate, a pH adjuster, a salt, a chelating agent, a mineral, a polymer, a surfactant, a protein stabilizer, an emulsifier, a preservative, an amino acid, an antioxidant, a protein, an organic solvent, a paraben, a bactericide, a fungicide, a vitamin, a preservative, a nutrient medium, or a combination thereof. The term "polarity" refers to the overall solvating ability (solvating power) of a solvent, which is determined by the effect of all possible nonspecific and specific intermolecular interactions between solute ions or molecules and solvent molecules, but excluding those interactions that result in a definite chemical transformation of the ions of the solute molecules (Chem. Rev., 1994, 94, 2319-2358). Solvent polarity can be predicted from their dielectric constant. Solvents with high dielectric constants are considered more polar, and solvents with low dielectric constants are considered less polar or nonpolar (< about 15).

[0245] In other embodiments, each of the other components independently represents from about 0.0001 to about 99% (w / v) of the first liquid, e.g., from about 0.0001 to about 90% (w / v), from about 0.0001 to about 50% (w / v), from about 0.0001 to about 10% (w / v), from about 0.0001 to about 1% (w / v), or from about 0.0001 to about 0.1% (w / v). In certain embodiments, the amount of additional compound, i.e., additive, present in the first liquid, second liquid, or medium is as shown in Table 2.

[0246] [Table 2]

[0247] In some embodiments, cohesive forces (e.g., interfacial tension) on the droplet surface in the second liquid drive the droplet to a spherical shape, which is maintained during drying. In other embodiments, the particle sphericity ranges from about 0.1 to about 1, e.g., at least about 0.2, about 0.4, about 0.6, or about 0.8. In certain embodiments, the process can yield uniform particles with high sphericity (about >0.9) and roundness or circularity. Methods for measuring particle sphericity include image analysis of scanning electron micrographs of the particles, where an average roundness is calculated based on the cross-sectional shape of the particle projected onto the image plane. Such roundness or circularity factors can be expanded to identify corresponding sphericities.

[0248] In other embodiments, the droplets have a core-shell morphology, in which a first liquid (the droplet "core") is surrounded by one or more concentric layers of additional liquids (the droplet "shells"), each of which may or may not be defined by a unique set of components and / or unique concentrations of components. Each shell liquid may be an aqueous liquid, an organic liquid, an oil, an ionic liquid, or a combination thereof, and contains one or more agents, e.g., therapeutic or diagnostic agents. The concentration of the agent, e.g., therapeutic or diagnostic agent, in the shell liquid may range from about 0.0001 to about 1000 mg / mL, e.g., from about 100 to about 900 mg / mL, from about 200 to about 800 mg / mL, from about 200 to about 700 mg / mL, from about 200 to about 600 mg / mL, or from about 300 to about 500 mg / mL. The shell liquid may further comprise, for example, carbohydrates, pH adjusters, salts, chelating agents, minerals, polymers, surfactants, amino acids, oligopeptides, biological additives, chemical additives, preservatives, antioxidants, parabens, bactericides, fungicides, vitamins, preservatives, analgesics, and / or nutrient media.

[0249] In some embodiments, a surfactant in the first liquid and / or shell liquid prevents droplet coalescence. In other embodiments, the oligopeptide additive, protein additive, and / or the agent itself, e.g., a therapeutic or diagnostic agent, acts as a surfactant. In other embodiments, one or more of the shell layers is a hydrogel, ionogel, organogel, or some combination thereof.

[0250] In certain embodiments, the droplets carry an electrical charge. As a fraction of the Rayleigh limit, the droplets may be charged, on average, from about 0 to about 1, e.g., from about 0.1 to about 1.0, from about 0.2 to about 1.0, from about 0.3 to about 1.0, from about 0.4 to about 1.0, or from about 0.5 to about 1.0. In some embodiments, the charge assists in reducing droplet coalescence and / or controlling various particle properties of interest. These include, but are not limited to, the morphology, surface chemistry, and crystallinity of selected components. The term "Rayleigh limit" refers to the specific charge, e.g., in units of coulombs per kilogram, corresponding to the point at which Coulombic repulsion overcomes the surface tension binding forces in the droplets, resulting in the detachment of charge from the droplets by Coulombic breakup or some other mechanism.

[0251] In some embodiments, the droplets of step a) are formed in a microfluidic device, e.g., the formed droplets are regularly spaced. The droplets are allowed to flow through the device for a time sufficient for particles to form.

[0252] In other embodiments, the second liquid has a density between the densities of the droplets and the particles. The droplets float on the second liquid, but the formed particles do not float on the second liquid. The first liquid evaporates, causing the droplets to dry. In certain embodiments, the second liquid has a density higher than the density of the droplets. The formed droplets and particles float on the second liquid. The first liquid evaporates, causing the droplets to dry. In yet other embodiments, the second liquid has a density lower than the density of the droplets, and the droplets do not float on the second liquid. The first liquid disperses in the second liquid, causing the droplets to dry.

[0253] Particle formation Particles as described herein can be formed by contacting droplets comprising a first liquid with a second liquid, which facilitates removal of the first liquid. In some embodiments, the droplets are formed in a separate medium and then contacted with the second liquid, for example, by dripping or spraying the droplets into or onto the second liquid. In other embodiments, the droplets are formed in the second liquid so that they immediately come into contact. Once the first liquid is removed, particle formation begins to occur when at least a portion of the components of the droplets begin to undergo precipitation or phase separation. In a preferred embodiment, the droplets are dried after contacting the droplets with the second liquid.

[0254] In some embodiments, particles are formed after the first liquid disperses throughout the second liquid, e.g., by a diffusion process. In other embodiments, the second liquid may have varying degrees of miscibility with the first liquid and may represent a weak or negligible solubilization medium for a component of the particle or a subset of the component of the particle, e.g., a therapeutic or diagnostic agent. The agent, e.g., a therapeutic or diagnostic agent, is typically less soluble in the second liquid than in the first liquid during the time frame or under the conditions of production, e.g., at least about 5, 10, 100, or about 1,000 times less soluble. In still other embodiments, the second liquid is an aqueous liquid, an organic liquid, an oil, an ionic liquid, or a combination thereof. The second liquid may further comprise carbohydrates, pH adjusters, salts, chelating agents, minerals, polymers, surfactants, amino acids, oligopeptides, biological additives, chemical additives, antiseptics, antioxidants, parabens, bactericides, fungicides, vitamins, preservatives, analgesics, nutrient media, or combinations thereof. Exemplary aqueous liquids may contain stabilizers, such as crowding agents. In certain embodiments, these solutions contain additives, such as salts (e.g., sodium chloride), sugars and sugar alcohols (e.g., sorbitol, dextran 40, dextran 6000, or trehalose), polymers (e.g., PEG3350, PEG300, PEG8000, PEG20k, Ficoll 400, Ficoll 70, or polyvinylpyrrolidone, such as povidone), proteins (e.g., human serum albumin or bovine serum albumin), or combinations thereof. In yet other embodiments in which the first and second liquids are aqueous, the particles are obtained by osmotic drying of the droplets. The second liquid used to dry the particles, in certain embodiments, comprises a high concentration of solute, for example, at least about 0.03 osmolal, at least about 0.2 osmolal, at least about 1.0 osmolal, or at least about 1.2 osmolal.

[0255] In other embodiments, a surfactant in the second liquid helps prevent droplet coalescence, hi certain embodiments, the oligopeptide additive, the protein additive, the agent itself, e.g., a therapeutic or diagnostic agent, or a combination thereof, acts as a surfactant.

[0256] The particle formation process is governed by the diffusion equation ∂c1(x,t) / ∂t=D 12 ▽ 2 This can be recognized by considering the dispersion of a first liquid in a second liquid as described by c1(x,t), where c1(x,t) is the concentration of the first liquid at location x and time t, and D 12 is the diffusivity of the first liquid in the second liquid. For spherically symmetric dispersion (gradient only along the spherical radius r), the differential relation is given by the boundary condition c1(r i ,t)=c 1,s and c1(r→∞,t)=c 1,0 Under the constraint of

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[0257] In another embodiment, the distribution of solutes within the droplets during the particle formation process is relevant: in spherically symmetric space, this distribution is described by (R. Vehring, J. Aerosol Sci., 2007, 38, 728-746):

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[0258] In some embodiments, Equation 8 provides a useful estimate of the radial distribution of droplet and particle components during the particle formation process. The surface concentration of the solute (R=1) is particularly important, as it can sometimes determine important aspects of the particle formation process. This is typically calculated by numerical integration of the denominator in Equation 8. However, for reasonably low Peclet numbers (Pe<20), this can be expressed as

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[0259] In other embodiments, the second liquid is dispersed throughout the droplets on the time scale and under the conditions of particle formation. Similar to Equation 3, the process is i ) with boundary conditions ∂c2 / ∂r(0,t)=0 and c2(r i ,t)=c 2,s Under the constraint of

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[0260] In some embodiments, the second liquid has a Fourier number (Fo) of less than about 1.500, which allows the droplets to dry in about 60 seconds. In other embodiments, the second liquid has a Fourier number (Fo) of less than about 1.000, which allows the droplets to dry in about 60 seconds. In still other embodiments, the second liquid has a Fourier number (Fo) of less than about 0.500, which allows the droplets to dry in about 60 seconds. In certain other embodiments, the second liquid has a Fourier number (Fo) of less than about 0.208, which allows the droplets to dry in about 5 seconds. One skilled in the art, when informed of the intended range for the Fourier number, can adjust the process parameters accordingly without undue burden.

[0261] In other embodiments as described herein, step b) further comprises lowering the temperature of the second liquid to within about ±30°C of the freezing point of the first liquid. In some embodiments, the boiling point of the second liquid at atmospheric pressure is from about 0°C to about 200°C. In certain embodiments, step b) further comprises lowering the temperature of the second liquid to a temperature within about 30°C of the freezing point of the first liquid. In certain other embodiments, the boiling point of the second liquid at atmospheric pressure is from about 0°C to about 200°C. In still other embodiments, the second liquid is a mixture of two or more liquids of different polarities. In certain preferred embodiments, the mixture includes liquids with different solubilities.

[0262] Ratio of Second Liquid to First Liquid: The ratio of the second liquid to the first liquid during the particle formation process may be manipulated to control the particle formation process embodiments described herein. For complete or near complete dispersion of the first liquid, the ratio of the second liquid to the first liquid, V=V2 / V1, is:

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[0263] The term "primary dehydration" refers to a step in which a droplet comprising a first liquid is placed in contact with a second liquid and dried or dehydrated by the second liquid, for example by dispersion and / or evaporation of the first liquid into the second liquid.

[0264] The term "secondary dehydration" refers to a post-processing step after removal of the first and second liquids, for example, by which the residual moisture and / or solvent content of the particles is modified. Exemplary methods of secondary dehydration include vacuum drying, freeze drying, fluidized bed drying, tray drying, belt drying, or slurry spray drying, with or without the application of heat. Secondary dehydration can also be used to remove wash liquids used to separate the particles from the second liquid. In preferred embodiments, the first and second liquids are removed through centrifugation, sieving, filtration, magnetic collection, solvent exchange, or decanting.

[0265] In certain embodiments, the methods as described herein include removing particles from the second liquid by centrifugation, sieving, filtration, magnetic collection, solvent exchange, inertial separation, hydrocyclone separation, or decanting.

[0266] In other embodiments, the method as described herein further comprises washing the particles after step d) with a washing fluid, such as an organic liquid, a supercritical fluid, a cryogenic liquid, or a combination thereof. In certain embodiments, the washing fluid is an organic liquid, a supercritical fluid, a cryogenic liquid, or a combination thereof.

[0267] Drying of the particles, e.g., removing the first and second liquids to produce dried particles, can be accomplished by methods described herein. These include, but are not limited to, warm gas evaporation, freeze-drying, critical point drying, emulsion solvent evaporation, emulsion solvent diffusion, or combinations thereof. In certain embodiments, the particles are further dried by freeze-drying or vacuum dehydration. In certain other embodiments, the residual amount of the first and second liquids in the particles after drying is about 0 to about 10 wt. %, e.g., about 0 to about 5 wt. %, or about 0 to about 3 wt. %, or preferably about 0 to about 1 wt. %. In still other embodiments, the particles have less than 10 wt. % of the first or second liquid remaining after drying.

[0268] In certain other embodiments, the method further comprises washing the particles with a third liquid. In a preferred embodiment, the third liquid is an organic solvent. The third liquid can also be removed by evaporation, vacuum dehydration, or freeze-drying, such as vacuum drying with or without the application of heat, freeze-drying, fluidized bed drying, tray drying, belt drying, or slurry spray drying. In a preferred embodiment, the particles are further dried by freeze-drying or vacuum dehydration.

[0269] In some embodiments, the particles are further dried using evaporation of a warm gas. In other embodiments, the particles are further dried by contacting the particles with a gas stream. In certain embodiments, the gas has a temperature of about -80 to about 200°C. In certain other embodiments, the gas has a temperature of about 10 to about 40°C. In still other embodiments, the gas has a relative humidity of about 0 to about 100%.

[0270] In certain embodiments, the particles may or may not contain residual first and / or second liquids as described herein.

[0271] Other forms of charge and control: As described herein, particles may form in the presence of an electric field. Particles generated in the presence of an electric field may have an average diameter equal to or less than the diameter of particles generated in the absence of an electric field. In certain embodiments, the particles comprise a net charge that substantially minimizes particle adhesion.

[0272] The droplets of the present disclosure can be formed in an electric field and optionally carry an electric charge. In certain embodiments, the medium in which the droplets are formed, e.g., the second liquid, is typically a dielectric medium. In some embodiments, the electric field and / or charge on the droplets are such that free charges and / or polar molecules migrate to the surface of the droplets of the first liquid via the Coulomb effect. The former phenomenon, localization of free charges at the interface between the first liquid and the dielectric medium in which the droplets are formed, creates a layer of surface charge. In other embodiments, such effects are exploited to affect the structure and / or surface properties of the droplets and / or particles. This includes using surface charge to achieve spherical particle morphology under conditions not otherwise readily attainable, i.e., high Peclet numbers. In still other embodiments, tailoring the first liquid to be polar near the surface of the droplets facilitates faster removal of the first liquid by the second liquid. This may also mitigate surface-related degradation events between agents, such as therapeutic or diagnostic agents, and reduce the amount of residual first liquid in the particles after drying relative to that which is typical in the absence of an electric field.

[0273] In some embodiments, the electric field and / or charge are such that free charges and / or polar molecules move to the surface of the droplet of the first liquid based on the Coulomb effect, and one or more components of the droplet, such as a therapeutic agent, a diagnostic agent, or any of various additives the droplet may contain, crystallize. Crystal nucleation of the agent or other droplet components can be controlled to obtain a desired polymorph (A. Ziabicki, L. Jarecki, Macromolecular Symposia, 1996, 104, 65-87). In other embodiments, crystallization proceeds in a preferred direction, such as along the line of the electric field.

[0274] Core-shell particles can also be generated from droplets containing the first liquid alone, i.e., in the absence of a shell liquid, through the mediation of a net charge or an electric field. In some embodiments, this is achieved by exploiting the tendency of certain polar molecules and free charges to align themselves at the surface of a droplet when the droplet carries a net charge and / or when an external electric field is applied. In other embodiments, this results in localization of therapeutic or diagnostic agents toward the core of the droplet or its surface, which can be preserved by dehydration. In certain embodiments, this involves the deterministic layering of various agents (e.g., therapeutic agents, diagnostic agents, additives) across the thickness of the particle. In still other embodiments, non-therapeutic components, such as salts (e.g., sodium chloride) or sugars (e.g., sucrose), are preferentially driven to the surface by the electric field, forming a thin shell around the particle, crystal, or otherwise. This shell may have a protective effect or provide a measure of control over pharmacokinetics. In certain other embodiments, portions of the droplet components are localized at the particle surface without necessarily forming a uniform or continuous shell.

[0275] In other embodiments, the particles are formed in the presence of an electric field. In some embodiments, the particles formed in the presence of an electric field have an average diameter equal to or less than the diameter of the particles produced in the absence of an electric field. In certain embodiments, the particles comprise a net electric charge.

[0276] Coalescence: Controlling the degree of coalescence during the particle formation process can be important for achieving a desired particle size distribution. Controlling coalescence can be achieved in several ways. These include the use of surfactants, droplet charging, controlled droplet spreading, mixing, or a combination thereof. In some embodiments, the latter method is preferred because it helps reduce the amount of surfactant required to stabilize the droplets over the timescale and under the conditions of particle formation. Such reduction can be advantageous for limiting the amount of surfactant that can be present in the particle, thereby enhancing the weight fraction of other components of the particle, including active agents, such as therapeutic or diagnostic agents. In other embodiments, the weight fraction of surfactant in the particle is about 0 to about 50%, e.g., about 0 to about 10%, about 0 to about 5%, about 0 to about 3%, about 0 to about 1%, about 0 to about 0.5%, about 0 to about 0.01%, or about 0 to about 0.001%. An exemplary method for quantifying the extent of coalescence involves measuring the volume-weighted mean particle size and comparing it to an estimated volume-weighted mean size based on the initial droplet size distribution, i.e., the droplet size distribution before coalescence occurs. The measured mean size is about 1 to about 5 times the estimated mean size, e.g., about 1 to about 3 times the estimated mean size, about 1 to about 2 times the estimated mean size, about 1 to about 1.5 times the estimated mean size, about 1 to about 1.2 times the estimated mean size, or about 1 to about 1.1 times the estimated mean size.

[0277] As described herein, the use of surfactants can be a method for stabilizing liquid-liquid systems. In some embodiments, droplets are stabilized on the timescale and under the conditions of particle formation by adding a suitable surfactant to the first and / or second liquid. With respect to the critical micelle concentration (CMC), the surfactant concentration is about 0.01 to about 100 times the CMC, e.g., about 0.1 to about 10 times the CMC, about 1 to about 5 times the CMC, or about 1 to about 3 times the CMC. In other embodiments, the surfactant concentration is about 0.0001 to about 100 mg / mL, e.g., about 0.001 to about 10 mg / mL, about 0.01 to about 10 mg / mL, or about 0.01 to about 1 mg / mL. As used herein, the term "critical micelle concentration" or "CMC" refers to the concentration of surfactant in a liquid above which micelles are formed and above which all additional surfactant added to the system is directed toward the micelles.

[0278] In some embodiments, the charge on the droplets reduces coalescence. In other embodiments, this effect is exploited as an alternative to adding surfactants to the first and / or second liquids. In certain embodiments, this effect complements the use of surfactants in the first and / or second liquids, reducing the concentration required for desired stabilization. With respect to the critical micelle concentration (CMC), the surfactant concentration is about 0 to about 10 times the CMC, e.g., about 0 to about 3 times the CMC, about 0 to about 1 time the CMC, about 0 to about 0.5 times the CMC, about 0 to about 0.1 times the CMC, about 0 to about 0.01 times the CMC, or about 0 to about 0.001 times the CMC. In certain other embodiments, the concentration of the surfactant is about 0 to about 10 mg / mL, e.g., about 0 to about 1 mg / mL, about 0 to about 0.1 mg / mL, about 0 to about 0.01 mg / mL, or about 0 to about 0.001 mg / mL.

[0279] In certain embodiments, droplets are formed in a device having at least one channel, e.g., a microfluidic device, where the properties of individual channels are manipulated to control the residence time of droplets in the channel. In some embodiments, droplets flow in a train and experience limited droplet-droplet interactions in the channel. When droplets and / or particles are collected from the channel or the droplet train is otherwise perturbed in a manner that enhances droplet-droplet interactions, the residence time can be designed such that coalescence is mitigated regardless of the surfactant content of the first or second liquid. In other embodiments, the residence time is on the order of or longer than the characteristic particle formation time from Equation 6, e.g., about 0.5 Fo * ~about 150Fo * , about 0.5Fo * ~approx. 15Fo * , about 0.5Fo * ~approx. 7.5Fo * , about 0.5Fo * ~approx. 4.5Fo * , about 0.5Fo * ~approx. 3.0Fo * or about 0.5Fo * ~approx. 1.5Fo * In certain other embodiments, particles are substantially formed during residence in the individual channels. In still other embodiments, the residence time is less than the characteristic particle formation time from Equation 6, but is still sufficient for the formation of primary particles (e.g., about 0.0001Fo * ~ approx. 0.5Fo * , approximately 0.001Fo * ~ approx. 0.5Fo * , about 0.01Fo * ~ approx. 0.5Fo * , about 0.05Fo * ~ approx. 0.5Fo * or about 0.1Fo * ~ approx. 0.5Fo * ) The proto-particles have surface properties, such as a thin layer of concentrated solute, that prevent coalescence despite potential droplet-droplet interactions that may occur during the particle formation process.

[0280] In some embodiments, the Peclet number Pem The solute m with initial concentration c is used to manipulate the required residence time for primary particle formation. m,0 into the droplet at a critical surface concentration c m * From Equation 8, when R=1, adhesion is effectively prevented.

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[0281] In other embodiments, coalescence is controlled even when the droplet generation frequency associated with the channel is very high. The droplet generation frequency associated with the channel can be about 1 Hz to about 10 MHz, e.g., about 10 Hz to about 10 MHz, about 100 Hz to about 10 MHz, about 1 kHz to about 10 MHz, or about 10 kHz to about 10 MHz. In certain embodiments, a suitable surfactant is added to the first liquid and / or the second liquid to further aid in controlling coalescence. With respect to the critical micelle concentration (CMC), the surfactant concentration is about 0 to about 10 times the CMC, e.g., about 0 to about 3 times the CMC, about 0 to about 1 times the CMC, about 0 to about 0.5 times the CMC, about 0 to about 0.1 times the CMC, about 0 to about 0.01 times the CMC, or about 0 to about 0.001 times the CMC. In certain other embodiments, the surfactant concentration is about 0 to about 10 mg / mL, e.g., about 0 to about 1 mg / mL, about 0 to about 0.1 mg / mL, about 0 to about 0.01 mg / mL, or about 0 to about 0.001 mg / mL. The use of agitation or movement of the second liquid, such as in a continuous stirred tank reactor or linear flow channel, can also be useful to reduce coalescence during drying. In preferred embodiments, the net charge substantially minimizes particle coalescence.

[0282] Sterility: Sterility is a critical aspect of pharmaceutical compositions because it affects the safety with which the composition can be administered. For example, many particle formulations, especially microparticle formulations, that have achieved sterility can be challenging because they are not compatible with common sterilization techniques, such as sterile filtration. The sterile filtration step typically involves a membrane through which only those components of the filtered liquid, e.g., 200 nm or less in size, can pass. Thus, particle formulations with solids greater than 200 nm in size are filtered rather than sterilized. In some embodiments, formulations of the present disclosure are subjected to an alternative process of terminal sterilization before use or administration. These sterilization protocols and the effectiveness of the process in reducing bioburden are based on regulatory guidelines, e.g., USP Chapter 10. <71> , European Pharmacopoeia Chapter, Sterility:2.6.1, 21 CFR 610.12, ICH Q4B ANNEX8(R1), ICH Q5A, etc. An exemplary method of demonstrating compliance involves incubating about 1 mL of drug product per container in an appropriate growth medium (soybean casein digest broth, tryptic soy broth, fluid thioglycollate broth) for a period of about 14 days to ensure there is no microbial growth in about 1 in about 1 billion units of drug product or about 1 in about 1 million units of drug product. As disclosed herein, a "sterile" formulation is aseptic or free of viable microorganisms and their spores.

[0283] In some embodiments, the terminal sterilization step involves gamma irradiation. In other embodiments, the terminal sterilization step involves at least about 2-4 log 10 The sterilization step required to inactivate viral microbial contaminants is about 10 kGy, about 20 kGy, about 40 kGy, about 60 kGy, or about 100 kGy. In certain embodiments, the particles include antioxidants or scavengers that mitigate the harmful effects of degradation products resulting from the sterilization step.

[0284] In other embodiments, the terminal sterilization step involves a transient heat treatment. In some embodiments, the formulation is exposed to a temperature of about 60 to about 200°C, e.g., about 60 to about 180°C, about 60 to about 160°C, about 60 to about 140°C, about 60 to about 130°C, about 60 to about 120°C, or about 60 to about 110°C. In certain embodiments, the exposure occurs for a period of about 1 to about 144 hours, e.g., about 1 to about 120 hours, about 1 to about 100 hours, about 1 to about 90 hours, about 1 to about 72 hours, about 1 to about 48 hours, about 1 to about 36 hours, or about 1 to about 24 hours. For example, dry heat sterilization can be carried out at a temperature of about 80°C for about 72 hours, about 160°C for about 2 hours, or about 170°C for about 1 hour. In certain other embodiments, pasteurization is carried out at about 60°C for about 10 hours.

[0285] In some embodiments, sterilization is ensured by using beta radiation, X-ray sterilization, steam sterilization, a solvent-detergent inactivation step, supercritical CO2-mediated sterilization, low pH hold, UV-C exposure, or ethylene oxide-mediated sterilization of the formulation. In other embodiments, the terminal sterilization step is performed at low temperatures of about -100 to about 60°C. In certain embodiments, the supercritical CO2 further comprises additives (e.g., hydrogen peroxide, water, acetic anhydride, etc.) intended to effectively inactivate microorganisms, including bacterial spores.

[0286] In other embodiments, the second liquid is selected so that its presence in the droplets during particle formation helps promote process sterility. In some embodiments, the second liquid is antimicrobial or contains such a compound contained within the particles. This compound may persist inside the particles even after the secondary drying step. Organic liquids that can be used as the second liquid with antimicrobial activity may include, but are not limited to, acetates (e.g., ethyl acetate) and alcohols (e.g., ethanol, phenol). The second liquid may also contain antimicrobial additives, such as phenolic substances, benzalkonium chloride, linalool, coumarin, peroxides, active chlorine, alkali, or combinations thereof.

[0287] In some embodiments, the use of nanofiltration membranes for inlet process streams, for example, for use on the first liquid and / or second liquid prior to particle formation, contributes to a reduction in the bioburden of the process. In other embodiments, a combination of preceding sterile measures is used to reach an appropriate bioburden level.

[0288] As described herein, the particles may be sterilized after formation, for example, by irradiation, pasteurization, freezing, or irradiation with gamma rays. In certain embodiments, the methods as described herein further include sterilizing the particles after removing the first and second liquids. In certain preferred embodiments, sterilization occurs by irradiation, pasteurization, or freezing. In preferred embodiments, irradiation is by gamma rays.

[0289] Control of particle properties Particle properties can be controlled by modulating the drying rate of the droplets, the Peclet number of the droplet's components, the concentration of the droplet's components, the particle formation kinetics following solute precipitation or phase separation within the droplets, the charge on the droplets, and / or the characteristics of the electric field within which the droplets may be positioned. In certain embodiments, modulation affects the particle's size, morphology, density, porosity, composition, and surface energy properties, establishes the distribution of components within the particle, and helps regulate important physicochemical properties that can be difficult to address when drying in air without a second liquid, such as in conventional spray drying. These properties include the particle's dissolution rate and flow characteristics (R. Vehring, Pharmaceutical Res., 2008, 25, 999-1022).

[0290] Methods described herein generally provide for controlling particle morphology, the methods including: a) providing a droplet comprising a first liquid and an agent; b) contacting the droplet with a second liquid under a specific Peclet number; c) drying the droplet; and d) removing the first and second liquids, wherein the specific Peclet number controls the particle morphology.

[0291] In one aspect, the present disclosure provides a method of controlling particle morphology, the method comprising: a) providing a droplet comprising a first liquid and an agent; b) contacting the droplet with a second liquid comprising a plasticizer having a particular Peclet number; c) drying the droplet; and d) removing the first and second liquids, wherein the plasticizer controls particle morphology.

[0292] In another aspect, the present disclosure provides a method of controlling particle morphology, the method comprising: a) providing a droplet comprising a first liquid and an agent; b) contacting the droplet with a second liquid; c) drying the droplet; and d) removing the first and second liquids, wherein the Peclet number of the second liquid controls the particle morphology.

[0293] As disclosed herein, the agent can be a therapeutic agent or a diagnostic agent. In certain embodiments, the therapeutic agent has a unit activity of about 0.5 to about 1.0. In certain preferred embodiments, the therapeutic agent is a therapeutic biological agent. In preferred embodiments, the therapeutic biological agent has a unit activity of about 0.5 to about 1.0.

[0294] In other embodiments, the first or second liquid further comprises a plasticizer that controls the morphology of the particles. Exemplary plasticizers include sucrose, xylitol, sorbitol, fructose, triglycerides, pectin, glycerol, triethyl citrate, ethyl acetate, citric acid, oleic acid, hydroxypropyl cellulose, methylpyrrolidone, polyethylene glycol, polypropylene glycol, polysorbate 80, diethyl phthalate and other phthalate derivatives, castor oil, triacetin, water, chlorpheniramine, 1-butyl-3-methylimidazolium dioctyl sulfosuccinate, hexyl acetate, water, 2-ethylhexyl acetate, triethyl citrate, dibutyl sebacate, benzyl alcohol, benzyl benzoate, dimethylacetamide, various aqueous liquids, organic liquids, oils, ionic liquids, polysaccharides, sugars, diols, polyols, fatty acids, fatty acid esters, esters, surfactants, or combinations thereof. In certain embodiments, the plasticizer is sucrose, xylitol, sorbitol, fructose, triglycerides, pectin, glycerol, triethyl citrate, ethyl acetate, citric acid, oleic acid, hydroxypropyl cellulose, methylpyrrolidone, polyethylene glycol, polypropylene glycol, polysorbate 20, polysorbate 60, polysorbate 80, fatty acid esters of sorbitol, diethyl phthalate and other phthalate derivatives, castor oil, triacetin, water, chlorpheniramine, 1-butyl-3-methylimidazolium dioctyl sulfosuccinate, hexyl acetate, 2-ethylhexyl acetate, triethyl citrate, dibutyl sebacate, benzyl alcohol, benzyl benzoate, dimethylacetamide, or a combination thereof. In preferred embodiments, the plasticizer is polysorbate 20, polysorbate 60, or polysorbate 80. In certain preferred embodiments, the plasticizer is polysorbate 20 or polysorbate 80. In certain other embodiments, the fatty acid ester of sorbitol is a sorbitan ester, such as span20, span40, span60, or span80.

[0295] In some embodiments, the particles may comprise less than 10% internal void space after removal of the first and second liquids, less than 5% internal void space after removal of the first and second liquids, less than 1% internal void space after removal of the first and second liquids, less than 0.1% internal void space after removal of the first and second liquids, or less than 0.01% internal void space after removal of the first and second ...

Claims

1. 1. A method of making a particle comprising a therapeutic biological agent, comprising: forming droplets of an aqueous first liquid, said first liquid including a therapeutic biological agent, said droplets being formed within a second liquid including an organic solvent; dehydrating the droplets in the second liquid to form particles comprising the therapeutic biological agent; A method comprising:

2. The method of claim 1 , wherein the droplets are formed in a microfluidic device or a homogenizer.

3. The method of claim 2 , wherein the droplets formed in the microfluidic device are regularly spaced in the microfluidic device.

4. The method of claim 1 , wherein dehydrating the droplet comprises contacting the second liquid with a dry material.

5. The method of claim 4 , wherein the dry material is a desiccant.

6. The method of claim 1 , further comprising drying the particles, wherein the plurality of particles has a first or second liquid content of less than 10% by weight after drying.

7. The method of claim 6 , wherein drying the particles comprises removing the first and second liquids.

8. 8. The method of claim 7, wherein the first and second liquids are removed by centrifugation, sieving, filtration, magnetic collection, solvent exchange, or decanting.

9. The method of claim 7, wherein the residual amount of the first and second liquids in the particles after drying is from about 0% to about 10% by weight.

10. The method of claim 7 , wherein drying the particles further comprises contacting the particles with a gas stream.

11. The method of claim 10, wherein the gas has a temperature of about -80°C to about 200°C.

12. The method of claim 10, wherein the gas is helium, air, nitrogen or argon.

13. The method of claim 7, wherein drying the particles further comprises using freeze-drying or vacuum dehydration.

14. The method of claim 1 further comprising removing particles from the second liquid.

15. 15. The method of claim 14, wherein the particles are removed from the second liquid by centrifugation, sieving, filtration, magnetic collection, solvent exchange, inertial separation, hydrocyclone separation, or decanting.

16. The method of claim 6 further comprising washing the dry particles with a washing fluid.

17. 17. The method of claim 16, wherein the cleaning fluid is an organic liquid, a supercritical fluid, a cryogenic liquid, or a combination thereof.

18. 10. The method of claim 1, wherein the therapeutic biological agent is an antibody.

19. 20. The method of claim 18, wherein the antibody is an IgG antibody.

20. 10. The method of claim 1, wherein the therapeutic biological agent in the particles has a unit activity of about 0.5 to about 1.

0.

21. The method of claim 7 , wherein the particles contain less than about 10% internal void space.

22. 8. The method of claim 7, wherein the particles have less than about 10% aggregates of the therapeutic biological agent.

23. 8. The method of claim 7, wherein the particles have greater than about 60% by weight of a therapeutic biological agent.

24. The method of claim 7, wherein the particles have a circularity of about 0.80 to about 1.

00.

25. 10. The method of claim 1, wherein the first liquid is water, 0.9% saline, lactated Ringer's solution, a buffered solution, dextrose 5%, or a combination thereof.

26. The method of claim 1 , wherein the first liquid is water.

27. 10. The method of claim 1, wherein the concentration of the therapeutic biological agent in the first liquid is from about 10 mg / mL to about 500 mg / mL.

28. The method of claim 1 , wherein the first liquid further comprises a surfactant.

29. 2. The method of claim 1, wherein the organic solvent is ethyl acetate or butyl acetate.

30. The method of claim 1 , wherein the droplets are formed in a container comprising tubing.

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