Albumin compositions and methods of making and using the same

Abstract

To provide an albumin composition having a defined fatty acid profile and a method for using the same.SOLUTION: The albumin compositions described herein are particularly suitable for use in cell culture methods, protein stabilization methods. The albumin compositions described herein may improve the viability and / or promote the growth of cells (e.g., mammalian cells) when the cells are cultured in media containing the albumin compositions. The albumin compositions described herein may improve the stability of a biological product when the biological product is in the presence of the albumin composition. Further provided herein is a method of formulating an albumin composition having a defined fatty acid profile as described herein.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to providing albumin compositions with defined fatty acid profiles and methods of using same. [Background technology]

[0002] Albumin is a family of globular, water-soluble proteins commonly found in plasma. Albumin is generally a transport protein capable of binding various ligands, such as fatty acids, enzymes, hormones, and trace elements. Albumin is commonly used in cell (e.g., stem cell) culture as well as in excipients (e.g., to stabilize biological products). The performance of various albumin compositions varies significantly, which may be partially due to the fatty acid composition of the composition. For example, the presence of some fatty acids has been reported to inhibit cell proliferation and / or viability. Furthermore, albumin compositions can vary significantly from batch to batch, reducing the reliability of the product. [Prior art documents] [Non-patent literature]

[0003] [Non-Patent Document 1] Karlin and Altschul, Proc. Natl. Acad. Sci. USA, 87:2264-2268 (1990) [Non-patent document 2] Altschul et al., J. Mol. Biol., 215:403-410 (1990) [Non-patent document 3] Karlin and Altschul, Proc. Natl. Acad. Sci. USA, 90:5873-5877 (1993) [Non-patent document 4] Altschul et al., Nucleic Acids Res., 25:3389-3402 (1997) [Non-patent document 5] Wootton and Federhen, Computers and Chemistry 17:149-163 (1993) Summary of the Invention [Problem to be solved by the invention]

[0004] The disclosure herein addresses a significant unmet need for albumin compositions with a defined and consistent fatty acid profile for use in cell culture processes and as excipients. [Means for solving the problem]

[0005] In one aspect, a composition is provided that comprises: a) an albumin polypeptide; b) one or more fatty acids having fewer than 18 carbon atoms present in a molar ratio to the albumin polypeptide ranging from about 0.02 to about 0.4; and c) one or more fatty acids having 18 or more carbon atoms present in a molar ratio to the albumin polypeptide ranging from about 0.03 to about 0.6, wherein the composition is substantially free of pentadecanoic acid (C15:0), margaric acid (C17:0), and / or heptadecenoic acid (C17:1 ω-7). In some instances, the one or more fatty acids having fewer than 18 carbon atoms comprise one or more fatty acids selected from the group consisting of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), and palmitoleic acid (C16:1). In some instances, the one or more fatty acids having fewer than 18 carbon atoms consist of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), and palmitoleic acid (C16:1). In some instances, the one or more fatty acids having 18 or more carbon atoms comprise one or more fatty acids selected from the group consisting of stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), and eicosadienoic acid (C20:2 ω-6). In some instances, the one or more fatty acids having 18 or more carbon atoms consist of stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), and eicosadienoic acid (C20:2 ω-6). In some cases, the one or more fatty acids having 18 or more carbon atoms further include one or more fatty acids selected from the group consisting of bishomo-gamma-linolenic acid (C20:3 ω-6), arachidonic acid (C20:4), docosatetraenoic acid (C22:4 ω-6), and docosahexaenoic acid (C22:6 ω-3).In some instances, (i) the one or more fatty acids having fewer than 18 carbon atoms consist of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), and palmitoleic acid (C16:1); and (ii) the one or more fatty acids having 18 or more carbon atoms consist of stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), eicosadienoic acid (C20:2 ω-6), bishomo-γ-linolenic acid (C20:3 ω-6), arachidonic acid (C20:4), docosatetraenoic acid (C22:4 ω-6), and docosahexaenoic acid (C22:6 ω-3). In some instances, (i) the one or more fatty acids having fewer than 18 carbon atoms consist of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), and palmitoleic acid (C16:1); and (ii) the one or more fatty acids having 18 or more carbon atoms consist of stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), and eicosadienoic acid (C20:2 ω-6). In some instances, (i) the one or more fatty acids having fewer than 18 carbon atoms consist of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), and palmitoleic acid (C16:1); and (ii) the one or more fatty acids having 18 or more carbon atoms consist of stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), eicosadienoic acid (C20:2 ω-6), bishomo-γ-linolenic acid (C20:3 ω-6), and arachidonic acid (C20:4). In some instances, the total molar ratio of fatty acids to albumin polypeptide is less than 1. In some cases, the composition is substantially free of alpha-linolenic acid (C18:3), gamma-linolenic acid (C18:3), arachidic acid (C20:0), eicosatrienoic acid (C20:3 ω-3), eicosapentaenoic acid (C20:5 ω-3), behenic acid (C22:0), docosapentaenoic acid (C22:5 ω-3), lignoceric acid (C24:0), and / or cerotic acid (C26:0).In some cases, both one or more fatty acids having fewer than 18 carbon atoms and one or more fatty acids having 18 or more carbon atoms are adsorbed to the albumin polypeptide.

[0006] In another embodiment, the composition comprises: a) an albumin polypeptide; b) lauric acid (C12:0) present in a molar ratio to the albumin polypeptide ranging from about 0.001 to about 0.008; c) myristic acid (C14:0) present in a molar ratio to the albumin polypeptide ranging from about 0.001 to about 0.022; d) palmitic acid (C16:0) present in a molar ratio to the albumin polypeptide ranging from about 0.02 to about 0.3; e) palmitoleic acid (C16:1) present in a molar ratio to the albumin polypeptide ranging from about 0.002 to about 0.03; f) stearic acid (C18:0) present in a molar ratio to the albumin polypeptide ranging from about 0.011 to about 0.2; g) oleic acid (C18:1) present in a molar ratio to the albumin polypeptide ranging from about 0.02 to about 0.3. ω-9); h) linoleic acid (C18:2) present in a molar ratio to albumin polypeptide ranging from about 0.0002 to about 0.12; and i) eicosadienoic acid (C20:2 ω-6) present in a molar ratio to albumin polypeptide ranging from about 0.0001 to about 0.002. In some cases, the composition further comprises bishomo-γ-linolenic acid (C20:3 ω-6) present in a molar ratio to albumin polypeptide ranging from about 0.0003 to about 0.002. In some cases, the composition further comprises arachidonic acid (C20:4) present in a molar ratio to albumin polypeptide ranging from about 0.001 to about 0.01. In some cases, the composition further comprises docosatetraenoic acid (C22:4 ω-6) present in a molar ratio to albumin polypeptide ranging from about 0.0009 to about 0.003. In some instances, the composition further comprises docosahexaenoic acid (C22:6 ω-3) present in a molar ratio to albumin polypeptide ranging from about 0.0003 to about 0.001. In some instances, the total molar ratio of fatty acids to albumin polypeptide is from about 0.06 to about 1.In some cases, the composition is substantially free of pentadecanoic acid (C15:0), margaric acid (C17:0), heptadecenoic acid (C17:1 ω-7), α-linolenic acid (C18:3), γ-linolenic acid (C18:3), arachidic acid (C20:0), eicosatrienoic acid (C20:3 ω-3), eicosapentaenoic acid (C20:5 ω-3), behenic acid (C22:0), docosapentaenoic acid (C22:5 ω-3), lignoceric acid (C24:0), and / or cerotic acid (C26:0).

[0007] In another aspect, provided is a composition comprising: a) an albumin polypeptide; and b) one or more fatty acids, the composition being substantially free of one or more of pentadecanoic acid (C15:0), margaric acid (C17:0), heptadecenoic acid (C17:1 ω-7), α-linolenic acid (C18:3), γ-linolenic acid (C18:3), arachidic acid (C20:0), eicosatrienoic acid (C20:3 ω-3), eicosapentaenoic acid (C20:5 ω-3), behenic acid (C22:0), docosapentaenoic acid (C22:5 ω-3), lignoceric acid (C24:0), and cerotic acid (C26:0), and wherein the total molar ratio of the one or more fatty acids to the albumin polypeptide is from about 0.05 to about 1.

[0008] In yet another aspect, a composition is provided comprising: a) an albumin polypeptide; and b) fatty acids, wherein the fatty acids consist of i) lauric acid (C12:0); ii) myristic acid (C14:0); iii) palmitic acid (C16:0); iv) palmitoleic acid (C16:1); v) stearic acid (C18:0); vi) oleic acid (C18:1 ω-9); vii) linoleic acid (C18:2); viii) eicosadienoic acid (C20:2 ω-6); ix) bishomo-γ-linolenic acid (C20:3 ω-6); x) arachidonic acid (C20:4); xi) docosatetraenoic acid (C22:4 ω-6); and xii) docosahexaenoic acid (C22:6 ω-3), wherein the total molar ratio of fatty acids to albumin polypeptide is less than about 1. In some instances, lauric acid (C12:0) is present in a molar ratio to albumin polypeptide of less than about 0.01; myristic acid (C14:0) is present in a molar ratio to albumin polypeptide of less than about 0.05; palmitic acid (C16:0) is present in a molar ratio to albumin polypeptide of less than about 0.5; palmitoleic acid (C16:1) is present in a molar ratio to albumin polypeptide of less than about 0.05; stearic acid (C18:0) is present in a molar ratio to albumin polypeptide of less than about 0.2; oleic acid (C18:1 ω-9) is present in a molar ratio to albumin polypeptide of less than about 0.5; linoleic acid (C18:2) is present in a molar ratio to albumin polypeptide of less than about 0.2; eicosadienoic acid (C20:2 bishomo-gamma-linolenic acid (C20:3 ω-6) is present in a molar ratio to albumin polypeptide of less than about 0.005; arachidonic acid (C20:4) is present in a molar ratio to albumin polypeptide of less than about 0.01; docosatetraenoic acid (C22:4 ω-6) is present in a molar ratio to albumin polypeptide of less than about 0.005; docosahexaenoic acid (C22:6 ω-3) is present in a molar ratio to albumin polypeptide of less than about 0.005; or any combination thereof.

[0009] In yet another aspect, a composition is provided comprising: a) an albumin polypeptide; and b) fatty acids, wherein the fatty acids consist of: i) lauric acid (C12:0); ii) myristic acid (C14:0); iii) palmitic acid (C16:0); iv) palmitoleic acid (C16:1); v) stearic acid (C18:0); vi) oleic acid (C18:1 ω-9); vii) linoleic acid (C18:2); and viii) eicosadienoic acid (C20:2 ω-6). In some instances, lauric acid (C12:0) is present in a molar ratio to albumin polypeptide of less than about 0.005; myristic acid (C14:0) is present in a molar ratio to albumin polypeptide of less than about 0.005; palmitic acid (C16:0) is present in a molar ratio to albumin polypeptide of less than about 0.05; palmitoleic acid (C16:1) is present in a molar ratio to albumin polypeptide of less than about 0.005; stearic acid (C18:0) is present in a molar ratio to albumin polypeptide of less than about 0.05; oleic acid (C18:1 ω-9) is present in a molar ratio to albumin polypeptide of less than about 0.05; linoleic acid (C18:2) is present in a molar ratio to albumin polypeptide of less than about 0.0005; eicosadienoic acid (C20:2 ω-6) is present in a molar ratio to albumin polypeptide of less than about 0.0005; or any combination thereof.

[0010] In yet another aspect, a composition is provided comprising: a) an albumin polypeptide; and b) fatty acids, wherein the fatty acids consist of: i) lauric acid (C12:0); ii) myristic acid (C14:0); iii) palmitic acid (C16:0); iv) palmitoleic acid (C16:1); v) stearic acid (C18:0); vi) oleic acid (C18:1 ω-9); vii) linoleic acid (C18:2); viii) eicosadienoic acid (C20:2 ω-6); ix) bishomo-γ-linolenic acid (C20:3 ω-6); and x) arachidonic acid (C20:4). In some instances, lauric acid (C12:0) is present in a molar ratio to albumin polypeptide of less than about 0.005; myristic acid (C14:0) is present in a molar ratio to albumin polypeptide of less than about 0.01; palmitic acid (C16:0) is present in a molar ratio to albumin polypeptide of less than about 0.1; palmitoleic acid (C16:1) is present in a molar ratio to albumin polypeptide of less than about 0.01; stearic acid (C18:0) is present in a molar ratio to albumin polypeptide of less than about 0.1; oleic acid (C18:1 ω-9) is present in a molar ratio to albumin polypeptide of less than about 0.5; linoleic acid (C18:2) is present in a molar ratio to albumin polypeptide of less than about 0.05; eicosadienoic acid (C20:2 bishomo-gamma-linolenic acid (C20:3 ω-6) is present in a molar ratio to albumin polypeptide of less than about 0.0005; arachidonic acid (C20:4) is present in a molar ratio to albumin polypeptide of less than about 0.005; or any combination thereof.

[0011] In any preceding aspect, the albumin polypeptide is of human or bovine origin. In any preceding aspect, the albumin polypeptide is a recombinant albumin polypeptide. In any preceding aspect, the recombinant albumin polypeptide is purified from bacteria, yeast, or rice. In any preceding aspect, the yeast is of the species Pichia pastoris, Saccharomyces cerevisiae, or Kluyveromyces lactis. In any preceding aspect, the rice is of the species Oryza sativa. In any preceding aspect, the albumin polypeptide is derived from plasma or serum. In any preceding aspect, the albumin polypeptide is a defatted albumin polypeptide.

[0012] In another aspect, a cell culture medium is provided, comprising the composition of any one of the preceding aspects and a basal medium. In some cases, the composition is present in the cell culture medium at a concentration of about 0.01% (w / w) to about 10% (w / w), or about 0.1 mg / mL to about 100 mg / mL.

[0013] In another aspect, a method is provided, comprising incubating biological cells in the cell culture medium of any one of the preceding aspects. In some cases, the biological cells are eukaryotic cells. In some cases, the eukaryotic cells are stem cells. In some cases, the stem cells are induced pluripotent stem (iPS) cells, embryonic stem (ES) cells, or mesenchymal stem cells (MSCs). In some cases, the eukaryotic cells are T cells. In some cases, the eukaryotic cells are neural cells. In some cases, the method results in an increase in viable cell density of the biological cells compared to biological cells cultured in the absence of the albumin polypeptide. In some cases, the increase in viable cell density is at least 10%.

[0014] In yet another aspect, provided is a method of stabilizing a protein, comprising incubating the protein in the presence of the composition of any one of the preceding aspects. In some cases, the composition is present at a concentration of about 0.01% (w / w) to about 20% (w / w), about 0.1 mg / mL to about 200 mg / mL, or a molar ratio relative to the protein of about 0.1 to about 10. In some cases, the protein is stabilized by at least about 80% compared to the protein in the absence of the composition.

[0015] In yet another aspect, a composition is provided comprising: a) an albumin polypeptide; and b) arachidonic acid (C20:4) present in a molar ratio relative to the total amount of fatty acids in the composition of at least about 0.9%.

[0016] In yet another aspect, a method for improving the function of an albumin polypeptide is provided, comprising incubating the albumin polypeptide with arachidonic acid (C20:4) in an amount sufficient to increase the molar ratio of arachidonic acid (C20:4) to the total amount of fatty acids in the composition to at least about 0.9%.

[0017] In yet another aspect, a method for improving the function of an albumin polypeptide is provided, comprising: a) passing a solution of albumin polypeptide through a ceramic hydroxyapatite resin to produce a flow-through containing the albumin polypeptide; and b) purifying the albumin polypeptide from the flow-through to obtain an albumin polypeptide with improved function. In some cases, the method further comprises, prior to a), delipidating the albumin polypeptide by incubating the solution with activated carbon and purifying the albumin polypeptide from the activated carbon. In some cases, the method further comprises, prior to a), incubating the solution with a chelating resin and purifying the albumin polypeptide from the chelating resin. In some cases, the chelating resin is Diaion CR20. In some cases, the method further comprises, prior to a), incubating the albumin polypeptide with one or more fatty acids. In some cases, the one or more fatty acids are selected from the group consisting of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), palmitoleic acid (C16:1), stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), eicosadienoic acid (C20:2 ω-6), bishomo-γ-linolenic acid (C20:3 ω-6), arachidonic acid (C20:4), docosatetraenoic acid (C22:4 ω-6), and docosahexaenoic acid (C22:6 ω-3). In some cases, the one or more fatty acids consist of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), palmitoleic acid (C16:1), stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), eicosadienoic acid (C20:2 ω-6), bishomo-γ-linolenic acid (C20:3 ω-6), arachidonic acid (C20:4), docosatetraenoic acid (C22:4 ω-6), and docosahexaenoic acid (C22:6 ω-3).In some cases, the one or more fatty acids consist of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), palmitoleic acid (C16:1), stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), and eicosadienoic acid (C20:2 ω-6). In some cases, the one or more fatty acids consist of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), palmitoleic acid (C16:1), stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), eicosadienoic acid (C20:2 ω-6), bishomo-γ-linolenic acid (C20:3 ω-6), and arachidonic acid (C20:4).

[0018] In another aspect, a composition is provided comprising a recombinant albumin polypeptide and one or more fatty acids in an amount in a molar ratio effective to promote at least a 35% increase in viable cell density of induced pluripotent stem (iPS) cells upon culturing iPS cells in the composition for a duration of 4 days, compared to a composition lacking the recombinant albumin polypeptide. In another aspect, a composition is provided comprising a recombinant albumin polypeptide and one or more fatty acids in an amount in a molar ratio effective to promote at least a 35% increase in viable cell density of mesenchymal stem cells (MSCs) upon culturing MSCs in the composition for a duration of 5 days, compared to a composition lacking the recombinant albumin polypeptide. In another aspect, a composition is provided comprising a recombinant albumin polypeptide and one or more fatty acids in an amount in a molar ratio effective to promote at least an 80% increase in viable cell density of T cells upon culturing T cells in the composition for a duration of 6 days, compared to a composition lacking the recombinant albumin polypeptide. In another aspect, compositions are provided comprising an albumin polypeptide and one or more fatty acids in amounts in a molar ratio effective to promote at least about an 80% increase in protein stabilization upon incubation of the protein in the composition for a duration of about 3000 seconds to about 6000 seconds, relative to a composition lacking the albumin polypeptide. In some instances, the composition is substantially free of growth factors.

[0019]

[0013] Additional aspects and advantages of the present disclosure will become readily apparent to those skilled in the art from the following detailed description, in which merely illustrative embodiments of the present disclosure are shown and described. As will be realized, the present disclosure is capable of other and different embodiments, and its several details are capable of modifications in various obvious respects, all without departing from the present disclosure. Accordingly, the drawings and description are to be regarded as illustrative in nature, and not as restrictive.

[0020] Incorporation by Reference All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.

[0021] The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings of which: [Brief explanation of the drawings]

[0022] [Figure 1] 1 depicts non-limiting examples of viable cell densities of induced pluripotent stem (iPS) cells grown in E8 medium supplemented with various albumins according to embodiments of the present disclosure. [Figure 2] 1 depicts non-limiting examples of viable cell densities of mesenchymal stem cells (MSCs) grown in chemically defined media supplemented with various albumins according to embodiments of the present disclosure. [Figure 3] 1 depicts non-limiting examples of viable cell numbers of T cells grown in chemically defined media supplemented with various albumins according to embodiments of the present disclosure. [Figure 4] 1 depicts non-limiting examples of absorbance at 360 nm of various albumin-stabilized insulin solutions according to embodiments of the present disclosure. [Figure 5]1 depicts non-limiting examples of viable cell densities of MSCs grown in chemically defined media supplemented with various albumins according to embodiments of the present disclosure. [Figure 6] 1 depicts non-limiting examples of viable cell densities of MSCs grown in chemically defined media supplemented with various albumins according to embodiments of the present disclosure. [Figure 7] 1 depicts non-limiting examples of viable cell densities of T cells grown in chemically defined media supplemented with various albumins according to embodiments of the present disclosure. [Figure 8] 1 depicts non-limiting examples of viable cell densities of iPS cells grown in Essential 8 Medium supplemented with various albumins according to embodiments of the present disclosure. [Figure 9] 1 depicts non-limiting examples of viable cell densities of T cells grown in chemically defined media supplemented with various albumins according to embodiments of the present disclosure. DETAILED DESCRIPTION OF THE INVENTION

[0023] While various embodiments of the present disclosure have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions may occur to those skilled in the art without departing from the present disclosure. It should be understood that various alternatives to the disclosed embodiments described herein may be employed.

[0024] As used in this specification and claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. For example, the term "a cell" includes a plurality of cells, including mixtures thereof.

[0025] As used herein, the term "about" a number refers to that number plus or minus 10% of that number. The term "about" a range refers to that range minus 10% of its lowest value and plus 10% of its highest value.

[0026] The terms "albumin" and "albumin polypeptide" are used interchangeably herein and generally refer to any protein or polypeptide that is a member of, related to, a variant of, a fragment of, a truncation of, and / or is derived from the albumin family of proteins. As used herein, "albumin" or "albumin polypeptide" may be a full-length albumin protein. As used herein, "albumin" or "albumin polypeptide" may be a wild-type albumin protein. As used herein, "albumin" or "albumin polypeptide" may be a variant of a wild-type albumin protein. For example, albumin or albumin polypeptide may comprise one or more variations compared to a wild-type albumin protein. The one or more variations may be one or more mutations. The one or more mutations may be one or more insertions, one or more deletions, and / or one or more substitutions compared to a wild-type albumin protein. As used herein, "albumin" or "albumin polypeptide" may be a truncated albumin protein. For example, albumin or albumin polypeptides may include truncations at the C-terminus, N-terminus, and / or internal truncations compared to wild-type albumin protein. As used herein, "albumin" or "albumin polypeptide" may be a fragment of albumin protein. Albumin polypeptides may be derived from any source. In some embodiments, albumin polypeptides are derived from blood (e.g., whole blood, plasma, serum). In some embodiments, albumin polypeptides are recombinant albumin. Recombinant albumin may be produced in host cells (e.g., yeast, bacteria, plant, mammalian cells, etc.).

[0027] As used herein, "albumin" or "albumin polypeptide" may comprise an amino acid sequence having at least about 50% sequence identity to a wild-type albumin protein. For example, "albumin" or "albumin polypeptide" as used herein may comprise an amino acid sequence having at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% sequence identity to a wild-type albumin protein.

[0028] In some embodiments, the albumin polypeptide is serum albumin or blood albumin. The serum albumin can be from any species. In some cases, the serum albumin is derived from a vertebrate. In some embodiments, the serum albumin is human serum albumin. In some embodiments, the serum albumin is bovine serum albumin. In some embodiments, the serum albumin is derived from blood.

[0029] In some instances, the albumin is wild-type human serum albumin. In some embodiments, "albumin" or "albumin polypeptide" as used herein comprises the amino acid sequence according to SEQ ID NO:1: (SEQ ID NO: 1)

[0030] In some embodiments, the albumin polypeptide comprises an amino acid sequence having at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% sequence identity to SEQ ID NO:1.

[0031] In some embodiments, the albumin polypeptide is bovine serum albumin. In some embodiments, the albumin polypeptide comprises the amino acid sequence according to SEQ ID NO:2: (SEQ ID NO: 2)

[0032] In some embodiments, the albumin polypeptide comprises an amino acid sequence having at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% sequence identity to SEQ ID NO:2.

[0033] Generally, "sequence identity" refers to the exact nucleotide-to-nucleotide or amino acid-to-amino acid correspondence of two polynucleotide or polypeptide sequences, respectively. Typically, techniques for determining sequence identity involve determining the nucleotide sequence of a polynucleotide and / or the amino acid sequence encoded thereby and comparing these sequences to a second nucleotide or amino acid sequence. Two or more sequences (polynucleotide or amino acid) can be compared by determining their "percent identity." The percent identity of two sequences, whether nucleic acid or amino acid, is the number of exact matches in the two aligned sequences divided by the length of the longer sequence, multiplied by 100. Percent identity may also be determined by comparing sequence information using, for example, the advanced BLAST computer program, including version 2.2.9, available from the National Institutes of Health. The BLAST program is based on the alignment method of Karlin and Altschul, Proc. Natl. Acad. Sci. USA, 87:2264-2268 (1990), and is described in Altschul et al., J. Mol. Biol., 215:403-410 (1990); Karlin and Altschul, Proc. Natl. Acad. Sci. USA, 90:5873-5877 (1993); and Altschul et al., Nucleic Acids Res., 25:3389-3402 (1997). The program may be used to determine percent identity over the entire length of the proteins being compared. For example, in the blastp program, default parameters are provided to optimize searches using short query sequences. The program also allows the use of a SEG filter to mask off segments of the query sequence as determined by the SEG program of Wootton and Federhen, Computers and Chemistry 17:149-163 (1993). The desired degree of sequence identity ranges from about 50% to 100% and integer values ​​therebetween.In general, the present disclosure encompasses sequences having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 98% sequence identity to any of the sequences provided herein.

[0034] The term "fatty acid" as used herein generally refers to a carboxylic acid having an aliphatic chain that can be either saturated or unsaturated. Fatty acids can be short-chain fatty acids, medium-chain fatty acids, long-chain fatty acids, or very long-chain fatty acids. Short-chain fatty acids can be fatty acids with an aliphatic tail of 5 or fewer carbons. Medium-chain fatty acids can be fatty acids with an aliphatic tail of 6 to 12 carbons. Long-chain fatty acids can be fatty acids with an aliphatic tail of 13 to 21 carbons. Very long-chain fatty acids can be fatty acids with an aliphatic tail of 22 or more carbons. Generally, fatty acids described herein are referred to by their common (or common) name and / or lipid number. As used herein, lipid numbers take the form C:D, where C is the number of carbon atoms in the fatty acid and D is the number of double bonds in the fatty acid.

[0035] The fatty acid may be a "saturated fatty acid," meaning a fatty acid that does not have a C=C double bond. Non-limiting examples of saturated fatty acids include propionic acid (C3:0), butyric acid (C4:0), valeric acid (C5:0), caproic acid (C6:0), enanthic acid (C7:0), caprylic acid (C8:0), pelargonic acid (C9:0), capric acid (C10:0), undecylic acid (C11:0), lauric acid (C12:0), tridecylic acid (C13:0), myristic acid (C14:0), pentadecylic acid (C15:0), and the like. :0), palmitic acid (C16:0), margaric acid (C17:0), stearic acid (C18:0), nonadecylic acid (C19:0), arachidic acid (C20:0), heneicosylic acid (C21:0), behenic acid (C22:0), tricosylic acid (C23:0), lignoceric acid (C24:0), pentacosylic acid (C25:0), cerotic acid (C26:0), carboceric acid acid (C27:0), montanic acid (C28:0), nonacosylic acid (C29:0), melissic acid (C30:0), hentriacontylic acid (C31:0), russellic acid (C32:0), psyllic acid (C33:0), gedaic acid (C34:0), ceroplastic acid (C35:0), hexatriacontylic acid (C36:0), heptatriacontylic acid (C37:0), octatriacontylic acid (C38:0), nonatriacontylic acid (C39:0), and tetracontylic acid (C40:0).

[0036] The fatty acid may be an "unsaturated fatty acid," which refers to a fatty acid having one or more C=C double bonds. Unsaturated fatty acids may be in either the cis or trans configuration. Non-limiting examples of unsaturated fatty acids include octenoic acid (C8:1), decenoic acid (C10:1), decadienoic acid (C10:2), lauroleic acid (C12:1), laurolinoleic acid (C12:2), myristovaccenic acid (C14:1), myristolinoleic acid (C14:2), myristolinolenic acid (C14:3), palmitolinolenic acid (C16:3), palmitidonic acid (C16:4), and the like. acid) (C16:4), α-linolenic acid (C18:3), stearidonic acid (C18:4), dihomo-α-linolenic acid (C20:3), eicosatetraenoic acid (C20:4), eicosapentaenoic acid (C20:5), clupanodonic acid (C22:5), docosahexaenoic acid (C22:6), 9,12,15,18,21-tetracosapentaenoic acid (C24:5), 6,9,12,15,18,21-tetracosahexaenoic acid (C24:6), myristoleic acid (C14:1) , palmitovaccenic acid (C16:1), α-eleostearic acid (C18:3), β-eleostearic acid (trans-C18:3), punicic acid (C18:3), 7,10,13-octadecatrienoic acid (C18:3), 9,12,15-eicosatrienoic acid (C20:3), β-eicosatetraenoic acid (C20:4), 8-tetradecenoic acid (C14:1), 12-octadecenoic acid (C18:1), linoleic acid (C18:2), linolelaidic acid acid) (trans-C18:2), gamma-linolenic acid (C18:3), calendic acid (C18:3), pinolenic acid (C18:3), dihomo-linoleic acid (C20:2), dihomo-gamma-linolenic acid (C20:3), arachidonic acid (C20:4), adrenic acid (C22:4), osbondic acid (C22:5), palmitoleic acid (C16:1), vaccenic acid (C18:1), rumenic acid (C18:2), paulic acid (C20:1), 7,10,13-eicosatrienoic acid (C20:3), oleic acid (C18:1), elaidic acid (trans-C18:1), gondoic acid (C20:1), erucic acid (C22:1), nervonic acid (C24:1), 8,11-eicosadienoic acid (C20:2), mead acid (C20:3), sapienic acid (C16:1), gadoleic acid (C20:1), 4-hexadecenoic acid (C16:1), petroselinic acid (C18:1), and 8-eicosenoic acid (C20:1).

[0037] The term "molar ratio" used herein generally refers to the moles of one substance relative to the moles of another substance.For example, the compositions provided herein can be described in terms of the mole ratio of fatty acid to albumin polypeptide (i.e., the moles of fatty acid relative to the moles of albumin polypeptide).

[0038] The term "substantially free," as used herein, generally refers to a composition that lacks a specified component or contains only trace amounts of the specified component. For example, a composition that is substantially free of a specific fatty acid may lack the fatty acid in the composition or may contain only trace amounts of the fatty acid in the composition. In some embodiments, the term "substantially free" can refer to a composition that has a level of the specified component that is below the level of detection by mass spectrometry. The term "substantially free" can also refer to a composition that contains, for example, a specified component in a molar ratio to albumin polypeptide that is less than 0.00005, less than 0.00001, less than 0.000005, less than 0.000001, or less than 0.0000005.

[0039] The term "blended" when used in connection with albumin polypeptides as described herein generally refers to albumin polypeptides that have been subjected to one or more processing steps, such as a delipidation step, a fat blending step, and / or a purification step.

[0040] The term "basal medium" and its plural forms (e.g., "basal media"), as used herein, generally refers to any medium capable of supporting the growth of cells (e.g., mammalian cells). Basal media provide standard inorganic salts, such as zinc, iron, magnesium, calcium, and potassium, as well as trace elements, vitamins, an energy source, a buffer system, and / or essential amino acids. Examples of basal media include, but are not limited to, Dulbecco's Modified Eagle's Medium (DMEM), DME / F12, Minimum Essential Medium (MEM), Basal Medium Eagle's (BME), RPMI 1640, F-10, F-12, α-Minimum Essential Medium (α-MEM), Glasgow Minimum Essential Medium (G-MEM), PF CHO (SAFC Biosciences), and Iscove's Modified Dulbecco's Medium. Basal media may be supplemented with various factors (e.g., one or more of the albumin compositions described herein) to enhance the growth and / or viability of cells cultured therein.

[0041] As used herein, the term "cell culture medium" and its plural forms (e.g., "cell culture media") generally refer to any nutrient solution for the maintenance, growth, propagation, and / or expansion of cells in an artificial in vitro environment outside of a multicellular organism or tissue. Cell culture media may be optimized for specific cell culture applications, including, for example, cell culture growth media formulated to promote cell growth or cell culture production media formulated to promote recombinant protein production. Cell culture media may include one or more of the albumin compositions described herein.

[0042] The term "defatted" when used with respect to the albumin polypeptides described herein generally refers to an albumin polypeptide composition that has been subjected to a "defatting" process to remove fatty acids. In some cases, the defatted albumin polypeptide is a "fatty acid-free" or "fatty acid-reduced" composition, i.e., a composition containing about 1% or less fatty acids. As used herein, "defatting" can refer to any method used to obtain defatted albumin polypeptide, including, but not limited to, charcoal treatment and the use of resin columns.

[0043] Disclosed herein are albumin compositions with defined fatty acid profiles and methods for using the same. The albumin compositions described herein are particularly suitable for use in cell culture methods and protein stabilization methods. The albumin compositions described herein can improve cell viability and / or promote proliferation when cells (e.g., mammalian cells) are cultured in a medium containing the albumin composition. The albumin compositions described herein can improve the stability of biological products when the biological product is in the presence of the albumin composition. Further provided herein are methods for formulating albumin compositions with defined fatty acid profiles as described herein.

[0044] Albumin Composition In one aspect, the disclosure provides a composition comprising: a) an albumin polypeptide; b) one or more fatty acids having fewer than 18 carbon atoms present in a molar ratio to the albumin polypeptide in the range of about 0.02 to about 0.4; and c) one or more fatty acids having 18 or more carbon atoms present in a molar ratio to the albumin polypeptide in the range of about 0.03 to about 0.6.

[0045] The one or more fatty acids having less than 18 carbon atoms are propionic acid (C3:0), butyric acid (C4:0), valeric acid (C5:0), caproic acid (C6:0), enanthic acid (C7:0), caprylic acid (C8:0), pelargonic acid (C9:0), capric acid (C10:0), undecylic acid (C11:0), lauric acid (C12:0), tridecylic acid (C13:0), myristic acid (C14:0), pentadecanoic acid (C15:0), palmitic acid (C16:0), hydroxybenzoic acid (C17:0), hydroxybenzoic acid (C18:0), hydroxybenzoic acid (C19:0), hydroxybenzoic acid (C20:0), hydroxybenzoic acid (C21:0), hydroxybenzoic acid (C22:0), hydroxybenzoic acid (C23:0), hydroxybenzoic acid (C24:0), hydroxybenzoic acid (C25:0), hydroxybenzoic acid (C26:0), hydroxybenzoic acid (C27:0), hydroxybenzoic acid (C28:0), hydroxybenzoic acid (C29:0), hydroxybenzoic acid (C30:0), hydroxybenzoic acid (C31:0), hydroxybenzoic acid (C32:0), hydroxybenzoic acid (C33:0), hydroxybenzoic acid (C34:0), hydroxybenzoic acid (C35:0), hydroxybenzoic acid (C36:0), hydroxybenzoic acid (C37:0), hydroxybenzoic acid (C38:0), hydroxybenzoic acid (C39 ... :0), margaric acid (C17:0), octenoic acid (C8:1), decenoic acid (C10:1), decadienoic acid (C10:2), lauroleic acid (C12:1), lauronolic acid (C12:2), myristovaccineic acid (C14:1), myristolinoleic acid (C14:2), myristolinolenic acid (C14:3), palmitrinolenic acid (C16:3), and palmitidonic acid (C16:4).

[0046] In some cases, the one or more fatty acids having fewer than 18 carbon atoms include one or more fatty acids selected from the group consisting of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), and palmitoleic acid (C16:1). In some cases, the one or more fatty acids having fewer than 18 carbon atoms consist of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), and palmitoleic acid (C16:1).

[0047] The one or more fatty acids having fewer than 18 carbon atoms may be present in the composition at a molar ratio to the albumin polypeptide ranging from about 0.02 to about 0.4. For example, the one or more fatty acids having fewer than 18 carbon atoms may be present at a molar ratio to the albumin polypeptide ranging from about 0.02, about 0.03, about 0.04, about 0.05, about 0.06, about 0.07, about 0.08, about 0.09, about 0.10, about 0.11, about 0.12, about 0.13, about 0.14, about 0.15, about 0.16, about 0.17, about 0.18, about 0.19, about 0.20, about 0.21 , about 0.22, about 0.23, about 0.24, about 0.25, about 0.26, about 0.27, about 0.28, about 0.29, about 0.30, about 0.31, about 0.32, about 0.33, about 0.34, about 0.35, about 0.36, about 0.37, about 0.38, about 0.39, or about 0.40. In some cases, one or more fatty acids having fewer than 18 carbons are attached to the albumin polypeptide.

[0048] The one or more fatty acids having 18 or more carbon atoms are stearic acid (C18:0), nonadecylic acid (C19:0), arachidic acid (C20:0), heneicosylic acid (C21:0), behenic acid (C22:0), tricosylic acid (C23:0), lignoceric acid (C24:0), pentacosylic acid (C25:0), cerotic acid (C26:0), carboselic acid (C27:0), montanic acid (C28:0), nonacosylic acid (C29:0), melissic acid (C30:0), hentriacontylic acid (C31:0), russellic acid (C32:0), russellic acid (C33:0), russellic acid (C34:0), russellic acid (C35:0), russellic acid (C36:0), russellic acid (C37:0), russellic acid (C38:0), russellic acid (C39:0), russellic acid (C40:0), russellic acid (C41:0), russellic acid (C42:0), russellic acid (C43:0), russellic acid (C44:0), russellic acid (C45:0), russellic acid (C46:0), russellic acid (C47:0), russellic acid (C48:0), russellic acid (C49:0), russellic acid (C49:0), russellic acid (C50:0), russellic acid (C51:0), russellic acid (C52:0), russellic acid (C53:0), :0), psyllic acid (C33:0), gedic acid (C34:0), ceroplastic acid (C35:0), hexatriacontylic acid (C36:0), heptatriacontylic acid (C37:0), octatriacontylic acid (C38:0), nonatriacontylic acid (C39:0), tetracontylic acid (C40:0), α-linolenic acid (C18:3), stearidonic acid (C18:4), dihomo-α-linolenic acid (C20:3), eicosatetraenoic acid (C20:4), eicosapentaenoic acid (C20:5), clupanodonic acid (C22:5), docosahexaenoic acid acid (C22:6), 9,12,15,18,21-tetracosapentaenoic acid (C24:5), 6,9,12,15,18,21-tetracosahexaenoic acid (C24:6), myristoleic acid (C14:1), palmitovaccenic acid (C16:1), α-eleostearic acid (C18:3), β-eleostearic acid (trans-C18:3), punicic acid (C18:3), 7,10,13-octadecatrienoic acid (C18:3), 9,12,15-eicosatrienoic acid (C20:3), β-eicosatetraenoic acid (C20:4), 8-tetracosapentaenoic acid (C20:4), α-ethoxybenzoic acid (C20:4), β ... Decenoic acid (C14:1), 12-octadecenoic acid (C18:1), linoleic acid (C18:2), linolelaidic acid (trans-C18:2), gamma-linolenic acid (C18:3), calendic acid (C18:3), pinolenic acid (C18:3), dihomo-linoleic acid (C20:2), dihomo-gamma-linolenic acid (C20:3), arachidonic acid (C20:4), adrenic acid (C22:4), osbondic acid (C22:5), palmitoleic acid (C16:1), vaccenic acid (C18:1), rumenic acid (C18:2), paulic acid (C20:1), 7,10,The fatty acids may include one or more fatty acids selected from the group consisting of 13-eicosatrienoic acid (C20:3), oleic acid (C18:1), elaidic acid (trans-C18:1), gondoic acid (C20:1), erucic acid (C22:1), nervonic acid (C24:1), 8,11-eicosadienoic acid (C20:2), mead acid (C20:3), sapienic acid (C16:1), gadoleic acid (C20:1), 4-hexadecenoic acid (C16:1), petroselinic acid (C18:1), and 8-eicosenoic acid (C20:1).

[0049] In some instances, the one or more fatty acids having 18 or more carbon atoms include one or more fatty acids selected from the group consisting of stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), and eicosadienoic acid (C20:2 ω-6). In some instances, the one or more fatty acids having 18 or more carbon atoms consist of stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), and eicosadienoic acid (C20:2 ω-6). In some cases, the one or more fatty acids having 18 or more carbon atoms further include one or more fatty acids selected from the group consisting of bishomo-gamma-linolenic acid (C20:3 ω-6), arachidonic acid (C20:4), docosatetraenoic acid (C22:4 ω-6), and docosahexaenoic acid (C22:6 ω-3).

[0050] The one or more fatty acids having 18 or more carbon atoms may be present in the composition at a molar ratio to the albumin polypeptide ranging from about 0.03 to about 0.6. For example, the one or more fatty acids having 18 or more carbon atoms may be present at a molar ratio to the albumin polypeptide ranging from about 0.03, about 0.04, about 0.05, about 0.06, about 0.07, about 0.08, about 0.09, about 0.10, about 0.11, about 0.12, about 0.13, about 0.14, about 0.15, about 0.16, about 0.17, about 0.18, about 0.19, about 0.20, about 0.21, about 0.22, about 0.23, about 0.24, about 0.25, about 0.26, about 0.27, about 0.28, about 0.29, about 0.30, about 0.31, about 0.32, about 0.33, about 0.34, about 0.35, about 0.36, about 0.37, about 0.38, about 0.39, about 100, about 101, about 102, about 103, about 104, about 105, about 106, about 107, about 108, about 109, about 110, about 111, about 112, about 113, about 114, about 115, about 116, about 117, about 118, about 119, about 120, about 121, about 122, about 123, about 124, about 125, about 126, about 127, about 128, about The fatty acid may be present in the composition in a molar ratio to the albumin polypeptide of about 0.32, about 0.33, about 0.34, about 0.35, about 0.36, about 0.37, about 0.38, about 0.39, about 0.40, about 0.41, about 0.42, about 0.43, about 0.44, about 0.45, about 0.46, about 0.47, about 0.48, about 0.49, about 0.50, about 0.51, about 0.52, about 0.53, about 0.54, about 0.55, about 0.56, about 0.57, about 0.58, about 0.59, or about 0.60. In some cases, one or more fatty acids having 18 or more carbons are attached to the albumin polypeptide.

[0051] In particular embodiments, the one or more fatty acids having fewer than 18 carbon atoms consist of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (16:0), and palmitoleic acid (C16:1); and the one or more fatty acids having 18 or more carbon atoms consist of stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), eicosadienoic acid (C20:2 ω-6), bishomo-γ-linolenic acid (C20:3 ω-6), arachidonic acid (C20:4), docosatetraenoic acid (C22:4 ω-6), and docosahexaenoic acid (C22:6 ω-3).

[0052] In particular embodiments, the one or more fatty acids having fewer than 18 carbon atoms consist of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), and palmitoleic acid (C16:1); and the one or more fatty acids having 18 or more carbon atoms consist of stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), and eicosadienoic acid (C20:2 ω-6).

[0053] In certain embodiments, the one or more fatty acids having fewer than 18 carbon atoms consist of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), and palmitoleic acid (C16:1); and the one or more fatty acids having 18 or more carbon atoms consist of stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), eicosadienoic acid (C20:2 ω-6), bishomo-γ-linolenic acid (C20:3 ω-6), and arachidonic acid (20:4).

[0054] In certain embodiments, the composition is substantially free of pentadecanoic acid (C15:0), margaric acid (C17:0), and / or heptadecenoic acid (C17:1 ω-7). In various aspects, the composition may be substantially free of one or more fatty acids selected from the group consisting of α-linolenic acid (C18:3), γ-linolenic acid (C18:3), arachidic acid (C20:0), eicosatrienoic acid (C20:3 ω-3), bishomo-γ-linolenic acid (C20:3 ω-6), arachidonic acid (C20:4), eicosapentaenoic acid (C20:5 ω-3), behenic acid (C22:0), docosatetraenoic acid (C22:4 ω-6), docosapentaenoic acid (C22:5 ω-3), docosahexaenoic acid (C22:6 ω-3), lignoceric acid (C24:0), or cerotic acid (C26:0).

[0055] In various aspects, the total molar ratio of fatty acids (e.g., one or more fatty acids having fewer than 18 carbon atoms and one or more fatty acids having 18 or more carbon atoms) to albumin polypeptide is less than about 1. For example, the total molar ratio of fatty acids to albumin polypeptide can be about 1, about 0.9, about 0.8, about 0.7, about 0.6, about 0.5, about 0.4, about 0.3, about 0.2, about 0.1, about 0.09, about 0.08, about 0.07, about 0.06, about 0.05, about 0.04, about 0.03, about 0.02, or about 0.01.

[0056] In another embodiment, the composition comprises: a) an albumin polypeptide; b) lauric acid (C12:0) present in a molar ratio to the albumin polypeptide ranging from about 0.001 to about 0.008; c) myristic acid (C14:0) present in a molar ratio to the albumin polypeptide ranging from about 0.001 to about 0.022; d) palmitic acid (C16:0) present in a molar ratio to the albumin polypeptide ranging from about 0.02 to about 0.3; e) palmitoleic acid (C16:1) present in a molar ratio to the albumin polypeptide ranging from about 0.002 to about 0.03; f) stearic acid (C18:0) present in a molar ratio to the albumin polypeptide ranging from about 0.011 to about 0.2; g) oleic acid (C18:1) present in a molar ratio to the albumin polypeptide ranging from about 0.02 to about 0.3. ω-9); h) linoleic acid (C18:2) present in a molar ratio to albumin polypeptide ranging from about 0.0002 to about 0.12; and i) eicosadienoic acid (C20:2 ω-6) present in a molar ratio to albumin polypeptide ranging from about 0.0001 to about 0.002.

[0057] In various embodiments, the lauric acid (C12:0) is present in a molar ratio to the albumin polypeptide ranging from about 0.001 to about 0.008. For example, the lauric acid (C12:0) can be present in a molar ratio to the albumin polypeptide of about 0.001, about 0.002, about 0.003, about 0.004, about 0.005, about 0.006, about 0.007, or about 0.008.

[0058] In various embodiments, myristic acid (C14:0) is present in a molar ratio to albumin polypeptide ranging from about 0.001 to about 0.022. For example, myristic acid (C14:0) may be present in a molar ratio to albumin polypeptide of about 0.001, about 0.002, about 0.003, about 0.004, about 0.005, about 0.006, about 0.007, about 0.008, about 0.009, about 0.010, about 0.011, about 0.012, about 0.013, about 0.014, about 0.015, about 0.016, about 0.017, about 0.018, about 0.019, about 0.020, about 0.021, or about 0.022.

[0059] In various embodiments, palmitic acid (C16:0) is present in a molar ratio to albumin polypeptide ranging from about 0.02 to about 0.3. For example, palmitic acid (C16:0) may be present in a molar ratio to albumin polypeptide of about 0.02, about 0.03, about 0.04, about 0.05, about 0.06, about 0.07, about 0.08, about 0.09, about 0.10, about 0.11, about 0.12, about 0.13, about 0.14, about 0.15, about 0.16, about 0.17, about 0.18, about 0.19, about 0.20, about 0.21, about 0.22, about 0.23, about 0.24, about 0.25, about 0.26, about 0.27, about 0.28, about 0.29, or about 0.30.

[0060] In various embodiments, palmitoleic acid (C16:1) is present in a molar ratio to albumin polypeptide ranging from about 0.002 to about 0.03. For example, palmitoleic acid (C16:1) may be present in a molar ratio to albumin polypeptide of about 0.002, about 0.003, about 0.004, about 0.005, about 0.006, about 0.007, about 0.008, about 0.009, about 0.010, about 0.011, about 0.012, about 0.013, about 0.014, about 0.015, about 0.016, about 0.017, about 0.018, about 0.019, about 0.020, about 0.021, about 0.022, about 0.023, about 0.024, about 0.025, about 0.026, about 0.027, about 0.028, about 0.029, or about 0.030.

[0061] In various embodiments, stearic acid (C18:0) is present in a molar ratio to albumin polypeptide ranging from about 0.011 to about 0.2. For example, stearic acid (C18:0) may be present in a molar ratio to albumin polypeptide of about 0.011, about 0.012, about 0.013, about 0.014, about 0.015, about 0.016, about 0.017, about 0.018, about 0.019, about 0.020, about 0.030, about 0.040, about 0.050, about 0.060, about 0.070, about 0.080, about 0.090, about 0.100, about 0.110, about 0.120, about 0.130, about 0.140, about 0.150, about 0.160, about 0.170, about 0.180, about 0.190, or about 0.200.

[0062] In various embodiments, the oleic acid (C18:1 ω-9) is present in a molar ratio to albumin polypeptide ranging from about 0.02 to about 0.3. For example, oleic acid (C18:1 omega-9) may be present in a molar ratio to albumin polypeptide of about 0.02, about 0.03, about 0.04, about 0.05, about 0.06, about 0.07, about 0.08, about 0.09, about 0.10, about 0.11, about 0.12, about 0.13, about 0.14, about 0.15, about 0.16, about 0.17, about 0.18, about 0.19, about 0.20, about 0.21, about 0.22, about 0.23, about 0.24, about 0.25, about 0.26, about 0.27, about 0.28, about 0.29, or about 0.30.

[0063] In various embodiments, linoleic acid (C18:2) is present in a molar ratio to albumin polypeptide ranging from about 0.0002 to about 0.12. For example, linoleic acid (C18:2) may be present in a molar ratio of about 0.0002, about 0.0003, about 0.0004, about 0.0005, about 0.0006, about 0.0007, about 0.0008, about 0.0009, about 0.0010, about 0.0020, about 0.0030, about 0.0040, about 0.0050, about 0.0060, about 0.0070, about 0.0080, about 0.0090, about 0.100, about 0.110, about 0.120, about 0.130, about 0.140, about 0.150, about 0.160, about 0.170, about 0.180, about 0.190, about 0.200, about 0.210, about 0.220, about 0.230, about 0.240, about 0.250, about 0.260, about 0.270, about 0.280, about 0.290, about 0.300, about 0.310, about 0.320, about 0.330, about 0.340, about 0.350, about 0.360, about 0.370, about 0.380, about 0.410, about 0.420, about 0.430, about 0.440, about 0.450, about 0.460, about 0.470, about The albumin polypeptide may be present in a molar ratio of about 0.0080, about 0.0090, about 0.0100, about 0.0200, about 0.0300, about 0.0400, about 0.0500, about 0.0600, about 0.0700, about 0.0800, about 0.0900, about 0.1000, about 0.1100, or about 0.1200.

[0064] In various embodiments, eicosadienoic acid (C20:2 ω-6) is present in a molar ratio to albumin polypeptide ranging from about 0.0001 to about 0.002. For example, eicosadienoic acid (C20:2 ω-6) may be present in a molar ratio to albumin polypeptide of about 0.0001, about 0.0002, about 0.0003, about 0.0004, about 0.0005, about 0.0006, about 0.0007, about 0.0008, about 0.0009, about 0.0010, about 0.0011, about 0.0012, about 0.0013, about 0.0014, about 0.0015, about 0.0016, about 0.0017, about 0.0018, about 0.0019, or about 0.0020.

[0065] In various embodiments, the composition may further comprise bishomo-gamma-linolenic acid (C20:3 ω-6) present in a molar ratio to albumin polypeptide ranging from about 0.0003 to about 0.002. For example, the bishomo-gamma-linolenic acid (C20:3 ω-6) may be present in a molar ratio to albumin polypeptide of about 0.0003, about 0.0004, about 0.0005, about 0.0006, about 0.0007, about 0.0008, about 0.0009, about 0.0010, about 0.0011, about 0.0012, about 0.0013, about 0.0014, about 0.0015, about 0.0016, about 0.0017, about 0.0018, about 0.0019, or about 0.0020.

[0066] In various embodiments, the composition may further comprise arachidonic acid (C20:4) present in a molar ratio to albumin polypeptide ranging from about 0.001 to about 0.01. For example, the arachidonic acid (C20:4) may be present in a molar ratio to albumin polypeptide of about 0.001, about 0.002, about 0.003, about 0.004, about 0.005, about 0.006, about 0.007, about 0.008, about 0.009, or about 0.010.

[0067] In various aspects, the composition may further comprise docosatetraenoic acid (C22:4 ω-6) present in a molar ratio to albumin polypeptide ranging from about 0.0009 to about 0.003. For example, docosatetraenoic acid (C22:4 ω-6) may be present in a molar ratio to albumin polypeptide of about 0.0009, about 0.0010, about 0.0011, about 0.0012, about 0.0013, about 0.0014, about 0.0015, about 0.0016, about 0.0017, about 0.0018, about 0.0019, about 0.0020, about 0.0021, about 0.0022, about 0.0023, about 0.0024, about 0.0025, about 0.0026, about 0.0027, about 0.0028, about 0.0029, or about 0.0030.

[0068] In various embodiments, the composition may further comprise docosahexaenoic acid (C22:6 ω-3) present in a molar ratio to albumin polypeptide ranging from about 0.0003 to about 0.001. For example, docosahexaenoic acid (C22:6 ω-3) may be present in a molar ratio to albumin polypeptide ranging from about 0.0003, about 0.0004, about 0.0005, about 0.0006, about 0.0007, about 0.0008, about 0.0009, or about 0.0010.

[0069] In various embodiments, the total molar ratio of fatty acids to albumin polypeptide is about 0.06 to about 1. For example, the total molar ratio of fatty acids to albumin polypeptide is about 0.06, about 0.07, about 0.08, about 0.09, or about 0.10, about 0.20, about 0.30, about 0.40, about 0.50, about 0.60, about 0.70, about 0.80, about 0.90, or about 1.00.

[0070] In various aspects, the composition is substantially free of pentadecanoic acid (C15:0), margaric acid (C17:0), heptadecenoic acid (C17:1 ω-7), α-linolenic acid (C18:3), γ-linolenic acid (C18:3), arachidic acid (C20:0), eicosatrienoic acid (C20:3 ω-3), eicosapentaenoic acid (C20:5 ω-3), behenic acid (C22:0), docosapentaenoic acid (C22:5 ω-3), lignoceric acid (C24:0), and / or cerotic acid (C26:0).

[0071] In another aspect, provided is a composition comprising: a) an albumin polypeptide; and b) one or more fatty acids, the composition being substantially free of one or more of pentadecanoic acid (C15:0), margaric acid (C17:0), heptadecenoic acid (C17:1 ω-7), α-linolenic acid (C18:3), γ-linolenic acid (C18:3), arachidic acid (C20:0), eicosatrienoic acid (C20:3 ω-3), eicosapentaenoic acid (C20:5 ω-3), behenic acid (C22:0), docosapentaenoic acid (C22:5 ω-3), lignoceric acid (C24:0), and cerotic acid (C26:0), and wherein the total molar ratio of the one or more fatty acids to the albumin polypeptide is from about 0.05 to about 1.

[0072] In another aspect, a composition is provided comprising: a) an albumin polypeptide; and b) fatty acids, wherein the fatty acids consist of: i) lauric acid (C12:0); ii) myristic acid (C14:0); iii) palmitic acid (C16:0); iv) palmitoleic acid (C16:1); v) stearic acid (C18:0); vi) oleic acid (C18:1 ω-9); vii) linoleic acid (C18:2); viii) eicosadienoic acid (C20:2 ω-6); ix) bishomo-γ-linolenic acid (C20:3 ω-6); x) arachidonic acid (C20:4); xi) docosatetraenoic acid (C22:4 ω-6); and xii) docosahexaenoic acid (C22:6 ω-3), wherein the total molar ratio of fatty acids to albumin polypeptide is less than about 1.

[0073] In various embodiments, the lauric acid (C12:0) is present in a molar ratio to the albumin polypeptide of less than about 0.01. For example, the lauric acid (C12:0) may be present in a molar ratio to the albumin polypeptide of about 0.001, about 0.002, about 0.003, about 0.004, about 0.005, about 0.006, about 0.007, or about 0.008, or about 0.009.

[0074] In various embodiments, myristic acid (C14:0) is present in a molar ratio to albumin polypeptide of less than about 0.05. For example, myristic acid (C14:0) may be present in a molar ratio to albumin polypeptide of about 0.001, about 0.002, about 0.003, about 0.004, about 0.005, about 0.006, about 0.007, about 0.008, about 0.009, about 0.010, about 0.020, about 0.030, or about 0.040.

[0075] In various embodiments, palmitic acid (C16:0) is present in a molar ratio to albumin polypeptide of less than about 0.5. For example, palmitic acid (C16:0) may be present in a molar ratio to albumin polypeptide of about 0.02, about 0.03, about 0.04, about 0.05, about 0.06, about 0.07, about 0.08, about 0.09, about 0.10, about 0.20, about 0.30, or about 0.40.

[0076] In various embodiments, palmitoleic acid (C16:1) is present in a molar ratio to albumin polypeptide of less than about 0.05. For example, palmitoleic acid (C16:1) may be present in a molar ratio to albumin polypeptide of about 0.002, about 0.003, about 0.004, about 0.005, about 0.006, about 0.007, about 0.008, about 0.009, about 0.010, about 0.020, about 0.030, or about 0.040.

[0077] In various embodiments, the stearic acid (C18:0) is present in a molar ratio to the albumin polypeptide of less than about 0.2. For example, stearic acid (C18:0) may be present in a molar ratio to albumin polypeptide of about 0.011, about 0.012, about 0.013, about 0.014, about 0.015, about 0.016, about 0.017, about 0.018, about 0.019, about 0.020, about 0.030, about 0.040, about 0.050, about 0.060, about 0.070, about 0.080, about 0.090, about 0.100, about 0.110, about 0.120, about 0.130, about 0.140, about 0.150, about 0.160, about 0.170, about 0.180, or about 0.190.

[0078] In various embodiments, the oleic acid (C18:1 ω-9) is present in a molar ratio to the albumin polypeptide of less than about 0.5. For example, the oleic acid (C18:1 ω-9) may be present in a molar ratio to the albumin polypeptide of about 0.02, about 0.03, about 0.04, about 0.05, about 0.06, about 0.07, about 0.08, about 0.09, about 0.10, about 0.20, about 0.30, or about 0.40.

[0079] In various embodiments, the linoleic acid (C18:2) is present in a molar ratio to the albumin polypeptide of less than about 0.2. For example, linoleic acid (C18:2) may be present in a molar ratio to albumin polypeptide of about 0.0002, about 0.0003, about 0.0004, about 0.0005, about 0.0006, about 0.0007, about 0.0008, about 0.0009, about 0.0010, about 0.0020, about 0.0030, about 0.0040, about 0.0050, about 0.0060, about 0.0070, about 0.0080, about 0.0090, about 0.0100, about 0.0200, about 0.0300, about 0.0400, about 0.0500, about 0.0600, about 0.0700, about 0.0800, about 0.0900, or about 0.1000.

[0080] In various embodiments, eicosadienoic acid (C20:2 ω-6) is present in a molar ratio to albumin polypeptide of less than about 0.005. For example, eicosadienoic acid (C20:2 ω-6) may be present in a molar ratio to albumin polypeptide of about 0.0001, about 0.0002, about 0.0003, about 0.0004, about 0.0005, about 0.0006, about 0.0007, about 0.0008, about 0.0009, about 0.0010, about 0.0020, about 0.0030, or about 0.0040.

[0081] In various embodiments, the bishomo-gamma-linolenic acid (C20:3 omega-6) is present in a molar ratio to the albumin polypeptide of less than about 0.005. For example, the bishomo-gamma-linolenic acid (C20:3 omega-6) may be present in a molar ratio to the albumin polypeptide of about 0.0003, about 0.0004, about 0.0005, about 0.0006, about 0.0007, about 0.0008, about 0.0009, about 0.0010, about 0.0020, about 0.0030, or about 0.0040.

[0082] In various embodiments, arachidonic acid (C20:4) is present in a molar ratio to albumin polypeptide of less than about 0.01. For example, arachidonic acid (C20:4) may be present in a molar ratio to albumin polypeptide of about 0.001, about 0.002, about 0.003, about 0.004, about 0.005, about 0.006, about 0.007, about 0.008, about 0.009, or about 0.010.

[0083] In various embodiments, docosatetraenoic acid (C22:4 ω-6) is present in a molar ratio to albumin polypeptide of less than about 0.005. For example, docosatetraenoic acid (C22:4 ω-6) may be present in a molar ratio to albumin polypeptide of about 0.0010, about 0.0020, about 0.0030, or about 0.0040.

[0084] In various embodiments, docosahexaenoic acid (C22:6 ω-3) is present in a molar ratio to albumin polypeptide of less than about 0.005. For example, docosahexaenoic acid (22:6 ω-3) may be present in a molar ratio to albumin polypeptide within the range of about 0.0003, about 0.0004, about 0.0005, about 0.0006, about 0.0007, about 0.0008, about 0.0009, or about 0.0010, about 0.0020, about 0.0030, or about 0.0040.

[0085] In yet another aspect, a composition is provided comprising: a) an albumin polypeptide; and b) fatty acids, wherein the fatty acids consist of: i) lauric acid (C12:0); ii) myristic acid (C14:0); iii) palmitic acid (C16:0); iv) palmitoleic acid (C16:1); v) stearic acid (C18:0); vi) oleic acid (C18:1 ω-9); vii) linoleic acid (C18:2); and viii) eicosadienoic acid (C20:2 ω-6).

[0086] In various embodiments, the lauric acid (C12:0) is present in a molar ratio to the albumin polypeptide of less than about 0.005. For example, the lauric acid (C12:0) may be present in a molar ratio to the albumin polypeptide of about 0.001, about 0.002, about 0.003, or about 0.004.

[0087] In various embodiments, myristic acid (C14:0) is present in a molar ratio to albumin polypeptide of less than about 0.005. For example, myristic acid (C14:0) may be present in a molar ratio to albumin polypeptide of about 0.001, about 0.002, about 0.003, or about 0.004.

[0088] In various embodiments, palmitic acid (C16:0) is present in a molar ratio to albumin polypeptide of less than about 0.05. For example, palmitic acid (C16:0) may be present in a molar ratio to albumin polypeptide of about 0.02, about 0.03, or about 0.04.

[0089] In various embodiments, palmitoleic acid (C16:1) is present in a molar ratio to albumin polypeptide of less than about 0.005. For example, palmitoleic acid (C16:1) may be present in a molar ratio to albumin polypeptide of about 0.002, about 0.003, or about 0.004.

[0090] In various embodiments, the stearic acid (C18:0) is present in a molar ratio to the albumin polypeptide of less than about 0.05. For example, the stearic acid (C18:0) may be present in a molar ratio to the albumin polypeptide of about 0.011, about 0.012, about 0.013, about 0.014, about 0.015, about 0.016, about 0.017, about 0.018, about 0.019, about 0.020, about 0.030, or about 0.040.

[0091] In various embodiments, the oleic acid (C18:1 ω-9) is present in a molar ratio to the albumin polypeptide of less than about 0.05. For example, the oleic acid (C18:1 ω-9) may be present in a molar ratio to the albumin polypeptide of about 0.02, about 0.03, or about 0.04.

[0092] In various embodiments, linoleic acid (C18:2) is present in a molar ratio to albumin polypeptide of less than about 0.0005. For example, linoleic acid (C18:2) may be present in a molar ratio to albumin polypeptide of about 0.0002, about 0.0003, or about 0.0004.

[0093] In various embodiments, eicosadienoic acid (C20:2 ω-6) is present in a molar ratio to albumin polypeptide of less than about 0.0005. For example, eicosadienoic acid (C20:2 ω-6) may be present in a molar ratio to albumin polypeptide of about 0.0001, about 0.0002, about 0.0003, or about 0.0004.

[0094] In another aspect, a composition is provided comprising: a) an albumin polypeptide; and b) fatty acids, wherein the fatty acids consist of: i) lauric acid (C12:0); ii) myristic acid (C14:0); iii) palmitic acid (C16:0); iv) palmitoleic acid (C16:1); v) stearic acid (C18:0); vi) oleic acid (C18:1 ω-9); vii) linoleic acid (C18:2); viii) eicosadienoic acid (C20:2 ω-6); ix) bishomo-γ-linolenic acid (C20:3 ω-6); and x) arachidonic acid (C20:4).

[0095] In various embodiments, the lauric acid (C12:0) is present in a molar ratio to the albumin polypeptide of less than about 0.005. For example, the lauric acid (C12:0) may be present in a molar ratio to the albumin polypeptide of about 0.001, about 0.002, about 0.003, or about 0.004.

[0096] In various embodiments, myristic acid (C14:0) is present in a molar ratio to albumin polypeptide of less than about 0.01. For example, myristic acid (C14:0) may be present in a molar ratio to albumin polypeptide of about 0.001, about 0.002, about 0.003, about 0.004, about 0.005, about 0.006, about 0.007, about 0.008, or about 0.009.

[0097] In various embodiments, palmitic acid (C16:0) is present in a molar ratio to albumin polypeptide of less than about 0.1. For example, palmitic acid (C16:0) may be present in a molar ratio to albumin polypeptide of about 0.02, about 0.03, about 0.04, about 0.05, about 0.06, about 0.07, about 0.08, or about 0.09.

[0098] In various embodiments, palmitoleic acid (C16:1) is present in a molar ratio to albumin polypeptide of less than about 0.01. For example, palmitoleic acid (C16:1) may be present in a molar ratio to albumin polypeptide of about 0.002, about 0.003, about 0.004, about 0.005, about 0.006, about 0.007, about 0.008, or about 0.009.

[0099] In various embodiments, the stearic acid (C18:0) is present in a molar ratio to the albumin polypeptide of less than about 0.1. For example, the stearic acid (C18:0) may be present in a molar ratio to the albumin polypeptide of about 0.01, about 0.020, about 0.030, about 0.040, about 0.050, about 0.060, about 0.070, about 0.080, or about 0.090.

[0100] In various embodiments, the oleic acid (C18:1 ω-9) is present in a molar ratio to the albumin polypeptide of less than about 0.5. For example, the oleic acid (C18:1 ω-9) may be present in a molar ratio to the albumin polypeptide of about 0.02, about 0.03, about 0.04, about 0.05, about 0.06, about 0.07, about 0.08, about 0.09, about 0.10, about 0.20, about 0.30, or about 0.40.

[0101] In various embodiments, the linoleic acid (C18:2) is present in a molar ratio to the albumin polypeptide of less than about 0.05. For example, the linoleic acid (C18:2) may be present in a molar ratio to the albumin polypeptide of about 0.0002, about 0.0003, about 0.0004, about 0.0005, about 0.0006, about 0.0007, about 0.0008, about 0.0009, about 0.0010, about 0.0020, about 0.0030, or about 0.0040.

[0102] In various embodiments, eicosadienoic acid (C20:2 ω-6) is present in a molar ratio to albumin polypeptide of less than about 0.0005. For example, eicosadienoic acid (C20:2 ω-6) may be present in a molar ratio to albumin polypeptide of about 0.0001, about 0.0002, about 0.0003, or about 0.0004.

[0103] In various embodiments, the bishomo-gamma-linolenic acid (C20:3 omega-6) is present in a molar ratio to the albumin polypeptide of less than about 0.0005. For example, the bishomo-gamma-linolenic acid (C20:3 omega-6) may be present in a molar ratio to the albumin polypeptide of about 0.0003 or about 0.0004.

[0104] In various embodiments, arachidonic acid (C20:4) is present in a molar ratio to albumin polypeptide of less than about 0.005. For example, arachidonic acid (C20:4) may be present in a molar ratio to albumin polypeptide of about 0.001, about 0.002, about 0.003, or about 0.004.

[0105] Further provided herein are albumin compositions having an increased molar ratio of arachidonic acid (C20:4). In certain embodiments, compositions are provided that include: a) an albumin polypeptide; and b) arachidonic acid (C20:4) present in a molar ratio relative to the total amount of fatty acids in the composition of at least about 0.9% (e.g., at least about 1.0%, at least about 2.0%, at least about 3.0%, at least about 4.0%, at least about 5.0%, at least about 6.0%, at least about 7.0%, at least about 8.0%, at least about 9.0%, at least about 10.0%, or more).

[0106] In some embodiments, the albumin polypeptide may be a full-length albumin polypeptide. In some embodiments, the albumin polypeptide may be a wild-type albumin polypeptide. In some cases, the wild-type albumin polypeptide is human serum albumin and has an amino acid sequence according to SEQ ID NO:1. In some cases, the wild-type albumin polypeptide is bovine serum albumin and has an amino acid sequence according to SEQ ID NO:2. In some embodiments, the albumin polypeptide is a variant of a wild-type albumin polypeptide. A variant albumin polypeptide may have one or more mutations (e.g., one or more insertions, one or more deletions, and / or one or more substitutions) compared to the wild-type albumin polypeptide. In some cases, a variant albumin polypeptide has one or more mutations (e.g., one or more insertions, one or more deletions, and / or one or more substitutions) compared to SEQ ID NO:1. In some cases, the variant albumin polypeptide has one or more mutations (e.g., one or more insertions, one or more deletions, and / or one or more substitutions) compared to SEQ ID NO:2. In some embodiments, the albumin polypeptide may be truncated (e.g., at the N-terminus, C-terminus, and / or internally truncated) compared to the wild-type albumin polypeptide. In some cases, the albumin polypeptide is truncated (e.g., at the N-terminus, C-terminus, and / or internally truncated) compared to SEQ ID NO:1. In some cases, the albumin polypeptide is truncated (e.g., at the N-terminus, C-terminus, and / or internally truncated) compared to SEQ ID NO:2. In some embodiments, the albumin polypeptide may be a fragment of the wild-type polypeptide. In some cases, the albumin polypeptide may be a fragment of an albumin polypeptide having an amino acid sequence according to SEQ ID NO:1. In some cases, the albumin polypeptide may be a fragment of an albumin polypeptide having an amino acid sequence according to SEQ ID NO:2.

[0107] In some embodiments, the albumin polypeptide may have an amino acid sequence that has at least about 50% sequence identity to the amino acid sequence of a wild-type albumin protein. For example, the albumin polypeptide may have an amino acid sequence that has at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% sequence identity to the amino acid sequence of a wild-type albumin protein. In some embodiments, the albumin polypeptide may have an amino acid sequence that has at least about 50% sequence identity to SEQ ID NO:1. For example, an albumin polypeptide may have an amino acid sequence having at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% sequence identity to SEQ ID NO: 1. In some embodiments, an albumin polypeptide may have an amino acid sequence having at least about 50% sequence identity to SEQ ID NO:2. For example, an albumin polypeptide may have an amino acid sequence having at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% sequence identity to SEQ ID NO:2.

[0108] The albumin polypeptide can be from any suitable species. In one embodiment, the albumin is from a human. In another embodiment, the albumin is from a bovine. In another embodiment, the albumin is from a porcine. The albumin polypeptide may be from any source. In some embodiments, the albumin polypeptide is derived from blood (e.g., whole blood, plasma, serum). In some embodiments, the albumin polypeptide is a recombinant albumin polypeptide. Recombinant albumin may be produced in any host cell, including, but not limited to, bacteria, yeast, fungi, plants, mammalian cells, and insect cells. In some embodiments, the albumin polypeptide may be recombinantly produced in bacteria. Without limitation, the bacteria may be of the species Escherichia coli. In some embodiments, the albumin polypeptide may be recombinantly produced in yeast. Without limitation, the yeast may be of the species Pichia pastoris, Saccharomyces cerevisiae, or Kluyveromyces lactis. In some embodiments, the albumin polypeptide may be recombinantly produced in a plant. Without limitation, the plant may be a plant of the species Oryza sativa (e.g., rice). In some embodiments, the albumin polypeptide may be recombinantly produced in a mammalian cell. Without limitation, the mammalian cell may be a CHO cell or a HEK293 cell. In some embodiments, the albumin polypeptide may be a defatted albumin polypeptide (e.g., that has been subjected to a delipidation process, e.g., by a method described herein). In some embodiments, the albumin polypeptide is commercially available albumin that has been defatted, e.g., according to a method described herein. In some cases, the albumin polypeptide compositions described herein (e.g., combinations of albumin polypeptides and fatty acid mixtures) are not found in nature (e.g., do not occur in nature).

[0109] In some embodiments, the albumin compositions described herein may be in a liquid formulation (e.g., a solution) or a solid formulation. The albumin compositions described herein may be provided in a liquid formulation at about 1% (w / w), about 5% (w / w), about 10% (w / w), about 15% (w / w), about 20% (w / w), or higher concentrations. The albumin compositions described herein may be provided in a buffer solution (e.g., Dulbecco's phosphate buffered saline (dPBS)). In some embodiments, the albumin compositions described herein may be in a dried form (e.g., lyophilized) that can be reconstituted (e.g., by the addition of a solution (e.g., dPBS)). The albumin compositions provided herein may be substantially free of one or more impurities. The albumin compositions provided herein may be substantially free of growth factors.

[0110] Further provided herein in various aspects are cell culture media, e.g., for culturing biological cells (e.g., mammalian cells). The cell culture media may comprise a basal medium and any of the albumin compositions described herein. The basal medium may be any basal medium, including, but not limited to, Dulbecco's Modified Eagle's Medium (DMEM), DME / F12, Minimum Essential Medium (MEM), Basal Medium Eagle's (BME), RPMI 1640, F-10, F-12, α-Minimum Essential Medium (α-MEM), Glasgow Minimum Essential Medium (G-MEM), PF CHO (SAFC Biosciences), and Iscove's Modified Dulbecco's Medium. It is understood that the type of basal medium selected will depend on the type of cells to be cultured therein.

[0111] The albumin composition may be present in the cell culture medium at any suitable concentration to achieve the desired result (e.g., improved growth and / or viability of cells cultured therein). In some embodiments, the albumin composition is present in the cell culture medium at a concentration in the range of about 0.02% (w / w) to about 10% (w / w). For example, any of the albumin compositions described herein may be present in an amount of about 0.02% (w / w), about 0.03% (w / w), about 0.04% (w / w), about 0.05% (w / w), about 0.06% (w / w), about 0.07% (w / w), about 0.08% (w / w), about 0.09% (w / w), about 0.1% (w / w), about 0.5% (w / w), about 1.0% (w / w), about 1.5% (w / w), about 2.0% (w / w), about 2.5% (w / w), It may be present in the cell culture medium at about 3.0% (w / w), about 3.5% (w / w), about 4.0% (w / w), about 4.5% (w / w), about 5.0% (w / w), about 5.5% (w / w), about 6.0% (w / w), about 6.5% (w / w), about 7.0% (w / w), about 7.5% (w / w), about 8.0% (w / w), about 8.5% (w / w), about 9.0% (w / w), about 9.5% (w / w), or about 10.0% (w / w).

[0112] In some embodiments, the albumin composition is present in the cell culture medium at a concentration ranging from about 0.2 mg / mL to about 100 mg / mL. For example, the albumin composition may be present in the cell culture medium at a concentration of about 0.2 mg / mL, about 0.5 mg / mL, about 1.0 mg / mL, about 5 mg / mL, about 10 mg / mL, about 15 mg / mL, about 20 mg / mL, about 25 mg / mL, about 30 mg / mL, about 35 mg / mL, about 40 mg / mL, about 45 mg / mL, about 50 mg / mL, about 55 mg / mL, about 60 mg / mL, about 65 mg / mL, about 70 mg / mL, about 75 mg / mL, about 80 mg / mL, about 85 mg / mL, about 90 mg / mL, about 95 mg / mL, or about 100 mg / mL.

[0113] The cell culture medium may further comprise one or more biological cells. Any type of biological cell may be cultured in the cell culture medium described herein. In some cases, the one or more biological cells are eukaryotic cells. In some cases, the eukaryotic cells are stem cells. Non-limiting examples of stem cells that may be cultured in the cell culture medium described herein include embryonic stem (ES) cells, induced pluripotent stem (iPS) cells, and mesenchymal stem cells (MSCs). In some cases, the eukaryotic cells are T cells. In some cases, the eukaryotic cells are neural cells.

[0114] In some embodiments, cells may exhibit increased cell viability, increased cell proliferation, or both when cultured in a cell culture medium as described herein (e.g., comprising an albumin composition described herein). In some embodiments, cells may exhibit an increase in viable cell density when cultured in a cell culture medium as described herein (e.g., comprising an albumin composition described herein). In some cases, cells may exhibit an increase in viable cell density of at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, or at least about 50% when cultured in a cell culture medium as described herein (e.g., comprising an albumin composition described herein). In any of the above embodiments, the increase in cell viability and / or cell proliferation is compared to cells cultured in the absence of an albumin composition described herein.

[0115] In another aspect, further provided herein is a composition comprising an albumin polypeptide and one or more fatty acids in an amount in a molar ratio effective to promote an increase in viable cell density of induced pluripotent stem (iPS) cells of at least about 35% (e.g., at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, or higher) upon culturing iPS cells in the composition for a duration of 4 days, compared to a composition lacking the albumin polypeptide. In some cases, the composition is added to a basal medium to provide a cell culture medium as described herein. In some cases, the albumin composition is provided in a cell culture medium at a concentration of about 0.2 mg / mL to about 2.0 mg / mL. In certain embodiments, the albumin composition is provided in a cell culture medium at a concentration of about 1.0 mg / mL. In some embodiments, the albumin composition is the albumin composition described in Example 2. A non-limiting example of culturing iPS cells in an albumin composition described herein and measuring viable cell density is provided in Example 3. In some embodiments, viable cell density is measured by counting the number of viable cells in the culture. In a non-limiting example, iPS cells may be cultured in Essential 8 medium supplemented with an albumin composition at about 1 mg / mL to about 1.5 mg / mL.

[0116] In another aspect, provided herein are compositions comprising an albumin polypeptide and one or more fatty acids in amounts in a molar ratio effective to promote an increase in viable cell density of mesenchymal stem cells (MSCs) of at least about 35% (e.g., at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, or higher) upon culturing the MSCs in the composition for a duration of 5 days compared to a composition lacking the albumin polypeptide. In some cases, the composition is added to a basal medium to provide a cell culture medium as described herein. In some cases, the albumin composition is provided in a cell culture medium at a concentration of about 0.2 mg / mL to about 1.5 mg / mL. In some embodiments, the albumin composition is the albumin composition described in Example 4. A non-limiting example of culturing MSCs in an albumin composition described herein and measuring viable cell density is provided in Example 5. In some embodiments, the viable cell density is measured by counting the number of viable cells in the culture. In a non-limiting example, MSCs may be cultured in MSC chemically defined medium supplemented with an albumin composition at a concentration of about 0.5 mg / mL to about 1.5 mg / mL.

[0117] In another aspect, provided herein are compositions comprising an albumin polypeptide and one or more fatty acids in amounts in a molar ratio effective to promote an increase in viable cell density of T cells by at least about 80% (e.g., at least about 85%, at least about 90%, at least about 95%, at least about 98%, or higher) upon culturing T cells in the composition for a duration of 6 days compared to a composition lacking the albumin polypeptide. In some cases, the albumin composition is added to a basal medium to provide a cell culture medium as described herein. In some cases, the albumin composition is provided in a cell culture medium at a concentration of about 0.5 mg / mL to about 100 mg / mL. In some embodiments, the albumin composition is the albumin composition described in Example 2. A non-limiting example of culturing T cells in an albumin composition described herein and measuring viable cell density is provided in Example 6. In some embodiments, the viable cell density is measured by counting the number of viable cells in the culture. In a non-limiting example, T cells may be cultured in T cell chemistry-defined medium supplemented with an albumin composition at a concentration of about 0.5 mg / mL to about 100 mg / mL.

[0118] In some embodiments, the albumin compositions described herein may be used to increase the stability of a biological product. As used herein, the term "biological product" includes proteins (including polypeptides and peptides), antibodies (or fragments or derivatives thereof), aptamers (or fragments or derivatives thereof), viruses (or fragments or derivatives thereof), vaccines, and biological cells (e.g., mammalian cells), among others. In some cases, a biological product may exhibit increased stability when in the presence of an albumin composition described herein. In some cases, a biological product may exhibit an increase in stability of at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or greater when in the presence of an albumin composition described herein. In any of the above embodiments, the increase in stability is compared to the biological product in the absence of an albumin composition described herein.

[0119] In some embodiments, the albumin composition may be present at a concentration in the range of about 0.01% (w / w) to about 20% (w / w) (e.g., when used to stabilize a biological product). For example, the albumin composition may be about 0.01% (w / w), about 0.02% (w / w), about 0.03% (w / w), about 0.04% (w / w), about 0.05% (w / w), about 0.06% (w / w), about 0.07% (w / w), about 0.08% (w / w), about 0.09% (w / w), about 0.1% (w / w), about 0.5% (w / w), about 1.0% (w / w), about 1.5% (w / w), about 2.0% (w / w), about 2.5% (w / w), about 3.0% (w / w), about 3.5% (w / w), about 4.0% (w / w), about 4.5% (w / w), about 5.0 ... The soluble solid may be present in a concentration of about 0.5% (w / w), about 6.0% (w / w), about 6.5% (w / w), about 7.0% (w / w), about 7.5% (w / w), about 8.0% (w / w), about 8.5% (w / w), about 9.0% (w / w), about 9.5% (w / w), about 10.0% (w / w), about 11.0% (w / w), about 12.0% (w / w), about 13.0% (w / w), about 14.0% (w / w), about 15.0% (w / w), about 16.0% (w / w), about 17.0% (w / w), about 18.0% (w / w), about 19.0% (w / w), or about 20.0% (w / w).

[0120] In some embodiments, the albumin composition may be present at a concentration in the range of about 0.1 mg / mL to about 200 mg / mL (e.g., when used to stabilize a biological product). For example, the albumin composition may be present at a concentration of about 0.1 mg / mL, about 0.5 mg / mL, about 1.0 mg / mL, about 5 mg / mL, about 10 mg / mL, about 15 mg / mL, about 20 mg / mL, about 25 mg / mL, about 30 mg / mL, about 35 mg / mL, about 40 mg / mL, about 45 mg / mL, about 50 mg / mL, about 55 mg / mL, about 60 mg / mL, about 65 mg / mL, about 70 mg / mL, about It may be present in a concentration of about 75 mg / mL, about 80 mg / mL, about 85 mg / mL, about 90 mg / mL, about 95 mg / mL, about 100 mg / mL, about 110 mg / mL, about 120 mg / mL, about 130 mg / mL, about 140 mg / mL, about 150 mg / mL, about 160 mg / mL, about 170 mg / mL, about 180 mg / mL, about 190 mg / mL, or about 200 mg / mL.

[0121] In some embodiments, the albumin composition may be present in a molar ratio relative to the biological product of about 0.1 to about 10. For example, the albumin composition may be present in a molar ratio relative to the biological product of about 0.1, about 0.5, about 1.0, about 1.5, about 2.0, about 2.5, about 3.0, about 3.5, about 4.0, about 4.5, about 5.0, about 5.5, about 6.0, about 6.5, about 7.0, about 7.5, about 8.0, about 8.5, about 9.0, about 9.5, or about 10.0.

[0122] In another aspect, provided herein are compositions comprising an albumin polypeptide and one or more fatty acids in amounts in a molar ratio effective to promote at least about an 80% increase in protein stabilization upon incubation of the protein in the composition for a duration of about 3000 seconds to about 6000 seconds, compared to a composition lacking the albumin polypeptide. In some cases, the albumin composition is present at a concentration of about 1 mg / mL to about 200 mg / mL. In some cases, the albumin composition is present at a molar ratio to protein of about 0.1 to about 10. In some embodiments, the albumin composition is an albumin composition as described in Example 8. In some cases, the protein is insulin, and the ability of the albumin composition to stabilize insulin aggregation is measured, for example, as described in Example 9.

[0123] Methods of formulating albumin compositions Further provided herein are methods for formulating the albumin compositions described herein. Generally, the methods involve one or more processing steps to remove the fatty acid composition of the albumin polypeptide (e.g., delipidation), one or more processing steps to re-formulate the fatty acid composition of the albumin polypeptide (e.g., fat blending), and / or one or more processing steps to purify and / or remove impurities from the albumin polypeptide.

[0124] In some embodiments, the method involves a delipidation step. In some cases, the delipidation step involves treating the albumin polypeptide with activated carbon. In other cases, the delipidation step involves ion exchange chromatography. In some embodiments, the delipidation step removes all or substantially all fatty acids from the albumin polypeptide. An albumin polypeptide after undergoing a delipidation step may be referred to as a "defatted" albumin polypeptide.

[0125] In some embodiments, the method involves a fat blending process. In some cases, the fat blending process involves incubating the defatted albumin polypeptide in a desired fatty acid mixture. The fatty acid mixture can be any mixture of fatty acids suitable to achieve the desired composition and molar ratio of fatty acids. The fatty acid mixture can be any mixture of fatty acids as described herein that is suitable to achieve the molar ratios and desired results as described herein. In some cases, the mixture of fatty acids comprises a mixture of one or more fatty acids selected from the group consisting of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), palmitoleic acid (C16:1); stearic acid (C18:0); oleic acid (C18:1), linoleic acid (C18:2), eicosadienoic acid (C20:2), bishomo-gamma-linolenic acid (C20:3), arachidonic acid (C20:4), docosatetraenoic acid (C22:4), and docosahexaenoic acid (C22:6).

[0126] In some embodiments, the methods may involve one or more steps of removing impurities from the albumin polypeptide and / or purifying the compositions described herein. In some cases, the methods may involve incubating the albumin polypeptide with a chelating resin (e.g., Diaion™ CR20).

[0127] In another aspect, a method for improving the function of an albumin polypeptide is provided, comprising: a) passing a solution of albumin polypeptide through a ceramic hydroxyapatite resin to produce a flow-through containing the albumin polypeptide; and b) purifying the albumin polypeptide from the flow-through to obtain an albumin polypeptide with improved function. The method may further comprise, prior to a), delipidating the albumin polypeptide by incubating the solution with activated carbon and purifying the albumin polypeptide from the activated carbon. The method may further comprise, prior to a), incubating the solution with a chelating resin and purifying the albumin polypeptide from the chelating resin. In some cases, the chelating resin is Diaion™ CR20. The method may further comprise, prior to a), incubating the albumin polypeptide with one or more fatty acids. In some cases, the one or more fatty acids are selected from the group consisting of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), palmitoleic acid (C16:1), stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), eicosadienoic acid (C20:2 ω-6), bishomo-γ-linolenic acid (C20:3 ω-6), arachidonic acid (C20:4), docosatetraenoic acid (C22:4 ω-6), and docosahexaenoic acid (C22:6 ω-3). In some cases, the one or more fatty acids consist of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), palmitoleic acid (C16:1), stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), eicosadienoic acid (C20:2 ω-6), bishomo-γ-linolenic acid (C20:3 ω-6), arachidonic acid (C20:4), docosatetraenoic acid (C22:4 ω-6), and docosahexaenoic acid (C22:6 ω-3).In some cases, the one or more fatty acids consist of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), palmitoleic acid (C16:1), stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), and eicosadienoic acid (C20:2 ω-6). In some cases, the one or more fatty acids consist of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), palmitoleic acid (C16:1), stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), eicosadienoic acid (C20:2 ω-6), bishomo-γ-linolenic acid (C20:3 ω-6), and arachidonic acid (C20:4).

[0128] How to use Further provided herein are methods for using the albumin compositions provided herein.In some cases, the albumin composition may be used to improve cell viability and / or proliferation (for example, by adding the albumin composition to a basal medium).In some cases, the albumin composition may be used to improve the stability of biological products.

[0129] In one aspect, a method for increasing cell viability, cell proliferation, or both is provided, the method comprising incubating cells in the presence of an albumin composition (e.g., in a cell culture medium) as described herein. In some cases, when cells are cultured in a cell culture medium as described herein (e.g., comprising an albumin composition described herein), they may exhibit an increase in viable cell density of at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, or at least about 50%. The increase in cell viability and / or cell proliferation may be compared to cells cultured in the absence of the albumin composition described herein.

[0130] The cell culture medium may comprise any basal medium and any of the albumin compositions described herein. The basal medium may be any basal medium, including, but not limited to, Dulbecco's Modified Eagle's Medium (DMEM), DME / F12, Minimum Essential Medium (MEM), Basal Medium Eagle's (BME), RPMI 1640, F-10, F-12, α-Minimum Essential Medium (α-MEM), Glasgow Minimum Essential Medium (G-MEM), PF CHO (SAFC Biosciences), and Iscove's Modified Dulbecco's Medium. It is understood that the type of basal medium selected will depend on the type of cells to be cultured therein.

[0131] The albumin composition may be present in the cell culture medium at any suitable concentration to achieve the desired result (e.g., improved growth and / or viability of cells cultured therein). In some embodiments, the albumin composition is present in the cell culture medium at a concentration in the range of about 0.01% (w / w) to about 10% (w / w). For example, any of the albumin compositions described herein may be present in an amount of about 0.01% (w / w), about 0.02% (w / w), about 0.03% (w / w), about 0.04% (w / w), about 0.05% (w / w), about 0.06% (w / w), about 0.07% (w / w), about 0.08% (w / w), about 0.09% (w / w), about 0.1% (w / w), about 0.5% (w / w), about 1.0% (w / w), about 1.5% (w / w), about 2.0% (w / w), about 2.5% (w / w), or about 3.0% (w / w). (w / w), about 3.0% (w / w), about 3.5% (w / w), about 4.0% (w / w), about 4.5% (w / w), about 5.0% (w / w), about 5.5% (w / w), about 6.0% (w / w), about 6.5% (w / w), about 7.0% (w / w), about 7.5% (w / w), about 8.0% (w / w), about 8.5% (w / w), about 9.0% (w / w), about 9.5% (w / w), or about 10.0% (w / w).

[0132] In some embodiments, the albumin composition is present in the cell culture medium at a concentration ranging from about 0.1 mg / mL to about 100 mg / mL. For example, the albumin composition may be present in the cell culture medium at a concentration of about 0.1 mg / mL, about 0.5 mg / mL, about 1.0 mg / mL, about 5 mg / mL, about 10 mg / mL, about 15 mg / mL, about 20 mg / mL, about 25 mg / mL, about 30 mg / mL, about 35 mg / mL, about 40 mg / mL, about 45 mg / mL, about 50 mg / mL, about 55 mg / mL, about 60 mg / mL, about 65 mg / mL, about 70 mg / mL, about 75 mg / mL, about 80 mg / mL, about 85 mg / mL, about 90 mg / mL, about 95 mg / mL, or about 100 mg / mL.

[0133] Any type of cell may be cultured in the cell culture medium described herein. In some cases, the cell may be a eukaryotic cell. In some cases, the eukaryotic cell is a stem cell. Non-limiting examples of stem cells that may be cultured in the cell culture medium described herein include embryonic stem (ES) cells, induced pluripotent stem (iPS) cells, and mesenchymal stem cells (MSCs). In some cases, the eukaryotic cell is a T cell. In some cases, the eukaryotic cell is a neural cell.

[0134] Further provided herein is a method for increasing the stability of a biological product, comprising incubating the biological product in the presence of an albumin composition as described herein. In some cases, the biological product may exhibit an increase in stability of at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or more when in the presence of an albumin composition as described herein. The increase in stability may be compared to the biological product in the absence of the albumin composition as described herein.

[0135] In some embodiments, the albumin composition may be present at a concentration in the range of about 0.01% (w / w) to about 20% (w / w) (e.g., when used to stabilize a biological product). For example, the albumin composition may be about 0.01% (w / w), about 0.02% (w / w), about 0.03% (w / w), about 0.04% (w / w), about 0.05% (w / w), about 0.06% (w / w), about 0.07% (w / w), about 0.08% (w / w), about 0.09% (w / w), about 0.1% (w / w), about 0.5% (w / w), about 1.0% (w / w), about 1.5% (w / w), about 2.0% (w / w), about 2.5% (w / w), about 3.0% (w / w), about 3.5% (w / w), about 4.0% (w / w), about 4.5% (w / w), about 5.0 ... The soluble solid may be present in a concentration of about 0.5% (w / w), about 6.0% (w / w), about 6.5% (w / w), about 7.0% (w / w), about 7.5% (w / w), about 8.0% (w / w), about 8.5% (w / w), about 9.0% (w / w), about 9.5% (w / w), about 10.0% (w / w), about 11.0% (w / w), about 12.0% (w / w), about 13.0% (w / w), about 14.0% (w / w), about 15.0% (w / w), about 16.0% (w / w), about 17.0% (w / w), about 18.0% (w / w), about 19.0% (w / w), or about 20.0% (w / w).

[0136] In some embodiments, the albumin composition may be present at a concentration in the range of about 0.1 mg / mL to about 200 mg / mL (e.g., when used to stabilize a biological product). For example, the albumin composition may be present at a concentration of about 0.1 mg / mL, about 0.5 mg / mL, about 1.0 mg / mL, about 5 mg / mL, about 10 mg / mL, about 15 mg / mL, about 20 mg / mL, about 25 mg / mL, about 30 mg / mL, about 35 mg / mL, about 40 mg / mL, about 45 mg / mL, about 50 mg / mL, about 55 mg / mL, about 60 mg / mL, about 65 mg / mL, about 70 mg / mL, about It may be present in a concentration of about 75 mg / mL, about 80 mg / mL, about 85 mg / mL, about 90 mg / mL, about 95 mg / mL, about 100 mg / mL, about 110 mg / mL, about 120 mg / mL, about 130 mg / mL, about 140 mg / mL, about 150 mg / mL, about 160 mg / mL, about 170 mg / mL, about 180 mg / mL, about 190 mg / mL, or about 200 mg / mL.

[0137] In some embodiments, the albumin composition may be present in a molar ratio relative to the biological product of about 0.1 to about 10. For example, the albumin composition may be present in a molar ratio relative to the biological product of about 0.1, about 0.5, about 1.0, about 1.5, about 2.0, about 2.5, about 3.0, about 3.5, about 4.0, about 4.5, about 5.0, about 5.5, about 6.0, about 6.5, about 7.0, about 7.5, about 8.0, about 8.5, about 9.0, about 9.5, or about 10.0. [Example]

[0138] The following examples are given for the purpose of illustrating various embodiments of the invention and are not intended to limit the invention in any manner. The examples herein, along with the methods described herein, are presently representative of preferred embodiments and are exemplary, and are not intended as limitations on the scope of the invention. Modifications therein and other uses encompassed by the spirit of the invention as defined by the scope of the claims will occur to those skilled in the art.

[0139] Example 1 Materials and Methods Recombinant human serum albumin was purchased from various sources: rHSA (S. cerevisiae) was from Sigma-Aldrich Inc. (A6608), rHSA (P. pastoris) was from Sigma-Aldrich (A7736), and rHSA (O. sativa) was from InVitria (Cellastim S, Optibumin). Plasma-derived human serum albumin, pHSA, was purchased from Taiwan Blood Services Foundation (TBSF). pHSA (Dialyzed): pHSA dialyzed against DPBS at least 1000 times using a laboratory centrifugal concentrator with a MWCO of 30 kD. DPBS was from Gibco (14190250). Fatty acid-free BSA was from Sigma-Aldrich (A8806).

[0140] Example 2 Preparation of Formulated Recombinant Human Serum Albumin, deAlbumin-I A 20% aqueous solution of 400 g of recombinant HSA expressed and purified from Pichia pastoris was incubated with 1.5 kg of Diaion™ CR20 chelating resin for 10 hours at room temperature. The resin was removed by filtration, and the solution was acidified to pH 3.5 with 1N hydrogen chloride and mixed with 134 g of activated charcoal. After 1 hour of gentle stirring, the solution was neutralized to pH 7.0 with 1N sodium hydroxide. GC-MS confirmed the removal of more than 95% of the total fatty acids from the input recombinant HSA feedstock. The rHSA and activated charcoal slurry was separated by centrifugation and filtration using a 0.45 μm cartridge filter. The resulting solution of recombinant HSA was diluted to 4–10% (w / w) HSA with purified water. The fatty acid mixture was dissolved in ethanol as described below in Table 1 and then added to the HSA solution, adjusting the final ethanol content to 5%. The solution was stirred at room temperature for 2 hours, concentrated to 20% w / w HSA, and diafiltered with at least 7 volumes of DPBS buffer to remove residual ethanol. The resulting HSA solution was passed through a column packed with 40 g of ceramic hydroxyapatite resin to obtain the final product.

[0141] [Table 1]

[0142] Example 3 Albumin performance assay with iPS cell culture Induced pluripotent stem cells (iPS cells, A18945) were cultured in Essential 8 medium (GIBCO, A1517001) supplemented with albumin at 1 mg / mL. 2 1 x 10 cells in an iMatrix-511-coated 6-well plate 5 iPS cells were seeded at 1000μg / well. The medium was changed daily. After 4 days of culture, the viable cell density of each group was determined by trypan blue assay.

[0143] The viable cell density data were plotted and are shown in Figure 1. Various recombinant human serum albumins (from different manufacturers) were also assayed side-by-side.

[0144] Example 4 Preparation of Formulated Recombinant Human Serum Albumin, deAlbumin-II A 20% aqueous solution of 400 g of recombinant HSA expressed and purified from Pichia pastoris was first acidified to pH 3.5 using 1N hydrogen chloride and mixed with 134 g of activated carbon. After 1 hour of gentle stirring, the solution was neutralized to pH 7.0 with 1N sodium hydroxide. GC-MS confirmed the removal of more than 95% of the total fatty acids from the input recombinant HSA feedstock. The rHSA and activated carbon slurry were separated by centrifugation and filtration using a 0.45 μm cartridge filter. The resulting solution of recombinant HSA was diluted to 4–10% (w / w) HSA with purified water. The fatty acid mixture, as described below in Table 2, was dissolved in ethanol and then added to the HSA solution, adjusting the final ethanol content to 5%. The solution was stirred at room temperature for 2 hours, concentrated to 20% w / w HSA, and diafiltered with at least 7 volumes of DPBS buffer to remove residual ethanol.

[0145] [Table 2]

[0146] Example 5 Performance assay of albumin with MSC cell culture Primary Human Adipose-Derived Stem Cells (ADSCs) (Lonza, PT-5006) were cultured in MesenPRO RS™ medium (Gibco 12746012) for two passages to 50%-70% confluency. ADSCs were then cultured at 7 x 10 3 cells / cm 2Cells were seeded into 6-well plates at a density of 0.5 mg / mL each (day 0). Each well of cells received MSC chemically defined medium supplemented with 0.5 mg / mL of various albumins. The composition of the MSC chemically defined medium is listed in Table 3 below. For each well, the medium was changed on day 3. After 5 days of culture, the viable cell density of each group was determined by trypan blue method. The viable cell density data were plotted and shown in Figure 2. Various recombinant human serum albumins (from various manufacturers) were also assayed side-by-side.

[0147] [Table 3]

[0148] Example 6 Albumin performance assay with T cell culture Human peripheral blood CD8+ T cells (Stemcell Technologies 70027) were thawed, expanded in Immunocult™ XF medium (Stemcell Technologies 10981) and 1X Pen-Strep (Gibco 15140122) for 8 days, and cryopreserved in CryoStor® CS10 (Stemcell Technologies 07930). Cells were thawed in Immunocult™ XF medium and cultured at 5 x 10 cells / ml in T cell chemistry-defined medium supplemented with CD3 / CD28 activators (Stemcell Technologies 10971) and various albumins. 4 After resuspending to 100 cells / mL, the cells were seeded into 12-well plates at 1 mL / well (day 0). On day 2, 0.5 mL of fresh medium was added on top of each well, and on day 5, an additional 0.75 mL of fresh medium was added on top. On day 6, the number of cells in each well was counted. The composition of the T cell chemistry-defined medium is listed in Table 4 below. The results are depicted in Figure 3.

[0149] [Table 4]

[0150] Example 7 Preparation of Formulated Bovine Serum Albumin deBSA-I A 20% aqueous solution of 10 g of BSA (Bioshop #ALB001) was first acidified to pH 3.5 with 1 N hydrogen chloride and mixed with 5 g of activated carbon. After gentle stirring at 400 rpm for 1 h, the solution was neutralized to pH 7.0 with 1 N sodium hydroxide. The BSA and activated carbon slurry was separated by filtration using a 0.2 μm filter. The resulting BSA solution was diluted to 4–10% (w / w) BSA with purified water. As described below in Table 5, a fatty acid mixture was dissolved in ethanol and then added to the BSA solution, adjusting the final ethanol content to 5%. The solution was stirred at room temperature for 2 h, concentrated to 20% w / w BSA, and exchanged three times with 10x volumes of DPBS buffer in a 30 kDa MWCO centrifugal concentrator (Sartorius #VS15T21). The formulated BSA was incubated overnight with two volumes of Diaion CRB30 resin and then filtered through a 0.2 μm filter to obtain deBSA-I.

[0151] Example 8 Preparation of Formulated Recombinant Albumin deAlbumin-III The same procedure as described above for the preparation of deBSA-I (Example 6) was used, except that 10 g of 20% recombinant HSA expressed and purified from Pichia pastoris was used.

[0152] [Table 5]

[0153] Example 9 Assay of insulin aggregation stabilization by various albumins Various albumin samples were diluted to 2 mg / mL in purified water. An assay standard solution containing 60 mM HEPES (pH 7.4), 300 mM NaCl, 30 mM 2-mercaptoethanol, and 0.7 mg / mL insulin was prepared. 100 μL of the assay standard solution was mixed with 100 μL of each albumin sample in a 96-well plate. The plate was incubated at 37°C in a plate reader, and the absorbance at 360 nm was measured every 30 seconds for 60 minutes.

[0154] The absorbance data at 360 nm (A360) of each group was obtained at a time point where the A360 of the control group (no albumin, labeled as (-)) was around 0.35. The data is depicted in Figure 4, and the efficiency of the albumin stabilization effect was calculated by the following formula and summarized in Table 6 below.

[0155] Stabilization efficiency % = 1 - (A360 of experimental group with albumin) / (A360 of (-) group)

[0156] [Table 6]

[0157] Example 10 Fatty acid composition of various albumins 500 μL of methanol and 25 μL of 1N HCl were added to 200 μL of aqueous solution of each 10 mg albumin sample, followed by two extractions with 1.5 mL of isooctane. The combined isooctane layers were evaporated to dryness. The residue was redissolved in 100 μL of 1% diisopropylethylamine in acetonitrile and reacted with 100 μL of 1% pentafluorobenzyl bromide, PFB (Sigma-Aldrich, St. Louis, MO) in acetonitrile at room temperature for 20 minutes. After removing the solvent by evaporation, the residue was dissolved in 1500 μL of isooctane, and 1 μL of fatty acid PFB esters were analyzed by an Agilent 6890y gas chromatograph (GC) coupled to an Agilent 5973N mass spectrometry (MS) operated in negative chemical ionization (NCI) mode. Samples were injected onto a Zebron ZB-1 column (15 m × 0.25 mm id, coated with 0.1 μm 100% dimethylpolysiloxane; Phenomenex, Torrance, CA) in pulsed (25 psi) splitless injection mode. Helium (0.9 mL / min) was used as the carrier gas. The GC oven temperature was programmed from 150 to 270 °C at 10 °C / min, increased to 310 °C at 20 °C / min, and held at 310 °C for 1 min. The injector and transfer line were maintained at 250 and 280 °C, respectively. Methane (99.99%) was used as the ionization gas with a source temperature of 150 °C. Data were acquired in selected ion monitoring (SIM) mode, and [MH] was measured. - The anions of fatty acids were monitored and the results are depicted in Table 8.

[0158] Example 11 Alterations in the fatty acid content of albumin tested individually in MSC and T cell cultures deAlbumin-IV was made using the same method as set forth in Example 1, except that each fatty acid listed in Table 5 was applied to 10 g of defatted albumin during the blending process. 10 g of each listed test albumin was blended with the individual fatty acid in the same manner as in Example 1, according to the following recipe (Table 7):

[0159] [Table 7]

[0160] 0.4 mg of deAlbumin-IV was mixed with 0.1 mg of each albumin in Table 6, and the resulting mixture was subjected to MSC cell culture testing as described in Example 4. After performing several rounds of testing with various combinations of Alb-1 to Alb-12 with deAlbumin-IV, it was concluded that Alb-10 was beneficial for the proliferation of T cells and MSCs across most combinations (see Figures 5 to 7).

[0161] Example 12 Ceramic hydroxyapatite improves cell culture performance of albumin The deAlbumin-IV and deAlbumin-II solutions were passed through columns packed with 10-20% w / w ceramic hydroxyapatite resin to obtain processed albumins labeled as alb-IV-CHT and alb-II-CHT. These albumins were tested for iPS and T cell culture performance according to the methods described in Examples 2 and 4, respectively. The results are shown in Figures 8 and 9.

[0162] [Table 8A]

[0163] [Table 8B]

[0164] While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.

Claims

1. a) albumin polypeptide; b) one or more fatty acids having fewer than 18 carbon atoms present in a molar ratio to said albumin polypeptide in the range of about 0.02 to about 0.4; and c) one or more fatty acids having 18 or more carbon atoms present in a molar ratio to said albumin polypeptide in the range of about 0.03 to about 0.6 A composition comprising: Substantially free of pentadecanoic acid (C15:0), margaric acid (C17:0), and / or heptadecenoic acid (C17:1 ω-7), composition.

2. 2. The composition of claim 1, wherein the one or more fatty acids having fewer than 18 carbon atoms comprise one or more fatty acids selected from the group consisting of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), and palmitoleic acid (C16:1).

3. 2. The composition of claim 1, wherein the one or more fatty acids having fewer than 18 carbon atoms consist of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), and palmitoleic acid (C16:1).

4. 4. The composition of claim 1, wherein the one or more fatty acids having 18 or more carbon atoms comprise one or more fatty acids selected from the group consisting of stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), and eicosadienoic acid (C20:2 ω-6).

5. 4. The composition of claim 1, wherein the one or more fatty acids having 18 or more carbon atoms consist of stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), and eicosadienoic acid (C20:2 ω-6).

6. 6. The composition of claim 1, wherein the one or more fatty acids having 18 or more carbon atoms further comprise one or more fatty acids selected from the group consisting of bishomo-gamma-linolenic acid (C20:3 ω-6), arachidonic acid (C20:4), docosatetraenoic acid (C22:4 ω-6), and docosahexaenoic acid (C22:6 ω-3).

7. 7. The composition of claim 1, wherein (i) the one or more fatty acids having fewer than 18 carbon atoms consist of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), and palmitoleic acid (C16:1); and (ii) the one or more fatty acids having 18 or more carbon atoms consist of stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), eicosadienoic acid (C20:2 ω-6), bishomo-γ-linolenic acid (C20:3 ω-6), arachidonic acid (C20:4), docosatetraenoic acid (C22:4 ω-6), and docosahexaenoic acid (C22:6 ω-3).

8. 7. The composition of any one of claims 1 to 6, wherein (i) the one or more fatty acids having fewer than 18 carbon atoms consist of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), and palmitoleic acid (C16:1); and (ii) the one or more fatty acids having 18 or more carbon atoms consist of stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), and eicosadienoic acid (C20:2 ω-6).

9. 7. The composition of claim 1, wherein (i) the one or more fatty acids having fewer than 18 carbon atoms consist of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), and palmitoleic acid (C16:1); and (ii) the one or more fatty acids having 18 or more carbon atoms consist of stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), eicosadienoic acid (C20:2 ω-6), bishomo-γ-linolenic acid (C20:3 ω-6), and arachidonic acid (C20:4).

10. 10. The composition of any one of claims 1 to 9, wherein the total molar ratio of fatty acids to albumin polypeptides is less than 1.

11. 11. The composition of any one of claims 1 to 10, which is substantially free of alpha-linolenic acid (C18:3), gamma-linolenic acid (C18:3), arachidic acid (C20:0), eicosatrienoic acid (C20:3 ω-3), eicosapentaenoic acid (C20:5 ω-3), behenic acid (C22:0), docosapentaenoic acid (C22:5 ω-3), lignoceric acid (C24:0), and / or cerotic acid (C26:0).

12. 12. The composition of any one of claims 1 to 11, wherein both the one or more fatty acids having fewer than 18 carbon atoms and the one or more fatty acids having 18 or more carbon atoms are adsorbed to the albumin polypeptide.

13. a) albumin polypeptide; b) lauric acid (C12:0) present in a molar ratio to said albumin polypeptide in the range of about 0.001 to about 0.008; c) myristic acid (C14:0) present in a molar ratio to said albumin polypeptide in the range of about 0.001 to about 0.022; d) palmitic acid (C16:0) present in a molar ratio to said albumin polypeptide in the range of about 0.02 to about 0.3; e) palmitoleic acid (C16:1) present in a molar ratio to said albumin polypeptide in the range of about 0.002 to about 0.03; f) stearic acid (C18:0) present in a molar ratio to said albumin polypeptide in the range of about 0.011 to about 0.2; g) oleic acid (C18:1 ω-9) present in a molar ratio to said albumin polypeptide in the range of about 0.02 to about 0.3; h) linoleic acid (C18:2) present in a molar ratio to said albumin polypeptide in the range of about 0.0002 to about 0.12; and i) eicosadienoic acid (C20:2 ω-6) present in a molar ratio to said albumin polypeptide in the range of about 0.0001 to about 0.002 A composition comprising:

14. 14. The composition of claim 13, further comprising bishomo-gamma-linolenic acid (C20:3 omega-6) present in a molar ratio to said albumin polypeptide in the range of about 0.0003 to about 0.

002.

15. 15. The composition of claim 13 or 14, further comprising arachidonic acid (C20:4) present in a molar ratio to said albumin polypeptide in the range of about 0.001 to about 0.

01.

16. 16. The composition of any one of claims 13 to 15, further comprising docosatetraenoic acid (C22:4 omega-6) present in a molar ratio to said albumin polypeptide in the range of about 0.0009 to about 0.

003.

17. 17. The composition of any one of claims 13 to 16, further comprising docosahexaenoic acid (C22:6 omega-3) present in a molar ratio to said albumin polypeptide in the range of about 0.0003 to about 0.

001.

18. 18. The composition of any one of claims 13 to 17, wherein the total molar ratio of fatty acids to albumin polypeptide is from about 0.06 to about 1.

19. 19. The composition of any one of claims 1 to 18, substantially free of pentadecanoic acid (C15:0), margaric acid (C17:0), heptadecenoic acid (C17:1 ω-7), α-linolenic acid (C18:3), γ-linolenic acid (C18:3), arachidic acid (C20:0), eicosatrienoic acid (C20:3 ω-3), eicosapentaenoic acid (C20:5 ω-3), behenic acid (C22:0), docosapentaenoic acid (C22:5 ω-3), lignoceric acid (C24:0), and / or cerotic acid (C26:0).

20. a) albumin polypeptide; and b) one or more fatty acids A composition comprising: substantially free of one or more of pentadecanoic acid (C15:0), margaric acid (C17:0), heptadecenoic acid (C17:1 ω-7), α-linolenic acid (C18:3), γ-linolenic acid (C18:3), arachidic acid (C20:0), eicosatrienoic acid (C20:3 ω-3), eicosapentaenoic acid (C20:5 ω-3), behenic acid (C22:0), docosapentaenoic acid (C22:5 ω-3), lignoceric acid (C24:0), and cerotic acid (C26:0); a total molar ratio of said one or more fatty acids to said albumin polypeptide of from about 0.05 to about 1; composition.

21. a) albumin polypeptide; and b) fatty acids A composition comprising: The fatty acid is i) lauric acid (C12:0); ii) myristic acid (C14:0); iii) palmitic acid (C16:0); iv) palmitoleic acid (C16:1); v) stearic acid (C18:0); vi) oleic acid (C18:1 ω-9); vii) Linoleic acid (C18:2); viii) eicosadienoic acid (C20:2 ω-6); ix) bishomo-γ-linolenic acid (C20:3 ω-6); x) arachidonic acid (C20:4); xi) docosatetraenoic acid (C22:4 ω-6); and xii) Docosahexaenoic acid (C22:6 ω-3) It consists of the total molar ratio of said fatty acids to said albumin polypeptide is less than about 1; composition.

22. the lauric acid (C12:0) is present in a molar ratio to the albumin polypeptide of less than about 0.01; the myristic acid (C14:0) is present in a molar ratio to the albumin polypeptide of less than about 0.05; the palmitic acid (C16:0) is present in a molar ratio to the albumin polypeptide of less than about 0.5; the palmitoleic acid (C16:1) is present in a molar ratio to the albumin polypeptide of less than about 0.05; the stearic acid (C18:0) is present in a molar ratio to the albumin polypeptide of less than about 0.2; the oleic acid (C18:1 ω-9) is present in a molar ratio to the albumin polypeptide of less than about 0.5; the linoleic acid (C18:2) is present in a molar ratio to the albumin polypeptide of less than about 0.2; the eicosadienoic acid (C20:2 ω-6) is present in a molar ratio to the albumin polypeptide of less than about 0.005; the bishomo-gamma-linolenic acid (C20:3 omega-6) is present in a molar ratio to the albumin polypeptide of less than about 0.005; the arachidonic acid (C20:4) is present in a molar ratio to the albumin polypeptide of less than about 0.01; the docosatetraenoic acid (C22:4 ω-6) is present in a molar ratio to the albumin polypeptide of less than about 0.005; the docosahexaenoic acid (C22:6 ω-3) is present in a molar ratio to the albumin polypeptide of less than about 0.005; or Any combination thereof 22. The composition of claim 21.

23. a) albumin polypeptide; and b) fatty acids A composition comprising: The fatty acid is i) lauric acid (C12:0); ii) myristic acid (C14:0); iii) palmitic acid (C16:0); iv) palmitoleic acid (C16:1); v) stearic acid (C18:0); vi) oleic acid (C18:1 ω-9); vii) linoleic acid (C18:2); and viii) Eicosadienoic acid (C20:2 ω-6) Consists of: composition.

24. the lauric acid (C12:0) is present in a molar ratio to the albumin polypeptide of less than about 0.005; the myristic acid (C14:0) is present in a molar ratio to the albumin polypeptide of less than about 0.005; said palmitic acid (C16:0) is present in a molar ratio to said albumin polypeptide of less than about 0.05; the palmitoleic acid (C16:1) is present in a molar ratio to the albumin polypeptide of less than about 0.005; the stearic acid (C18:0) is present in a molar ratio to the albumin polypeptide of less than about 0.05; the oleic acid (C18:1 ω-9) is present in a molar ratio to the albumin polypeptide of less than about 0.05; the linoleic acid (C18:2) is present in a molar ratio to the albumin polypeptide of less than about 0.0005; the eicosadienoic acid (C20:2 ω-6) is present in a molar ratio to the albumin polypeptide of less than about 0.0005; or Any combination thereof 24. The composition of claim 23.

25. a) albumin polypeptide; and b) fatty acids A composition comprising: The fatty acid is i) lauric acid (C12:0); ii) myristic acid (C14:0); iii) palmitic acid (C16:0); iv) palmitoleic acid (C16:1); v) stearic acid (C18:0); vi) oleic acid (C18:1 ω-9); vii) Linoleic acid (C18:2); viii) eicosadienoic acid (C20:2 ω-6); ix) bishomo-γ-linolenic acid (C20:3 ω-6); and x) Arachidonic acid (C20:4) Consists of: composition.

26. the lauric acid (C12:0) is present in a molar ratio to the albumin polypeptide of less than about 0.005; the myristic acid (C14:0) is present in a molar ratio to the albumin polypeptide of less than about 0.01; the palmitic acid (C16:0) is present in a molar ratio to the albumin polypeptide of less than about 0.1; the palmitoleic acid (C16:1) is present in a molar ratio to the albumin polypeptide of less than about 0.01; the stearic acid (C18:0) is present in a molar ratio to the albumin polypeptide of less than about 0.1; the oleic acid (C18:1 ω-9) is present in a molar ratio to the albumin polypeptide of less than about 0.5; the linoleic acid (C18:2) is present in a molar ratio to the albumin polypeptide of less than about 0.05; the eicosadienoic acid (C20:2 ω-6) is present in a molar ratio to the albumin polypeptide of less than about 0.0005; the bishomo-gamma-linolenic acid (C20:3 omega-6) is present in a molar ratio to the albumin polypeptide of less than about 0.0005; the arachidonic acid (C20:4) is present in a molar ratio to the albumin polypeptide of less than about 0.005; or Any combination thereof 26. The composition of claim 25.

27. 27. The composition of any one of claims 1 to 26, wherein the albumin polypeptide is of human or bovine origin.

28. 28. The composition of any one of claims 1 to 27, wherein the albumin polypeptide is a recombinant albumin polypeptide.

29. 29. The composition of claim 28, wherein the recombinant albumin polypeptide is purified from bacteria, yeast, or rice.

30. 30. The composition of claim 29, wherein the yeast is of the species Pichia pastoris, Saccharomyces cerevisiae, or Kluyveromyces lactis.

31. 30. The composition of claim 29, wherein the rice is of the species Oryza sativa.

32. 28. The composition of any one of claims 1 to 27, wherein the albumin polypeptide is derived from plasma or serum.

33. 33. The composition of any one of claims 1 to 32, wherein the albumin polypeptide is a delipidated albumin polypeptide.

34. 34. A cell culture medium comprising the composition of any one of claims 1 to 33 and a basal medium.

35. 35. The cell culture medium of claim 34, wherein the composition is present in the cell culture medium at a concentration of from about 0.01% (w / w) to about 10% (w / w), or from about 0.1 mg / mL to about 100 mg / mL.

36. 36. A method comprising incubating biological cells in the cell culture medium of claim 34 or 35.

37. 37. The method of claim 36, wherein the biological cell is a eukaryotic cell.

38. 38. The method of claim 37, wherein the eukaryotic cell is a stem cell.

39. 39. The method of claim 38, wherein the stem cells are induced pluripotent stem (iPS) cells, embryonic stem (ES) cells, or mesenchymal stem cells (MSCs).

40. 37. The method of claim 36, wherein the eukaryotic cell is a T cell.

41. 37. The method of claim 36, wherein the eukaryotic cell is a neuronal cell.

42. 42. The method of any one of claims 36 to 41, which results in an increase in viable cell density of the biological cells compared to biological cells cultured in the absence of the albumin polypeptide.

43. 43. The method of claim 42, wherein the increase in viable cell density is at least 10%.

44. 34. A method for stabilizing a protein, the method comprising incubating said protein in the presence of a composition according to any one of claims 1 to 33.

45. 45. The method of claim 44, wherein the composition is present at a concentration of about 0.01% (w / w) to about 20% (w / w), about 0.1 mg / mL to about 200 mg / mL, or in a molar ratio of about 0.1 to about 10 relative to the protein.

46. 46. ​​The method of claim 44 or 45, wherein the protein is stabilized by at least about 80% compared to the protein in the absence of the composition.

47. a) albumin polypeptide; and b) arachidonic acid (C20:4) present in a molar ratio of at least about 0.9% relative to the total amount of fatty acids in the composition Including, composition.

48. A method for improving the function of an albumin polypeptide, comprising: incubating said albumin polypeptide with a sufficient amount of arachidonic acid (C20:4) to increase the molar ratio of arachidonic acid (C20:4) to the total amount of fatty acids in the composition to at least about 0.9%. Including, method.

49. A method for improving the function of an albumin polypeptide, comprising: a) passing a solution of said albumin polypeptide through a ceramic hydroxyapatite resin to produce a flow-through comprising said albumin polypeptide; and b) purifying the albumin polypeptide from the flow-through to obtain an albumin polypeptide with improved functionality. Including, method.

50. 50. The method of claim 49, further comprising, prior to a), delipidating the albumin polypeptide by incubating the solution with activated charcoal and purifying the albumin polypeptide from the activated charcoal.

51. 51. The method of claim 49 or 50, further comprising, prior to a), incubating the solution with a chelating resin and purifying the albumin polypeptide from the chelating resin.

52. 52. The method of claim 51, wherein the chelating resin is Diaion CR20.

53. 53. The method of any one of claims 49 to 52, further comprising, prior to a), incubating the albumin polypeptide with one or more fatty acids.

54. 54. The method of claim 53, wherein the one or more fatty acids are selected from the group consisting of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), palmitoleic acid (C16:1), stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), eicosadienoic acid (C20:2 ω-6), bishomo-γ-linolenic acid (C20:3 ω-6), arachidonic acid (C20:4), docosatetraenoic acid (C22:4 ω-6), and docosahexaenoic acid (C22:6 ω-3).

55. 54. The method of claim 53, wherein the one or more fatty acids consist of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), palmitoleic acid (C16:1), stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), eicosadienoic acid (C20:2 ω-6), bishomo-γ-linolenic acid (C20:3 ω-6), arachidonic acid (C20:4), docosatetraenoic acid (C22:4 ω-6), and docosahexaenoic acid (C22:6 ω-3).

56. 54. The method of claim 53, wherein the one or more fatty acids consist of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), palmitoleic acid (C16:1), stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), and eicosadienoic acid (C20:2 ω-6).

57. 54. The method of claim 53, wherein the one or more fatty acids consist of lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), palmitoleic acid (C16:1), stearic acid (C18:0), oleic acid (C18:1 ω-9), linoleic acid (C18:2), eicosadienoic acid (C20:2 ω-6), bishomo-γ-linolenic acid (C20:3 ω-6), and arachidonic acid (C20:4).

58. A composition comprising a recombinant albumin polypeptide and one or more fatty acids in a molar ratio effective to promote at least a 35% increase in viable cell density of induced pluripotent stem (iPS) cells upon culturing the iPS cells in the composition for a duration of 4 days, compared to a composition lacking the recombinant albumin polypeptide.

59. A composition comprising a recombinant albumin polypeptide and one or more fatty acids in a molar ratio effective to promote at least a 35% increase in viable cell density of mesenchymal stem cells (MSCs) upon culturing the MSCs in the composition for a duration of 5 days compared to a composition lacking the recombinant albumin polypeptide.

60. A composition comprising a recombinant albumin polypeptide and one or more fatty acids in amounts in a molar ratio effective to promote at least an 80% increase in viable cell density of T cells upon culturing the T cells in the composition for a duration of 6 days compared to a composition lacking the recombinant albumin polypeptide.

61. A composition comprising an albumin polypeptide and one or more fatty acids in amounts in a molar ratio effective to promote at least about an 80% increase in protein stabilization upon incubation of the protein in the composition for a duration of about 3000 seconds to about 6000 seconds, compared to a composition lacking the albumin polypeptide.

62. 62. The composition of any one of claims 58 to 61, which is substantially free of growth factors.

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