Flocculant Functionalized Separation Media

Patent metadata

Field	Value
Publication number	US-2021170389-A1
Application number	US-201916970444-A
Country	US
Kind code	A1
Filing date	Feb 27, 2019
Priority date	Feb 27, 2018
Publication date	Jun 10, 2021
Grant date	—

Abstract

Provided herein are compositions, methods and uses that relate to or result from providing separation media having at least one flocculant ligand covalently attached to a base surface or support, and the separation and/or purification of biological molecules using the separation media of the present disclosure. Certain embodiments provide separation media which under certain modes of operation, enhance the separation of the molecule of interest from impurities.

First claim

1 . A separation medium comprising:
a. a base surface; and b. at least one flocculant ligand covalently attached to the base surface.

2 . The separation medium of claim 1 wherein the at least one flocculant ligand is selected from the group consisting of cationic, anionic, non-ionic and natural flocculants.

3 . The separation medium of claim 2 , wherein the anionic flocculant further comprises an unsubstituted or substituted aliphatic carboxylic acid, an unsubstituted or substituted aromatic carboxylic acid, an unsubstituted or substituted aliphatic sulfonic acid, an unsubstituted or substituted aliphatic acrylic acid, an unsubstituted or substituted aliphatic thiosulfate, an unsubstituted or substituted aliphatic phosphonic acid, or an unsubstituted or substituted aliphatic phosphoric a fatty acid.

4 . The separation medium of claim 3 , further wherein the unsubstituted or substituted aliphatic groups are either linear or branched, and optionally, comprise one or more double bonds.

5 . (canceled)

6 . The separation medium of claim 4 , wherein the unsubstituted or substituted aliphatic group is a C 1 -C 8 aliphatic acid.

7 . The separation medium of claim 4 , wherein the unsubstituted or substituted aliphatic group is a C 9 -C 30 aliphatic acid.

8 . The separation medium of claim 1 , wherein the separation medium is contacted with a solution, a feed or an eluent that comprises one or more ligate species, under operating conditions that allow the binding of at least one ligate species from the solution, feed or eluent, to the separation medium, wherein the one or more ligate species is a mixture of biological substances, wherein the mixture of biological substances comprises a target molecule and at least one impurity.

9 .- 10 . (canceled)

11 . The separation medium of claim 8 , wherein the impurity is an aggregate, and the target molecule is either a monomeric antibody, a therapeutic peptide or protein, a virus or viral particle, a particular variant of a peptide or protein or antibody or virus or viral particle, or a nucleic acid.

12 .- 23 . (canceled)

24 . The separation medium of claim 1 , wherein the separation medium selectively binds an antibody monomer, and wherein the separation medium has a separation factor (α) of less than 1.

25 . The separation medium of claim 24 , wherein the antibody monomer is separated from one or more aggregates with the separation medium having a separation factor (α) of at least about 0.1 to about 0.9.

26 . (canceled)

27 . The separation medium of claim 2 , wherein cationic flocculant ligand is either a primary aliphatic amine, a secondary aliphatic amine, a tertiary aliphatic amine, an aliphatic imine, an aliphatic hydrazide, an imidazole, an aliphatic oxime, an aliphatic hydrazine, an aliphatic hydrazone, a linear polyethyl amine, a polyethyleneimine, a heterocyclic quaternary ammonium, and a cationic polyelectrolyte.

28 . The separation medium of claim 2 , wherein the cationic flocculant ligand is selected from the group consisting of tris(2-aminoethyl)amine, tris(3-aminopropyl)amine, polydiallyldimethylammonium chloride (PolyDADMAC) and poly(N,N-dimethylpiperidinium chloride), poly(N-vinylpyrrolidone) (PVP), copolymers of poly(ethyleneimine), and quaternary aminated polyacrylates.

29 . The separation medium of claim 1 , wherein the separation medium can selectively bind to an impurity, and/or, wherein the impurity is a nucleic acid, and/or, wherein the charged variant are is a glycosylated, a glycated, an oxidized, a deaminated, an acidic, a basic, a phosphorylated, a sialylated or a N-terminal acetylated form.

30 . (canceled)

31 . The separation medium of claim 2 , wherein the at least one non-ionic flocculant ligand is either a styrene, substituted styrene, polymeric styrenes, an uncharged aliphatic, an uncharged branched aliphatic, a hydrophobic polyester, a hydrophilic polyester, a polyacrylamide, a poly(ethylene oxide), or a copolymer thereof.

32 . The separation medium of claim 2 , wherein the flocculant natural ligand is a polysaccharide, an amino, imino, ammonium, sulfonium or phosphonium functionalized polysaccharide, a collagen, an anionic protein, a cationic protein, a chitosan, an ininglass, guar gum, a cationic protein from Moringa oleifera seeds or Strychnos potatorum seeds, or an alginate.

33 . The separation medium of claim 1 , wherein the base surface includes but is not limited to: comprises a resin, bead, sphere, particle, microcarrier, membrane, web, bag, bioreactor, tube, plate, array, flat surface, filter, fiber or a fabric.

34 . The separation medium of claim 1 , wherein the base surface is porous, nonporous, microporous, woven, non-woven, polymeric, non-polymeric, fibrous or winged, and the base surface comprises a material selected from ceramics, glass, metal, silica, synthetic polymeric materials such as styrenic, acrylate, acrylamide, acrylamide containing one or more polymerizable vinyl groups, polymeric monoliths, natural polymers, such as cellulose, lignocellulose or their derivatives, agarose, and a combination of any of these materials.

35 .- 44 . (canceled)

45 . A chromatography system comprising:
a column, and, enclosed within the column, a separation medium according to claim 1 ; and a pump for passing a liquid through the separation medium at a velocity of about 50 to about 1000 cm/hr, wherein the liquid is a solution, an eluent, or a feed comprising one or more biological substances.

46 . (canceled)

47 . A method of separating a monomer and least one aggregate comprising;
i. providing a separation medium according to claim 1 ; and ii. passing a solution, an eluent, or a feed comprising one or more biological substances through the separation medium at a rate sufficient to allow at least one soluble molecule to bind to the separation medium.

48 .- 57 . (canceled)

58 . The method of purifying a protein of interest from a solution, an eluent, or a feed comprising:
i. providing a separation medium according to claim 1 ; and ii. passing the solution, eluent, or feed comprising the protein of interest and one or more impurities through the separation medium at a rate sufficient to allow the one or more impurities to bind to the separation medium.

59 .- 64 . (canceled)

Assignees

• Life Technologies Corp

Inventors

• Tomlinson Andrew
• Buchanan Matthew
• Flook Kelly
• Galipeau Kevin

Classifications

• B01J39/26

  Cation exchangers for chromatographic processes · CPC title

• B01J41/20

  Anion exchangers for chromatographic processes · CPC title

• C07K1/16

  by chromatography · CPC title

• B01D15/362 Primary

  Cation-exchange · CPC title

• C07K1/18

  Ion-exchange chromatography · CPC title