Coated media for chromatography

• US 5,030,352 A
• Filed: 01/25/1990
• Issued: 07/09/1991
• Est. Priority Date: 01/25/1990
• Status: Expired due to Term

• Current Assignee: Purdue Research Foundation (The Trustees of Purdue University (d/b/a Purdue University))
• Original Assignee: Purdue Research Foundation (The Trustees of Purdue University (d/b/a Purdue University))
• Inventors: Yang, Yan B., Cook, Steven E., Regnier, Fred E., Varady, Laszlo
• Primary Examiner(s): Spitzer, Robert
• Application Number:
  US07/469,956
• Time in Patent Office: 530 Days
• Field of Search: 55/386, 210/656, 210/198.2, 210/198.3, 210/502.1, 428/406, 428/422, 428/458, 428/473.5, 525/54.1, 427/220, 427/301
• US Class Current: 210/502.1
• CPC Class Codes: B01D 2323/30 Cross-linking
• CPC Class Codes: B01D 67/0088 Physical treatment with compounds, e.g. swelling, coating or impregnation
• CPC Class Codes: B01D 67/00931 by introduction of specific groups after membrane formation, e.g. by grafting
• CPC Class Codes: B01D 67/00933 by addition of a layer chemically bonded to the membrane
• CPC Class Codes: B01J 20/321 consisting of a polymer obtained by reactions involving only carbon to carbon unsaturated bonds
• CPC Class Codes: B01J 20/3212 consisting of a polymer obtained by reactions otherwise than involving only carbon to carbon unsaturated bonds
• CPC Class Codes: B01J 20/3272 Polymers obtained by reactions otherwise than involving only carbon to carbon unsaturated bonds
• CPC Class Codes: B01J 20/3276 Copolymers
• CPC Class Codes: B01J 20/3282 Crosslinked polymers
• CPC Class Codes: B01J 20/3285 Coating or impregnation layers comprising different type of functional groups or interactions, e.g. different ligands in various parts of the sorbent, mixed mode, dual zone, bimodal, multimodal, ionic or hydrophobic, cationic or anionic, hydrophilic or hydrophobic
• CPC Class Codes: B01J 20/3287 Layers in the form of a liquid
• CPC Class Codes: B01J 39/17 containing also inorganic materials, e.g. inert material coated with an ion-exchange resin
• CPC Class Codes: B01J 39/20 obtained by reactions only involving unsaturated carbon-to-carbon bonds
• CPC Class Codes: B01J 41/14 obtained by reactions only involving unsaturated carbon-to-carbon bonds
• CPC Class Codes: Y10T 428/2996 Glass particles or spheres
• CPC Class Codes: Y10T 428/31544 Addition polymer is perhalogenated

Abstract

Disclosed are pellicular materials useful in contact with proteins and as chromatography media. Rigid, inert, hydrophobic, polymeric materials are exposed to a solution comprising a solute defining plural interspersed hydrophobic and hydrophilic domains. The solute molecules adsorb onto the surface of the substrate by hydrophobic - hydrophobic interaction with their hydrophilic domains extending outwardly away from the surface into the solution. The molecules are then crosslinked in place to produce a solvent and pH resistant coating which presents a hydrophilic surface sufficient to mask hydrophobic regions therebeneath. The multidomain adsorbing compounds may comprise reactive groups which subsequently can be derivatized using conventional techniques to produce chromatography materials useful for conducting affinity, size exclusion, cationic exchange, anionic exchange, neutral hydrophobic interaction, and other forms of chromatographic separations.

Claims

1. A method of producing a hydrophilic coating on a hydrophobic surface comprising the steps of:

contacting said hydrophobic surface with a liquid phase, hydrophilic with respect to said surface, containing a solvated compound, molecules of said solvated compound comprising a hydrophobic domain covalently and flexibly bonded to a hydrophilic domain, and a crosslinkable site, thereby to deposit molecules of said compound on said hydrophobic surface whereby said molecules of said compound are oriented with said hydrophobic domains proximate to and adsorbed on said hydrophobic surface and with said hydrophilic domains extending outwardly from said hydrophobic surface into said liquid phase; and

crosslinking molecules adjacent one another at said crosslinkable sites to produce an adherent film on said surface.

2. The method of claim 1 wherein at least one of said domains comprises derivatizable moieties, the method comprising the additional step of derivatizing said adherent film.

3. The method of claim 1 wherein said compound is a polymer comprising plural hydrophobic domains said plural hydrophilic domains.

4. The method of claim 1 wherein the liquid phase is formulated such that said compound is sparingly soluble therein to promote partition of hydrophobic domains of said compound onto said hydrophobic surface.

5. The method of claim 1 wherein the crosslinking step is effected by reacting a subset of said hydrophobic domains.

6. The method of claim 1 wherein the crosslinking step is effected by reacting a subset of said hydrophilic domains.

7. The method of claim 1 wherein the molecules of said solvated compound have a hydrophilic domain of a molecular dimension sufficient to mask said hydrophobic surface.

8. The method of claim 1 wherein said hydrophilic domains comprise moieties comprising hydroxyl groups or ether linkages.

9. The method of claim 1 wherein the hydrophobic domains comprise moieties comprising halides or hydrocarbons.

10. The method of claim 1 wherein said compound comprises a multipolymer wherein said hydrophobic domains comprise plural linked hydrophobic monomers and said hydrophilic domains comprise plural linked hydrophilic monomers.

11. The method of claim 1 wherein said compound is a copolymer wherein blocks comprise said hydrophobic domain and said hydrophilic domain.

12. The method of claim 1 wherein said hydrophobic surface defines a chromatography matrix.

13. The method of claim 1 wherein said hydrophobic surface is an organic polymeric material stable from pH2 to pH12.

14. The method of claim 1 wherein said hydrophobic surface comprises polystyrene.

15. The method of claim 1 wherein said hydrophobic surface comprises divinylbenzene crosslinked polystyrene.

16. The method of claim 1 wherein said compound is a monomer, said method comprising the additional step of polymerizing said monomer in place on said surface.

17. The method of claim 1 wherein said hydrophilic domain contains epoxy groups and said crosslinking step is effected by reacting said epoxy groups on adjacent molecules.

18. The method of claim 1 wherein said crosslinking step is effected by reacting a halide group on one of said molecules with a hydroxyl group on another.

19. The method of claim 1 wherein said crosslinking step is effected by reacting a crosslinking compound comprising plural reactive groups with reactive groups on said molecules.

20. Hydrophilic material useful in contact with a protein solution to inhibit denaturation comprising a hydrophobic substrate and, adhered thereto by hydrophobic interaction, an enveloping, continuous film, the individual interaction an enveloping, continuous film, the individual molecules of said film comprising a hydrophobic domain covalently and flexibly bonded to a hydrophilic domain, and a crosslinkable site, wherein such molecules are adsorbed onto said surface by hydrophobic-hydrophobic interaction and said hydrophilic domain are disposed outwardly therefrom, said individual molecules being crosslinked at said crosslinkable site.

21. The material of claim 20 wherein said hydrophilic domains have a surface density within said film sufficient to mask said hydrophobic domains and said hydrophobic substrate.

22. The material of claim 20 wherein said hydrophobic substrate comprises an organic polymeric material.

23. The material of claim 20 wherein said hydrophobic substrate comprises polystyrene.

24. The material of claim 23 wherein said substrate comprises polystyrene crosslinked with divinylbenzene.

25. The material of claim 20 wherein said hydrophobic substrate comprises a member selected from the group consisting of a microparticulate chromatography medium and a microporous membrane.

26. The material of claim 20 further comprising moieties covalently linked to said film disposed outwardly from said surface selected from the group consisting of cationic groups, anionic groups, hydrocarbon groups, groups capable of metal chelation, binding proteins, and combinations thereof.