Covalently bonded coatings

• US 5,503,933 A
• Filed: 02/25/1994
• Issued: 04/02/1996
• Est. Priority Date: 02/25/1994
• Status: Expired due to Term

• Current Assignee: Perseptive Biosystems Incorporated (Ecovyst, Inc.), Purdue Research Foundation (The Trustees of Purdue University (d/b/a Purdue University))
• Original Assignee: Perseptive Biosystems Incorporated (Ecovyst, Inc.), Purdue Research Foundation (The Trustees of Purdue University (d/b/a Purdue University))
• Inventors: Mu, Ning, Regnier, Fred E., Afeyan, Noubar B.
• Primary Examiner(s): LUSIGNAN, MICHAEL R
• Application Number:
  US08/201,906
• Time in Patent Office: 767 Days
• Field of Search: 210/198.2, 210/502.1, 210/635, 210/638, 210/656, 427/220, 427/221, 428/406, 428/407, 530/413, 530/417
• US Class Current: 428/407
• CPC Class Codes: B01D 15/08 Selective adsorption, e.g. chromatography
• CPC Class Codes: B01J 20/261 obtained by reactions only involving carbon to carbon unsaturated bonds macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds per se C08F
• CPC Class Codes: B01J 20/264 derived from different types of monomers, e.g. linear or branched copolymers, block copolymers, graft copolymers
• CPC Class Codes: B01J 20/265 modified or post-treated polymers polymer carriers or substrates subjected to further impregnating or coating B01J20/3208
• CPC Class Codes: B01J 20/267 Cross-linked polymers
• CPC Class Codes: B01J 20/28004 Sorbent size or size distribution, e.g. particle size
• CPC Class Codes: B01J 20/28078 Pore diameter
• CPC Class Codes: B01J 20/286 Phases chemically bonded to a substrate, e.g. to silica or to polymers
• CPC Class Codes: B01J 20/3092 Packing of a container, e.g. packing a cartridge or column of chromatography columns B01D15/206
• CPC Class Codes: B01J 20/321 consisting of a polymer obtained by reactions involving only carbon to carbon unsaturated bonds
• CPC Class Codes: B01J 20/3217 Resulting in a chemical bond between the coating or impregnating layer and the carrier, support or substrate, e.g. a covalent bond
• CPC Class Codes: B01J 20/327 Polymers obtained by reactions involving only carbon to carbon unsaturated bonds
• CPC Class Codes: B01J 20/3282 Crosslinked polymers
• CPC Class Codes: B01J 2220/54 Sorbents specially adapted for analytical or investigative chromatography
• CPC Class Codes: B01J 2220/58 Use in a single column
• CPC Class Codes: Y10T 428/2998 including synthetic resin or polymer

Abstract

Disclosed are hydrophilic coatings covalently bound to hydrophobic surfaces as well as methods for their production. To form the coated surfaces, a compound is provided which comprises a hydrophobic domain including an unsaturated group and a hydrophilic domain. Also provided is a hydrophobic surface comprising unsaturated groups. Molecules of the compound are adsorbed onto the hydrophobic surface, and the unsaturated groups in the hydrophobic domains of molecules of the compound then are covalently crosslinked to the unsaturated groups on the hydrophobic surface by a free radical reaction. In one embodiment, hydrophilic coatings may be covalently attached to hydrophobic polymers such as divinylbenzene crosslinked polystyrene. Hydrophilic coatings covalently attached to hydrophobic surfaces are obtained which are stable, which may be readily derivatized, and which are useful in a wide range of chromatography applications.

Claims

1. A method of producing a hydrophilic coating covalently linked to a hydrophobic surface of a polymer, the method comprising the steps of:

a) providing a high surface area polymer having a hydrophobic surface comprising unsaturated groups;

b) providing a first compound, free of poly(glycidyl methacrylate), comprising a hydrophobic domain covalently and flexibly bonded to a hydrophilic domain, the hydrophobic domain comprising an unsaturated group;

c) contacting the hydrophobic surface with a liquid phase, hydrophilic with respect to the surface, containing solvated molecules of the first compound, thereby to deposit molecules of the compound on the hydrophobic surface oriented with the hydrophobic domains proximate to and adsorbed on the surface and with the hydrophilic domains extending outwardly from the surface into the liquid phase; and

d) covalently crosslinking the unsaturated group on the hydrophobic domain of molecules of the first compound to the unsaturated groups on the hydrophobic surface to produce a hydrophilic coating covalently bound to the surface of the polymer, wherein the hydrophilic domains present in the coating substantially shield the hydrophobic surface of the polymer.

2. The method of claim 1 wherein the compound provided in step b) comprises plural hydrophilic domains and plural hydrophobic domains.

3. The method of claim 1 wherein said crosslinking step (d) is a free radical reaction.

4. The method of claim 3 wherein the first compound is modified polyvinylalcohol.

5. The method of claim 3 further comprising:

e) derivatizing the first compound after step d) to produce a hydrophilic moiety.

6. The method of claim 5 wherein the hydrophilic moiety is selected from the group consisting of hydroxyl, aidehyde, carboxylic acid, sulfonic acid and quaternary amine.

7. The method of claim 5 wherein the hydrophilic coated polymer obtained in step e) defines a chromatography matrix.

8. The method of claim 5 wherein the hydrophilic coating covalently bound to the hydrophobic surface is stable from pH 2 to pH 12.

9. The method of claim 5 wherein the covalent coating comprises a fimbriate structure with sufficient hydrophilic groups to mask the hydrophobic domains and the hydrophobic surface.

10. The method of claim 1 wherein the polymer comprises divinylbenzene crosslinked polystyrene.

11. The method of claim 1 wherein the unsaturated groups on the hydrophobic surface of the polymer titrate to at least about 0.23 mmol/g of polymer.

12. The method of claim 5 wherein the first compound further comprises a reactive group, and wherein step (e) comprises:

covalently reacting the reactive group on the first compound to a second compound comprising a hydrophilic moiety.

13. The method of claim 12 wherein the reactive group on the first compound comprises a hydroxyl, the second compound comprises glycidol;

and

wherein step (e) comprises polymerizing plural molecules of glycidol to the reactive group of the first compound, thereby to produce a covalent coating comprising plural hydroxyls.

14. The method of claim 13 further comprising, in step (e), oxidizing hydroxyls of the covalently bound coating, thereby to produce carboxylic acid groups.

15. The method of claim 12 wherein the reactive group on the first compound comprises a hydroxyl, and wherein the second compound comprises an epihalohydrin;

and

wherein step (e) comprises;

reacting the epihalohydrin with the reactive group of the first compound to produce a terminal halogen on the covalent coating; and

reacting the terminal halogen on the covalent coating with an amine to produce a quaternary amine on the covalent coating.

16. A method of producing a hydrophilic coating covalently attached to a hydrophobic surface of a polymer, the method comprising the steps of:

a) providing a high surface area polymer having a hydrophobic surface comprising unsaturated groups;

b) providing a first compound, free of methacrylic acid, comprising a hydrophobic domain covalently and flexibly bonded to a hydrophilic domain, the hydrophobic domain comprising an unsaturated group;

c) contacting the hydrophobic surface with a liquid phase, hydrophilic with respect to the surface, containing solvated molecules of the first compound, thereby to deposit molecules of the compound on the hydrophobic surface oriented with the hydrophobic domains proximate to and adsorbed on the surface and with the hydrophilic domains extending outwardly from the surface into the liquid phase; and

d) covalently crosslinking the unsaturated group on on the hydrophobic domain of molecules of the first compound to the unsaturated groups on the hydrophobic surface by a free radical reaction to produce a hydrophilic coating covalently bound to the surface of the polymer, wherein the hydrophilic domains present in the coating substantially shield the hydrophobic surface of the polymer.

17. The method of claim 16 further comprising:

(e) derivatizing the first compound to produce a hydrophilic moiety.

18. The method of claim 17 wherein the hydrophilic moiety is selected from the group consisting of carboxylic acid, sulfonic acid and quaternary amine.

19. The method of claim 17 wherein the first compound is poly(glycidyl methacrylate).

20. The method of claim 19 wherein step (e) includes the step of reacting the poly(glycidyl methacrylate) coating with a second compound selected from the group consisting of acrylic acid, 2-acrylamido-2-methyl-1-propanesulfonic acid, 3-(methacryloyl-amino)propyl! trimethylammonium chloride, 2-acrylamidoglycolic acid, itaconic acid and ethyl vinyl ketone.

21. The method of claim 20 wherein steps d) and e) are conducted simultaneously.

22. A hydrophilic material useful in contact with a protein solution to inhibit denaturation thereof, said material comprising a high surface area polymer having a hydrophobic surface, and a hydrophilic coating, covalently bound to said hydrophobic surface, said hydrophilic material being formed by the process of:

a) providing a high surface area polymer having a hydrophobic surface comprising unsaturated groups, and a compound, free of poly(glycidyl methacrylate), or free of methacrylic acid, comprising a hydrophobic domain covalently and flexibly bonded to a hydrophilic domain, the hydrophobic domain comprising an unsaturated group;

b) contacting the hydrophobic surface with a liquid phase, hydrophilic with respect to the surface, containing solvated molecules of the compound, thereby to deposit molecules of the compound on the hydrophobic surface oriented with the hydrophobic domains proximate to and adsorbed on the surface and with the hydrophilic domains extending outwardly from the surface into the liquid phase; and

c) covalently crosslinking the unsaturated group on on the hydrophobic domain of molecules of the compound to the unsaturated groups on the hydrophobic surface by a free radical reaction to produce a hydrophilic coating covalently bound to the surface of the polymer.

23. The material of claim 22 wherein the compound is modified polyvinylalcohol free of poly(glycidyl methacrylate)

24. The material of claim 22 wherein the compound is poly(glycidyl methacrylate) free of methacrylic acid.

25. The material of claim 22 wherein said polymer comprises polystyrene crosslinked with divinylbenzene.

26. The material of claim 22 wherein step (c) further comprises derivatizing the compound to produce a hydrophilic moiety.

27. The material of claim 26 wherein said hydrophilic coating comprises sufficient hydrophilic groups to to mask said hydrophobic domains and said hydrophobic surface.

28. The material of claim 26 wherein said hydrophilic moiety is selected from the group consisting of hydroxyl, aidehyde, carboxylic acid, quaternary amine and sulfonic acid.

29. A hydrophilic material useful in contact with a protein solution to inhibit denaturation thereof, said material comprising:

a hydrophobic surface enveloped by a hydrophilic coating, covalently bound to said surface, said coating comprising a fimbriate structure comprising a multiplicity of elongate polymer chains extending from said surface and comprising plural hydrophilic groups, said chains being present at a surface density sufficient effectively to mask said hydrophobic surface and substantially to eliminate hydrophobic interaction between a said protein solution and areas of said hydrophobic surface.

30. The material of claim 29 wherein said hydrophobic surface comprises particulate polystyrene crosslinked with divinyl benzene.

31. The material of claim 29 wherein said hydrophilic groups are selected from the group consisting of hydroxyl, carboxylic acid, quaternary amine, and sulfonic acid.