Synthetic coating for cell culture

Patent metadata

Field	Value
Publication number	US-2016222247-A1
Application number	US-201514970618-A
Country	US
Kind code	A1
Filing date	Dec 16, 2015
Priority date	Mar 17, 2011
Publication date	Aug 4, 2016
Grant date	—

Abstract

A method for coating a surface of a cell culture article includes dissolving a polymer having a covalently attached polypeptide in an aqueous solution to produce a polymer solution. The polymer is formed from monomers selected to form a polymer having a linear backbone, wherein the polymer is crosslink free. The weight percentage of the polypeptide relative to the polymer conjugated to the polypeptide is sufficiently high to render the polymer conjugated to the polypeptide water soluble. The aqueous solution is substantially free of organic solvents. The method further includes (i) disposing the polymer solution on the surface of the cell culture article to produce a coated article; and (ii) subjecting the coated article to sufficient heat or electromagnetic radiation to attach the polymer conjugated to a polypeptide to the surface of the cell culture article.

First claim

1 . A method for coating a surface of a cell culture article, comprising:
dissolving a polymer having a covalently attached polypeptide in an aqueous solution to produce a polymer solution,
wherein the polymer is formed from monomers selected to form a polymer having a linear backbone, wherein the polymer is crosslink free,
wherein the weight percentage of the polypeptide relative to the polymer conjugated to the polypeptide is sufficiently high to render the polymer conjugated to the polypeptide water soluble,
wherein the aqueous solution is substantially free of organic solvents;
disposing the polymer solution on the surface of the cell culture article to produce a coated article; and subjecting the coated article to sufficient heat or electromagnetic radiation to attach the polymer conjugated to a polypeptide to the surface of the cell culture article.

2 . The method of claim 1 , wherein a substantially similar polymer that it not conjugated to the polypeptide is insoluble in water at 25° C.

3 . The method of claim 1 , wherein the weight percentage of the polypeptide relative to the polymer conjugated to the polypeptide is greater than 40%.

4 . The method of claim 1 , wherein the weight percentage of the polypeptide relative to the polymer conjugated to the polypeptide is greater than 60%.

5 . The method of claim 1 , wherein the polypeptide is a cell adhesive polypeptide.

6 . The method of claim 1 , wherein the polypeptide comprises an RGD sequence.

7 . The method of claim 1 , wherein the polypeptide is a selected from the group of a vitronectin polypeptide, a collagen polypeptide, of a laminin polypeptide, a bone sialoprotein polypeptide, and a fibronectin polypeptide.

8 . The method of claim 1 , wherein the polypeptide is a vitronectin polypeptide.

9 . The method of claim 1 , wherein the polymer is formed from at least one monomer comprising a conjugated polypeptide.

10 . The method of claim 9 , wherein the at least one monomer comprising a conjugated polypeptide is methacrylic acid.

11 . The method of claim 1 , wherein the polymer is formed from polymerization of (i) methacrylic acid conjugated to the polypeptide and (ii) hydroxyethylmethacrylate.

12 . The method of claim 1 , wherein the polymer is formed from polymerization of a (i) monomer comprising a methacrylic acid functional group and (ii) hydroxyethmethacrylate.

13 . The method of claim 1 , wherein the polymer conjugated to the polypeptide has a molecular weight of between 10 kilodaltons and 1000 kilodaltons.

14 . The method of claim 1 , wherein the polymer solution comprises between 0.1 mg/ml and 0.5 mg/ml of the polymer conjugated to the polypeptide.

15 . The method of claim 1 , wherein the polymer solution comprises between 0.2 mg/ml and 0.3 mg/ml of the polymer conjugated to the polypeptide.

16 . The method of claim 1 , wherein subjecting the coated article to sufficient heat or electromagnetic radiation comprises incubating the coated article at a temperature of 37° C. or greater.

17 . The method of claim 1 , wherein subjecting the coated article to sufficient heat or electromagnetic radiation comprises incubating the coated article at a temperature of 60° C. or greater.

18 . The method of claim 1 , wherein subjecting the coated article to sufficient heat or electromagnetic radiation comprises subjecting the coated article to UV radiation.

19 . The method of claim 1 , wherein the surface of the substrate has a water contact angle between 12° and 85°.

20 . The method of claim 1 , wherein the surface of the substrate has a water contact angle between 25° and 70°.

21 . The method of claim 1 , wherein the surface of the substrate has a water contact angle between 30° and 60°.

22 . The method of claim 1 , wherein the surface of the substrate is a plasma treated polystyrene surface.

23 . A coated cell culture article produced according to a method for coating a surface of a cell culture article, comprising:
dissolving a polymer conjugated to a polypeptide in an aqueous solution to produce a polymer solution;
wherein the weight percentage of the polypeptide relative to the polymer conjugated to the polypeptide is sufficiently high to render the polymer conjugated to the polypeptide water soluble;
wherein if the polymer were not conjugated to the polypeptide, the polymer would be insoluble in water at 25° C. and wherein the aqueous solution is substantially free of organic solvents;
disposing the polymer solution on the surface of the cell culture article to produce a coated article; and subjecting the coated article to sufficient heat or electromagnetic radiation to attach the polymer conjugated to a polypeptide to the surface of the cell culture article.

24 . A composition comprising:
an aqueous solution; and a polymer conjugated to a polypeptide dissolved in the aqueous solution;
wherein the polymer is formed from monomers selected to form a polymer having a linear backbone, wherein the polymer is crosslink free,
wherein a substantially similar polymer that it not conjugated to the polypeptide is insoluble in water at 37° C., and
wherein the weight percentage of the polypeptide relative to the polymer conjugated to the polypeptide is sufficiently high to render the polymer conjugated to the polypeptide water soluble, and
wherein the composition is substantially free of organic solvents.

25 . The composition of claim 24 , wherein the polymer is formed from polymerization of (i) methacrylic acid conjugated to the polypeptide and (ii) hydroxyethylmethacrylate.

Assignees

• Corning Inc

Inventors

• Frutos Anthony Glenn
• Henry David
• Walerack Corinne

Classifications

• C09D189/00 Primary

  Coating compositions based on proteins; Coating compositions based on derivatives thereof (foodstuff preparations A23J3/00) · CPC title

• C08F220/286

  and containing polyethylene oxide in the alcohol moiety, e.g. methoxy polyethylene glycol (meth)acrylate · CPC title

• C09D151/00 Primary

  Coating compositions based on graft polymers in which the grafted component is obtained by reactions only involving carbon-to-carbon unsaturated bonds (based on ABS polymers C09D155/02); Coating compositions based on derivatives of such polymers · CPC title

• B05D3/06

  by exposure to radiation (B05D3/02 takes precedence {; plasma treatment B05D3/141}) · CPC title

• C08F220/20

  of polyhydric alcohols or phenols {, e.g. 2-hydroxyethyl (meth)acrylate or glycerol mono-(meth)acrylate} · CPC title