Methods for producing surfaces that resist non-specific protein binding and cell attachment
View Patent ↗A method is disclosed herein for treating a polymeric surface to resist non-specific binding of biomolecules and attachment of cells. The method includes the steps of: imparting a charge to the polymeric surface to produce a charged surface; exposing the charged surface to a nitrogen-rich polymer to form a polymerized surface; exposing the polymerized surface to an oxidized polysaccharide to form an aldehyde surface; and exposing the aldehyde surface to a reducing agent. Advantageously, a method is provided which produces surfaces that resist non-specific protein binding and cell attachment and that avoids the use of photochemical reactions or prior art specially designed compounds.
1. A method for treating a polymeric surface to resist non-specific binding of biomolecules and attachment of cells, the method comprising the steps of:
a) imparting a positive charge to the polymeric surface to produce a positively charged surface;
b) exposing said positively charged surface to a linker to produce an active surface;
c) exposing the active surface to a nitrogen-rich polymer to form a polymerized surface;
d) exposing the polymerized surface to an aldehyde-bearing polymer to form an aldehyde surface; and
e) exposing the aldehyde surface to a reducing agent;
wherein the linker is selected from the group consisting of dialdehydes, diesters, diimidoesters, NHS-esters, hydrazides, carbodiimides, aryl azides, and heterobifunctional linkers having an ester on one end and an aldehyde on the other end.
2. The method of claim 1 , wherein the step of exposing the aldehyde surface to the reducing agent further includes exposing the aldehyde surface to an amine-terminated polymer selected from the group consisting of amine-terminated hydrocarbyl polymers and amine-terminated polyethers, and then exposing the surface to the reducing agent.
3. The method of claim 2 , wherein the amine-terminated polyether is amine-terminated polyethylene glycol.
4. The method of claim 1 , wherein the step of imparting the charge to the polymeric surface includes disposing the polymeric surface into a substantially gas-free chamber, introducing a gas into the chamber, and exciting the gas to produce the charged surface.
5. The method of claim 4 , wherein the exciting of the gas includes subjecting the gas to radiofrequency excitation.
6. The method of claim 4 , wherein the gas is ammonia gas.
7. The method of claim 1 , wherein the linker is glutaric aldehyde.
8. The method of claim 1 , wherein the nitrogen-rich polymer is selected from the group consisting of: a polyalkylenimine and a polymeric amino acid.
9. The method of claim 8 , wherein the nitrogen-rich polymer is selected from the group consisting of: polyethylenimine and poly-lysine.
10. The method of claim 1 , wherein the aldehyde-bearing polymer is an oxidized polysaccharide.
11. The method of claim 10 , wherein the oxidized polysaccharide is selected from the group consisting of: oxidized amylose, oxidized amylopectin, oxidized cellulose, oxidized chitin, oxidized guaran, oxidized glucomannan, and oxidized dextran.
12. The method of claim 1 , wherein the polymeric surface is a polystyrene surface.
13. The method of claim 1 , wherein the polymeric surface is a surface of a multiwell plate.
14. The method of claim 1 , wherein the reducing agent is sodium borohydride.
15. The method of claim 1 , wherein the linker is a dialdehyde having the structure
wherein R is a C 2 to C 30 alkylenyl.
16. A surface treated by the method of claim 1 .
17. The surface of claim 16 , wherein the surface is the surface of a multiwell plate.