Hierarchical films having ultra low fouling and high recognition element loading properties

Patent metadata

Field	Value
Publication number	US-10031138-B2
Application number	US-201313747290-A
Country	US
Kind code	B2
Filing date	Jan 22, 2013
Priority date	Jan 20, 2012
Publication date	Jul 24, 2018
Grant date	Jul 24, 2018

Abstract

Hierarchical films with structurally regulated functionalities through the integration of two-dimensional and three-dimensional structures to achieve ultra low nonspecific binding and high loading of molecular recognition elements, and methods for making and using the films.

First claim

The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows:

1. A surface-anchored polymer assembly, comprising:
(a) a nonfouling first layer having a first layer density, wherein the first layer is attached to a substrate and comprises a plurality of polycarboxybetaine polymers; and
(b) a second layer having a second layer density, wherein the second layer comprises a plurality of polycarboxybetaine polymers each having carboxy groups and a plurality of recognition elements attached to the carboxy groups such that the recognition elements are attached throughout the entire second layer, and wherein the second layer is attached to the first layer,
wherein the first layer density is greater than the second layer density.

2. The surface-anchored polymer assembly of claim 1 , further comprising one or more additional layers attached to the second layer.

3. The surface-anchored polymer assembly of claim 1 , wherein the first layer has a fibrinogen binding level less than about 30 ng/cm 2 .

4. The surface-anchored polymer assembly of claim 1 , wherein the first layer has a lysozyme binding level less than about 30 ng/cm 2 .

5. The surface-anchored polymer assembly of claim 1 , wherein the second layer is a crosslinked layer.

6. The surface-anchored polymer assembly of claim 1 , wherein the first layer polymers are grafted from the substrate.

7. The surface-anchored polymer assembly of claim 1 , wherein the first layer polymers are grafted to the substrate.

8. The surface-anchored polymer assembly of claim 1 , wherein the second layer polymers are grafted from the first layer polymers.

9. The surface-anchored polymer assembly of claim 1 , wherein the second layer polymers are grafted to the first layer polymers.

10. The surface-anchored polymer assembly of claim 1 , wherein the polycarboxybetaine polymers have the formula:
PB[L 1 -(N + (R 2 )(R 3 ))-L 2 -AO 2 − ] n (X − ) n (M + ) n
wherein
PB is the polymer backbone having n pendant zwitterionic groups;
R 2 and R 3 are independently selected from the group consisting of hydrogen, alkyl, and aryl, or taken together with the nitrogen to which they are attached form a cationic center;
L 1 is a linker that covalently couples the cationic center [N + (R 2 )(R 3 )] to the polymer backbone;
L 2 is a linker that covalently couples the anionic center [A(═O)—O − ] to the cationic center;
A is C;
M + is an optional counter ion associated with the (A=O)O − anionic center;
X − is an optional counter ion associated with the cationic center; and
n is an integer from 1 to about 10,000.

11. The surface-anchored polymer assembly of claim 1 , wherein the substrate is selected from metal and metal oxide surfaces, ceramic surfaces, synthetic and natural polymeric surfaces, glass surfaces, fiber glass surfaces, silicon/silica surfaces, carbon-based material surfaces, cell surfaces, or macromolecule surfaces (protein, DNAs, lipids).

12. The surface-anchored polymer assembly of claim 1 , wherein the substrate is the surface of a diagnostic device, a medical device, a separation device, a targeting delivery carrier, a scaffold, or a marine coating.

13. The surface-anchored polymer assembly of claim 1 , wherein the recognition element is selected from the group consisting of a peptide, a protein, a nucleic acid, and a small molecule.

14. The surface-anchored polymer assembly of claim 1 , wherein the recognition element is selected from the group consisting of an antibody, an antibody fragment, a DNA, and an RNA.

15. A method for determining the presence of an analyte in a sample, comprising:
(a) contacting a sample with a surface-anchored polymer assembly of claim 1 , wherein the recognition element has a specific binding affinity for the analyte; and
(b) interrogating the surface-anchored polymer assembly to determine whether the analyte has bound to the surface-anchored polymer assembly.

16. A hierarchical surface-anchored polymer assembly, comprising:
a first layer comprising a plurality of first polycarboxybetaine polymers, wherein each first polycarboxybetaine polymer has a first end and a second end, wherein the first end of each first polycarboxybetaine polymer is attached to a surface;
a second layer comprising a plurality of second polycarboxybetaine polymers, wherein each second polycarboxybetaine polymer has a first end and a second end, wherein the first end of each second polycarboxybetaine polymer is attached to the second end of a first polycarboxybetaine polymer, and wherein fewer than all of the plurality of first polycarboxybetaine polymers has a second polycarboxybetaine polymer attached to the second end; and
a plurality of recognition elements attached to each of the plurality of second polycarboxybetaine polymers such that the recognition elements are attached throughout the entire second layer.

17. A hierarchical surface-anchored polymer assembly produced by a process comprising:
grafting a plurality of first polycarboxybetaine polymers from a surface to provide a plurality of first polymers having a first end attached to the surface and a second end distal from the surface;
capping less than all of the plurality of first polycarboxybetaine polymers at the second ends to prevent further polymerization from the second ends of a portion of the plurality of first polycarboxybetaine polymers, thereby providing a first polycarboxybetaine polymer layer with a first layer density;
grafting a plurality of second polycarboxybetaine polymers from the second end of less than all of the plurality of first polycarboxybetaine polymers grafted from the surface, thereby providing a second polycarboxybetaine polymer layer with a second layer density that is less than the first layer density; and
attaching a plurality of recognition elements throughout the second polycarboxybetaine polymer layer.

Assignees

• Univ Washington Through Its Center For Commercialization

Inventors

• Jiang Shaoyi
• Brault Jr Norman David
• Sundaram Harihara S
• Huang Chun-Jen
• Yu Qiuming
• Li Yuting

Classifications

• G01N21/553

  and using surface plasmons (fluorescence excitation G01N21/648; enhanced Raman G01N21/658) · CPC title

• G01N33/54393 Primary

  Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding · CPC title