Polymer brushes for immobilizing molecules to a surface and having water-soluble or water-dispersible segments therein and probes bonded thereto

a technology of polymer brushes and probes, which is applied in the field of polymer brushes, can solve the problems of affecting the accessibility of biological molecules, difficult to attach probe molecules to surfaces, and impede so as to optimize the accessibility of probes of a given size or type, the effect of optimizing the attachment of probe molecules

Inactive Publication Date: 2005-07-21
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0006] Accordingly, a need to continues to exist for a polymer brush, as well as a process for the preparation thereof, having polymer chains of a controlled molecular architecture (i.e., composition, functionality, molecular weight, polydispersity, etc.), as well as spacing or grafting density on the substrate surface, such that the attachme

Problems solved by technology

Attaching probe molecules to surfaces is often difficult because the surfaces lack functional groups that are uniquely reactive in an aqueous system or are readily accessible to the probe molecules as a result of factors such as surface crowding or steric hindrance.
This problem becomes particularly acute as the number of functional groups per unit area of surface increases.
Here, too, factors such as steric hindrance may hamper accessibility.
Molecular crowding (i.e., density) becomes a critical issue as well, particularly in systems where fluorescence qu

Method used

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  • Polymer brushes for immobilizing molecules to a surface and having water-soluble or water-dispersible segments therein and probes bonded thereto
  • Polymer brushes for immobilizing molecules to a surface and having water-soluble or water-dispersible segments therein and probes bonded thereto
  • Polymer brushes for immobilizing molecules to a surface and having water-soluble or water-dispersible segments therein and probes bonded thereto

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0185] The following example illustrates one approach for the preparation of a monomer suitable for use in the present invention for polymer brush preparation.

I. Synthesis of N-Methyl,N-(2-hydroxyethyl)acrylamide

A. Synthesis of N-Methyl,N-2-(1-Trimethylsiloxy)ethylacrylamide

[0186]

[0187] An ovendried 4 L three-necked flask equipped with an overhead mechanical stirrer, a 250 mL dropping funnel and an adapter to an argon line was charged under an atmosphere of argon with 113 g (1.5 mol, 121 mL) 2-(methylamino)ethanol (1), commercially available from Sigma-Aldrich, 1500 mL of anhydrous dichloromethane, and 334 g (460 mL, 3.3 mol) of triethylamine. The solution was chilled to ca. 0° C. (icebath) and 171 g (1.58 mol, 200 mL) of chlorotrimethylsilane (TMSCl) were added dropwise. Upon completion of the exothermic reaction (ca. 1 h), the reaction mixture is cooled again to ca. 0° C. (icebath) and acryloyl chloride (2) (136 g, 1.5 mol, 122 mL) were added dropwise and the reaction mixture w...

example 2

[0202] This example describes the preparation of a surface-bound copolymer having a target molecular weight of 50,000 daltons and 10 mol % of N-methyl-N-(2-hydroxyethyl)-acrylamide and 90 mol % N,N-dimethylacrylamide. Three fused silica wafers and one silicon wafer were used in this experiment. It was assumed that each surface contained hydroxyl functionalities typically in the picomole per square inch range. In order to add an initiator-control ageht adduct to each surface hydroxyl functionality, 5×10−4 mol of the trichlorosilyl-substituted initiator-control agent adduct was used per 2×3 in. wafer. Thus, the surface in this example is considered to have a fraction of chain initiator to the total number of reactive sites on the substrate surface of about 1.

[0203] Unbound initiator-control agent adduct (1.9 g) was dissolved in 291 mL of N,N-dimethylacrylamide, 37.9 g of N-methyl-N-(2-hydroxyethyl)acrylamide, 32 mg of α-hydridonitroxide, and 30 mL of water. The reaction vessel was se...

example 3

[0204] This example describes the preparation of a surface-bound copolymer brush having a target molecular weight of 50,000 daltons and 25 mol % incorporation of N-methyl-N-(2-hydroxyethyl)acrylamide.

[0205] Substantially following Example 2, three fused silica wafers and one silicon wafer were modified with the trichlorosilyl-substituted initiator / control agent adduct to provide a fraction of chain initiator to the total number of surface functionalities of about 1. 1.9 g of unbound initiator-control agent adduct was dissolved in 232 mL of N,N-dimethylacrylamide, 90.8 g of N-methyl-N-(2-hydroxyethyl)acrylamide, 32 mg of α-hydridonitroxide, and 30 mL of water. The reaction vessel was sealed under argon and heated at 130° C. for 48 h. After the polymerization reaction, the wafers were placed in a DMF bath and heated at 50° C. for 12 h to remove non-covalently attached polymer that was then analyzed by SEC, as described above. After washing with water and acetone, the wafers were air ...

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Abstract

Sensors for determining the presence and concentration of bio-molecules in a biological sample are provided in the form of polymer brushes, which comprise a substrate having a surface that is modified with a water-dispersible or water-soluble polymer segment having functional groups that bind probes. The method of synthesis of such sensors preferably includes use of controlled free radical polymerization techniques, and in particular the use of an iniferter initiator, which allows for controlled architecture polymers to modify the surface of the substrate. In this manner functional groups in the polymer chain are removed from the surface, which allows for solution chemistry to be more realistically reproduced with the benefits of a solid bound probe.

Description

CROSS-REFERENCE TO RELATED APPLICATION [0001] This application is a continuation of, and claims priority from, U.S. application Ser. No. 09 / 911,683 (filed Jul. 24, 2001), which is a continuation of, and claims priority from, U.S. application Ser. No. 09 / 609,461 (filed Jul. 3, 2000), which is a continuation-in-part of U.S. application Ser. Nos. 09 / 347,606, 09 / 347,607, 09 / 347,608 and 09 / 347,609 (all of which were filed on Jul. 2, 1999 and which are now abandoned) and which claims the benefit of priority therefrom as well as from U.S. Provisional Application Nos. 60 / 146,936 (filed Jul. 31, 1999) and 60 / 177,879 (filed Jan. 24, 2000). All of these related applications are incorporated by reference herein.BACKGROUND [0002] This invention relates to a polymer brush that features a polymer layer on a substrate surface, the polymer layer being composed of a number of polymer chains each of which include a water-soluble or water-dispersible segment having two termini, one terminus being free ...

Claims

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Application Information

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IPC IPC(8): C08F4/00C08F220/54C08F220/58G01N27/447G01N33/543
CPCC08F4/00C08F220/54Y10T436/10G01N27/44747G01N33/54353C08F220/58Y10T428/8305
Inventor KLAERNER, GERRITNIELSEN, RALPH B.MANSKY, PAULBENOIT, DIDIERCHARMOT, DOMINIQUEJANDELEIT, BERNDMAZZOLA, LAURA T.
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