Dynamic thermoresponsive nanoparticles for stabilization of enzymes at high temperatures

• US 9,121,017 B2
• Filed: 05/07/2013
• Issued: 09/01/2015
• Est. Priority Date: 11/13/2009
• Status: Active Grant

• Current Assignee: ICx Agentase
• Original Assignee: Flir Detection, Inc. (Teledyne Technologies Incorporated)
• Inventors: Walker, Jeremy P., Leech, Anna M.
• Primary Examiner(s): Ware, Debbie K
• Assistant Examiner(s): Srivastava, Kailash C
• Application Number:
  US13/888,771
• Publication Number:
  US 20130244301A1
• Time in Patent Office: 847 Days
• US Class Current: 1/1
• CPC Class Codes: A61K 31/74 Synthetic polymeric materials
• CPC Class Codes: A61K 47/543 Lipids, e.g. triglycerides; Polyamines, e.g. spermine or spermidine
• CPC Class Codes: A61K 47/58 obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. poly[meth]acrylate, polyacrylamide, polystyrene, polyvinylpyrrolidone, polyvinylalcohol or polystyrene sulfonic acid resin
• CPC Class Codes: C12N 11/082 obtained by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: C12N 11/087 Acrylic polymers
• CPC Class Codes: C12N 11/098 formed in the presence of the enzymes or microbial cells
• CPC Class Codes: C12N 9/96 Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
• CPC Class Codes: C12Y 101/03004 Glucose oxidase (1.1.3.4)
• CPC Class Codes: C12Y 301/08001 Aryldialkylphosphatase (3.1.8.1), i.e. paraoxonase

Abstract

The present invention provides a thermoresponsive nanoparticle useful for the stabilization of enzymes in environments having a temperature greater than thirty degrees Centigrade. The thermoresponsive nanoparticle has (a) a functionalized enzyme conjugate having one or more enzymes or biological catalysts, the enzymes or biological catalysts are modified with palmitic acid N-hydroxysuccinimide ester and acryclic acid N-hydroxysuccinimide ester, and (b) a thermally responsive polymer, wherein the functionalized enzyme conjugate is encapsulated within the thermally responsive polymer. A nanocatalyst is provided that has one or more proteins. The proteins are covalently immobilized and encapsulated within a thermally responsive polymer shell. The proteins are one or more enzymes or biological catalysts. A method for protecting the proteins is also set forth.

Claims

1. A method for protecting nanocatalysts comprising one or more proteins from environmental conditions, said method comprising:

immobilizing one or more nanocatalysts comprising the proteins and completely encapsulating said one or more immobilized nanocatalysts comprising the proteins within one or more thermally responsive polymeric nanoparticles by raising the temperature above the lower critical solution temperature of said thermally responsive polymeric nanoparticles during an initiation process wherein said nanoparticles undergo a conformational change when heated whereby protecting said nanocatalysts comprising the proteins from temperatures greater than 30 degrees Centigrade.

2. The method of claim 1, wherein said one or more proteins is/are at least one enzyme or biological catalyst.

3. The method of claim 2 further including grafting at least one moiety to a surface of said at least one enzyme to bind to said polymeric nanoparticles for forming modified enzymes.

4. The method of claim 3, wherein said at least one moiety is a vinyl group.

5. The method of claim 3 further including subjecting said modified enzymes to a polymerization process.

6. The method of claim 5 further including employing a free-radical addition polymerization process or a living radical polymerization process.

7. The method of claim 6, wherein said free-radical addition polymerization process is selected from the group consisting of an emulsion polymerization process and a non-emulsion polymerization process.

8. The method of claim 7, wherein said free-radical addition polymerization process optionally includes at least one thermally responsive comonomer selected from the group consisting of N-isopropylacrylamide, poly (isopropyl-N-vinylpyrolidone), and combinations thereof.