Enzyme-encapsulated nanoparticle platform

Patent metadata

Field	Value
Publication number	US-2016243262-A1
Application number	US-201415022551-A
Country	US
Kind code	A1
Filing date	Sep 17, 2014
Priority date	Sep 17, 2013
Publication date	Aug 25, 2016
Grant date	—

Abstract

Disclosed are methods, systems, and devices for implementing nanoparticles to encapsulate biomolecules such as enzymes. In one aspect, a nanoparticle device includes a shell structure including an internal layer structured to enclose a hollow interior region and include one or more holes penetrating through the internal layer, and an external layer formed of a porous material around the internal layer; and an enzyme contained within the interior region of the shell structure, the enzyme having entered the shell structure through the one or more holes and incapable of passing through the external layer, in which the pores are of a size that prevents the enzyme to pass through the pores while permitting substances smaller than the pore size to pass through the pores.

First claim

What is claimed is:

1 . A nanoparticle for catalyzing an analyte, comprising:
a shell structure including an internal layer and an external layer, the internal layer structured to enclose an interior region and structured to form one or more holes penetrating through the internal layer, and the external layer formed of a porous material around the internal layer; an enzyme contained within the interior region of the shell structure, wherein the internal layer is structured to form the one or more holes sized to allow the enzyme to pass through the internal layer; and a biochemical cofactor corresponding to the enzyme, wherein the biochemical cofactor is contained in the interior region and structured to bind to the enzyme, wherein the porous material of the external layer is structured to prevent the enzyme from passing through the external layer while permitting an analyte smaller than the enzyme to pass through the external layer, wherein the enzyme is structured to catalyze the analyte.

2 . The nanoparticle as in claim 1 , wherein the enzyme includes an apoenzyme, wherein the cofactor binds to the apoenzyme to form a holoenzyme capable of catalyzing the analyte.

3 . The nanoparticle as in claim 1 , further comprising a charged material layer formed in the holes to provide an electrostatic force to further contain the biochemical cofactor in the interior region.

4 . The nanoparticle as in claim 1 , wherein the enzyme contained within the shell structure includes methioninase, the cofactor includes pyroxidal-5′-phosphate (PLP), and the analyte includes methionine.

5 . The nanoparticle as in claim 4 , wherein the nanoparticle is configured to be deployed to a biological tissue within an organism through the blood stream, wherein the shell structure inhibits antibodies and other substances that degrade methioninase from entering the interior region while permitting the methionine to enter into the interior region through the pores and catalytically interact with the contained methioninase.

6 . The nanoparticle as in claim 4 , wherein the nanoparticle is configured to increase the catalysis of methionine based on reduced loss of the PLP in the blood stream.

7 . The nanoparticle as in claim 1 , further comprising a ligand molecule conjugated to the shell structure, the ligand molecule having an affinity to a receptor molecule of the biological tissue to attract the shell structure to the biological tissue.

8 . The nanoparticle as in claim 1 , wherein the nanoparticle is configured to a size capable of in vivo injection including in a range between 100 nm to 500 nm.

9 . The nanoparticle as in claim 1 , further comprising a paramagnetic material in the shell structure.

10 . The nanoparticle as in claim 1 , wherein the biological system includes a living organism including a non-human animal or a human being.

11 . An ultrasound-interactive nanoparticle sensor device for detecting reactive oxidative species, comprising:
a nanoparticle structured to include a shell structure including an internal layer and an external layer, the internal layer enclosing a hollow interior region and structured to form one or more holes penetrating through the internal layer, and the external layer formed of a porous material arranged around the internal layer; and an enzyme encapsulated within the interior region of the shell structure, wherein the internal layer is structured to form the one or more holes sized to allow the enzyme to pass through the internal layer, and wherein the enzyme is structured to catalyze a reactive oxidative species (ROS) to decompose and produce oxygen, wherein the enzyme-encapsulated nanoparticle is configured to produce microbubbles from the oxygen produced by decomposition of the ROS within the nanoparticles, and wherein the produced microbubbles are configured to cause a change in a returned acoustic waveform carrying information on the microbubbles responsive to an application of an ultrasonic acoustic energy.

12 . The device as in claim 11 , further comprising nanoscale structures protruding from the interior surface of the internal layer toward the hollow interior region, wherein the nanoscale structures initiate cavitation of oxygen bubbles based on the applied ultrasonic acoustic energy.

13 . The device as in claim 11 , wherein the shell structure is configured to prevent proteases from the biological system that are chemically interactive with the enzyme from entering the interior region.

14 . The device as in claim 11 , wherein the ROS includes hydrogen peroxide.

15 . The device as in claim 11 , wherein the enzyme includes catalase.

16 . The device as in claim 11 , wherein the external layer of the nanoparticle includes a nanoporous surface to provide a nucleation site for oxygen microbubble formation.

17 . The device as in claim 11 , wherein the enzyme-encapsulated nanoparticle is configured to be deployed in a biological system including a living organism, and the produced microbubbles are configured to react to the applied ultrasonic acoustic energy directed at a particular region of the biological system including a depth up to 20 cm in a biological tissue of the living organism.

18 . The device as in claim 17 , wherein the produced microbubbles are configured to react to the applied ultrasonic acoustic energy pulsed to elicit nonlinear oscillations of the microbubbles affecting the returned acoustic energy to detect the ROS in the biological tissue.

19 . The device as in claim 17 , wherein the nanoparticle includes a ligand molecule conjugated to the shell structure, the ligand molecule having an affinity to a receptor molecule of the biological tissue to attract the shell structure to the biological tissue.

20 . The device as in claim 11 , wherein the nanoparticle is configured to a size capable of in vivo injection including in a range between 100 nm to 500 nm.

21 . The device as in claim 11 , wherein the nanoparticle includes a paramagnetic material in the shell structure.

22 . The device as in claim 11 , wherein the enzyme-encapsulated nanoparticle is configured to be deployed in the biological system including a non-human animal or a human being.

23 . A nanoparticle sensor device for detecting analyte, comprising:
enzyme-encapsulated nanoparticles capable of being injected into a biological system, the enzyme-encapsulated nanoparticles structured to include:
a shell structure including an internal layer and an external layer, the internal layer enclosing a hollow interior region and structured to form one or more holes penetrating through the internal layer, and the external layer formed of a porous material arranged around the internal layer,
an enzyme contained within the interior region of the shell structure, wherein the internal layer is structured to form the one or more holes sized to allow the enzyme to pass through the internal layer, wherein the external layer is structured to prevent the enzyme from passing through the external layer but allow an analyze smaller than an enzyme to pass through the external layer, and wherein the enzyme is structured to catalyze the analyte that enters the interior region, and
a fluorophore attached to the shell structure and configured to emit an optical fluorescent signal based at least on the concentration of a chemical reactant or chemical product of a catalytic interaction of at least the enzyme and the analyte;
a light source to direct an ex

Assignees

• Univ California
• Devacell Inc

Inventors

• Ortac Inanc
• Esener Sadik C
• Yayla Ibrahim Gokce
• Messmer Bradley

Classifications

• A61K38/50

  acting on carbon-nitrogen bonds, other than peptide bonds (3.5), e.g. asparaginase · CPC title

• C12Q1/30

  involving catalase · CPC title

• C12Q1/54

  involving glucose or galactose · CPC title

• A61B5/6831

  Straps, bands or harnesses · CPC title

• A61B5/1455

  using optical sensors, e.g. spectral photometrical oximeters · CPC title