Method of stabilizing molecules without refrigeration using water soluble polymers and applications thereof in performing chemical reactions

Patent metadata

Field	Value
Publication number	US-11067569-B2
Application number	US-201916274616-A
Country	US
Kind code	B2
Filing date	Feb 13, 2019
Priority date	Nov 8, 2013
Publication date	Jul 20, 2021
Grant date	Jul 20, 2021

Abstract

The present application is directed to methods of performing chemical reactions, including multi-step chemical reactions in which two or more of the reagents in the chemical reaction are incorporated or entrapped in a solid polymeric structure comprising pullulan. In certain embodiments, the chemical reaction or multi-step reaction serves as a sensor. Accordingly the present application is also directed to sensors for performing the methods of the application. In certain embodiments, at least one of the reagents is a biomolecule and the sensor is a biosensor. In certain other embodiments, the solid polymeric structure comprising pullulan and the reagents for performing a chemical reaction form a convenient device for performing a chemical reaction.

First claim

The invention claimed is:

1. A device comprising two or more reagents entrapped in the same solid polymeric structure or in separate solid polymeric structures wherein the solid polymeric structure or structures are prepared from an aqueous solution comprising about 10% (w/v) to about 13% (w/v) of pullulan and at least one of the two or more reagents is selected from one or more different reagents that are a protein, antibody, peptide, nucleic acid, phage, antidote and vaccine, or at least one of the two or more reagents is a material from a microorganism, or at least one of the two or more reagents is an inorganic or an organic molecule.

2. The device of claim 1 , wherein the two or more reagents are entrapped in separate solid polymeric structures and the solid polymeric structures are stacked in layers on top of each other.

3. The device of claim 1 , wherein the two or more reagents are for performing a chemical reaction and are entrapped in separate solid polymeric structures and the solid polymeric structures are in the shape of a pill, tablet or capsule, with separate polymeric structures forming separate layers surrounding each other wherein a layer on the outside of the pill, tablet or capsule comprises reagents that must react first in the chemical reaction and the remaining layers are arranged internally to the outside layer in an order that corresponds to an order required to perform the chemical reaction.

4. The device of claim 1 , wherein the two or more reagents are entrapped in the same solid polymeric structure.

5. The device of claim 1 , wherein one or more of the solid polymeric structures are cast onto a substrate comprising at least one of the two or more reagents for a chemical reaction.

6. The device of claim 1 , wherein at least one of the two or more reagents is a material from a microorganism.

7. The device of claim 1 , wherein at least one of the two or more reagents is an inorganic or an organic molecule.

8. The device of claim 7 , wherein the inorganic molecule is selected from one or more different reagents that are an inorganic acid, an inorganic base, an alkaline earth metal carbonate, an alkaline earth metal sulfate, an alkali metal carbonate, an alkali metal sulfate and a metal complex.

9. The device of claim 7 , wherein the organic molecule is selected from one or more different molecules that are indoxyl acetate, luciferin, lactate, acetaldehyde, chelating agents, NTPs (ATP, GTP, UTP and CTP) and dNTPs (dATP, dGTP, dTTP and dCTP).

10. The device of claim 1 , wherein the two or more reagents are in separate polymeric structures and the two or more reagents that are in separate polymeric structures comprise reagents to perform the Simon's test for secondary amines.

11. The device of claim 10 , wherein a top or first layer in a stacked sensor, or a first or outer layer of a pill, tablet or capsule, comprises acetaldehyde and a second or inner layer comprises sodium nitroprusside and sodium carbonate.

12. A method of performing a single step or multi-step chemical reaction comprising:
a) combining two or more reagents for the reaction, either separately or together, with an aqueous pullulan solution to provide reagent pullulan solutions or a reagent pullulan solution, respectively, wherein the aqueous pullulan solution comprises about 10% (w/v) to about 13% (w/v) of pullulan;
b) drying the reagent pullulan solutions or the reagent pullulan solution to provide solid polymeric structures or a solid polymeric structure, respectively; and
c) if the two or more reagents are in separate solid polymeric structures in b), then treating the solid polymeric structures under conditions to dissolve the solid polymeric structures for the two or more reagents to interact in a chemical reaction; or
d) if the two or more reagents are together in the solid polymeric structure in b), then treating the solid polymeric structure under conditions to dissolve the solid polymeric structures for the two or more reagents to interact in a chemical reaction, wherein the solid polymeric structures are or the solid polymeric structure is the component(s) of the device of claim 1 .

13. The method of claim 12 , wherein, the two or more reagents are in separate solid polymeric structures and the solid polymeric structures are stacked in layers on top of each other.

14. The method of claim 13 , wherein the layers are stacked in an order that corresponds to an order required to perform the chemical reaction.

15. The method of claim 12 , wherein the two or more reagents are in separate solid polymeric structures and the solid polymeric structures are in the shape of a pill, tablet or capsule, with each separate polymeric structure forming separate layers surrounding each other wherein a layer on the outside of the pill, tablet or capsule comprises reagents that must react first in the chemical reaction and the remaining layers are arranged internally to the outside layer in an order that corresponds to an order required to perform the chemical reaction.

16. The method of claim 15 , wherein timing of dissolution of the layers is controlled to allow sufficient reaction time for each step of the chemical reaction.

17. The method of claim 16 , wherein the timing is controlled by one or more of thickness of the layers and pullulan concentration.

18. The method of claim 12 , wherein the two or more reagents are in the same solid polymeric structure.

19. The method of claim 12 , wherein the two or more reagents are in separate solid polymeric structures and the solid polymeric structures are each dissolved in a single water or buffer solution to release the two or more reagents for interaction in the chemical reaction.

Assignees

• Univ Mcmaster

Inventors

• Filipe Carlos
• Brennan John D
• Pelton Robert
• Jahanshahi-Anbuhi Sana
• Li Yingfu

Classifications

• G01N2333/922

  Ribonucleases (RNAses); Deoxyribonucleases (DNAses) · CPC title

• G01N21/78

  producing a change of colour · CPC title

• C12Q1/66

  involving luciferase · CPC title

• G01N33/5436 Primary

  with ligand physically entrapped within the solid phase (liposomes G01N33/5432; immunological test elements G01N33/54386) · CPC title

• G01N2021/7786

  Fluorescence · CPC title