SENSOR FOR DETECTING SACCHARIDE AND MANUFACTURING METHOD THEREOF AND DETECTION METHOD OF GLYCATED HEMOGLOBIN USING THE SAME

• US 20160109370A1
• Filed: 03/11/2015
• Published: 04/21/2016
• Est. Priority Date: 10/21/2014
• Status: Active Grant

• Current Assignee: Ajou University Industry, LG Electronics, Inc. (LG Corporation)
• Original Assignee: Ajou University Industry, LG Electronics, Inc. (LG Corporation)
• Inventors: KU, Yunhee, KIM, Moosub, YANG, Yongju, SONG, Seung Yeon, LEE, Kang Sun, YOON, Hyun Chul, HAN, Yong Duk, PARK, Yoo Min
• Granted Patent: US 9,897,920 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: G01N 21/78 producing a change of colour
• CPC Class Codes: G01N 2400/00 Assays, e.g. immunoassays or enzyme assays, involving carbohydrates
• CPC Class Codes: G03F 7/027 Non-macromolecular photopolymerisable compounds having carbon-to-carbon double bonds, e.g. ethylenic compounds G03F7/075 takes precedence
• CPC Class Codes: G03F 7/2002 with visible light or UV light, through an original having an opaque pattern on a transparent support, e.g. film printing, projection printing; by reflection of visible or UV light from an original such as a printed image

Abstract

The present disclosure relates to a sensor for detecting saccharide and manufacturing method thereof and detection method of glycated hemoglobin using the same wherein the sensor for detecting saccharide includes a reactive layer synthesized with hydrogel having a boronic acid-modified 3D mesh structure. A selective and sensitive detection of the glycated protein including glucose or glycated hemoglobin (HbA_1c) in blood through high level of signal may be enabled, and the durability of the sensor for detecting saccharide may be enhanced.

Claims

1. A sensor for detecting saccharide, the sensor comprising:

a reactive layer synthesized with boronic acid-modified hydrogel; and

a porous membrane substrate fixed with the reactive layer.

2. The sensor of claim 1, wherein the saccharide is a saccharide portion of glycated protein or glucose.

3. The sensor of claim 2, wherein the glycated protein is glycated hemoglobin (HbA_1c).

4. The sensor of claim 1, wherein the hydrogel is synthesized by a photo-polymerization reaction with a polymer monomer, a polymer cross-linker, and boronic acid in the porous membrane substrate.

5. The sensor of claim 4, wherein the polymer monomer is an acrylamide derivative, the polymer cross-linker is an acrylamide derivative having two N functional groups (N,N′

-), and the boronic acid is a boronic acid having acryl groups.

6. The sensor of claim 5, wherein the polymer monomer is N-(2-hydroxyethyl)acrylamide, and the polymer cross-linker is N,N′

-methylene-bisacrylamide.

7. The sensor of claim 5, wherein the boronic acid is 3-(acrylamido)phenylboronic acid.

8. The sensor of claim 1, wherein the porous membrane substrate is a substrate using a membrane for a LFA (Lateral Flow Assay) formed of glass fibers.

9. A method for manufacturing the sensor of claim 1, the method comprising:

injecting a polymer monomer, a polymer cross-linker, boronic acid, and a photo-polymerization reaction initiator into the porous membrane substrate;

patterning the porous membrane substrate with a photomask;

irradiating UV (ultraviolet) onto the patterned porous membrane substrate;

washing the UV-irradiated porous membrane substrate; and

drying the washed porous membrane substrate.

10. The method of claim 9, wherein the polymer monomer is an acrylamide derivative, the polymer cross-linker is an acrylamide derivate having two N functional groups (N,N′

-), and the boronic acid is a boronic acid having acryl groups.

11. The method of claim 9, wherein:

the polymer monomer is N-(2-hydroxyethyl)acrylamide,

the polymer cross-linker is N,N′

-methylene-bisacrylamide,

the boronic acid is 3-(acrylamido)phenylboronic acid,

the N-(2-hydroxyethyl)acrylamide, the N,N′

-methylene-bisacrylamide, the 3-(acrylamido)phenylboronic acid and the photo-polymerization reaction initiator are mixed in a dimethyl sulfoxide solvent, and

with respect to the dimethyl sulfoxide solvent, a weight ratio of the N-(2-hydroxyethyl)acrylamide is 8.00˜

10.00%(w/v), a weight ratio of the N,N′

-methylene-bisacrylamide is 0.40˜

0.50%(w/v), a weight ratio of the 3-(acrylamido)phenylboronic acid is 0.10˜

2.00%(w/v), and a weight ratio of the photo-polymerization reaction initiator is 0.05˜

0.14%(w/v).

12. The method of claim 9, wherein the porous membrane substrate is a substrate using a membrane for a LFA (Lateral Flow Assay) formed of glass fibers.

13. A method for detecting glycated hemoglobin, the method comprising:

mixing and reacting glycated hemoglobin and horseradish peroxidase on the sensor of claim 1;

washing non-reactive protein after the reacting of the mixed glycated hemoglobin and horseradish peroxidase;

inducing a color development by dripping a chromogen working solution including a chromogen and a substrate with respect to the horseradish peroxidase onto the sensor;

digitizing a degree of the color development; and

outputting the digitized degree corresponding to a concentration of the detected glycated hemoglobin.

14. The method of claim 13, wherein the substrate is hydrogen peroxide, and the chromogen is any one selected from the group consisting of 3,3′

,5,5′

-tetramethylbenzidine, o-phenylenediamine dihydrochloride, 2,2′

-azinobis[3-ethylbenzothiazoline-6-sulfonic acid]-diammonium salt, 3,3′

-diaminobenzidine tetrahydrochloride, SAT-3 (analog of o-tolidine), N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3-methoxyaniline sodium salt dihydrate, N-ethyl-N-(3-sulfopropyl)-3-methoxyaniline sodium salt monohydrate, N-ethyl-N-(3-sulfopropyl)aniline, sodium salt, N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline sodium salt, N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline sodium salt), N,N-Bis(4-sulfobutyl)-3,5-dimethylaniline disodium salt, N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethylaniline sodium salt monohydrate, N,N-Bis(4-sulfobutyl)-3-methylaniline disodium salt, N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3-methylaniline sodium salt dihydrate and N-ethyl-N-(3-sulfopropyl)-3-methylaniline sodium salt monohydrate.

15. The method of claim 13, wherein the mixing and reacting the glycated hemoglobin and the horseradish peroxidase comprises:

dropping 1 to 100 μ

l of a solution, which is prepared by mixing and resolving the horseradish peroxidase and the glycated hemoglobin at a volumetric ratio of 1;

1 in a PBS (phosphate-buffered saline) solution to a concentration of 1 to 10 μ

g/ml, onto the sensor; and

reacting the dropped solution for 10 to 600 seconds.