METHOD OF MEASURING GLYCOSYLATED PROTEIN PROPORTION BY AC IMPEDANCE METHOD

• US 20130233727A1
• Filed: 03/06/2013
• Published: 09/12/2013
• Est. Priority Date: 03/06/2012
• Status: Active Grant

• Current Assignee: National Central University
• Original Assignee: National Central University
• Inventors: TSAI, Jang-Zern, CHEN, Ching-Jung, WEI, Yung-Nien
• Granted Patent: US 9,080,939 B2
• Time in Patent Office: Days
• US Class Current: 205/780.5
• CPC Class Codes: G01N 27/02 by investigating impedance
• CPC Class Codes: G01N 27/026 Dielectric impedance spectroscopy electrochemical impedance spectroscopy for measuring corrosion G01N17/02
• CPC Class Codes: G01N 33/49 Blood chemical methods for determining blood cell populations G01N33/5094; chemical analysis of blood groups or blood types G01N33/80

Abstract

A method of measuring a glycosylated protein proportion by an AC impedance method includes passing an AC voltage or AC to two inspecting electrodes; covering a solution containing glycosylated protein completely onto the inspecting electrodes; measuring an impedance value between the electrodes and calculating the difference between the impedance value and an impedance value of a solution without glycosylated protein by an AC impedance inspection device; comparing the impedance difference with AC impedance difference/glycosylated protein concentration to obtain the proportion of the glycosylated protein in the solution containing glycosylated protein. The solution without glycosylated protein includes a salt solution and a non-glycosylated protein; the proportion of glycosylated protein in the solution containing glycosylated protein refers to the proportion of glycosylated protein in the total quantity of protein in the salt solution; and the proportion of glycosylated hemoglobin of the total quantity of hemoglobin in the salt solution.

Claims

1. A method of measuring a glycosylated protein proportion by an AC impedance method, comprising the steps of:

passing an alternating current (AC) voltage or an alternating current (AC) to a pair of inspecting electrodes, and covering the inspecting electrodes completely by a solution containing a glycosylated protein, and obtaining an impedance value between the electrodes through an AC impedance inspection device;

calculating the impedance difference between the impedance value and an impedance value of a solution without the glycosylated protein;

and comparing the impedance difference with a relation of AC impedance difference/glycosylated protein concentration to obtain the proportion of the glycosylated protein in a solution containing the glycosylated protein.

2. The method of measuring a glycosylated protein proportion by an AC impedance method according to claim 1, wherein the relation of AC impedance difference/glycosylated protein concentration is created by obtaining impedance values of a solution A without glycosylated protein and a solution B without glycosylated protein by the AC impedance inspection device after glycosylated proteins in different concentrations are dissolved therein, and obtaining, an impedance difference between the solutions A and B and a curve of the impedance difference after glycosylated proteins of different concentrations are dissolved in the solution without glycosylated protein, and using the curve to obtain the relation of AC impedance difference/glycosylated protein concentration required for the inspection.

3. The method of measuring a glycosylated protein proportion by an AC impedance method according to claim 1, wherein the step passing an AC voltage or AC to the inspecting electrodes to obtain impedance values of a solution containing glycosylated protein and a solution without glycosylated protein further comprises the step of performing a full-spectrum scan of the AC voltage or AC, and the spectral region falls within a range from 1 Hz to 1 MHz.

4. The method of measuring a glycosylated protein proportion by an AC impedance method according to claim 3, wherein the step of passing an AC voltage or AC to the inspecting electrodes further comprises the step of obtaining an impedance value within the spectral region at a constant frequency band to compare and analyze the glycosylated protein proportion and the impedance difference.

5. The method of measuring a glycosylated protein, proportion by an AC impedance method according to claim 1, wherein the solution without glycosylated protein includes a salt solution and a non-glycosylated protein.

6. The method of measuring a glycosylated protein proportion by an AC impedance method according to claim 5, wherein the salt solution is diluted by deionized water in advance, and the dilution percentage by volume is greater than 0.01% and smaller than 90%.

7. The method of measuring a glycosylated protein proportion by an AC impedance method according to claim 5, wherein the total quantity of the glycosylated protein and the non-glycosylated protein are defined as the total quantity of protein, and the proportion of the glycosylated protein in a solution containing glycosylated protein refers the proportion of the glycosylated protein in the total quantity of protein contained in the salt solution.

8. The method of measuring a glycosylated protein proportion by an AC impedance method according to claim 7, wherein the glycosylated protein is glycosylated hemoglobin, and the non-glycosylated protein is non-glycosylated hemoglobin, and the total quantity of the glycosylated hemoglobin and the non-glycosylated hemoglobin is defined as total quantity of hemoglobin, and the proportion of the glycosylated protein in the total quantity of protein refers to the proportion of the glycosylated hemoglobin in the total quantity of hemoglobin contained in the salt solution.

9. The method of measuring a glycosylated protein proportion by an AC impedance method according to claim 8, wherein the total quantity of hemoglobin contained in the salt solution has a concentration controlled to be greater than 0.001 mg/ml and smaller than 1 mg/ml.

10. The method of measuring a glycosylated protein proportion by an AC impedance method according to claim 9, wherein the proportion of glycosylated hemoglobin in the total quantity of hemoglobin falls within a range from 0% to 10%.

11. The method of measuring a glycosylated protein proportion by an AC impedance method according to claim 1, wherein the glycosylated protein is glycosylated hemoglobin, and the solution containing glycosylated protein is a solution containing glycosylated hemoglobin, and the solution without glycosylated protein includes a salt solution and a non-glycosylated hemoglobin, and the total quantity of the glycosylated hemoglobin and the non-glycosylated hemoglobin is defined as the total quantity of hemoglobin, and the proportoin of the glycosylated protein in a solution containing glycosylated protein refers to the proportion of the glycosylated hemoglobin in the total quantity of hemoglobin contained in the salt solution.

12. The method of measuring a glycosylated protein proportion by an AC impedance method according to claim 11, wherein the proportion of glycosylated hemoglobin in the total quantity of hemoglobin falls within a range from 0% to 10%.

13. The method of measuring a glycosylated protein proportion by an AC impedance method according to claim 11, wherein the relation of AC impedance difference/glycosylated protein concentration is a relation of AC impedance difference/glycosylated hemoglobin concentration created by obtaining impedance values of a solution A without glycosylated hemoglobin and a solution B without glycosylated hemoglobin by the AC impedance inspection device after glycosylated hemoglobins in different concentrations are dissolved therein, and obtaining an impedance difference between the solutions A and B and a curve of the impedance difference after glycosylated hemoglobins of different concentrations are dissolved in the solution without glycosylated hemoglobin, and using the curve to obtain the relation of AC impedance difference/glycosylated hemoglobin concentration required for the inspection.

14. The method of measuring a glycosylated protein proportion by an AC impedance method according to claim 11, wherein the step of passing an AC voltage or AC to the inspecting electrodes to obtain impedance values of a solution containing glycosylated hemoglobin and a solution without any glycosylated hemoglobin further comprises the step of performing a full-spectrum scan of the AC voltage or AC, and the spectral region falls within a range from 1 Hz to 1 MHz.