Electrochemical detection method for high concentration denatured bovine serum albumin

A bovine serum albumin and detection method technology, applied in the field of electrochemical detection of high-concentration denatured bovine serum albumin, can solve the problems of troublesome sample processing, high test cost, and complex data analysis, and achieve high cost and low detection limited and cumbersome operation

Active Publication Date: 2019-07-26
SOUTH CHINA NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the common methods for analysis of protein secondary structure and denatured protein secondary structure include X-ray diffraction, circular dichroism, fluorescence spectroscopy, ultraviolet-visible spectroscopy and Fourier transform infrared spectroscopy, etc., but these The method generally has high test costs, complex data analysis, and troublesome sample processing

Method used

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  • Electrochemical detection method for high concentration denatured bovine serum albumin
  • Electrochemical detection method for high concentration denatured bovine serum albumin
  • Electrochemical detection method for high concentration denatured bovine serum albumin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Preparation of different concentrations of denatured BSA solutions containing MB

[0022] (1) Use an acidity meter to prepare 0.02mol L of pH 5.0-9.0 -1 Trishydroxymethylaminomethane / 0.1mol·L - 1 NaCl buffer solution, and prepare 10mmol / L MB solution, 10mol / L urea solution and 1mmol / L BSA solution;

[0023] (2) Use a pipette gun to measure 20 μL of MB solution (10 mmol / L) into the sample tube, add different volumes of BSA solution (1 mmol / L) to the sample tube in turn, and then add 1600 μL to each sample tube in turn Urea solution (10mol / L);

[0024] (3) Shake and mix at room temperature to obtain 2000 μL of mixed solution, which contains 100 μmol / L MB and 8 mol / L urea, and the concentrations of BSA are 0, 5, 10, 20, 50 and 100 μmol / L, respectively. Multiple parallel denatured BSA solutions containing MB with different concentrations were stored in the refrigerator and reacted overnight for 15 hours.

Embodiment 2

[0025] Example 2: Linear sweep voltammetry test of different concentrations of denatured BSA solutions containing MB

[0026] The conductive glass ITO electrode cut in advance is immersed in the solution prepared in Example 1 as the working electrode, the titanium sheet is used as the counter electrode, and the saturated calomel electrode is used as the reference electrode, which are respectively connected to 3 electrode joints of the electrochemical workstation, Carry out linear sweep voltammetry test, measure oxidation peak current density linear change curve with denatured BSA solution concentration;

[0027] The result is as figure 1 As shown, with the increase of denatured BSA concentration, the oxidation peak current density of MB increased significantly. Take the oxidation peak current density as the ordinate and the concentration of denatured BSA as the abscissa to draw a standard curve, i.e. pa =23.17+0.41C 变性BSA , there is a good linear relationship between the ox...

Embodiment 3

[0028] Example 3: Practical application of electrochemical detection method based on high-concentration denatured BSA

[0029] To the practical application after the establishment of the method of the linear sweep voltammetry test of the denatured BSA solution of different concentrations containing MB in Example 2, the specific operation steps are as follows:

[0030] The MB solution containing 100 μmol / L denatured BSA prepared in Example 1 was added to 400 μL milk sample, and the linear sweep voltammetry test was carried out according to the linear change curve of the oxidation peak current density of MB with the concentration of denatured BSA established in Example 2 , to measure denatured BSA in milk samples.

[0031] The result is as Figure 4 As shown, the peak current density difference between milk with 100 μmol / L denatured BSA solution and pure milk is 39.14 μA / cm -2 . And as figure 2 As shown in the standard curve, the difference in peak current density between 1...

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Abstract

The invention belongs to the technical field of sensors and discloses an electrochemical detection method for high concentration denatured bovine serum albumin. The method induces BSA denaturation with urea, simultaneously adds a certain proportion of MB, transfers the MB to a container having a constant area of indium tin oxide (ITO) electrode after reacting for a certain period of time, and performs linear sweep voltammetry test. The oxidation peak current density of MB shows a good linear relationship with the amount of denatured BSA. The method not only has the characteristics of simple operation, low cost and good biocompatibility, but also observes BSA denaturation by adding MB visualization, realizes detection of high concentration denaturing BSA, and has broad application prospectin the field of bio-sensing electrochemical detection.

Description

technical field [0001] The invention belongs to the technical field of sensors, and in particular relates to an electrochemical detection method for high-concentration denatured bovine serum albumin. Background technique [0002] Serum albumin is an important component in blood and has two main functions, one is binding function and the other is carrying function. In the life process of organisms, serum albumin also plays the role of maintaining osmotic pressure, eliminating free radicals and anticoagulation. BSA is highly similar in structure and function to human serum albumin (HSA), so it is usually used as a model protein for research. Only when the protein is in a naturally folded state can it be biologically active. Therefore, when the protein is denatured, that is, the body is under unhealthy conditions, such as liver disease, etc., the binding ability and transport ability of serum albumin will be affected. Therefore, by urea-induced denaturation of BSA, as a model...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/48
CPCG01N27/48
Inventor 颜紫乔张雨薇李红
Owner SOUTH CHINA NORMAL UNIVERSITY
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