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High-flux protein titration method based on moving reaction boundary electrophoresis

A technology of interfacial electrophoresis and mobile reaction, which is applied in the field of high-throughput protein titration, and can solve the problems of long time required for determination, low throughput, and the range of ionic strength in the range of applied voltage is not given.

Inactive Publication Date: 2013-06-26
SHANGHAI JIAO TONG UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, this prior art does not elaborate on the functional relationship between MRB moving speed and protein concentration, and does not give the range of applied voltage and the range of ionic strength of the background electrolyte solution. In addition, the titration device involved can only perform single-tube detection. The throughput is low, the operation is more complicated, and the measurement takes a long time

Method used

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  • High-flux protein titration method based on moving reaction boundary electrophoresis
  • High-flux protein titration method based on moving reaction boundary electrophoresis
  • High-flux protein titration method based on moving reaction boundary electrophoresis

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Embodiment 1

[0046] Such as figure 1 Shown, the present embodiment adopts alkali solution titration:

[0047] First, inject gel into the electrophoresis tube containing biological samples, so that the protein molecules to be detected are uniformly fixed in the electrophoresis tube through the gel; -300V voltage, making the anion OH in the alkaline solution - Meet with the free acidic residues of the protein and undergo a neutralization reaction to form MRB, and the MRB moves to the anode; during titration, add an indicator to the electrophoresis tube to indicate the position of the MRB at different times for visual observation; after that, The standard curve of MRB moving speed and protein concentration was established by using the functional relationship between MRB moving speed and protein concentration, and the determined protein concentration was calculated based on the obtained MRB moving speed.

[0048] The functional relationship between the moving speed of the MRB and the protein...

Embodiment 2

[0071] In this embodiment, the alkali titration in Example 1 is replaced by acid titration, that is, the acid solution injected into the anolyte tank is applied with a voltage range of 100-300V, so that the cation H in the acid solution + It meets the free basic residues of the protein and undergoes a neutralization reaction to form an MRB. The MRB moves from the anode to the cathode. According to the functional relationship between the protein concentration and the distance moved by the interface in different biological samples in the same acid solution at the same time, the protein is drawn. For the standard curve of the concentration, the unknown concentration of the protein is calculated on the standard curve according to the moving distance of the MRB formed by the unknown concentration of the protein and the acid per unit time.

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Abstract

The invention relates to a moving reaction boundary electrophoresis based on high-flux protein titration method of an electrochemical method analyzing material. The high-flux protein titration method comprises the following steps of: uniformly fixing protein molecules to be detected into an electrophoretic tube through gel by injecting the gel into the electrophoretic tube with a biological sample contained; then carrying out alkali titration; and finally establishing a standard curve of MRB (Moving Reaction Boundary) movement speed and protein concentration by utilizing the functional relation of the MRB movement speed and the protein concentration to obtain the MRB movement speed, and calculating to obtain the definite protein concentration. The high-flux protein titration method disclosed by the invention can be used for fast detecting the total protein quantity and is high in anti-interference capacity.

Description

technical field [0001] The invention relates to a method for analyzing materials by an electrochemical method, in particular to a high-throughput protein titration method based on moving reaction boundary (MRB) electrophoresis for detecting protein content in biological samples. Background technique [0002] Protein is one of the main nutrients in foods such as milk. Detecting the total protein content in food is of great significance for measuring the nutritional value of food, determining the price of commodities, and evaluating the quality of food materials (Moore, J.C., DeVries, J.W., Lipp, M., Griffiths, J.C., Abernethy, D.R., 2010, Comper. Rev. Food Sci. Food Saf., 9, 330-357.). And protein content analysis has an important application in clinical medical diagnosis (Kangxi Xiong, clinical electrophoresis, People's Health Publishing House, 2006). The Kjeldahl method has become one of the standard methods for protein content in various biological samples (Kjeldahl, J.Z...

Claims

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Application Information

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IPC IPC(8): G01N27/44
Inventor 曹成喜王后禹郭成叶颜健唐芸芸尹晓阳樊柳荫
Owner SHANGHAI JIAO TONG UNIV
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