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Method for detecting apoptosis degree of cells in solution to be tested

A solution and cell technology, applied in the field of electrochemical detection, can solve the problem of expensive reagents, avoid the use of expensive reagents, overcome the dependence on expensive instruments, and reduce the cost of detection

Inactive Publication Date: 2014-09-17
SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method relies on large-scale instruments, and the reagents used are very expensive, so it is still necessary to develop some easy-to-operate and low-cost detection methods

Method used

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  • Method for detecting apoptosis degree of cells in solution to be tested
  • Method for detecting apoptosis degree of cells in solution to be tested
  • Method for detecting apoptosis degree of cells in solution to be tested

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0025] The space-occupying molecule used in this example is mercaptohexanol, the cell used is MCF-7, and the working electrode used is a gold electrode.

[0026] 1.1. Preparation of peptide-modified electrodes

[0027] a. Grinding the gold electrode, the steps include grinding with sandpaper, grinding with aluminum oxide slurry of different particle sizes, ultrasonication with ethanol / water, soaking in 50% nitric acid, electrochemical cleaning with 0.5M sulfuric acid, etc.

[0028] b. Soak the gold electrode in 0.1mM peptide solution, react overnight, and then wash the electrode with deionized water.

[0029] c. Then soak the electrode in 0.1mM mercaptohexanol, react for 30min, and then clean the electrode with deionized water.

[0030] 1.2. Cell culture

[0031] MCF-7 cells were cultured in DMEM medium (10% serum) at a temperature of 37°C and a carbon dioxide concentration of 5%. When the cells were cultured to the exponential phase, the cells were digested and washed for l...

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Abstract

The invention discloses a method for detecting an apoptosis degree of cells in a solution to be tested. The method comprises the steps of designing and synthesizing one section of polypeptide sequence, firmly modifying the polypeptide sequence on the surface of a working electrode, orderly arranging the polypeptide sequence through space occupying molecules, identifying the property of phosphatidylserine on surfaces of apoptotic cells in the solution to be tested through the polypeptide specificity, and fixing the apoptotic cells on the surface of the electrode. Because one layer of apoptotic cells is covered on the surface of the electrode, the electrochemical response of the working electrode is correspondingly changed; through comparing degrees of electrochemical signal changes before and after detection, the apoptosis degree of the cells in a sample to be tested can be detected. The method prevents from using the specific binding of an antigen and an antibody, thus the detection cost can be effectively lowered.

Description

technical field [0001] The invention relates to the technical field of electrochemical detection, in particular to an electrochemical detection method of cell apoptosis based on a polypeptide recognition system. Background technique [0002] Cell death can be divided into necrosis and apoptosis according to the mechanism, inducement, scope of death, morphological characteristics, and biochemical characteristics. Apoptosis is a manifestation of programmed cell death, which induces the pre-existing death program in cells by in vivo and external factors, leading to active cell death, which is different from necrosis in terms of morphology and biochemical characteristics, and the process is controlled by genes. Spontaneous and orderly death will not cause the body's inflammatory response, and can maintain the stability of the internal environment of the human body. It plays a very important role in the normal growth and development of the entire organism, the regulation of the i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26G01N27/327
Inventor 缪鹏王弼陡殷建韩坤唐玉国
Owner SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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