Electrochemical immune sensor for phosphating protein

A phosphoprotein, electrochemical technology, applied in the direction of material electrochemical variables, scientific instruments, instruments, etc., can solve the problems of unfavorable on-site detection of grass-roots use and popularization, to avoid sample processing process, good reproducibility, good selection sexual effect

Inactive Publication Date: 2011-11-23
HUAZHONG NORMAL UNIV
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  • Abstract
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Problems solved by technology

Although the method is accurate and can achieve high-throughput detection, it includes multiple incubation and washing steps, and the subsequent spectrophotometric detection also requires a special substrate color reagent, and requires expensive instruments, professional laboratories and Well-trained professional and technical personnel are not conducive to on-site testing and the promotion and popularization of grassroots use

Method used

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  • Electrochemical immune sensor for phosphating protein
  • Electrochemical immune sensor for phosphating protein
  • Electrochemical immune sensor for phosphating protein

Examples

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Embodiment 1

[0029] Example 1 Phospho-p53 protein 15 electrochemical immunoassay

[0030] Add 10 μL of 1.0 M hydrochloric acid to the centrifuge tube to make the Pb 2+ Released from the inner cavity of LPA; then add 50 μL of acetic acid buffer solution containing 0.5 mg / L Bi, the concentration of acetic acid buffer solution is 0.2M, pH is 4.6, after mixing for 2 minutes, the magnetic nanoparticles (MPs) are magnetically separated; 50 μL of supernatant was dropped on the surface of the printed electrode, enriched at -0.9V for 2 minutes, and then recorded the SWV curve from -0.9-0.3V, the potential increment was 4mV, the amplitude was 25mV, and the frequency was 15Hz; the lead ion was obtained at -0.56V Stripping voltammetry peak. By detecting Pb encapsulated in lead phosphate-apoferritin (LPA) 2+ Determination of the anodic dissolution signal of the phosphoprotein phospho-p53 in samples 15 content. The experimental results are as image 3 .

Embodiment 2

[0032] Add 25 μL of phospho-p53 with different concentrations (0.02, 0.05, 0.1, 0.2, 0.5, 1.0, 2.0, 5.0, 10, 20 ng / mL) into ten 1.5 mL plastic centrifuge tubes 15 . Formation of LPA-p53 after immune reaction 15 Ab 2 -CNS / phospho-p53 15 / MPs-Ab 1 immune complexes, and then performed electrochemical immunoassays, the experimental results are as follows Figure 4 .

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Abstract

The invention relates to an electrochemical immune sensor for phosphating protein. In the electrochemical immune sensor, a magnetic nanometer-antibody composition MPs-p53<15>Ab1 is used as an adsorbent to separate the phosphating protein, a carbon nanometer sphere CNS is used as a carrier, lead phosphate-apoferritin (LPA) and a phosphating protein antibody p5315Ab2 are modified on the surface of the CNS, and thus, LPA-p53<15>Ab2-CNS nanometer complexing agent is prepared for detecting signal amplification. According to the sandwich immunoassay principle, the content of the phosphating protein to be detected is in proportion to the captured LPA-p5315Ab2-CNS, and the content of lead ions encapsulated in LPA is detected according to a dissolving volt-ampere method to measure the concentration of the phosphating protein in a sample. The method has the advantages of simplicity and convenience for operation and simple and efficient separation, and the advantage that an integral immunologic reaction is finished in an epoxy (EP) tube; a large number of LPA signal molecules are introduced by taking the CAN as the carrier, and each LPA molecule contains a large number of Pb<2+>, so the sensitivity of detection is improved; and the dissolved Pb<2+> is used as a detection signal without adding an enzyme substrate, so the electrochemical immune sensor is a reagent-free sensor. In the electrochemical immune sensor for the phosphating protein, the linear concentration range of the phosphating protein phosphor-p53<15> is from 0.02 to 20 ng mL<-1>, and the detection limit is 0.01 ng mL<-1>.

Description

technical field [0001] The present invention relates to LPA-p53 15 Ab 2 -Preparation method of CNS nanocomposite material, and determination of phospho-p53 in samples by combining highly sensitive anodic stripping voltammetry and nanocarrier amplification technology 15 concentration. Background technique [0002] P53 protein is the most important molecule in regulating cell growth in the human body and monitoring DNA replication during cell division. It is an important tumor suppressor gene, which can prevent cancer and help cells to repair defects in genes. However, when the human body is exposed to ultraviolet light or chemical carcinogens, p53 cannot perform the above functions, and loses control over cell proliferation, often leading to cancerous cells. It is reported that in more than half of human cancer cells, the tumor suppressor p53 protein is damaged in different forms and degrees. Human p53 protein is composed of 393 amino acid residues and has 4 structural do...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543G01N27/48
Inventor 杜丹
Owner HUAZHONG NORMAL UNIV
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