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Kit for detecting low-abundance and low-molecular-weight protein spectrum

A low molecular weight, protein profiling technology, applied in the field of kits for the direct detection of low abundance and low molecular weight protein profiles in complex samples, can solve the problems of interference affecting mass spectrometry identification, masking, and complicated steps.

Active Publication Date: 2014-05-14
HANGZHOU WELL HEALTHCARE TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has cumbersome steps, multi-step separation and elution operations may lose trace amounts of low-molecular-weight proteins, and requires quite large detection equipment
[0004] At present, the main difficulties in the analysis of low-abundance and low-molecular-weight proteins are: 1) easy to be lost and degraded by other high-molecular-weight proteases during the separation process; 2) easy to be covered by information of other high-molecular-weight proteins during mass spectrometry identification; 3 ) are more susceptible to the interference of small molecules such as salts and surfactants, which affect mass spectrometry identification, etc.

Method used

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  • Kit for detecting low-abundance and low-molecular-weight protein spectrum
  • Kit for detecting low-abundance and low-molecular-weight protein spectrum
  • Kit for detecting low-abundance and low-molecular-weight protein spectrum

Examples

Experimental program
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Effect test

Embodiment 1

[0021] Example 1 Preparation of Porous Silicon

[0022] Take the P-type boron-doped silicon chip (100) crystal form, fix it in the electrolytic cell, add 0.5ml ethanol, 2-3ml hydrofluoric acid with a mass concentration of 40% as the electrolyte, use the silicon chip as the anode, and the platinum electrode as the cathode , carry out DC electrolytic etching and stripping, set the current intensity at 10~80mA, and the etching time is 5 minutes. After etching and stripping the porous silicon layer, rinse it with ethanol and ultrasonically pulverize it for 15 minutes, and then dry it with nitrogen. , the pore diameter of the obtained porous silicon particles is 5-20nm;

[0023] The porous silicon particles were subjected to ozone oxidation treatment for 20 minutes to form surface silicon-oxygen bonds;

[0024] Soak the oxidized porous silicon particles in a mixed solution of aminosilylating reagent and ethanol solvent with a volume concentration of 1% to react for 1 hour, then dr...

Embodiment 2

[0026] Embodiment 2 Separation device 1

[0027] see figure 1 , is an embodiment of a separation device of the present invention. In the figure, 1 is the porous silicon particle, and 2 is the microcolumn. The porous silicon particle is placed in the microcolumn to realize the separation and enrichment of specific proteins.

Embodiment 3

[0028] Embodiment three Separation device two

[0029] see figure 2 , is another embodiment of the separation device of the present invention. In the figure, 1 is the porous silicon particle, and 3 is the centrifuge tube. The porous silicon particle is placed in the centrifuge tube to achieve the separation and enrichment of specific proteins.

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Abstract

The invention discloses a kit for detecting low-abundance and low-molecular-weight protein spectrums. The kit comprises a kit box; a separator, a protein separation and enrichment material, a diluting buffer solution and a cleaning buffer solution are arranged in the kit body; and the protein separation and enrichment material is disposed in the separator and are porous silicon particles, the channel diameter of the material is 1-50nm, and the channel surface of the material has a chemically modified group which is an anion exchange group, a cation exchange group, a hydrophobic group or a polar group. The kit reaches an effect of one-step separation, enrichment and detection of low-molecular-weight proteins in a serum sample through preparing the porous silicon particles in the invention. The kit disclosed in he invention has the advantages of simplicity, portability, and accurate and fast detection method.

Description

technical field [0001] The invention belongs to the technical field of chemical determination, in particular to a kit for directly detecting low-abundance and low-molecular-weight protein profiles in complex samples. Background technique [0002] In recent years, with the introduction of the concept of population proteomics and its application in practical research, the distance between experimental research and clinical application will be further shortened. Since changes in the content and type of proteins in human body fluids caused by diseases are important indicators for the early diagnosis and evaluation of various diseases in clinical practice, the application of modern biotechnology to find specific protein changes related to diseases is a current research hotspot. However, so far there are very few biomarkers that can be used for routine clinical detection. Theoretically, because of the large molecular weight of protein, it is difficult to enter the body fluid circ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/64G01N1/40
Inventor 邬建敏谈洁赵伟洁
Owner HANGZHOU WELL HEALTHCARE TECH CO LTD
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