Concentration and demineralization purification treatment method of biological samples

A biological sample purification treatment technology, applied in the field of biochemical analysis, can solve problems such as analyte loss

Inactive Publication Date: 2012-06-27
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method requires a large amount of matrix solution to drain salt, which will lead to the loss of so

Method used

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  • Concentration and demineralization purification treatment method of biological samples
  • Concentration and demineralization purification treatment method of biological samples
  • Concentration and demineralization purification treatment method of biological samples

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Single crystal silicon [n type, (100)] was treated with oxygen plasma at a power of 100 W for 3 minutes to make the surface of the silicon wafer hydrophilic. Then use high-purity water to ultrasonically clean the surface of the substrate three times, each time for 4 minutes, so that the surface is thoroughly cleaned and hydroxylated.

[0047] Take 0.1 μL of polymethyl methacrylate (PMMA, Sigma-Aldrich) and drop-coat it on the surface of the treated silicon wafer to form a polymer barrier layer with a thickness of about 60 μm and a radius of 800 μm. The silicon surface substrate (hydrophilized) was attached to a clean upper surface dish, and 1 μL of fluorine-containing silicon coupling reagent (heptadecafluorodecyltrimethoxysilane, Sigma-Aldrich) was dropped on the surface with a pipette gun. The lower surface plate is fastened to the upper surface plate, and heated and grown at 250°C for 3 hours, so that fluorine-containing silane is grown on the entire surface except f...

Embodiment 2

[0052] The glass sheet was treated with oxygen plasma at a power of 100 W for 3 minutes to make the surface of the glass sheet hydrophilic. Then use high-purity water to ultrasonically clean the surface of the substrate three times, each time for 4 minutes, so that the surface is thoroughly cleaned and hydroxylated.

[0053]0.1 μL of polymethyl methacrylate (PMMA, Sigma-Aldrich) was drop-coated on the surface of the treated glass slide to form a polymer barrier layer with a thickness of about 60 μm and a radius of 800 μm.

[0054] Subsequently, the glass surface substrate (hydrophilized) containing the baffle was attached to a clean upper surface dish, and 1 μL of fluorine-containing silicon coupling reagent (heptadecafluorodecyltrimethoxysilane, Sigma -Aldrich) was dropped on the lower surface dish, buckled the upper surface dish, and heated and grown at 250°C for 3 hours, so that fluorine-containing silane was grown on the entire surface except for the PMMA coverage area, wh...

Embodiment 3

[0058] The quartz plate was treated with oxygen plasma at a power of 100W for 3 minutes to make the surface of the quartz plate hydrophilic. Then use high-purity water to ultrasonically clean the surface of the substrate three times, each time for 4 minutes, so that the surface is thoroughly cleaned and hydroxylated.

[0059] 0.1 μL of polymethyl methacrylate (PMMA, Sigma-Aldrich) was drop-coated on the surface of the treated quartz plate to form a polymer barrier layer with a thickness of about 60 μm and a radius of 800 μm.

[0060] Subsequently, the quartz surface substrate (hydrophilized) containing the baffle was attached to a clean upper surface dish, and 1 μL of fluorine-containing silicon coupling reagent (heptadecafluorodecyltrimethoxysilane, Sigma -Aldrich) was dropped on the lower surface dish, buckled the upper surface dish, and heated and grown at 250°C for 3 hours, so that fluorine-containing silane was grown on the entire surface except for the PMMA coverage area...

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Abstract

The invention belongs to the field of biochemistry analysis technology, and specifically relates to a one-step concentration and demineralization purification treatment method of biological samples by the use of patterning surface. The patterning surface can be widely applied in pretreatment and detection of biological samples. The method comprises the following steps of: constructing sealing patterning surface of hydrophobic-hydrophilic-hydrophobic region on substrate; dropping a polypeptide or protein solution onto the patterning surface prepared by the above step, naturally drying at room temperature to obtain dried sample points; dropping a substrate solution onto the above dried sample points, and naturally drying at room temperature and forming uniform cocrystallization of the sample and substrates on the patterning surface, so as to realize the concentration and demineralization purification treatment of biological samples. By the use of the patterning surface constructed in the invention, in MALDI-TOF MS detection, signal noise ratio, sensitivity and salt tolerance are significantly improved. The method has a great practical application value.

Description

technical field [0001] The invention belongs to the technical field of biochemical analysis, and specifically relates to a treatment method for one-step enrichment and desalination purification of biological samples by using a patterned surface. The patterned surface can be widely used in the pretreatment and detection of biological samples. Background technique [0002] Matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an effective soft ionization method with high sensitivity for the analysis and detection of biological samples in proteomics. However, in the process of mass spectrometry, the detection sensitivity and reproducibility of mass spectrometry will be reduced due to the uncontrollable sample deposition area and the inhomogeneous co-crystallization of matrix and sample. In addition, salts and other reagents present in biological samples can also seriously interfere with the crystallization of samples and matrices, result...

Claims

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Application Information

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IPC IPC(8): G01N1/40G01N1/34
Inventor 吕男王燕东曾周芳国新华
Owner JILIN UNIV
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