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Protein chromatographic separation platform and application thereof

A chromatographic separation and protein technology, applied in material separation, analytical materials, measurement devices, etc., can solve the problem of high sample complexity, and achieve the effect of increasing the number of identifications, improving the reproducibility, and improving the accuracy of quantitative analysis.

Active Publication Date: 2017-05-31
深圳深维超生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the complexity of the sample after 2D separation is still high

Method used

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  • Protein chromatographic separation platform and application thereof
  • Protein chromatographic separation platform and application thereof
  • Protein chromatographic separation platform and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0048] As shown in Figure 1 (A), a protein chromatographic separation platform includes a proteome reactor 1 and a liquid chromatography-mass spectrometer 2 (Orbitrap Fusion, Thermo Fisher Scientific, USA); wherein, the proteome reactor 1 includes a standard 200μL tip 3, strong anion exchange membrane 4 (3M Empore, USA) and C 18 Film 5 (3M Empore, USA); C 18 The membrane 5 is filled in the lower end of the pipette tip 3, and the strong anion exchange membrane 4 is filled in the lower end of the pipette tip 3 and is located at C 18 on membrane 5.

[0049] As shown in Figure 1(B), during operation, place the support block 6 on the upper end of the 1.5mL collection tube 7, place the proteome reactor 1 on the collection tube 7 through the support block 6, and put the collection tube 7 into the centrifuge In 8, the protein solution or reagent can flow through the proteome reactor through centrifugation to complete the pre-enrichment of protein, enzymatic hydrolysis, desalination ...

Embodiment 2

[0055] As shown in Figure 1 (A), a protein chromatographic separation platform includes a proteome reactor 1 and a liquid chromatography-mass spectrometer 2 (Orbitrap Fusion, Thermo Fisher Scientific, USA); wherein, the proteome reactor 1 includes a standard 200 μL pipette tip 3, strong anion exchange membrane 4 (3M Empore, USA) and C18 membrane 5 (3M Empore, USA); C18 membrane 5 is filled in the lower end of pipette tip 3, and strong anion exchange membrane 4 is filled in The lower end of the tip 3 of the liquid gun is located on the C18 film 5 .

[0056] As shown in Figure 1(B), during operation, place the support block 6 on the upper end of the 1.5mL collection tube 7, place the proteome reactor 1 on the collection tube 7 through the support block 6, and put the collection tube 7 into the centrifuge In 8, the protein solution or reagent can flow through the proteome reactor through centrifugation to complete the pre-enrichment of protein, enzymatic hydrolysis, and strong an...

Embodiment 3

[0062]As shown in Figure 1 (A), a protein chromatographic separation platform includes a proteome reactor 1 and a liquid chromatography-mass spectrometer 2 (Orbitrap Fusion, Thermo Fisher Scientific, USA); wherein, the proteome reactor 1 includes a standard 200μL tip 3, strong anion exchange membrane 4 (3M Empore, USA) and C 18 Film 5 (3M Empore, USA); C 18 The membrane 5 is filled in the lower end of the pipette tip 3, and the strong anion exchange membrane 4 is filled in the lower end of the pipette tip 3 and is located at C 18 on membrane 5.

[0063] As shown in Figure 1(B), during operation, place the support block 6 on the upper end of the 1.5mL collection tube 7, place the proteome reactor 1 on the collection tube 7 through the support block 6, and put the collection tube 7 into the centrifuge In 8, the protein solution or reagent can flow through the proteome reactor through centrifugation to complete the operations of protein pre-enrichment, enzymatic hydrolysis, str...

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Abstract

The invention relates to a protein chromatographic separation platform and application thereof. The platform comprises a proteome reactor (1) and a liquid phase chromatography-mass spectrometer (2), wherein the proteome reactor (1) comprises a pipette head (3), strong negative-ion exchange resin (4) and a solid-phase extraction membrane (5); the lower end of the pipette head (3) is filled with the solid-phase extraction membrane (5); the strong negative-ion exchange resin (4) is filled at the lower end of the pipette head (3) and is positioned on the solid-phase extraction membrane (5). The protein chromatographic separation platform provided by the invention integrates pre-concentration, reduction, alkylation, enzymolysis, strong negative ion exchange and grading of polypeptides, high pH value reverse phase grading and low pH value liquid-phase chromatography separation, has one-dimensional separation, two-dimensional separation and three-dimensional separation working modes, and can be applied to large-scale authentication and quantitative proteomics of proteins in a small amount of cells, tissues or blood samples.

Description

technical field [0001] The invention relates to the technical field of qualitative and quantitative proteomics, in particular to a protein chromatographic separation platform and its application, in particular to a protein sample pretreatment and three-dimensional chromatographic separation platform and its application. Background technique [0002] In proteomics research, the protein in the sample is enzymatically hydrolyzed into polypeptides, and then used for liquid chromatography-mass spectrometry analysis to obtain protein information. This separation and identification method is currently the most widely used proteomics research method. [0003] The composition of protein samples is complex, and pre-fractionation of samples before liquid chromatography-mass spectrometry analysis is an effective method to reduce sample complexity and increase the number of protein identifications. Two-dimensional chromatographic separation is commonly used, including strong cation excha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/06
CPCG01N30/06
Inventor 田瑞军陈文东
Owner 深圳深维超生物科技有限公司
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