Sample pretreatment method for multi-omics analysis, and application thereof

A sample preparation and sample technology, applied in the field of sample preparation for multi-omics analysis, can solve the problems of high transportation and storage costs, high pollution, differences, etc., and achieve good reproducibility, high sensitivity, and high accuracy. sexual effect

Active Publication Date: 2020-10-13
SOUTH UNIVERSITY OF SCIENCE AND TECHNOLOGY OF CHINA
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  • Application Information

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Problems solved by technology

[0006] In the current analysis technology, milliliter-level plasma samples are collected in the hospital and transported to the mass spectrometry laboratory for proteomics and metabolomics analysis, but the liquid chromatography-mass spectrometry analysis procedures of these two omics are different, so Separate analysis is often required, which inevitably leads to discrepancies in the results of the two analyses.
In addition, the storage stability of liquid plasma samples is poor, and the cost of transportation and storage is high, which brings certain limitations to subsequent omics analysis; while dry plasma samples are generally collected through blood collection paper, such dry samples have There is a high risk of contamination, and there are also problems such as low-abundance proteins and metabolites that are difficult to analyze and detect

Method used

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  • Sample pretreatment method for multi-omics analysis, and application thereof
  • Sample pretreatment method for multi-omics analysis, and application thereof
  • Sample pretreatment method for multi-omics analysis, and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0090] A sample pretreatment method for multi-omics analysis, specifically comprising the following steps:

[0091] (1) Preparation and sample loading of the sample pretreatment device:

[0092] The structural schematic diagram of the sample pretreatment device is as follows: figure 1 As shown, among them, 1 is the tip of the pipette, 2 is the solid phase extraction membrane, 3 is the C18 resin, and 4 is the ion exchange resin; the preparation method is as follows: 3 pieces of C18 membranes are sequentially loaded into the 200 μL pipette tip (U.S. 3M Empore), 0.5mg C18 resin particles (particle diameter 20 μm, Dr.Maisch GmbH, Germany), 2 mg ion exchange resin particles (the mass ratio of SCX / SAX is a mixed ion exchange resin of 1:1, and the particle diameter is 20 μm , U.S. AppliedBiosystems), to obtain the sample pretreatment device;

[0093] Use 80 μL of methanol and 40 μL of 10 mM ammonium bicarbonate solution (pH=7.4) to wash, activate and equilibrate the materials fille...

Embodiment 2

[0107] The difference between this example and Example 1 is that the filling amount of ion exchange resin particles in the sample pretreatment device is 1 mg; other components of the sample pretreatment device, sample pretreatment method, mass spectrometry detection method and mass spectrometry data analysis method All the same as in Example 1.

Embodiment 3

[0109] The difference between this example and Example 1 is that the filling amount of ion exchange resin particles in the sample pretreatment device is 3 mg; other components of the sample pretreatment device, sample pretreatment method, mass spectrometry detection method and mass spectrometry data analysis method All the same as in Example 1.

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Abstract

The invention provides a sample pretreatment method for multi-omics analysis, and application thereof. The sample pretreatment method comprises the following steps: (1) adding a to-be-detected sampleto a sample pretreatment device, centrifuging, and loading the obtained sample to obtain a loaded sample; (2) eluting the sample to obtain an eluate and an eluted sample, and collecting the eluate toobtain a metabonomics analysis sample; and (3) carrying out enzymolysis on the eluted sample, transferring the sample to a solid-phase extraction membrane, eluting, and collecting the obtained eluateto obtain a proteomics analysis sample. According to the sample pretreatment method, single-drop blood collection, dry sample transportation, enrichment of a to-be-detected object and preparation of the multi-omics analysis sample can be integrally carried out, and the same liquid phase mass spectrometry device is adopted to carry out mass spectrometry on a plurality of omics samples from a singledrop of plasma, such that the process from sample pretreatment to analysis result obtaining can be controlled within 6 h so as to achieve rapid, high-sensitivity and high-accuracy analysis detection.

Description

technical field [0001] The invention belongs to the technical field of biological analysis and detection, and in particular relates to a sample pretreatment method for multi-omics analysis and an application thereof. Background technique [0002] Mass spectrometry (MS) is a spectroscopic analysis method with high sensitivity, strong specificity and fast analysis speed. Mass spectrometry can obtain rich structural information in one analysis. Combining mass spectrometry with different separation techniques is an analytical science A breakthrough in the method, especially the combination of liquid chromatography and mass spectrometry, has become a reliable means of separating and identifying the composition and structure of complex mixtures, and has great significance in many fields such as biomedicine, public safety and environmental monitoring. Wide range of applications. In the field of biomedicine, omics research methods based on mass spectrometry technology provide power...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/06G01N30/08G01N30/88
CPCG01N30/06G01N30/08G01N30/88G01N2030/027G01N2030/062G01N2030/8822G01N2030/8831
Inventor 田瑞军高伟娜张巧云苏怡然
Owner SOUTH UNIVERSITY OF SCIENCE AND TECHNOLOGY OF CHINA
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