Mass spectrometry detecting method for serum polypeptides

A serum peptide and mass spectrometry detection technology, applied in the field of mass spectrometry detection of serum peptidomics, can solve the problems of few reports in the field of peptidomics

Inactive Publication Date: 2019-05-07
SOUTH UNIVERSITY OF SCIENCE AND TECHNOLOGY OF CHINA
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

At present, many DIA-based mass spectrometry detection methods have been developed in the field of proteomics and metabolomics, but there are few reports in the field of peptidomics.

Method used

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  • Mass spectrometry detecting method for serum polypeptides
  • Mass spectrometry detecting method for serum polypeptides
  • Mass spectrometry detecting method for serum polypeptides

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Experimental program
Comparison scheme
Effect test

Embodiment

[0062] sample preparation

[0063] Take 40μL of serum in a 1.5mL centrifuge tube, add 200μL of 25mM ammonium bicarbonate solution containing 20% ​​acetonitrile for dilution, vortex for 10s to break the adsorption between protein and polypeptide, then add 480μL of acetonitrile, vortex for 10s , let it stand for 10 minutes to precipitate high-abundance proteins in serum. The sample was centrifuged at 4C°C, 000×g for 10min, the supernatant was absorbed, and freeze-dried. The freeze-dried sample was reconstituted with 100 μL aqueous solution containing 1% formic acid, and the reconstituted solution was desalted with a C18 desalting column. The final sample was redissolved with 18 μL of an aqueous solution containing 0.1% formic acid, and 2 μL of a polypeptide retention time correction standard (IRT) was added for testing.

[0064] sample analysis

[0065] In order to investigate the performance of the method, the same serum sample was collected three times using the traditional ...

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Abstract

The invention provides a mass spectrometry detecting method for a serum polypeptides, comprising sample preparation, data-independent mass spectrometry acquisition, and data processing. According to the mass spectrometry detecting method for the serum polypeptides, a data-independent mass spectrometry acquisition method for serum peptidomics samples is developed, and the parent ion distribution ofthe sample by pre-analytical experiment on serum samples is obtained, thereby obtaining an optimal variable isolation window. Each condition and each step cooperate with each other by optimizing thepreparation method of the polypeptideomics sample, the mass spectrometry acquisition method, and the data processing method, and finally, the number of the detection of the polypeptide, the reproducibility of the identification, and the quantitative accuracy are significantly improved. The method of the invention has the characteristics of high throughput, high reproducibility, and high accuracy,and is suitable for analysis of large-scale clinical serum samples and detection of disease markers, thereby having wide application prospects and great market value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a mass spectrometry detection method of serum peptidomics. Background technique [0002] Peptidomics is a science that studies the composition, structure, function, and changes of endogenous polypeptides and low-molecular-weight proteins in the body. It is considered to be a bridge between proteomics and metabolomics. As an emerging subject, recent studies have shown that endogenous peptides have great potential as drugs themselves, drug targets, and disease markers. Serum, as an important clinical fluid, contains a wealth of disease marker information. Peptides with small molecular weight can be freely secreted into the extracellular space, penetrate the adventitia of blood vessels, and enter the blood circulation. Research can study tumors from a new perspective, find tumor markers, and provide an important basis for early diagnosis, prognosis and dynamic monitoring of tumors. [0...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
Inventor 林琳
Owner SOUTH UNIVERSITY OF SCIENCE AND TECHNOLOGY OF CHINA
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