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Method for simultanuously enriching desalting and appraising micro protein or polypeptide solution

A trace protein and enrichment technology, applied in the field of biochemical analysis and identification, can solve problems such as limited desalting ability, and achieve the effects of high-efficiency desalination, strong desalination ability and low cost

Inactive Publication Date: 2006-08-23
FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the most commonly used solvent evaporation method will also enrich the impurities such as salt while concentrating the sample, while the traditional desalination method uses the C 18 Packing ZipTip TM Nozzle desalting or on-target elution of samples with deionized water, but both methods have limited desalting capabilities

Method used

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  • Method for simultanuously enriching desalting and appraising micro protein or polypeptide solution
  • Method for simultanuously enriching desalting and appraising micro protein or polypeptide solution
  • Method for simultanuously enriching desalting and appraising micro protein or polypeptide solution

Examples

Experimental program
Comparison scheme
Effect test

example 1-3

[0028] The enrichment and MALDI TOF / MS determination of example 1-3 silicon oxide nanoparticles to protein solution

[0029] Take 1ml of protein sample solution with a certain concentration and add 2μL of 10mg / mL silica nano-suspension respectively, shake at 37°C for 30min; of suspension.

[0030] Take 0.5 μL of the aforementioned suspension and drop it on a MALDI rake plate, then add an equal volume of 5 mg / mL α-CHCA saturated solution (0.1% TFA, 50% acetonitrile aqueous solution), and perform mass spectrometry analysis after the crystallization is dried. The mass spectrometer used is MALDITOF / TOF (4700 Proteomics Analyzer, Applied Biosystems); the laser is Nd-YAG laser, the wavelength is 355nm, the laser pulse frequency is 200Hz; the accelerating voltage is 20kV, and the positive ion mode linear TOF detection.

example 4-5

[0031] Example 4-5 Silica Nanoparticles Anti-salt Enrichment and MALDI TOF / MS Determination of High Salt Protein Samples

[0032] Take 1ml of a certain concentration of protein sample solution containing 8M sodium chloride or 4M urea, add silicon oxide nano-suspension according to the method of Example 1-3, centrifuge, apply the sample, and perform linear TOF detection in positive ion mode.

example 6

[0033] Example 6 Traditional on-board desalting

[0034] First mix the sample and the matrix and spot it on a stainless steel spot plate, and place it in the air to dry naturally. Use a pipette to take 1.5 μL of deionized water containing 0.1% TFA and put it on the sample point. After 3-5 seconds, absorb the solution at the sample point, and repeat this three times.

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Abstract

This invention relates to a method for synchronously enriching and desalting minim albumens or polypeptide solution and verifying them directly, which takes SiO nm particles as the sorbent to adsorb proteins and their enzyme polypeptides efficiently and overcomes interference of high concentration salt.Since SiO nm particles are compatible with MALDI-MS very well and MALDI-TOF / MS analysis can be done directly to the adsorbed samples without elution to avoid the loss.

Description

technical field [0001] The invention belongs to the technical field of biochemical analysis and identification, and specifically relates to a method for synchronously enriching and desalting a trace protein or polypeptide solution and directly identifying it. Background technique [0002] Matrix-assisted laser desorption ionization / mass spectrometry (MALDI / MS) has become an effective tool in proteomics research due to its fast analysis speed, high sensitivity, and easy operation. However, a large amount of salt or other non-protein components (added during sample pre-treatment) in the analyzed sample will greatly reduce the sensitivity of mass spectrometry, which is called the suppression effect in mass spectrometry research. An effective way to avoid the suppression effect is to desalt the sample. In addition, in the process of proteomics research, a large number of proteins that undertake important life activities of organisms are low-abundance proteins, and their extreme...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/08G01N30/72B01J20/281
Inventor 于锡娟张亚红王晓燕唐颐杨芃原
Owner FUDAN UNIV
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