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MALDI steel target for conducting single-step enrichment and desalination on biological sample and preparation method of MALDI steel target

A biological sample and steel target technology, applied in the field of analysis and testing, can solve the problems of cumbersome production steps, high cost, uneconomical, etc., and achieve the effects of improving analysis signal and sensitivity, prolonging incubation and drying time, and increasing sample volume

Active Publication Date: 2016-05-11
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still several problems in this patterned sample construction technique: 1) The fabrication of patterned silicon wafer target is somewhat cumbersome, time-consuming and uneconomical
In addition, the silicon wafer needs to be pasted horizontally on the steel target with double-sided tape for target analysis; 2) Since the fluorinated silicon wafer target is not stored for a long time, it is necessary to prepare the target plate before each use; 3) Since the surface patterning process is based on a silicon substrate, it is difficult to achieve large-scale applications
In addition, a common problem in the methods of jia and zeng et al. is that when the matrix is ​​placed on the prepared target plate, when the salt is effluxed, a large amount of matrix is ​​also effluxed.
The important thing is that after the first step of sample deposition process, the salt and matrix can be effectively separated, however, when the second step of matrix spot coating process, because the matrix is ​​in contact with the salt and has a strong binding effect, Part of the salt will recombine with the matrix and cover or dope into the sample-matrix co-crystal, resulting in attenuation of the signal
In order to overcome the shortcomings of cumbersome fabrication steps, high cost, time-consuming, matrix efflux, and salt recombination of silicon-based patterned targets, a simple, fast, high-throughput, and effective enrichment and desalination method is urgently needed. Method for MALDI-MS Analysis of Peptide / Protein Analytes

Method used

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  • MALDI steel target for conducting single-step enrichment and desalination on biological sample and preparation method of MALDI steel target
  • MALDI steel target for conducting single-step enrichment and desalination on biological sample and preparation method of MALDI steel target
  • MALDI steel target for conducting single-step enrichment and desalination on biological sample and preparation method of MALDI steel target

Examples

Experimental program
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Effect test

Embodiment 1

[0031] like figure 1 In the steps shown, take a clean MALDI steel target, and apply glycerol to the center of the target. The volume of the glycerol spot is 2.5 μL, and the diameter of the glycerol spot is 2.5 mm. Spray the glycerol spot or directly apply it with a pipette. Paraffin wax solution, the solvent of the paraffin wax solution is n-hexane, the concentration of the paraffin wax solution is 25g / L, the solvent of the paraffin wax solution evaporates rapidly around the glycerol point and forms a thin and uniform solid annular hydrophobic paraffin layer; the glycerin point is rinsed with distilled water Remove, expose the steel target, and then apply the hydrophobic material solution to the central area of ​​the bare steel. The hydrophobic adsorption material is polystyrene, and the solvent is chloroform. The concentration of polystyrene chloroform solution is 15g / L, and the volume is 0.4μL. The solvent evaporates quickly, forming a very smooth hydrophobic adsorption mate...

Embodiment 9

[0036] Adjust the sample to 1fmol / μL equal concentration polypeptide mixed solution (polypeptides are respectively bradykinin 1-7, P1; angiotensin II, P2; angiotensin I, P3; substance P, P4; bombesin, P5; hormone substrate, P6; corticotropin 1-17, P7;) and 50fmol / μL protein mixed solution of equal concentration (respectively containing: insulin, insulin; cytochrome C, cyto; ubiquitin protein, ubi; myoglobin , myo;) to evaluate the enrichment efficiency of the steel target of the present invention to trace polypeptide samples. The steel target of the present invention and the traditional steel target obtained in Example 1 were used as sample target plates. For the steel target of the present invention, 5 μL of sample solution was added dropwise, incubated at room temperature and dried, and then 0.3 μL of matrix solution (0.4 mg / mL α-cyano-4-hydroxycinnamic acid, solvent 30% acetonitrile / 0.1% trifluoroacetic acid aqueous solution) in the center of the polymer layer. For the tr...

Embodiment 10

[0038] The sample was adjusted to a 50 fmol / μL polypeptide mixture solution added with 200 mM ammonium bicarbonate salt, and the steel target prepared in Example 1 and the traditional steel target were used as sample target plates. Other conditions embodiment 9 is consistent, and the result obtained is as attached Figure 5a and 5b shown. In addition, ziptipC18 was used as a sample enrichment and desalination device to compare the use effect of the steel target of the present invention. The operation of ZiptipC18 is as follows: first, wet the ZiptipC18 column with 100% acetonitrile solution, and then use 0.1% trifluoroacetic acid aqueous solution Equilibrate the column; then draw 10 μL of salt-containing sample solution, drain the liquid, and repeat the operation 10 times; then draw 0.1% trifluoroacetic acid aqueous solution for the removal of salt, and repeat the operation 3 times; then draw 10 μL of 0.1% trifluoroacetic acid / 50 % acetonitrile, and drain the liquid into a c...

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Abstract

The invention provides an MALDI steel target for conducting single-step enrichment and desalination on a biological sample. A hydrophobic adsorption material round layer is arranged in the center of a target spot of the steel target, annular uncoated steel surrounds the hydrophobic adsorption material round layer, and an annular paraffin layer surrounds the uncoated steel. The preparation method comprises the following steps: 1, taking 0.2-3.0 microlitre of glycerin, and putting the glycerin in the center of the target spot of the MALDI steel target in a dripping mode, wherein the diameter of a glycerin point is 0.6-2.8 mm; 2, covering the glycerin point with a paraffin solution, so that a thin and uniform annular paraffin layer is formed quickly, and flushing away the glycerin, so that uncoated steel is obtained at the original glycerin point position; 3, covering the central area of the uncoated steel in the annular paraffin layer with a hydrophobic adsorption material solution in a dripping mode, and reserving an annular gap between the central area and an inner ring of the paraffin layer. The MALDI steel target and the preparation method thereof have the advantages of being easy and quick to operate, low in cost, high in sensitivity and applicable to high-flux sample analysis.

Description

technical field [0001] The invention belongs to the technical field of analysis and testing, and in particular relates to a MALDI steel target used for one-step enrichment and desalination of biological samples and a preparation method thereof. Background technique [0002] At present, matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) has become an indispensable technique for analyzing peptides / proteins in biological samples. However, the detection of low-abundance peptides / proteins is still a big challenge, because not only the concentration of these analytes in biological samples is very low, but also the high concentration of salt and other reagents in biological samples will greatly suppress the mass spectrometry signal and reduce the spectrum. Band reproducibility. [0003] There are many off-target methods for pre-separation and purification of samples, such as micro-volume C18 chromatography columns, high-performance liquid chromatography, and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/08
CPCG01N30/08
Inventor 国新华王晟
Owner JILIN UNIV
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