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Method for Enrichment, Desalting and Identification of Glycopeptides Based on Boron-Containing Comb Polymer Patterned Target Plate

A patterned surface and enrichment technology, applied in the field of biochemical analysis, can solve problems such as analyte loss

Active Publication Date: 2021-04-23
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method requires a large amount of matrix solution to drain salt, which will lead to the loss of some analytes; in addition, this method also needs to strictly control the deposition volume of the sample solution

Method used

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  • Method for Enrichment, Desalting and Identification of Glycopeptides Based on Boron-Containing Comb Polymer Patterned Target Plate
  • Method for Enrichment, Desalting and Identification of Glycopeptides Based on Boron-Containing Comb Polymer Patterned Target Plate
  • Method for Enrichment, Desalting and Identification of Glycopeptides Based on Boron-Containing Comb Polymer Patterned Target Plate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Such as image 3 As shown, the monocrystalline silicon [n type, (100)] is placed in concentrated sulfuric acid / H with a volume ratio of 7:3 2 o 2 In the mixed solution, soak the silicon wafer at 80°C for 1h. Then put in acetone, chloroform, ethanol, and water for 5 minutes so that the surface is thoroughly cleaned and hydroxylated;

[0043] Dip the PMMA toluene solution that concentration is 7.5% (wt %) with capillary tube, then contact and be drip-coated on the substrate, form the PMMA dot that radius is about 800 μ m on the surface of substrate; Substrate and 1H, 1H, 2H , 2H-perfluorodecyltrichlorosilane (FDTS) molecules were heated together in a closed container and kept at 250°C for 3h. The hydrophilic region outside the PMMA barrier is modified with a fluorine-containing hydrophobic monolayer;

[0044] Put the modified silicon wafer into acetone, chloroform, ethanol, and water in turn for 5 minutes to remove the PMMA barrier layer on the surface, so as to form ...

Embodiment 2

[0050] Adjust the sample solution to different ratios of horseradish peroxidase and BSA enzymatic hydrolysis products, respectively use the hydrophobic-hydrophilic-hydrophobic annular patterned surface and the traditional stainless steel target plate as the sample carrier, other conditions and implementation Same as Example 1, the obtained mass spectrum is as follows Figure 6 shown.

Embodiment 3

[0052] Adjust the sample solution to add sodium chloride (1M), ammonium bicarbonate (100mM) 118.5fmolμL -1Polypeptide mixed solution, using the hydrophobic-hydrophilic-hydrophobic annular patterned surface and the traditional stainless steel target plate as the sample carrier, and other conditions are the same as in Example 1. The obtained mass spectrum is as follows Figure 7 shown.

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Abstract

A method for enrichment, desalination and identification of glycopeptides based on a boron-containing comb-shaped polymer patterned target plate, which uses PPA-EAPBA comb-shaped molecular brushes to form a patterned surface for one-step enrichment and desalination purification of biological samples This patterned surface can be widely used in the pretreatment and detection of biological samples. The steps are: synthesizing a comb-shaped molecular brush containing boric acid PPA-EAPBA; constructing a closed patterned surface with hydrophobic-hydrophilic-hydrophobic regions on the substrate; dropping the polypeptide or protein solution on the patterned surface prepared in the above steps, and letting it dry naturally Dry to obtain dry sample points; drop the matrix solution on the above dried sample points, and after natural drying, a uniform co-crystal of the sample and the matrix is ​​formed on the patterned surface, thereby realizing the enrichment and desalination of biological samples Purification treatment. Utilizing the patterned surface constructed by the present invention, in MALDI-TOF MS detection, the signal-to-noise ratio, sensitivity and salt tolerance are all significantly improved, which has great practical application value.

Description

technical field [0001] The invention belongs to the technical field of biochemical analysis, and specifically relates to a PPA-EAPBA comb-shaped molecular brush and a treatment method for one-step enrichment and desalination purification of biological samples by using the PPA-EAPBA comb-shaped surface to form a patterned surface. This patterned surface can be widely used Applied to the pretreatment and detection of biological samples. Background technique [0002] Matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an effective soft ionization method with high sensitivity for the analysis and detection of biological samples in proteomics. However, in the process of mass spectrometry, the detection sensitivity and reproducibility of mass spectrometry will be reduced due to the uncontrollable sample deposition area and the inhomogeneous co-crystallization of matrix and sample. In addition, salts and other reagents present in biologic...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/531G01N33/53
CPCG01N33/5306G01N33/531G01N33/6851G01N2560/00G01N2570/00
Inventor 丁寅徐雷刘丽章福平
Owner NANJING UNIV
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