Method for enriching glycosylated peptide by utilizing mass spectrum target plate
A glycosylation and target plate technology, applied in the field of biochemical analysis, can solve the problems of easy loss of micro samples, numerous steps, and inability to directly connect with MALDI-MS, so as to avoid sample loss, reduce elution steps, and reduce sample loss Effect
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0025] Such as figure 1 As shown, first cut a commercial or self-made gold-plated single crystal silicon wafer (thickness: 1mm) into a silicon wafer with the same or slightly smaller sample point size as the MALDI target plate, wash the surface three times with ethanol and water respectively, and then The aforementioned silicon wafer was immersed in an ethanol solution containing 11-mercapto-1-undecanol, shaken overnight, and then the surface was washed three times with ethanol and water respectively to remove excess 11-mercapto-1-undecanol. In this step, a layer of organic long-chain molecules with hydroxyl ends is covered on the gold surface through the self-assembly process of mercapto groups on the gold surface. Then immerse the above-mentioned silicon chip in the N, N-dimethylformamide solution of succinic anhydride and 4-dimethylaminopyridine, and heat to 60 degrees Celsius for 3 hours to react. Thereby, the hydroxyl groups at the ends of the organic chains can be conve...
Embodiment 2
[0027] Such as figure 2 As shown, the silicon chip grafted with boronic acid groups on the surface (hereinafter referred to as silicon chip) can realize the enrichment of glycosylated peptides through two simple steps. First, spot the peptide mixed solution containing glycosylated peptides and 50mM ammonium bicarbonate on the above-mentioned silicon wafer. Due to the effect of surface tension, the peptide mixed solution (containing 50mM ammonium bicarbonate) droplets can be well maintained on the silicon wafer surface. In order to prevent the volatilization of the solvent in the droplet, the silicon wafer was placed in an airtight container filled with water and allowed to stand for 1 hour. Due to the interaction between boronic acid groups and sugar chains, glycosylated peptides are immobilized on the surface of silicon wafers. If the volume of the peptide mixture solution is too large to be maintained on the surface of the silicon wafer by surface tension, the peptide mix...
Embodiment 3-5
[0029] Three samples were used, each with a volume of 100 microliters, and the concentrations were 0.4ng / ul asialofetus, 0.8ng / ul horseradish peroxidase and 0.4ng / ul fetuin trypsinization solution to investigate the selective enrichment ability of this method for glycosylated peptides. Using the boric acid-modified silicon chip made according to Example 1, the above-mentioned three samples were enriched respectively according to the method described in Example 2, and the MALDI mass spectrogram of it is as follows image 3 shown. It can be clearly seen from the figure that only glycosylated peptides and their fragment peaks are detected, but no signals of non-glycosylated peptides appear.
PUM
| Property | Measurement | Unit |
|---|---|---|
| thickness | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More