Preparation method of magnetic amphiphilic metal organic framework material for enriching glycopeptides
A metal-organic framework and sugar-enrichment technology, which is applied in peptide preparation methods, organic chemistry, and alkali metal compounds, can solve the problems of limited hydrophilicity and low efficiency of enriching glycopeptides, and achieve increased specific surface area and simplified The effect of glycopeptide enrichment process and mild reaction conditions
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Embodiment 1
[0025] Example 1: Enrichment of Glycopeptides in HRP and IgG Enzymatic Hydrolyzate Using Magnetic Double Hydrophilic Metal-Organic Framework Materials
[0026] (1) Preparation process of magnetic amphiphilic metal-organic framework materials
[0027] (a) Fe 3 o 4 @SiO 2 The preparation process of nanoparticles: Weigh Fe 3 o 4 Put 1.0 g of nanoparticles in a three-necked flask, add 50 mL of deionized water and 150 mL of absolute ethanol, and ultrasonically disperse them evenly for 15 min. Then add 5mL NH 3 ·H 2 O was reacted under alkaline conditions for 10 min. Finally, a mixed solution of 2 mL tetraethyl orthosilicate and 50 mL absolute ethanol was added dropwise, and stirred at room temperature for 8 h. Under the assistance of ultrasound, Fe can be obtained by the following steps 3 o 4 @SiO 2 Washing: water (2 times, each time 100mL, time 5min, except NH 3 ·H 2 O), 1M HCl solution (3 times, 50 mL each time, 5 min, hydroxylation), water (4 times, 100 mL each time...
Embodiment 2
[0041] Example 2: Enrichment of Glycopeptides in Human Serum Enzymatic Hydrolyzate Using Magnetic Double Hydrophilic Metal-Organic Framework Materials
[0042] (1) Magnetic double hydrophilic metal-organic framework material MUiO-66-NH 2 / PA preparation process: with (1) in Example 1.
[0043](2) Preparation of human serum enzymatic hydrolyzate: First, 10 μL of mixed human serum was washed with denaturing buffer containing 8M urea (50 mM NH 4 HCO 3 ) was diluted to 100 μL. The mixture was then treated with dithiothreitol (DTT, 5 μL, 200 mM) and iodoacetamide (IAA, 20 μL, 400 mM), respectively. The reaction mixture was then washed with 50 mM NH 4 HCO 3 The aqueous solution was diluted to 1 mL, and trypsin (w / w=1:25) was added to incubate at 37° C. for 18 h. After the enzymatic hydrolysis was completed, 2 μL of formic acid solution was added to make the solution acidic to terminate the enzymatic hydrolysis, and it was stored in a -20°C refrigerator for frozen storage until...
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