Sialic acid oligosaccharide-magnetic nanozyme and its preparation method and application
A sialic acid oligosaccharide, magnetic nanotechnology, applied in the preparation of sugar derivatives, chemical instruments and methods, sugar derivatives, etc., can solve the problems of complicated operation and high price
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Embodiment 1
[0068] Embodiment 1, the preparation of sialooligosaccharide-magnetic nanozyme
[0069] 1. Preparation of Magnetic Nanozymes
[0070] Add polystyrenesulfonic acid-maleic acid copolymer sodium salt (PSSMA3:1, 1g) into 20ml ethylene glycol and stir to dissolve, then add ferric chloride hexahydrate (0.54g) and anhydrous sodium acetate (1g), Stir until completely dissolved. Then the above solution was transferred to a 25 ml polytetrafluoroethylene-lined high-pressure hydrothermal reaction kettle, and the reaction kettle was placed in an oven preheated to 200° C. for heating and reaction for 10 hours. Take out the reaction kettle and cool to room temperature naturally, separate the magnetic microspheres by magnetic separation, wash with pure water to remove unreacted reactants and by-products, and dry in vacuum to prepare ferric oxide microspheres (Fe 3 o 4 ).
[0071] Ferric oxide microspheres (average particle size about 200nm) (0.2mg) and potassium ferrocyanide (0.2mg) were ...
Embodiment 2
[0103] Example 2. Enrichment and isolation of influenza virus with sialooligosaccharide-magnetic nanozyme
[0104] Add 50 μl of an aqueous solution containing influenza virus to 200 μl of PBS buffer, then add 100 μl of 1 mg / ml sialooligosaccharide-magnetic nanozyme solution, and incubate at 4° C. for 8 hours. The sialic acid oligosaccharide-magnetic nanozyme with influenza virus was separated by magnetic separation, and washed three times with PBST to remove unreacted reactants and by-products. Finally, PBST (0.05% Tween-20) was used to dissolve the sialooligosaccharide-magnetic nanozyme enriched in influenza virus to obtain a solution of sialooligosaccharide-magnetic nanozyme enriched in influenza virus.
Embodiment 3
[0105] Example 3, detection of influenza virus with α2,3 binding characteristics (taking VieH5N1 as an example)
[0106] Different concentrations of VieH5N1 influenza virus aqueous solutions were prepared, and the HA titers were 0, 1, 2, 4, 8, and 16, respectively.
[0107] Experimental group No. 1: 50 μl of each concentration of influenza virus aqueous solution was added to 200 μl of PBS buffer, and then 100 μl of 1 mg / ml 3’SLac-magnetic nanozyme was added to each tube, and incubated at 4°C for 8 hours. The 3'SLac-magnetic nanozyme enriched in influenza virus was separated by magnetic separation, and washed 3 times with PBST to remove unreacted reactants. Finally, 800 μl of PBST buffer solution was used to dissolve the 3'SLac-magnetic nanozyme enriched in influenza virus to obtain the No. 1 experimental group solution.
[0108] Experimental group No. 2: 50 μl of each concentration of influenza virus aqueous solution was added to 200 μl of PBS buffer, and then 100 μl of 1 mg / ...
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