Superparamagnetism composite nanosphere with protein molecular imprinting as well as preparation method and application thereof
A magnetic composite nanometer and superparamagnetic technology, which is applied in the preparation methods of peptides, chemical instruments and methods, organic chemistry, etc. Narrow distribution, simple process and good magnetic responsiveness
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Embodiment 1
[0052] Example 1 Preparation of Superparamagnetic Composite Nanospheres with Protein Molecular Imprints
[0053] (1) Prepare polymethyl methacrylate magnetic composite nanospheres with reference to the method described in the Chinese invention patent with the publication number CN 102070864 A (wherein the superparamagnetic nanoparticles used can be triiron tetroxide, gamma trioxide Iron or the like has at least one type of superparamagnetic nanoparticles, and can also be at least one type of ferromagnetic nanoparticles doped with metal elements such as manganese, cobalt or zinc to increase the saturation magnetization);
[0054] (2) Evenly disperse the polymethyl methacrylate magnetic composite nanospheres prepared in step (1) in 4 mL of tris buffer solution (10 mM, pH 8) or PBS, add protein as a template molecule, and probe under ice bath Sonicate for 5-15 minutes, then magnetically stir for 0.5-3 hours at room temperature;
[0055] (3) Add dopamine, sonicate the probe in ...
Embodiment 2
[0058] Example 2 Preparation of hydroxyl-functionalized polymethyl methacrylate magnetic composite nanospheres Fe by improved microemulsion polymerization 3 o 4 / PMMA
[0059] 1) Superparamagnetic Fe was prepared following the pyrolysis method described in JACS 2004, 126, 273-279 3 o 4 Nanoparticles;
[0060] 2) Take 69 mgFe 3 o 4 Nanoparticles are uniformly dispersed in n-hexane to form a magnetic fluid;
[0061] 3) Dissolve 130 μL triethylene glycol (TEG) and 13 μL Tween-80 (tween-80) in 5 mL water;
[0062] 4) Ferrofluid and methyl methacrylate monomer (MMA) are added to the above aqueous solution at a ratio of 1:2;
[0063] 5) Ultrasonic the above mixture probe for 10 min to form a microemulsion;
[0064] 6) The temperature was raised to 80°C, and ammonium persulfate (APS) was added to initiate the polymerization reaction.
[0065] 7) React for 5 h under mechanical stirring at 300 rpm at 80°C;
[0066] 8) Under an external magnetic field, using Fe 3 o 4 Super...
Embodiment 3
[0067] Example 3 Preparation of Superparamagnetic Composite Nanospheres with Molecular Imprint of Lysozyme
[0068] 1) Take 10 mg of the hydroxyl-functionalized superparamagnetic Fe prepared in Example 2 3 o 4 / PMMA composite nanospheres were uniformly dispersed in 4mL tris-buffer (10 mM, pH 8);
[0069] 2) Add 2 mg lysozyme as template protein;
[0070] 3) The above mixture was sonicated by the probe in an ice bath for 10 min at an ultrasonic intensity of 30-60%, and then magnetically stirred at room temperature for 2 h;
[0071] 4) Add 4 mg dopamine;
[0072] 5) The above mixture was sonicated by the probe in an ice bath for 10 minutes at an ultrasonic intensity of 30-60%, and then magnetically stirred at room temperature for 5 hours;
[0073] 6) Under an external magnetic field, using Fe 3 o 4 / PMMA / PDA (denoted as MIPNs) superparamagnetism, discarding uncomplexed Fe 3 o 4 Nanoparticles of polydopamine and buffer solution; then fully washed Fe with a mixed solutio...
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