Method for immobilizing urokinase by employing magnetic tragacanth microspheres
A technology of gum tragacanth and urokinase, which is applied in biomedical engineering and biological fields, can solve the problems of poor biocompatibility of magnetic microspheres, unsatisfactory magnetic response, and complicated operation, and achieves uniform particle size and strong magnetic response. , good dispersion effect
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Embodiment 1
[0018] ① Preparation of magnetic tragacanth particles
[0019] Dissolve 0.5g of tragacanth in 40ml of re-distilled water, transfer it into a four-necked flask equipped with a stirrer, a condenser and a nitrogen inlet, and add 60mL of 0.15mol / L ferrous chloride solution and 0.010% ferric chloride solution dropwise successively. 10 mL of hydrogen oxide solution, stirred (400 r / min), 40 mL of 3 mol / L potassium hydroxide solution was added dropwise, and at the same time under nitrogen protection, the reaction was carried out in a boiling water bath for 4.5 h.
[0020] ②Fixation of urokinase
[0021] Weigh 0.2g of magnetic tragacanth particles, add 18mg of urokinase, shake at 4°C for 6h, fully wash with re-distilled water, and separate the mixture in a magnetic field with a strength of 100Gs until no enzyme protein is washed out, freeze-vacuum-dry to obtain magnetic Immobilized urokinase.
Embodiment 2
[0023] ① Preparation of magnetic tragacanth particles
[0024] Dissolve 1.2g of tragacanth in 40ml of redistilled water, transfer it into a four-necked flask equipped with a stirrer, a condenser and a nitrogen inlet, and add 0.20mol / L ferrous chloride solution 70mL and 0.012% ferric chloride solution dropwise successively. Hydrogen oxide solution 10mL, stirred (360r / min), dropwise added 45ml of 3.5mol / L sodium hydroxide solution, while nitrogen protection, reacted in a boiling water bath for 3h.
[0025] ②Fixation of urokinase
[0026] Weigh 0.1 g of magnetic tragacanth particles, treated with 0.2% glutaraldehyde for 1 h, and then cross-linked with 7 mg of urokinase in boric acid buffer (pH 8.8), shaken at room temperature for 8 h, and washed with distilled water. Separate in a magnetic field of 120Gs until enzyme-free protein washes out.
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