Novel magnetic target gene transmission system and preparation thereof
A gene delivery system and magnetic targeting technology, used in gene therapy, genetic material components, medical preparations with non-active components, etc., can solve the problem of unsatisfactory effect of target gene to target cells, and achieve to promote collapse and improve expression. , the effect of improving stability
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Embodiment 1
[0030] 1. Add 0.5M Fe 2+ and Fe 3+ Solution, mixed at a molar ratio of 1:2, with a total volume of 30ml. At 75°C, under nitrogen protection, add dropwise to NH 3 -NH 4 In the Cl buffer solution (pH=10, total volume 15ml), 10ml of concentrated ammonia water was added at the same time, and the reaction was terminated after 90min of magnetic stirring. After magnetic separation and distilled water washing, the product was redispersed in 1.0M sodium citrate solution, and mechanically stirred at 60°C for 12h. After the product is washed by magnetic separation, it is made into a 30 mg / ml water-based magnetic fluid.
[0031] 2. Secondary surface modification of magnetic fluid with silane coupling agent. Disperse 2ml of magnetic fluid in an alcohol-water mixture with a volume ratio of 4:1, then add 5ml of ammonia water and 0.01ml of tetraethylorthosilicate, and stir mechanically at 30°C for 12h. The product was magnetically separated, washed with anhydrous ethanol, acetone and an...
Embodiment 2
[0043] 1. Add 0.5M Fe 2+ and Fe 3+ Solution, mixed at a molar ratio of 1:1, with a total volume of 30ml. At 55°C, under nitrogen protection, add dropwise to NH 3 -NH 4 In the Cl buffer solution (pH=9, total volume 15ml), 10ml of concentrated ammonia water was added at the same time, and the reaction was terminated after magnetic stirring for 60min. After magnetic separation and distilled water washing, the product was redispersed in 1.0M sodium citrate solution, and mechanically stirred at 60°C for 12h. After the product is washed by magnetic separation, it is made into a 30 mg / ml water-based magnetic fluid.
[0044] 2. Secondary surface modification of magnetic fluid with silane coupling agent. Disperse 2ml of magnetic fluid in an alcohol-water mixture with a volume ratio of 4:1, then add 5ml of ammonia water and 0.01ml of tetraethylorthosilicate, and stir mechanically at 30°C for 10h. After the product was magnetically separated, washed with absolute ethanol, acetone a...
Embodiment 3
[0049] 1. Add 0.5M Fe 2+ and Fe 3+ Solution, mixed at a molar ratio of 1:1, with a total volume of 30ml. At 60°C, under the protection of nitrogen, add dropwise to NH 3 -NH 4 In the Cl buffer solution (pH = 11, total volume 15 ml), 10 ml of concentrated ammonia water was added at the same time, and the reaction was terminated after magnetic stirring for 60 min. After magnetic separation and distilled water washing, the product was redispersed in 1.0M sodium citrate solution, and mechanically stirred at 60°C for 12h. After the product is washed by magnetic separation, it is made into a 30 mg / ml water-based magnetic fluid.
[0050] 2. Secondary surface modification of magnetic fluid with silane coupling agent. Disperse 2ml of magnetic fluid in an alcohol-water mixture with a volume ratio of 4:1, then add 5ml of ammonia water and 0.01ml of tetraethylorthosilicate, and stir mechanically at 30°C for 12h. After the product was magnetically separated, washed with absolute ethan...
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