A multifunctional nano-biological material transfection reagent with the effect of gene therapy for Parkinson's disease, its preparation method and application
A technology for Parkinson's disease and biomaterials, applied in the field of multifunctional nano-biomaterial transfection reagents and its preparation, can solve the problems of Parkinson's disease and non-existing nano-biomaterials, and achieve the improvement and treatment of Parkinson's disease symptoms, less side effects, and improved curative effect
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Embodiment 1
[0067] Embodiment 1 Preparation of multifunctional nano-biological material transfection reagent of the present invention
[0068] Proceed as follows:
[0069] S1. Preparation of photoactive NGF
[0070] S11. According to the volume ratio of 1:4, add dimethylformamide (DMF) to phosphate buffer solution (PBS solution, pH=7.4), then add 800 μg N-succinimide ester; add 60 μg NGF to 26 mL In the above-mentioned phosphate buffer solution (PBS solution, pH=7.4) containing N-succinimide ester and dimethylformamide (DMF), stir and react for 50 h in an ice bath at 4°C and in the dark;
[0071] S12. After the synthesis is completed, use ultrafiltration centrifuge tubes (MiliporeMolecut II, 10KNa) at a speed of 4000rpm / min, centrifuge for 30min to purify the generated azidophenyl derivatives, which are photoactive NGF (AzPhNGF), and freeze-dry spare;
[0072] S13. Before use, add 50 mL of PBS solution to dissolve and adjust to the desired concentration of 1 ng / μL.
[0073] S2. Preparat...
Embodiment 2
[0078] Embodiment 2 Preparation of multifunctional nano-biological material transfection reagent of the present invention
[0079] Proceed as follows:
[0080] S1. Preparation of photoactive NGF
[0081] S11. According to the volume ratio of 1:5, add dimethylformamide (DMF) to phosphate buffer solution (PBS solution, pH=7.4), then add 800 μg N-succinimide ester; add 60 μg NGF to 27 mL In the above-mentioned phosphate buffer solution (PBS solution, pH=7.4) containing N-succinimide ester and dimethylformamide (DMF), stir and react for 55 hours in an ice bath at 4°C and in the dark;
[0082] S12. After the synthesis is completed, use ultrafiltration centrifuge tubes (MiliporeMolecut II, 10KNa) to centrifuge for 40 minutes at a speed of 3500 rpm / min to purify the generated azidophenyl derivatives, which are photoactive NGF (AzPhNGF), and freeze-dry spare;
[0083] S13. Before use, add 50 mL of PBS solution to dissolve and adjust to the desired concentration of 1 ng / μL.
[0084...
Embodiment 3
[0089] Embodiment 3 Preparation of multifunctional nano-biological material transfection reagent of the present invention
[0090] Proceed as follows:
[0091] S1. Preparation of photoactive NGF
[0092] S11. According to the volume ratio of 1:4.5, add dimethylformamide (DMF) to phosphate buffer solution (PBS solution, pH=7.4), then add 850 μg N-succinimide ester; add 60 μg NGF to 30 mL In the above-mentioned phosphate buffer solution (PBS solution, pH=7.4) containing N-succinimide ester and dimethylformamide (DMF), stir and react for 60 h in an ice bath at 4°C and in the dark;
[0093] S12. After the synthesis is completed, use ultrafiltration centrifuge tubes (MiliporeMolecut II, 10KNa) to centrifuge for 30 minutes at a speed of 4500 rpm / min to purify the generated azidophenyl derivatives, which are photoactive NGF (AzPhNGF), and freeze-dry spare;
[0094] S13. Before use, add 50 mL of PBS solution to dissolve and adjust to the desired concentration of 1 ng / μL.
[0095] ...
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