A functionalized dendrimer-encapsulated gold nanoparticle/pd-l1 siRNA complex and its preparation and application
A technology of dendrimers and amine dendrimers, which is applied in the field of multifunctional polymer nanocarriers and their preparation and application, to achieve good application potential, mild experimental conditions, and good gene transfection ability
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Embodiment 1
[0049] (1) Weigh 50 mg of the fifth-generation polyamidoamine dendrimer (PAMAM) and dissolve it in 50 mL of ultrapure water. Weigh M-SCM-2000 (78 mg) and dissolve it in 7.8 mL of ultrapure water. Ultrasonic for 15 minutes Then quickly added to G5.NH 2 In the solution, the reaction was stirred at room temperature for 4 hours to obtain a crude G5-PEG product.
[0050] (2) 687 μL HAuCl was added dropwise to the G5-PEG obtained above 4 (30mg / mL), after stirring and mixing in an ice bath for 20 minutes, excess pre-cooled NaBH was added 4 The solution (5.67 mg, 1 mg / mL) was reacted for 4 hours to obtain {G5-PEG-(Au 0 ) 25 }crude product.
[0051] (3) The {G5-PEG-(Au 0 ) 25 } Dissolve in pH 8 phosphate buffer (25 mL), then dropwise add 680 μL of fluorescamine solution (4 mg / mL, dissolved in methanol), react in a dark, 40°C water bath and stir for 30 minutes, then use the interception A dialysis bag with a molecular weight of 5000 was dialyzed against the aqueous solution for 3...
Embodiment 2
[0053] Take 6 mg of {G5-PEG-(Au 0 ) 25 -F} dissolve in 600 μL of D 2 O, the solution was sonicated for 10 min, and then the H NMR spectrum of the material was measured using an NMR spectrometer. NMR results such as figure 2 shown, the proton peak at chemical shift 2.25-3.4 ppm represents G5.NH 2 The methylene proton peak of 3.63ppm is (M-SCM-2000) repeating unit -CH 2 CH 2 - The proton peaks of each G5.NH are calculated from the ratio of their integrated areas 2 There are 16.8 M-SCM-2000 molecules connected to it, which is close to its feeding molar ratio G5.NH 2 : M-SCM-2000 (1:20). The proton peak at 7.2-7.6ppm is attributed to the proton on the benzene ring of fluorescein, and the proton peak at 8.63ppm is attributed to the amino group (-NH) of fluorescein and the dendrimer. 2 ) protons on -COOH generated after the reaction, according to the ratio of their integral areas, calculate each G5.NH 2 There are 3.8 fluorescamine molecules connected to it, which is close ...
Embodiment 3
[0055] The {G5-PEG-(Au 0 ) 25 } Dissolved in ultrapure water, prepared into a solution with a concentration of 200 μg / mL, and detected the UV absorption value in the range of 300nm-800nm by UV-Vis spectrophotometry. UV-Vis spectrophotometric detection results are as follows image 3 As shown, {G5-PEG-(Au 0 ) 25 } There is an absorption peak around 520 nm, indicating the successful encapsulation of gold nanoparticles.
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