Wnt protein\zif-8 nanocomplex, its preparation method and application
A nanocomposite, ZIF-8 technology, applied in the direction of peptide/protein components, drug combinations, pharmaceutical formulations, etc., can solve the problems of ZIF-8 disintegration speed, strong cytotoxicity, etc. Live, enhance the effect of stability
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[0048] The embodiment of the present invention also provides a preparation method of Wnt protein\ZIF-8 nanocomplex, including: Wnt protein\ZIF-8 nanocomplex is synthesized in situ so that Wnt protein is loaded on the ZIF-8 zeolite imidazole ester skeleton.
[0049] Firstly, preparing an imidazole ligand solution: dissolving imidazole materials to form a raw solution containing imidazole materials, and then mixing the original solution with Wnt protein raw materials to form an imidazole ligand solution containing Wnt protein.
[0050] Further, the Wnt protein raw material includes Wnt protein; preferably, the Wnt protein raw material also includes a protective protein; preferably, the protective protein is albumin, preferably BSA; preferably, the Wnt protein raw material is Wnt protein and A mixture formed by the protective protein; the preparation of the mixture includes: mixing Wnt protein and BSA according to the mass ratio of 1:1-1:9.
[0051] That is to say, the Wnt prote...
Embodiment 1
[0061] The present embodiment provides a preparation method of Wnt protein\ZIF-8 nanocomplex, comprising:
[0062] (1) Dissolve 259mg (3.15mmol) of 2-methylimidazole in 0.9mL of enzyme-free water, stir well to dissolve (temperature 30°C, 400rpm), and obtain solution A;
[0063] (2) Dissolve 250 micrograms of Wnt3a in 250 micrograms of BSA solution to form a Wnt3a / BSA mixture;
[0064] (3) Add 500 micrograms of Wnt3a / BSA mixture to solution A and stir thoroughly for 10 minutes (30°C, 400rpm) to obtain solution B;
[0065] (4) Dissolve 13.4mg (0.045mmol) of zinc nitrate hexahydrate in 0.1mL of enzyme-free water, stir and dissolve fully (30°C, 400rpm) to obtain solution C;
[0066] (5) Add solution C to solution B and stir thoroughly for 10 minutes (30°C, 400rpm) to obtain white suspension D;
[0067] (6) Centrifuge the suspension D for 20min (4000rpm), discard the supernatant to obtain the precipitate E; add 1mL of absolute ethanol to the precipitate E, resuspend, centrifuge f...
Embodiment 2- Embodiment 5
[0069] Embodiment 2-Example 5 provides a preparation method of Wnt protein\ZIF-8 nanocomplex and the preparation method of Wnt protein\ZIF-8 nanocomplex provided in Example 1 is basically the same operation, the difference lies in the raw materials and specific Operating conditions vary as follows:
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