ZIF-8 polypeptide composite nano preparation and preparation method thereof
A ZIF-8, nano-formulation technology, applied in the field of biomedicine and pharmaceutical preparations, can solve problems such as undeveloped, and achieve the effect of high polypeptide loading rate
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[0031] like figure 1 As shown, the preparation method of the ZIF-8 polypeptide composite nano-preparation comprises: the polypeptide with a low isoelectric point at a concentration of 100-1000 μg / ml, a zinc salt solution with a concentration of 10mM, and a dimethylimidazole solution with a concentration of 350mM-700mM Mix evenly, let stand at room temperature for 4-6 hours, and obtain ZIF-8 / polypeptide composite nano-preparation after purification and store in freezer.
[0032] As a preferred method, the purification method: centrifuge to remove the supernatant, centrifuge again after reconstitution, and cycle multiple times; the mixing methods of the polypeptide with low isoelectric point, zinc salt solution, and dimethylimidazole solution include: magnetic stirring, mixing, One or more of mechanical stirring mixing, hand shaking mixing, ultrasonic mixing, shaker mixing, oscillator mixing, and static diffusion mixing.
[0033] As an example, the zinc salt includes: zinc nitr...
Embodiment 1
[0040] Embodiment 1: the preparation method of ZIF-8 / P1 composite nano-preparation, specifically prepare through the following steps:
[0041] Configure the solution:
[0042] Prepare 10mM zinc nitrate mother solution: use a pipette gun to draw 0.5ml solution from zinc nitrate standard aqueous solution (10mM) and add it to a 1.5ml centrifuge tube.
[0043] Preparation of 350mM dimethylimidazole mother solution: Weigh 287.3mg of dimethylimidazole into a 15ml centrifuge tube with a microanalytical balance, add 10ml of deionized water to obtain a 350mM ligand solution.
[0044]Prepare 1mg / ml polypeptide mother solution: dissolve 1mg P1 in 1ml PBS buffer to obtain 1mg / ml P1 solution;
[0045] 100 μl of P1 solution was mixed with 500 μl of 350 mM dimethylimidazole ligand solution, followed by the addition of 500 μl of 10 mM zinc nitrate solution. The above mixed solution was thoroughly mixed with ultrasound, and then left to stand at room temperature for 6 hours. After the react...
Embodiment 2
[0046] Embodiment 2: The preparation method of ZIF-8 / Cy3-P1 composite nano-preparation, specifically prepared by the following steps:
[0047] Configuration solution:
[0048] Prepare 10mM zinc nitrate mother solution: use a pipette gun to draw 0.5ml solution from zinc nitrate standard aqueous solution (10mM) and add it to a 1.5ml centrifuge tube.
[0049] Preparation of 700mM dimethylimidazole mother solution: Weigh 574.5mg of dimethylimidazole into a 15ml centrifuge tube with a microanalytical balance, add 10ml of deionized water to obtain a 700mM ligand solution.
[0050] Prepare 1mg / ml polypeptide mother solution: dissolve 1mg Cy3-P1 in 1ml PBS buffer to obtain 1mg / ml Cy3-P1 solution. 100 μl of Cy3-P1 solution was mixed with 500 μl of 700 mM dimethylimidazole ligand solution, and then 500 μl of 10 mM zinc nitrate solution was added. The above mixed solution was thoroughly mixed with ultrasound, and then left to stand at room temperature for 6 hours. After the reaction w...
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