HPV vaccine modified material for highly responsive anti-tumor immunotherapy and preparation method thereof
An anti-tumor immunity and modified material technology is applied in the field of HPV vaccine modified materials for highly responsive anti-tumor immunotherapy and its preparation, which can solve the problems of low response rate of therapy and large toxic and side effects.
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Embodiment 1
[0039] [Example 1] Preparation of HPV modified material (siRNA1@HPVP)
[0040] 1. Disassembly of HPV vaccine protein
[0041]Dithiothreitol (DTT) was used as a reducing agent to cut the disulfide bond in the HPV vaccine protein to disassemble the HPV vaccine protein assembly. The specific preparation process is as follows: Disperse 5 mg of vaccine protein assembly in an EP tube filled with 5 mL of PB buffer solution (pH 8.2, containing 0.166 M NaCl), slowly add dithiothreitol solution dropwise to make the reducing agent The final concentration was 2 mM, and the mixed solution was placed on a constant temperature shaker at 25°C for 1 hour, and the obtained disassembled vaccine protein solution was placed at 4°C for use.
[0042] 2. Load siRNA and reassemble HPV vaccine protein
[0043] Preparation of PEGylated pseudovirus nanoparticles loaded with siRNA (siRNA@HPVP): Add 50 μg of PEG to the mixed solution of the disassembled vaccine protein 2000 - NHS and 2.5 mg of siRNA1 se...
Embodiment 2
[0045] [Example 2] Preparation of HPV modified material (siRNA2@HPVP)
[0046] The HPV vaccine protein solution disassembled in this example is from Example 1.
[0047] 1. Load siRNA2 and reassemble HPV vaccine protein
[0048] Preparation of PEGylated pseudovirus nanoparticles loaded with siRNA2 (siRNA2@HPVP): Add 50 μg of PEG to the disassembled vaccine protein mixture solution 2000 - NHS and 2.5 mg of siRNA2 sequence. The above mixed solution was dialyzed against PB buffer solution (pH 8.2, containing 0.5M NaCl). The dialysate was changed every 6 hours, and the HPV vaccine modified nanomaterials were obtained after 24 hours.
Embodiment 3
[0049] [Example 3] Preparation of HPV modified material (siRNA3@HPVP)
[0050] The HPV vaccine protein solution disassembled in this example is from Example 1.
[0051] 1. Load siRNA3 and reassemble HPV vaccine protein
[0052] Preparation of PEGylated pseudovirus nanoparticles loaded with siRNA3 (siRNA3@HPVP): Add 50 μg of PEG to the mixed solution of the disassembled vaccine protein 2000 - NHS and 2.5 mg of siRNA3 sequence. The above mixed solution was dialyzed against PB buffer solution (pH 8.2, containing 0.5M NaCl). The dialysate was changed every 6 hours, and the HPV vaccine modified nanomaterials were obtained after 24 hours.
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