Bioactive protein for preventing and treating human papilloma virus infection as well as preparation method and application thereof
A technology of biologically active protein and human papillomavirus, which is applied in the field of medicine, can solve the problems of affecting the effect of anti-HPV virus, poor modification effect of lactoferrin, and large steric hindrance, so as to prevent the expansion of infection, low cost, The effect of small steric hindrance
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Embodiment 1
[0036] Example 1 Preparation of acid anhydrided lactoferrin
[0037] In this example, the preparation of acid-anhydrided lactoferrin includes the following steps: at a temperature of 25°C, dissolving 4-hydroxyphthalic anhydride in dimethyl sulfoxide to obtain a saturated 4-hydroxyphthalic anhydride solution; Dissolve ferritin in 0.1M trisodium phosphate solution to obtain a protein solution with a final concentration of 30 mg / mL; add the above-mentioned 4-hydroxyphthalic anhydride solution into the protein solution, stir and mix, and the final concentration of 4-hydroxyphthalic anhydride in the reaction system It was 60mM, and left to react for 2 hours; to obtain acid anhydride lactoferrin; use 0.45uM filter membrane to filter and sterilize, and then use pH 7.2 phosphate buffer for ultrafiltration replacement purification, and the molecular weight cut-off of the ultrafiltration membrane used is 10KD; 0.45uM microporous membrane filter to sterilize, add the protective agent man...
Embodiment 2
[0039] Embodiment 2 prepares acid anhydride lactoferrin
[0040] Dissolve 4-hydroxyphthalic anhydride in dimethyl sulfoxide at a temperature of 35°C to obtain a saturated 4-hydroxyphthalic anhydride solution; dissolve lactoferrin in 0.1M trisodium phosphate solution to obtain a final concentration of 50mg / mL protein solution; add the above-mentioned 4-hydroxyphthalic anhydride solution into the protein solution, stir and mix, the final concentration of 4-hydroxyphthalic anhydride in the reaction system is 80mM, and let it stand for 2 hours to obtain acid anhydrided lactoferrin; Use a 0.45uM filter membrane to sterilize by suction and then use pH 7.2 phosphate buffer for ultrafiltration replacement purification. The molecular weight cut-off of the ultrafiltration membrane used is 10KD; Use a freeze dryer to freeze-dry, collect the freeze-dried powder, and obtain the acid-anhydrided lactoferrin freeze-dried powder.
[0041] The protein freeze-drying step includes the following ...
Embodiment 3
[0042] Example 3 Preparation of acid anhydrided lactoferrin
[0043] At a temperature of 50°C, 4-hydroxyphthalic anhydride was dissolved in dimethyl sulfoxide to obtain a saturated 4-hydroxyphthalic anhydride solution; lactoferrin was dissolved in 0.1M trisodium phosphate solution to obtain a final concentration of 80mg / mL protein solution; add the above-mentioned 4-hydroxyphthalic anhydride solution into the protein solution, stir and mix, the final concentration of 4-hydroxyphthalic anhydride in the reaction system is 80mM, let it stand for 2 hours to react; obtain acid anhydrided lactoferrin; Use a 0.45uM filter membrane to sterilize by suction and then use a pH 7.2 phosphate buffer for ultrafiltration replacement purification. The molecular weight cut-off of the ultrafiltration membrane used is 10KD; Freeze-drying is carried out by a freeze dryer, and the freeze-dried powder is collected to obtain the freeze-dried powder of acid anhydrided lactoferrin.
[0044] The protei...
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