PAP (pokeweed antiviral protein) freeze-dried powder compounding agent and preparation method of thereof
An anti-virus and protein freezing technology, applied in the field of medicine, can solve the problems of volatile inactivation, affecting the anti-viral function of drugs, etc., and achieve the effects of treating HPV virus infection, good clinical application prospects, and easy storage.
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Embodiment 1
[0072] 1. Preparation of natural pokeweed antiviral protein
[0073] (1) Pretreatment: grind pokeweed plant and protein extract in a ratio of 1:4 (w / v) to grind and homogenate, filter, adjust the pH of the supernatant to 3-6, and overnight at 4°C;
[0074](2) Rough extraction: centrifuge to get the supernatant, add solid ammonium sulfate to a final concentration of 40%, leave it overnight at room temperature, and carry out the first salting out; collect the supernatant by centrifugation, add solid ammonium sulfate to a final concentration of 70%, and place it at room temperature Overnight, carry out the second salting out; centrifuge to get the precipitate, which is PAP crude protein, store it at -20°C, and set aside;
[0075] (3) Purification: PAP crude protein is dissolved in phosphate buffer, gel filtration, ion exchange column chromatography and ultrafiltration technology are used to purify pokeweed antiviral protein, concentrated and vacuum freeze-dried.
[0076] 2. Prep...
Embodiment 2
[0087] 1. Preparation of natural pokeweed antiviral protein
[0088] (1) Pretreatment: grind pokeweed plant and protein extract in a ratio of 1:5 (w / v) to grind and homogenate, filter, adjust the supernatant to PH3-6, and overnight at 4°C;
[0089] (2) Rough extraction: centrifuge to get the supernatant, add solid ammonium sulfate to a final concentration of 45%, leave it overnight at room temperature, and carry out the first salting out; collect the supernatant by centrifugation, add solid ammonium sulfate to a final concentration of 75%, and place it at room temperature Overnight, carry out the second salting out; centrifuge to get the precipitate, which is PAP crude protein, store it at -20°C, and set aside;
[0090] (3) Purification: PAP crude protein is dissolved in phosphate buffer, gel filtration, ion exchange column chromatography and ultrafiltration technology are used to purify pokeweed antiviral protein, concentrated and vacuum freeze-dried.
[0091] 2. Preparation...
Embodiment 3
[0102] 1. Preparation of natural pokeweed antiviral protein
[0103] (1) Pretreatment: grind pokeweed plant and protein extract in a ratio of 1:3 (w / v) to grind and homogenate, filter, adjust the supernatant to PH3-6, and overnight at 4°C;
[0104] (2) Rough extraction: centrifuge to get the supernatant, add solid ammonium sulfate to a final concentration of 55%, leave it overnight at room temperature, and carry out the first salting out; collect the supernatant by centrifugation, add solid ammonium sulfate to a final concentration of 85%, and place it at room temperature Overnight, carry out the second salting out; centrifuge to get the precipitate, which is PAP crude protein, store it at -20°C, and set aside;
[0105] (3) Purification: PAP crude protein is dissolved in phosphate buffer, gel filtration, ion exchange column chromatography and ultrafiltration technology are used to purify pokeweed antiviral protein, concentrated and vacuum freeze-dried.
[0106] 2. Preparation...
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