Novel snake venom polypeptide, preparation and application thereof
A technology of snake venom polypeptide and crude venom, applied in the field of biochemistry, can solve the problem of less anti-tumor and achieve good anti-tumor effect
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[0047] method:
[0048] 1) 20 mg of the crude venom of Agkistrodon akistrodon was fully dissolved in 500 μl of ultrapure water, centrifuged at 5000 rpm for 5 min at 4°C to remove insoluble substances, and the supernatant was taken for later use.
[0049] 2) Use a diameter of 5 μm and a pore size of 300 Reversed-phase silica gel C18 granules were used as a stationary phase filler to make a C18 reverse-phase preparative column (20×300mm) for high performance liquid chromatography. The chromatographic mobile phase is 0.1% trifluoroacetic acid and acetonitrile, the flow rate is 7ml / min, and the detection wavelength is 210nm. On the HPLC chromatography system, 0.1% trifluoroacetic acid + acetonitrile is used as the mobile phase, and 0.1% trifluoroacetic acid is eluted The gradient is from 100% to 20%, the gradient of acetonitrile elution is from 0 to 80%, the elution time is 120min, the separation temperature is 25°C, and the components with absorbance of 0.5-2.0 are collected at...
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