A red ray cartilage polypeptide angiogenesis inhibitory factor
An angiogenesis inhibition and peptide technology, applied in the direction of peptides, can solve the problems of low content, difficult drug development, and large molecular weight of active substances, and achieve the effect of advanced purification process, high accuracy, and significant activity
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[0020] A preparation method of red ray cartilage polypeptide angiogenesis inhibitor. The preparation process is as follows: red ray cartilage-tissue fragmentation-guanidine hydrochloride extraction-ultrafiltration-cell membrane stationary phase adsorption-HPLC fingerprint spectrum comparative analysis-angiogenesis inhibitor.
[0021] 1) Preparation of cell membrane suspension: take vascular endothelial growth factor specific receptor (VEGFR) highly expressed liver cancer cell Bel-7402 in the logarithmic growth phase, using MEM medium (containing 10% fetal bovine serum), At 37℃, 5% CO 2 Culture in an incubator, when the cell count reaches 10 7 At this time, the cells were digested with trypsin, centrifuged at 4°C and 1000 r / min for 10 min, the precipitate was taken and washed twice with physiological saline, and then added to 50 mM Tris-HCl solution and sonicated for 30 min to disrupt the cells. Centrifuge the suspension at 1000r / min and 4°C for 5min. Take the supernatant and centr...
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