Purification method of recombinant herpes simplex virus
A technology of herpes simplex virus and purification method, which is applied in the field of purification of recombinant herpes simplex virus, can solve the problems of high price, high cost, and difficult promotion, and achieve the effect of no virus damage, low cost, and good purification effect
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[0062] I. Virus Proliferation
[0063] Use a 10 mL sterile pipette to draw 6 mL of PBS from Vero cells with a cell density of 80%-90% to wash the cells three times; o C adsorption for 1 hour; add virus growth solution and maintenance solution to the cell bottle, and culture it in a 37 ℃ constant temperature incubator until 75-100% of the cells have CPE lesions. Harvest. Before harvesting, the cells can be placed in a -70 ℃ refrigerator and frozen Melt 2 to 3 times, 4 o Centrifuge at 6000g for 20 minutes, remove the precipitate, and collect 50 mL of virus supernatant.
[0064] II. Virus Concentration
[0065] Add 5% PEG8000 to the supernatant, 4 o C stirring for 6 hours or overnight, 4 o Centrifuge at 13500g for 2 hours; dissolve the precipitate with TNMC buffer, and add n-dodecyl-β-D-maltoside to the solution to a concentration of 20mM to promote the dissolution of the precipitate, and finally make it centrifuged to purify the virus sample The final volume is 1% of that b...
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