PRV (Porcine pseudorabies virus) purification method
A technology of porcine pseudorabies virus and purification method, which is applied in the fields of antiviral agents, recovery/purification, and pharmaceutical formulations, and can solve the problems of high side reaction rate, poor uniformity, and poor immune effect of porcine pseudorabies live vaccines, and achieve immune effects Good, uniform, low cost effect
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Embodiment 1
[0057] 1Operation process
[0058] 1.1 Pre-processing
[0059] 1.1.1 Assembly of the system
[0060] Under sterile conditions, the clarification and purification system and the concentration system are assembled according to the assembly requirements. A sterile 0.65μm, intact hollow fiber microfiltration membrane can be installed in the clarification system, and a sterile hollow fiber ultrafiltration membrane with a pore size of 300KD can be installed in the concentration system.
[0061] 1.1.2 System Integrity Detection
[0062] The pressure hold method checks the integrity of the purification system.
[0063] 1.1.3 System processing
[0064] Cleaning and Sterilization:
[0065] Fill the feed tank of the purification system with 0.5M NaOH solution, soak for 20 minutes, turn on the circulation pump at 300 rpm, and clean and sterilize the system for 30 minutes.
[0066] Washing and flux detection:
[0067] After the sterilization, drain the NaOH solution in the system. ...
Embodiment 2
[0110] 1) Add sterile Tween 20 into the porcine pseudorabies virus (PRV) virus solution at a ratio of 10% (v / v), and shake it;
[0111] 2) Inject the virus liquid after the oscillation treatment in step 1 into the feeding tank of the microfiltration clarification and purification system, turn on the circulation pump, and after a certain period of circulation, filter through a 0.45 μm hollow fiber microfiltration column, and collect the permeate for use;
[0112] 3) Add sterile 0.1M PBS buffer solution into the retentate in the feeding tank at a ratio of 1:1 (v / v), resuspend, turn on the circulation pump to circulate for a certain period of time, collect the permeate, and obtain the first Wash the filtrate and save it at 4-10°C for later use;
[0113] 4) Add sterile 0.1M PBS buffer solution into the retentate in the feeding tank at a ratio of 1:1 (v / v), resuspend, turn on the circulation pump to circulate for a certain period of time, collect the permeate, and obtain the second ...
Embodiment 3
[0121] 1) Add sterile Tween 20 into the porcine pseudorabies virus (PRV) virus solution at a ratio of 0.5% (v / v), and shake at 800rpm for 10min;
[0122] 2) Inject the virus liquid after shaking treatment in step 1 into the feeding tank of the microfiltration clarification and purification system, turn on the circulating pump to circulate at 400rpm for 30min, and then microfiltration through a 0.65μm hollow fiber microfiltration column to collect the permeate stand-by;
[0123] 3) Add sterile 0.1M PBS buffer solution to the retentate in the feeding tank at a ratio of 1:1 (v / v), resuspend, turn on the circulation pump and circulate at 400rpm for 30min, collect the permeate, The first washing filtrate was obtained, and stored at 4-10°C for later use;
[0124] 4) Add sterile 0.1M PBS buffer solution to the retentate in the feeding tank at a ratio of 1:1 (v / v), resuspend, turn on the circulation pump and circulate at 400rpm for 30min, collect the permeate, Obtain the second wash...
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