Method for extracting rabies virus
A rabies virus and molecular sieve technology, applied in the direction of microorganism-based methods, viruses, virus/bacteriophage, etc., can solve the problems of finished product preparation obstacles, restricted scale, and inability to industrialize production, achieve small fluctuations in antigen content, break production bottlenecks, The effect of rationalization of the production process
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Embodiment 1
[0026] 1 Use a hollow fiber ultrafiltration column with a molecular weight cut-off of 750KD or 500KD to concentrate the virus harvest liquid and remove some impurities
[0027] First, 10 L of the hollow fiber ultrafiltration column was equilibrated with 10 mmol / L PBS-0.15M NaCl buffer solution (pH 7.6). 40L virus harvest liquid was loaded and concentrated, the hollow fiber ultrafiltration column model was UFP-750-E6A, and the membrane area was 2800cm 2 , with a molecular weight cut-off of 750,000 Daltons. After the virus harvest solution was concentrated to 0.4L, 10 mL of the virus concentrate and filtrate were sampled respectively. Then use 10mmol / L PBS-0.15M NaCl buffer solution (pH7.6) to add and elute 8L of equilibrium liquid. Collect a sample of the filtered submembrane liquid every 0.4 L, and take 10 mL of the virus concentrated liquid sample at the same time. Detection of the protein concentration of the collected fractions showed that the removal rate of impurity pr...
Embodiment 2
[0063] 1. Utilize the ultrafiltration membrane with a molecular weight cut-off of 750KD or 500KD to concentrate the virus harvest liquid and remove some impurities
[0064] The operation process and parameters are the same as in Step 1 of Embodiment 1, so they are omitted.
[0065] 2. Extract the sample obtained in step 1 with molecular sieve gel 6% cross-linking degree high-flow agarose gel filtration (SepHarose 6FF)
[0066] Prepare one chromatographic column as mentioned in Example 1 or cut the chromatographic column into four in series to form quadruple columns, then each chromatographic column is 26×175mm, and the volume of each column is 32.5mL. It is best to use four columns connected in series to form a quadruple column.
[0067] The rest of the operation process is the same as Step 3 of Embodiment 1, so it is omitted.
[0068] 3. Extract the sample obtained in step 2 with anion exchange gel high flow rate tetravalent amino agarose (Q SepHarose FF)
[0069] The oper...
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