Purification method and application of recombinant Sendai virus-like particles
A technology of Sendai virus and purification method, applied in the field of virus biology, can solve the problems of low purification efficiency, difficult to remove impurities, etc., and achieve the effect of simple operation and high recovery rate
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Embodiment 1
[0035] Embodiment 1 Purification method of recombinant Sendai virus-like particles
[0036] A method for purifying recombinant Sendai virus-like particles, comprising the steps of:
[0037] S1, cell lysis: collect the cells containing the recombinant Sendai virus-like particles, add the cell lysate, the components of the cell lysate are as follows: 50mM Tris-HCl buffer (pH 7.4), 150mM NaCl, 1% TritonX-100 , 0.5% SDS, 0.5% deoxycholate, 4% β-mercaptoethanol and 1mM EDTA, after crushing, a cell lysate is obtained, and the crushing is carried out by a high-pressure homogeneous method, and the specific conditions are as follows: Break the cells 4-6 times under certain conditions;
[0038] S2. Ion-exchange chromatography: use anion-exchange chromatography column (one of Capto-DEAE anion-exchange column or Capto-Q anion-exchange column) to purify the cell lysate obtained in step S1, the cell lysate The specific method for purification is as follows: Use 50mM Tris-HCl buffer (pH 8....
Embodiment 2
[0051] Example 2 Performance Test of Recombinant Sendai Virus-Like Particles
[0052] The purity and yield of the recombinant Sendai virus-like particles prepared in Example 1 and Comparative Examples 1-5 were tested, and the results are shown in Table 1 below:
[0053] Table 1 Performance test results of recombinant Sendai virus-like particles
[0054] Purity (%) of recombinant Sendai virus-like particles Yield (%) of recombinant Sendai virus-like particles Example 1 95 87 Comparative example 1 80 75 Comparative example 2 65 59 Comparative example 3 84 79 Comparative example 4 85 80 Comparative example 5 83 77
[0055] As can be seen from the performance test results in Table 1, the purity of the recombinant Sendai virus-like particles prepared in Example 1 is 95%, and the yield is 87%. Perform ion-exchange chromatography, then carry out hydroxyapatite chromatography, and finally carry out affinity chromatography, a...
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