Optimization of purification process for freeze-dried attenuated live hepatitis A vaccine
10.13200/j.cnki.cjb.004553
- VernacularTitle:冻干甲型肝炎减毒活疫苗纯化工艺的优化
- Author:
LI Qiurui
- Publication Type:Journal Article
- Keywords:
Hepatitis A virus(HAV);
Attenuated live hepatitis A vaccine;
Purification;
Process optimization
- From:
Chinese Journal of Biologicals
2025;38(09):1101-1104+1110
- CountryChina
- Language:Chinese
-
Abstract:
Objective To optimize the purification process of freeze-dried attenuated live hepatitis A vaccine to improve vaccine quality and safety.Methods The extraction times(six times) and centrifugal forces(2 990, 4 070, 5 316, 6 728 × g)of hepatitis A virus(HAV) harvests were optimized, and the virus titers were detected under different extraction times and centrifugal forces. The extracted solution was homogenized and the trichloromethane residue was measured pre-ultrafiltration. The trichloromethane residue, protein content, virus titer and virus content below the membrane were detected after ultrafiltration concentration with a 300 KD molecular weight cutoff(MWCO) membrane. The bulk solutions were prepared by two processes, sampled, and detected for the virus titer, protein content and trichloromethane residue.Results With the increase of extraction times, the virus titer of the supernatant showed a decreasing trend. The average virus titers of the supernatant in the first four extractions exceeded 7. 00 lgCCID_(50)/mL, while the fifth and sixth extractions fell below this threshold. The extraction times of virus harvest were selected as four times. The average virus titers of the bulk solutions prepared under different centrifugation conditions in the four groups ranged from 8. 11 to 8. 39 lgCCID_(50)/mL with no statistically significant difference(F = 1. 319, P = 0. 334). Using 300 KD membrane for ultrafiltration concentration, the virus detection below the membrane was negative. The trichloromethane residues decreased significantly(t = 4. 975, P = 0. 037),and the virus titers and protein contents in bulk solutions were all within the qualification criteria. There was no significant difference in virus titers and trichloromethane residues between the original and optimized processes(t =-0. 970 and 2. 306,P = 0. 369 and 0. 082, respectively), but the trichloromethane residue of the optimized process was lower than that of the original process. The protein levels of bulk solutions prepared by the two processes were stable, well below the internal control standards.Conclusion The purification process of the freeze-dried attenuated live hepatitis A vaccine was successfully optimized, further improving the quality of the vaccine.
