Mechanism and process optimization of ultrafiltration of residual DNA in rabies vaccine
10.7644/j.issn.1674-9960.2014.12.012
- VernacularTitle:超滤截留狂犬疫苗残留DNA的工艺优化和机制探讨
- Author:
Ziwei LIU
;
Wei ZHAO
;
Huan XIONG
;
Kai WANG
;
Ju GUO
;
Zhen LIU
;
Heng ZHANG
- Publication Type:Journal Article
- Keywords:
Vero cell;
DNA residues;
ultrafiltration;
protamine
- From:
Military Medical Sciences
2014;(12):968-971
- CountryChina
- Language:Chinese
-
Abstract:
Objective To control residual DNA by optimizing methodology during the production of rabies vaccine using Vero cells as a vector .Methods The antigen recovery rate was assessed by linked immunosorbent assay-sandwich technique while the residual DNA was detected by DNA probe hybridization method .Antigen recovery and removal of DNA were the main indexes for evaluateing ultrafiltration , the vital part of rabies vaccine production .Three key factors in ultrafiltration were assessed: selection of membrane packages , ultrafiltration pressure and the concentration ratio .Then protamine was used to pretreat ultrafiltrates .Based on the two indicators mentioned above , the effect of protamine pretreat-ment on the ultrafiltrate was evaluated .Results and Conclusion The optimum condition of ultrafiltration was obtained on the basis of the general antigen recovery rate , DNA removal rate and actual production .The primary parameters of ultrafil-tration were as follows:7.5 ×105 ultrafiltration membrane packages, 20 times concentrated, 15 psi ultrafiltration pressure. After pretreatment with protamine , ultrafiltration has proved to be a molecular sieve in intercepting DNA ,while protamine can tangle the fragmented DNA and form a larger molecular segment , which is believed to be more conducive to ultrafiltra-tion interception .
