摘要
目的利用响应面法优化14型肺炎链球菌荚膜多糖的超滤工艺。方法选择跨膜压力、切向流量及多糖浓度为自变量,膜包处理能力为响应值,采用Box-Behnken设计分析各自变量及其交互作用对膜包处理能力的影响;利用Design Expert软件对数据进行回归分析,得到回归方程的预测模型,取3批14型肺炎链球菌荚膜多糖水溶液,对优化的工艺参数进行验证;按照最佳超滤工艺对3批14型肺炎链球菌荚膜多糖水溶液进行等体积透析,确定超滤终点;并对纯化的荚膜多糖进行各项检定。结果优化的14型肺炎链球菌荚膜多糖超滤工艺为:跨膜压力12.8 Psi,切向流量66 L/h,多糖浓度0.86 g/L,最适透析倍数为5倍,在该条件下,膜包处理能力的理论值为91.7 g/(h·m2),实际值为91.9 g/(h·m2)。3批纯化的14型肺炎链球菌荚膜多糖水溶液的多糖组分均符合《欧洲药典》7.0版相关规定,均未检出残余脱氧胆酸钠和乙醇,3批多糖的相对分子质量均大于420 000,抗原性良好,且抗原物质成分无差异。结论响应面法优化的14型肺炎链球菌荚膜多糖超滤工艺显著提高了工作效率,降低了生产成本,为14型肺炎链球菌荚膜多糖的规模化生产提供了实验依据。
Objective To optimize the ultrafihration process for capsular polysaccharide of Streptococcus pneumoniae type 14 by response surface methodology. Methods The performance of membrane was investigated with respect to three factors including transmembrane pressure （TMP）, cross-flow rate and polysaccharide concentration. The effects of various factors and their interactions on the performance of membrane were also investigated by Box-Behnken Design. The pre- dictive model of regression equation was analyzed by Design Expert software. The optimized process parameters were veri- fied by using three batches of capsular polysaceharide of S. pneumonia type 14. Three batches of the capsular polysac- charide were subjected to equivoluminal dialysis by the optimal ultrafiltration process, based on which the end point of ul- trafiltration was determined. Overall control tests were performed on the purified capsular polysaceharide. Results The optimal TMP, cross-flow rate, polysaceharide concentration and diavolume for ultrafihration were 12. 8 Psi, 66 L/h, 0. 86 g/L and 5 respectively. Under such a condition, the predictive performance of membrane was 91. 7 g/（h ·m2）, while the actual value was 91.9 g/（h·m2）. All the polysaccharide components in three batches of capsular polysaceha- ride of S. pneumonia type 14 met the relevant requirements in European Pharmacopoeia （7. 0 edition）, while the relative molecular masses were more than 420 000, and no residual sodium deoxyeholate or ethanol was detected. The three batches of capsular polysaecharide showed good antigenicity, of which the antigenic components showed no difference. Conclusion By using the uhrafihration process for capsular polysaccharide of S. pneumonia type 14 optimized by re- sponse surface methodology, the working efficiency increased significantly, while the production cost decreased, which provided an experimental data for large-scale production of the capsular polysaeeharide.
出处
《中国生物制品学杂志》
CAS
CSCD
2013年第10期1477-1482,共6页
Chinese Journal of Biologicals
关键词
肺炎链球菌
多糖
超滤
响应面
Streptococcus pneumonia
Polysaccharide
Uhrafihration
Response surface methodology
