摘要
目的应用DOE试验优化破伤风抗毒素（tetanus antitoxin,TAT）生产中胃酶消化马血浆过程的工艺参数。方法以破伤风类毒素免疫后的马血浆为原料,通过AKTA avant25中UNICORN 6.0软件设计4因子2水平全因子DOE试验模型,并以抗体F（ab′）2含量为评价指标,对影响胃酶消化的主要因子即胃酶加量、消化反应pH值、反应温度和反应时间进行优化。结果胃酶消化马血浆的最适工艺参数为：胃酶加量为13~15 U/ml,消化反应pH值为3.0~3.1,反应温度为35~37℃,反应时间为90~120 min,在该条件下进行6次稳定性试验,制备的TAT抗体F（ab′）2含量平均值达83.1%,标准偏差为2.47%。结论 DOE试验优化了TAT生产中胃酶消化马血浆过程的工艺参数,建立了有效的消化工艺,为降低过敏反应,提高TAT制品的质量提供了实验依据。
Objective To optimize the technological parameters for digestion of equine plasma with pepsin during pro-duction of tetanus antitoxin（TAT）by DOE. Methods DOE model with four factors at two levels was designed by the UNICORN 6. 0 software in AKTA avant25 using the equine plasma immunized with TAT as material. The main influencing factors of digestion with pepsin,including pepsin amount as well as pH value,temperature and time for digestion were optimized using F（ab′）2content as an index for evaluation. Results The optimal pepsin amount for digestion of equine plasma was 13 ~ 15 U / ml,while the optimal pH value,temperature and time for digestion were 3. 0 ~ 3. 1,35 ~ 37 ℃and 90 ~ 120 min respectively. Six stability tests were performed under the optimal condition,and the mean F（ab′）2content was 83. 1%,with a standard deviation of 2. 47%. Conclusion The technological parameters for digestion of equine plasma with pepsin during production of TAT was optimized by DOE,and an effective digestion process was developed,which provided an experimental basis for the improvement of quality of TAT.
出处
《中国生物制品学杂志》
CAS
CSCD
2014年第7期959-963,共5页
Chinese Journal of Biologicals
