层析法纯化马破伤风免疫球蛋白F(ab')_(2)的质量研究  

Study on preparation technology of equine tetanus immunoglobulin F(ab’)2

作　　者：梁凌宇 包正琦 段丽娟 杨凯 马江 任正祥 戴于栋 高建军 LIANG Ling-yu;BAO Zheng-qi;DUAN Li-juan;YANG Kai;MA Jiang;REN Zheng-xiang;DAI Yu-dong;GAO Jian-jun(Serum Department,Lanzhou Institute of Biological Products Co.,Ltd.,Center for Gansu Provincial Vaccine Engineering Research,Lanzhou 730046,Gansu Province,China)

机构地区：[1]兰州生物制品研究所有限责任公司血清室、甘肃省疫苗工程技术研究中心,甘肃兰州730046

出　　处：《微生物学免疫学进展》2023年第1期31-35,共5页Progress In Microbiology and Immunology

摘　　要：目的 建立一种在现行硫酸铵盐析工艺基础上,采用疏水层析和离子交换层析制备马破伤风免疫球蛋白F(ab’)_(2)的层析纯化工艺,以进一步提高该类产品的纯度及比活性,改善其安全性。方法 通过3种不同工艺(传统硫酸铵工艺、传统硫酸铵工艺+一步阴离子交换层析、传统硫酸铵盐析+疏水层析+阴离子交换层析)纯化制备马破伤风免疫球蛋白F(ab’)_(2),然后检测各产物的比活性、纯度、聚合物含量等。结果 采用硫酸铵盐析工艺结合疏水层析和离子交换层析制备的马破伤风免疫球蛋白F(ab’)_(2),比活性均>100 000 IU/g,分子排阻高效液相色谱法检测F(ab’)_(2)含量>97.0%,完整IgG和聚合物含量之和降至0.5%以下,产品质量大幅提升。结论 建立的马破伤风免疫球蛋白F(ab’)_(2)的层析纯化工艺能有效地提高该类产品的纯度、比活性等关键质量,改善产品的安全性,且工艺可控性较好,易于生产放大,适用于马破伤风免疫球蛋白F(ab’)_(2)的规模化生产。Objective On the basic ammonium sulfate precipitation, the hydrophobic and ion-exchange chromatography were added to further improve the purity and specific activity of this product and improve its safety. Methods The equine tetanus immunoglobulin F(ab’)2were purified through three different processes(traditional ammonium sulfate process, traditional ammonium sulfate process + one-step anion exchange chromatography and traditional ammonium sulfate chromatography + hydrophobic chromatography + anion exchange chromatography), and then were detected for the product specific activity, purity, polymer content, etc. Results The quality was greatly improved for the product prepared by ammonium sulfate salt analysis combined with hydrophobic chromatography and ion exchange chromatography, with specific activity > 100 IU/mg, the content of F(ab’)2was > 97.0% by molecular exclusion high performance liquid chromatography testing, the sum of complete IgG and polymer was reduced to less than 0.5%. Conclusion The established chromatographic purification process of equine tetanus immunoglobulin F(ab’)2can effectively improve the key quality attributes, and can be used for large-scale production.

关 键 词：马破伤风免疫球蛋白F(ab’)_(2) 破伤风抗毒素 疏水层析 离子交换层析 硫酸铵盐析 免疫马血清 

分 类 号：Q816[生物学—生物工程]