猪瘟DNA疫苗制备工艺研究

Study on Preparative Protocol of DNA Vaccine Against Classical Swine Fever

猪瘟DNA疫苗制备工艺的建立是其走向产业化 ,从而在猪瘟防制中发挥作用所需解决的关键问题之一。以猪瘟DNA疫苗质粒pVAXE2IL2转化大肠杆菌后 ,转入发酵罐发酵培养。将菌体悬浮裂解和中和后 ,经澄清和浓缩处理 ,上阴离子交换色谱和体积排阻色谱进行纯化 ,首次提纯出符合药学规格的猪瘟DNA疫苗 ,且整个工艺回收率达 64 5 3% 。

DNA immunization is one of the stratigies in control of classic swine fever (CSF),a viral disease. Here we reported a chromotagraphy_based technology in production of DNA vaccine plasmid with high yield and purity.The experimental DNA vaccine encoding CSFV E2 protein and IL_2 in one plasmid was amplified in Escherichia coli strain of DH5α. After fermentation in complex medium the plasmid DNA was pre_processed and purified first with anion exchange chromatography followed by size exclusion chromatography. The final results showed that the quality of purified plasmid DNA met the standards for preparation of pharmaceutical grade plasmid DNA.