中国兽药杂志2026，Vol.60Issue(7)：35-40,6.DOI:10.11751/ISSN.1002-1280.2026.7.05
羊棘球蚴EG95重组蛋白的CHO细胞表达及免疫原性分析
Expression of Echinococcus granulosus EG95 Recombinant Protein in CHO Cells and Immunogenicity Analysis
摘要
Abstract
This study utilized the CHO eukaryotic expression system to express the EG95 recombinant protein of Echinococcus granulosus and analyzed its safety and immunogenicity in lambs.The codon-optimized and truncated EG95 gene was ligated into the pEE12.4 eukaryotic expression vector and transfected into CHO cells via electroporation.The stable CHO cell lines which expressing the EG95 recombinant protein were selected using MSX pressure screening.The expression supernatant was purified by affinity chromatography.SDS-PAGE revealed that the apparent molecular weight of the recombinant protein matched the theoretical prediction,and Western blot further confirmed its specific recognition by anti-His antibodies.Subsequently,the safety and immunogenicity of the vaccine prepared from the purified protein were investigated.The results demonstrated successful soluble expression of the EG95 recombinant protein in the CHO eukaryotic system,achieving a purified yield of 2 mg/mL relative to the initial culture volume.SDS-PAGE and Western blot confirmed that the target protein matched the expected molecular size.Vaccination of 2～4-month-old lambs,no clinical symptoms or injection site abnormalities were observed in lambs,and the temperature remained normal,indicating excellent safety of the EG95-based vaccine.After vaccinating lambs,the antibodies of the lamb can turn positive 14 days after the first immunization,which confirms its strong immunogenicity.This study provides a foundation for the the development of genetically engineered subunit vaccines for ovine echinococcosis.关键词
羊棘球蚴病/CHO真核表达系统/EG95重组蛋白/安全性/免疫原性Key words
ovine echinococcosis/CHO eukaryotic expression system/EG95 recombinant protein/safety/immunogenicity分类
生物科学引用本文复制引用
侯真真,张华伟,王斌,罗修鑫,徐高原..羊棘球蚴EG95重组蛋白的CHO细胞表达及免疫原性分析[J].中国兽药杂志,2026,60(7):35-40,6.基金项目
湖北省农业微生物产业发展重大专项"家畜重要疫病新型疫苗创制"(NYWSWZX2025-2027-3) （NYWSWZX2025-2027-3）
