文章摘要
血管内皮生长因子单链抗体在大肠埃希菌中的高效表达
High Level Expression of pTA anti VEGF scFv in Escherichia coli
  
DOI：doi:10.3969/j.issn.1005-7021.2019.02.008
中文关键词: 血管内皮生长因子单链抗体  条件优化  大肠埃希菌  表达
英文关键词: pTA-anti-VEGF-scFv  condition optimization  Escherichia coli  expression
基金项目:辽宁省教育厅高等学校创新团队项目（LT2014023）
作者单位
张冀原 沈阳药科大学，辽宁 沈阳 110016 
陈羽 沈阳药科大学，辽宁 沈阳 110016 
林涵 沈阳药科大学，辽宁 沈阳 110016 
何晴 沈阳药科大学，辽宁 沈阳 110016 
徐威 沈阳药科大学，辽宁 沈阳 110016 
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中文摘要:
      以血管内皮生长因子单链抗体(pTA-anti-VEGF-scFv)为研究对象，考察不同培养基配方和培养条件对血管内皮生长因子单链抗体表达量的影响。经均匀设计试验，确定工程菌R22-4高效表达重组抗体的最佳培养基为以2×YT培养基为基础，添加质量分数为0.3%葡萄糖、1.0%酪蛋白胨、0.4%玉米浆、0.133% KH2PO4、0.159 6% K2HPO4、0.532% NaCl、0.01%微量元素、0.04%维生素B1；诱导条件为30 ℃、0.1 mmol/L IPTG(异丙基硫代半乳糖苷，Isopropyl β-D-Thiogalactoside)诱导5 h。经ELISA (酶联免疫吸附测定，enzyme linked immunosorbent assay) 检测，血管内皮生长因子单链抗体的表达量是原始对照组的2.27倍，实现了血管内皮生长因子单链抗体在大肠埃希菌中的高效表达。
英文摘要:
      A vascular endothelial growth factor (VEGF) was used as research object to investigate the influence of different media and culture conditions on the expression of pTA-anti-VEGF-scFv. Based on the uniform design experiment, the optimal growth media for high level expression of recombinant antibodies in engineered strain R22-4 was found to be 2×YT, adding 0.3% glucose, 1.0% casein peptone, 0.4% corn starch, 0.133% KH2PO4, 0.159 6% K2HPO4, 0.532% NaCl, 0.01% trace elements, 0.04% vitamin B1. Induction conditions was 30 ℃, 0.1 mmol/L IPTG to induce for 5 h. ELISA detected results demonstrated that the expression of pTA-anti-VEGF-scFv was 2.27 times higher than that of original control group,and the high expression of vascular endothlial growth factor single chain achieved in Escherichia coli.
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