利用植物质体转基因技术高效表达抗人源白介素6单链抗体  

High-level expression of anti-interleukin-6 single chain variable fragment through plastid transformation technology

作　　者：吴梦婷 王海涛 张淼 许文博 张望 李圣纯 WU Mengting;WANG Haitao;ZHANG Miao;XU Wenbo;ZHANG Wang;LI Shengchun(State Key Laboratory of Biocatalysis and Enzyme Engineering,School of Life Sciences,Hubei University,Wuhan 430062,Hubei,China)

机构地区：[1]湖北大学生命科学学院省部共建生物催化与酶工程国家重点实验室,湖北武汉430062

出　　处：《生物工程学报》2022年第6期2269-2280,共12页Chinese Journal of Biotechnology

基　　金：国家自然科学基金(32071477);湖北省引智创新示范基地(2019BJH021)。

摘　　要：白介素6 (interleukin-6, IL-6)是与包括癌症在内的多种疾病相关的一种多功能细胞因子,免疫疗法利用抗人源IL-6抗体来治疗相关疾病取得了很好的治疗效果。植物作为生物反应器可以有效降低药用蛋白的生产成本,同时积累功能蛋白。通过基因枪轰击和再生筛选,得到了2个在烟草质体中表达了鼠源抗人源IL-6单链抗体(single chain variable fragment, scFv)的独立株系,并用Southern blotting鉴定了质体转化烟草的同质化状态。抗人源IL-6 scFv基因在质体转基因烟草中成功转录和翻译,功能性抗人源IL-6 scFv在质体转基因烟草叶片中的含量占到总可溶性蛋白的1%,达到41 mg/kg鲜重。另外,质体转基因烟草的表型与野生型烟草相比并没有显著差异,它们具有相似的生长速率、成熟植株的株高以及果荚数目。抗人源IL-6 scFv的高表达量也表明了利用质体转基因植物低成本生产scFv的潜力。Interleukin-6(IL-6) is a pleiotropic cytokine which participates in the pathogenesis of a variety of clinical disorders, including many kinds of cancers. Anti-IL-6 antibody was proved to be useful for the immunotherapy of various inflammatory diseases. Plants are low-cost platforms for producing specific proteins of therapeutic interest. Two dependent transplastomic tobacco lines expressing murine anti-IL-6 single chain variable fragment(scFv) were generated after bombardment and regeneration, homoplasmy was then verified by Southern blotting analysis. The anti-IL-6 scFv gene was successfully expressed at both transcriptional and translational levels in transplastomic tobacco plants. Functional anti-IL-6 scFv accumulated to 1% of total soluble proteins, namely 41 mg/kg fresh weight. There was no obvious phenotypic difference between the wild-type and the transplastomic tobacco plants, including the growth rate, the height of mature plants and the number of siliques. The high-level expression of anti-IL-6 scFv indicates the potential for cost-effective production of scFV using transplastomic plants.

关 键 词：质体转基因技术 白介素6 单链抗体 植物生物反应器 免疫疗法 

分 类 号：R392-33[医药卫生—免疫学]