摘要
瞬时基因表达(TGE)是一种省去筛选高表达细胞株步骤,快速获得重组蛋白质的技术。昆虫细胞作为一种广泛使用的蛋白表达系统,在国内外关于它的瞬时基因表达的研究比较少。这篇文章进行了以昆虫SF9细胞在悬浮培养条件下进行瞬时表达eGFP和肿瘤坏死因子受体融合蛋白(TNFR-Fc)的条件优化,以期为外源蛋白在昆虫细胞中瞬时表达打下基础。经优化后,转染效率达到37%,重组TNFR-Fc达到700μg/mL。
Transient Gene Expression （TGE） is a fast approach for r-protein production in the absence of selection of high-expression cell line. There are few reports about TGE in insect cell although insect cell line SF9 is a widely used in gene expression system. In this study, the factors influencing transfection efficiency, including amount of DNA, ratio of DNA and PEI, and cell density were optimized to express eGFP and TNFR-Fc in insect cell Sf9 in suspension. Under the optimized conditions,37% of the transfection efficiency and 700μg/L recombinant TNFR-Fc were obtained.
出处
《药物生物技术》
CAS
2014年第4期321-324,共4页
Pharmaceutical Biotechnology
基金
National major drug discovery "twelfth five-year" project "biological medicine drug patents expire large varieties of transformation" (No.2012ZX09202-301)
Strategic emerging industry's core technology in guangdong province-antibody scale preparation of key technology R&D and tumor therapeutic antibody drugs (2012A080800008)
Special major science and technology in guangdong province-development of a class of drugs for the treatment of malignant tumors of the anti AGR2 humanized monoclonal antibody 18A4HU (No.2012A080202014)
Research development and Innovation Fund Project of Jinan University-CHO cell genome epigenetic mechanisms of regulation and its application in animal cell engineering (No.21612110)
