The Jenner Institute, The Henry Wellcome Building for Molecular Physiology, Nuffield Department of Medicine, University of Oxford, Oxford, UK.
Division of Structural Biology, Wellcome Centre for Human Genetics, University of Oxford, Oxford, UK.
Methods Mol Biol. 2020;2142:103-112. doi: 10.1007/978-1-0716-0581-3_9.
Zika virus (ZIKV) is an emerging mosquito-borne flavivirus, which has recently caused global epidemics with its association with congenital Zika syndrome such as severe microcephaly. The recombinant ZIKV envelope (Env) glycoprotein is useful for immunological applications such as serodiagnosis of ZIKV infection and for monitoring immune responses in preclinical and clinical ZIKV vaccine developments. In this chapter, we describe the optimization of production of Zika virus envelope glycoprotein in Human Embryonic Kidney (HEK 293T) cells by small-scale expression followed by large-scale protein production. Small-scale expression of HEK 293T cells allows screening of a large number of vectors simultaneously to select the vectors with best secretory profiles for scale-up in Expi293 mammalian system to maximize the protein yield followed by purification for research and clinical applications.
Zika 病毒（ZIKV）是一种新兴的蚊媒黄病毒，最近因其与先天性 Zika 综合征（如严重小头畸形）的关联而在全球范围内引发了流行。重组 ZIKV 包膜（Env）糖蛋白可用于免疫应用，如 Zika 病毒感染的血清学诊断，以及监测临床前和临床 ZIKV 疫苗开发中的免疫反应。在本章中，我们描述了通过小规模表达 followed by 大规模蛋白生产来优化人胚肾（HEK 293T）细胞中 Zika 病毒包膜糖蛋白的生产。HEK 293T 细胞的小规模表达允许同时筛选大量载体，以选择具有最佳分泌谱的载体，以便在 Expi293 哺乳动物系统中进行放大，以最大限度地提高蛋白产量，然后进行纯化，用于研究和临床应用。
