Transient Gene Expression in Suspension HEK293-EBNA1 Cells
Transient Gene Expression in Suspension HEK293-EBNA1 Cells
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DOI：
10.1007/978-1-4939-8730-6_1
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发表时间：
2018-01-01
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影响因子：
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通讯作者：
Durocher, Yves
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文献类型：
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作者：
L'Abbe, Denis;Bisson, Louis;Durocher, Yves
在人胚肾293（HEK 293）细胞中的瞬时基因表达是用于快速生产大量重组蛋白（r-蛋白）的既定方法。毫克至克量的r-蛋白通常可以在转染后不到10天内获得。在这一章中，我们描述了一个简单而强大的转染悬浮生长的人胚肾293细胞的过程中使用两种市售的无血清培养基和聚乙烯亚胺作为转染试剂。本章提供了组氨酸标记的重组蛋白和两种单克隆抗体的生产和纯化的例子。
Transient gene expression in human embryo kidney 293 (HEK293) cells is an established approach for the rapid production of large amounts of recombinant proteins (r-proteins). Milligram to gram quantities of r-proteins can be typically obtained within less than 10 days following transfection. In this chapter, we describe a simple and robust transfection process of suspension-growing human embryo kidney 293 cells using two commercially available serum-free media and polyethylenimine as the transfection reagent. This chapter provides examples for the production and purification of a his-tagged recombinant protein and two monoclonal antibodies.