An Optimized Method for the Production Using PEI, Titration and Neutralization of SARS-CoV Spike Luciferase Pseudotypes
An Optimized Method for the Production Using PEI, Titration and Neutralization of SARS-CoV Spike Luciferase Pseudotypes
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DOI：
10.21769/bioprotoc.2514
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发表时间：
2017-08-20
期刊：
影响因子：
0.8
通讯作者：
Temperton, Nigel
中科院分区：
文献类型：
--
作者：
Carnell, George;Grehan, Keith;Temperton, Nigel
概述的方案代表了使用广泛可用的转染试剂PEI在慢病毒载体核心上生成具有野生型SARS-CoV刺突蛋白的高滴度病毒伪型颗粒的成本效益，快速和可靠的方法。该方案针对6孔板转染进行了优化；然而，它可以很容易地根据应用扩展到不同的产量。该方案具有多种优点，包括使用现成的试剂，一致的高伪病毒产生相对发光单位（RLU）滴度和快速生成新型冠状病毒伪病毒，用于研究菌株变异，向性和免疫原性/血清患病率。
The protocol outlined represents a cost-effective, rapid and reliable method for the generation of high-titre viral pseudotype particles with the wild-type SARS-CoV spike protein on a lentiviral vector core using the widely available transfection reagent PEI. This protocol is optimized for transfection in 6-well plates; however it can be readily scaled to different production volumes according to application. This protocol has multiple benefits including the use of readily available reagents, consistent, high pseudotype virus production Relative Luminescence Units (RLU) titres and rapid generation of novel coronavirus pseudotypes for research into strain variation, tropism and immunogenicity/sero-prevalence.