Transient transfection and purification of SARS-CoV-2 spike protein from mammalian cells.

Journal Article (Journal Article)

SARS-CoV-2 spike (S) protein ectodomain purification can be challenging, with engineered and natural variations often resulting in lower yields. Here, we present a detailed transfection and purification protocol for the SARS-CoV-2 S ectodomain. We describe how to trace protein yields during purification using highly sensitive and characteristic changes in S ectodomain intrinsic fluorescence upon thermal denaturation. Additionally, we detail several optimized aspects of the purification including timing and temperature. This protocol facilitates consistent, high-quality preparations of the SARS-CoV-2 S ectodomain. For complete details on the use and execution of this protocol, please refer to Stalls et al. (2022), Gobeil et al. (2022), Edwards et al. (2021), and Henderson et al. (2020).

Full Text

Duke Authors

Cited Authors

  • Stalls, V; Janowska, K; Acharya, P

Published Date

  • September 16, 2022

Published In

Volume / Issue

  • 3 / 3

Start / End Page

  • 101603 -

PubMed ID

  • 35983170

Pubmed Central ID

  • PMC9283606

Electronic International Standard Serial Number (EISSN)

  • 2666-1667

Digital Object Identifier (DOI)

  • 10.1016/j.xpro.2022.101603

Language

  • eng

Conference Location

  • United States