Your browser doesn't support javascript.
Transient transfection and purification of SARS-CoV-2 spike protein from mammalian cells.
Stalls, Victoria; Janowska, Katarzyna; Acharya, Priyamvada.
  • Stalls V; Duke Human Vaccine Institute, Durham, NC 27710, USA. Electronic address: victoria.stalls@duke.edu.
  • Janowska K; Duke Human Vaccine Institute, Durham, NC 27710, USA. Electronic address: katarzyna.janowska@duke.edu.
  • Acharya P; Duke Human Vaccine Institute, Durham, NC 27710, USA; Department of Surgery, Duke University, Durham, NC 27710, USA. Electronic address: priyamvada.acharya@duke.edu.
STAR Protoc ; 3(3): 101603, 2022 09 16.
Article in English | MEDLINE | ID: covidwho-1937316
ABSTRACT
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).
Subject(s)
Keywords

Full text: Available Collection: International databases Database: MEDLINE Main subject: Spike Glycoprotein, Coronavirus / COVID-19 Limits: Animals / Humans Language: English Journal: STAR Protoc Year: 2022 Document Type: Article

Similar

MEDLINE

...
LILACS

LIS


Full text: Available Collection: International databases Database: MEDLINE Main subject: Spike Glycoprotein, Coronavirus / COVID-19 Limits: Animals / Humans Language: English Journal: STAR Protoc Year: 2022 Document Type: Article