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