High-yield production in Escherichia coli and convenient purification of a candidate vaccine against SARS-CoV-2.

ABSTRACT:

Objectives

The aim of the present work was to identify a time-saving, effective, and low-cost strategy to produce in Escherichia coli a protein chimera representing a fusion anti-SARS-CoV-2 candidate vaccine, consisting of immunogenic and antigenic moieties.

Results

We overexpressed in E. coli BL21(DE3) a synthetic gene coding for CRM197-RBD, and the target protein was detected in inclusion bodies. CRM197-RBD was solubilized with 1 % (w/v) of the anionic detergent N-lauroylsarcosine (sarkosyl), the removal of which from the protein solution was conveniently accomplished with Amberlite XAD-4. The detergent-free CRM197-RBD was then separated from contaminating DNA using polyethylenimine (PEI), and finally purified from PEI by salting out with ammonium sulfate. Structural (CD s

SUBMITTER: Maltoni G 

PROVIDER: S-EPMC9512991 | biostudies-literature | 2022 Nov

REPOSITORIES: biostudies-literature