A DNA sequence encoding SARS-CoV-2 RBD Omicron BA.5 was designed with His-tag at the N-terminal for protein purification. The synthesized gene was inserted into pET15b at the NdeI/BamH1 with ampicillin resistance (Supplementary Figure S1a ). The RBD was expressed in E. coli T7 SHuffle cells and purified from inclusion bodies as explained elsewhere [18 (link)]. The RBD proteins tagged by 6-Histidines were purified using denaturing open nickel-nitrilotriacetic acid (Ni-NTA) (Wako, Japan) chromatography. The proteins were further purified using reversed-phase (RP)-HPLC. The purified RBD were lyophilized and stored at –30°C.
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Purification of SARS-CoV-2 Omicron BA.5 RBD
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Corresponding Organization :
Other organizations : Tokyo University of Agriculture and Technology, Yokohama City University
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Variable analysis
independent variables
- Protein expression system (E. coli T7 SHuffle cells)
- Protein purification yield
- Protein purity
- Plasmid vector (pET15b)
- Antibiotic resistance (ampicillin)
- Protein tag (His-tag)
- Protein purification methods (denaturing open nickel-nitrilotriacetic acid (Ni-NTA) chromatography, reversed-phase (RP)-HPLC)
- Protein storage conditions (-30°C)
- Positive control: Not explicitly mentioned.
- Negative control: Not explicitly mentioned.
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