Volume - 13 | Issue-1
Volume - 13 | Issue-1
Volume - 13 | Issue-1
Volume - 13 | Issue-1
Volume - 13 | Issue-1
Severe Acute Respiratory Syndrome coronavirus-2 (SARS-CoV-2) is the virus responsible for causing COVID-19. The virus has a homotrimer S protein, which is a glycoprotein essential to the early stage of infection. The Spike protein has 1273 amino acids and a molecular weight of approximately 142.6 kDa. In this study, the Spike protein was expressed and purified from Escherichia coli using pGBW-m4046887, which was purchased from Addgene. Due to codon bias in the conventional Escherichia coli BL21 (DE3) strain, pGBW-m4046887 was transformed in Escherichia coli BL21-CodonPlus (DE3)-RP and grown in colonies with 50 μg/mL of chloramphenicol at 30℃. The Spike protein was induced using 0.5 mM IPTG overnight at 18℃, and the expressed protein was purified using immobilized metal affinity chromatography (IMAC) since it contains a 6xHis tag at the C-terminus end. After protein induction, samples were analyzed using SDS-PAGE, and predominant protein bands were observed between 100 and 130 kDa. However, protein at the expected molecular weight was not observed, which could be attributed to codon bias. In the future, protein expression could be performed in the eukaryotic expression system to overcome this.