The successful expression of a potent recombinant scFvantibody against the SARS-CoV-۲

Background and Aim : There are many platforms for expression of recombinant proteins. Amongthis, Escherichia coli has been successfully used for production of recombinant proteins since۱۹۸۰s, due to the ease of manipulation, low costs, the rapid growth, etc. In this regard and basedon the importance of antibodies against the SARS-CoV-۲, the main goal of the current study wasstudy on the expression of a recombinant scFv against the SARS-CoV-۲'s RBD which recentlydesigned based on the in-depth bioinformatics studies.Methods : For study on the anti-RBD scFv expression, the expression of pET۲۸a (+)-scFv vectorwas performed using E. coli BL۲۱. The time-course studies of induction by ۰.۵mM and ۱mM ofIPTG was done and periplasmic proteins' extraction was performed. The results were analyzedusing SDS-PAGE and recombinant protein expression was verified using western blotting (WB).The protein's purification and study on its efficiency is doing.Results : Comparing the expression results in various times after induction showed the bestinduction results in presence of ۱mM IPTG and the successful expression in all hours ranging from۲-۲۴hrs after induction. The WB analysis showed the specific single-band of anti-RBD scFv(~۳۰kDa) on the nitrocellulose membrane.Conclusion : The production of high quality recombinant therapeutical proteins is essential fortheir usage for humankinds. In this study, we successfully produced a potent recombinant scFv byE. coli BL۲۱ expression system in relatively high amounts and good stability.

کلیدواژه ها:

SARS-CoV-۲ ، recombinant scFv antibody ، RBD ، Escherichia coli ، Expression ، Western blotting