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Development of an Optimized Process for Functional Recombinant SARS-CoV-2 Spike S1 Receptor-Binding Domain Protein Produced in the Baculovirus Expression Vector System
Boumaiza, M.; Chaabene, A.; Akrouti, I.; Ben Zakour, M.; Askri, H.; Salhi, S.; Ben Hamouda, W.; Marzouki, S.; Benabdessalem, C.; Ben Ahmed, M.; Trabelsi, K.; Rourou, S. Development of an Optimized Process for Functional Recombinant SARS-CoV-2 Spike S1 Receptor-Binding Domain Protein Produced in the Baculovirus Expression Vector System. Trop. Med. Infect. Dis.2023, 8, 501.
Boumaiza, M.; Chaabene, A.; Akrouti, I.; Ben Zakour, M.; Askri, H.; Salhi, S.; Ben Hamouda, W.; Marzouki, S.; Benabdessalem, C.; Ben Ahmed, M.; Trabelsi, K.; Rourou, S. Development of an Optimized Process for Functional Recombinant SARS-CoV-2 Spike S1 Receptor-Binding Domain Protein Produced in the Baculovirus Expression Vector System. Trop. Med. Infect. Dis. 2023, 8, 501.
Boumaiza, M.; Chaabene, A.; Akrouti, I.; Ben Zakour, M.; Askri, H.; Salhi, S.; Ben Hamouda, W.; Marzouki, S.; Benabdessalem, C.; Ben Ahmed, M.; Trabelsi, K.; Rourou, S. Development of an Optimized Process for Functional Recombinant SARS-CoV-2 Spike S1 Receptor-Binding Domain Protein Produced in the Baculovirus Expression Vector System. Trop. Med. Infect. Dis.2023, 8, 501.
Boumaiza, M.; Chaabene, A.; Akrouti, I.; Ben Zakour, M.; Askri, H.; Salhi, S.; Ben Hamouda, W.; Marzouki, S.; Benabdessalem, C.; Ben Ahmed, M.; Trabelsi, K.; Rourou, S. Development of an Optimized Process for Functional Recombinant SARS-CoV-2 Spike S1 Receptor-Binding Domain Protein Produced in the Baculovirus Expression Vector System. Trop. Med. Infect. Dis. 2023, 8, 501.
Abstract
To map Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV 2) spreading and evaluate immune responses variations against this virus, it was essential to locally set up efficient serological tests. The SARS-CoV2 immunogenic proteins were very expensive and not affordable mainly for Low and Middle Income Countries (LMICs). For this purpose, the commonly used antigen, Re-ceptor-Binding Domain (RBD) of Spike S1 protein (S1RBD), was produced using the the Baculovirus Expression Vector System (BEVS) . During the current study, the expression of S1RBD was monitored using western blot under different culture conditions. Different parameters were studied: Multiplicity Of Infection (MOI), cell density at infection and harvest time. Hence, optimal conditions for efficient S1RBD production were identified: MOI 3; cell density at infection 2-3x 106 cells/mL and time post-infection (tpi or harvest time) of 72h and 72-96 h successively for expression in shake-flasks and 7L-bioreactor. A high production yield of S1RBD varying between 4mg and 70 mg per liter of crude cell culture supernatant was achieved, respectively, in shake-flasks and in 7L-bioreactor. Moreover, the produced S1RBD showed an excellent antigenicity potential against COVID-19 patient sera evaluated by western blot. Thus, additional serological assays, such as ELISA, were developed using the purified S1RDB (published by other research groups).
Keywords
Multiplicity of infection; cell density at infection; SARS-CoV-2; Spike S1 glycoprotein; Receptor binding domain; Sf9 cells
Subject
Biology and Life Sciences, Biology and Biotechnology
Copyright:
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