Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Development of an Optimized Process for Functional Recombinant SARS-CoV-2 Spike S1 Receptor-Binding Domain Protein Produced in the Baculovirus Expression Vector System

Mohamed Boumaiza
ORCID logo ,
Amani Chaabene
,
Ines Akrouti
,
Meriem Ben Zakour
,
Hana Askri
,
Said Salhi
,
Chaouki Benabdessalem
ORCID logo ,
Melika Ben Ahmed
ORCID logo ,
Khaled Trabelsi
,
Samia Rourou
*
Version 1 : Received: 22 August 2023 / Approved: 22 August 2023 / Online: 23 August 2023 (07:53:03 CEST)

A peer-reviewed article of this Preprint also exists.

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: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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