Expression and Purification of Functional SARS-CoV-2 RBD in E. coli for Therapeutic and Diagnostic Purposes DOI Creative Commons
Hajarossadat Ghaderi, Alireza Shoari, Shima Salehi

et al.

Research Square (Research Square), Journal Year: 2023, Volume and Issue: unknown

Published: Jan. 17, 2023

Abstract SARS-CoV-2 causes a severe respiratory disease known as COVID-19 and is responsible for global viral pandemic. The receptor binding domain (RBD) located on the spike protein (S), which dedicated identifying to angiotensin converting enzyme 2 (ACE2) receptor. RBD an important target development of virus neutralizing antibodies, vaccines, inhibitors. In this study, recombinant was expressed in E. coli BL21 (DE3) purified well its activity determined. Purification conducted by Ni-NTA column. ELISA flow cytometry assays were evaluate ability different anti-RBD antibodies native ACE2 HEK293A cells, respectively. results showed that produced human sera could bind commercial antibody. Also, analysis able surface cells. Our outcomes displayed strain has biological can be used antigen diagnosis kits vaccines tool screening drugs against SASR-CoV-2.

Language: Английский

Enhancement of exogenous protein stability in AcMNPV by overexpressing lef5 gene during passaging DOI Creative Commons
Jie Pei,

Rui-Lun Liu,

Li W

et al.

Applied Microbiology and Biotechnology, Journal Year: 2025, Volume and Issue: 109(1)

Published: March 24, 2025

The baculovirus insect cell expression system is pivotal for exogenous protein expression. However, serial passages of in cells often result defective virus generation and a rapid decline expression, limiting its wider application. Previous studies have shown that the late factor 5 (lef5) from other baculoviruses can enhance stability genes Autographa californica multiple nucleopolyhedrovirus (AcMNPV). In this study, we engineered diverse recombinant strains AcMNPV to express enhanced green fluorescent (eGFP) enterovirus 71 virus-like particles (EV71-VLPs), co-expressed with P1 3CD proteins EV71. We investigated influence lef5 overexpression, regulated by various promoters, on their expressions. Notably, overexpression significantly improved eGFP EV71-P1 gene expressions during Sf9 cells. Specifically, driven op166 promoter was more beneficial enhancing complex proteins, such as EV71-VLPs. This study underscores importance maintaining systems, particularly producing VLPs enterovirus. • stabilizes system. Promoter choice affects lef5-mediated stability. crucial like

Language: Английский

Citations

0
Effective expression and characterization of the receptor binding domains in SARS-CoV-2 Spike proteins from original strain and variants of concern using Bombyx mori nucleopolyhedrovirus in silkworm DOI
Akira Tsukamoto,

Lee Jae Man,

Kosuke Oyama

et al.

Protein Expression and Purification, Journal Year: 2024, Volume and Issue: 218, P. 106450 - 106450

Published: Feb. 22, 2024

Language: Английский

Citations

2
Diverse Approaches to Express Recombinant Spike Protein: A Comprehensive Review DOI

Jk Nithya Shree,

T Premika,

S Sharlin

et al.

Protein Expression and Purification, Journal Year: 2024, Volume and Issue: 223, P. 106556 - 106556

Published: Nov. 1, 2024

Language: Английский

Citations

1
Improved Expression of SARS-CoV-2 Spike RBD Using the Insect Cell-Baculovirus System DOI Creative Commons

Joaquín Poodts,

Ignacio Smith,

Joaquín Manuel Birenbaum

et al.

Viruses, Journal Year: 2022, Volume and Issue: 14(12), P. 2794 - 2794

Published: Dec. 15, 2022

Insect cell-baculovirus expression vector system is one of the most established platforms to produce biological products, and it plays a fundamental role in context COVID-19 emergency, providing recombinant proteins for treatment, diagnosis, prevention. SARS-CoV-2 infection mediated by interaction spike glycoprotein trimer via its receptor-binding domain (RBD) with host's cellular receptor. As RBD required many applications, pandemic important meet challenge producing high amount (rRBD). For this reason, present study, we developed process based on Sf9 insect cells improve rRBD yield. was recovered from supernatant infected easily purified metal ion affinity chromatography, yield 82% purity higher than 95%. Expressed under novel chimeric promoter (polh-pSeL), after purification 21.1 ± 3.7 mg/L, which highest performance described cell lines. Finally, successfully used an assay detect specific antibodies serum samples. The efficient strategy herein has potential high-quality line diagnostic purpose.

Language: Английский

Citations

5
Expression and purification of SARS-CoV-2 receptor binding domain in Escherichia coli for diagnostic and therapeutic purposes DOI Creative Commons
Hajarossadat Ghaderi, Alireza Shoari, Shima Salehi

et al.

Research in Pharmaceutical Sciences, Journal Year: 2024, Volume and Issue: 19(5), P. 500 - 508

Published: Sept. 1, 2024

SARS-CoV-2 causes a severe respiratory disease known as COVID-19 and is responsible for global viral pandemic. The receptor binding domain (RBD) located on the spike protein, which identifies binds to angiotensin-converting enzyme 2 (ACE2) receptor. RBD an important target developing virus-neutralizing antibodies, vaccines, inhibitors.

Language: Английский

Citations

0
Expression and Purification of Functional SARS-CoV-2 RBD in E. coli for Therapeutic and Diagnostic Purposes DOI Creative Commons
Hajarossadat Ghaderi, Alireza Shoari, Shima Salehi

et al.

Research Square (Research Square), Journal Year: 2023, Volume and Issue: unknown

Published: Jan. 17, 2023

Abstract SARS-CoV-2 causes a severe respiratory disease known as COVID-19 and is responsible for global viral pandemic. The receptor binding domain (RBD) located on the spike protein (S), which dedicated identifying to angiotensin converting enzyme 2 (ACE2) receptor. RBD an important target development of virus neutralizing antibodies, vaccines, inhibitors. In this study, recombinant was expressed in E. coli BL21 (DE3) purified well its activity determined. Purification conducted by Ni-NTA column. ELISA flow cytometry assays were evaluate ability different anti-RBD antibodies native ACE2 HEK293A cells, respectively. results showed that produced human sera could bind commercial antibody. Also, analysis able surface cells. Our outcomes displayed strain has biological can be used antigen diagnosis kits vaccines tool screening drugs against SASR-CoV-2.

Language: Английский

Citations

0