| Autor: |
Mozafar Mohammadi, Ramezan Ali Taheri, Peyman Bemani, Mohammad Sadegh Hashemzadeh, Gholamreza Farnoosh, Razieh Amini |
| Rok vydání: |
2022 |
| Předmět: |
|
| Zdroj: |
Protein and peptide letters. 29(7) |
| ISSN: |
1875-5305 |
| Popis: |
Background: SARS-CoV-2 uses angiotensin-converting enzyme 2 (ACE2) as a receptor for entering the host cells. Production of the ACE2 molecule is important because of its potency to use as a blocker and therapeutic agent against SARS-CoV-2 for the prophylaxis and treatment of COVID-19. Objective: The recombinant human ACE2 (rhACE2) is prone to form an inclusion body when expressed in the bacterial cells. Method: We used the SUMO tag fused to the rhACE2 molecule to increase the expression level and solubility of the fusion protein. Afterward, the freeze-thawing method plus 2 M urea solubilized aggregated proteins. Subsequently, the affinity of solubilized rhACE2 to the receptor binding domain (RBD) of the SARS-CoV-2 spike was assayed by ELISA and SPR methods. Results: SUMO protein succeeded in increasing the expression level but not solubilization of the fusion protein. The freeze-thawing method could solubilize and recover the aggregated fusion proteins significantly. Also, ELISA and SPR assays confirmed the interaction between solubilized rhACE2 and RBD with high affinity. Conclusion: The SUMO tag and freeze-thawing method would be utilized for high-level expression and solubilization of recombinant rhACE2 protein. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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