Protein Scaffold‐Based Multimerization of Soluble ACE2 Efficiently Blocks SARS‐CoV‐2 Infection In Vitro and In Vivo

Kayabölen, Alişan; Akcan, Uğur; Özturan, Doğancan; Polat, Hivda Ulbeği; Şahin, Gizem Nur; Degirmenci, Nareg Pinarbasi; Bayraktar, Canan; Söyler, Gizem; Sarayloo, Ehsan; Nurtop, Elif; Özer, Berna; Güney-Esken, Gülen; Barlas, Tayfun; Yildirim, Ismail Selim; Doğan, Özlem; Karahüseyinoğlu, Serçin; Lack, Nathan A.; Kaya, Mehmet; Albayrak, Cem; Can, Füsun; Solaroğlu, İhsan; Bagcı-Önder, Tugba

doi:10.1002/advs.202201294

Cited by 13 publications

(25 citation statements)

References 81 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…At 10 dpi, 7 of the ten animals in the control group had significant weight loss (> 25%) and were euthanized in the final days of the experiment because three mice in this group were dying. Vaccinated animals completed the experiment without losing weight 29 . 12 days after the challenge, lung tissues of all animals were harvested at necropsy and analyzed.…”

Section: Resultsmentioning

confidence: 99%

Process development for an effective COVID-19 vaccine candidate harboring recombinant SARS-CoV-2 delta plus receptor binding domain produced by Pichia pastoris

Kalyoncu

Yılmaz

Kuyucu

et al. 2023

Sci Rep

View full text Add to dashboard Cite

Recombinant protein-based SARS-CoV-2 vaccines are needed to fill the vaccine equity gap. Because protein-subunit based vaccines are easier and cheaper to produce and do not require special storage/transportation conditions, they are suitable for low-/middle-income countries. Here, we report our vaccine development studies with the receptor binding domain of the SARS-CoV-2 Delta Plus strain (RBD-DP) which caused increased hospitalizations compared to other variants. First, we expressed RBD-DP in the Pichia pastoris yeast system and upscaled it to a 5-L fermenter for production. After three-step purification, we obtained RBD-DP with > 95% purity from a protein yield of > 1 g/L of supernatant. Several biophysical and biochemical characterizations were performed to confirm its identity, stability, and functionality. Then, it was formulated in different contents with Alum and CpG for mice immunization. After three doses of immunization, IgG titers from sera reached to > 106 and most importantly it showed high T-cell responses which are required for an effective vaccine to prevent severe COVID-19 disease. A live neutralization test was performed with both the Wuhan strain (B.1.1.7) and Delta strain (B.1.617.2) and it showed high neutralization antibody content for both strains. A challenge study with SARS-CoV-2 infected K18-hACE2 transgenic mice showed good immunoprotective activity with no viruses in the lungs and no lung inflammation for all immunized mice.

show abstract

Section: Resultsmentioning

confidence: 99%

Process development for an effective COVID-19 vaccine candidate harboring recombinant SARS-CoV-2 delta plus receptor binding domain produced by Pichia pastoris

Kalyoncu

Yılmaz

Kuyucu

et al. 2023

Sci Rep

View full text Add to dashboard Cite

show abstract

“…This encouraged experts to produce a stable mutant dimeric and a homo-trimeric variant of sACE2 with preserved peptidase activity but higher binding affinity to RBD (about 600 pM for the dimeric and 60 pM for the trimeric sACE2) to hinder mACE2-RBD complexification [ 257 , 259 ]. In order to block S proteins on each SARS-CoV-2 particle with high efficiency, multimerized sACE2-fusion proteins have been manufactured through molecular engineering [ 260 ].…”

Section: Sace2: Increasing Sars-cov-2 Infectivity?mentioning

confidence: 99%

S Protein, ACE2 and Host Cell Proteases in SARS-CoV-2 Cell Entry and Infectivity; Is Soluble ACE2 a Two Blade Sword? A Narrative Review

2023

View full text Add to dashboard Cite

Since the spread of the deadly virus SARS-CoV-2 in late 2019, researchers have restlessly sought to unravel how the virus enters the host cells. Some proteins on each side of the interaction between the virus and the host cells are involved as the major contributors to this process: (1) the nano-machine spike protein on behalf of the virus, (2) angiotensin converting enzyme II, the mono-carboxypeptidase and the key component of renin angiotensin system on behalf of the host cell, (3) some host proteases and proteins exploited by SARS-CoV-2. In this review, the complex process of SARS-CoV-2 entrance into the host cells with the contribution of the involved host proteins as well as the sequential conformational changes in the spike protein tending to increase the probability of complexification of the latter with angiotensin converting enzyme II, the receptor of the virus on the host cells, are discussed. Moreover, the release of the catalytic ectodomain of angiotensin converting enzyme II as its soluble form in the extracellular space and its positive or negative impact on the infectivity of the virus are considered.

show abstract

“…In the treated group, histological examination of lung tissues revealed healing and restoration of normal tissue components (Figure 3C). [35][36][37] Lung sections were analyzed after staining with hematoxylin and eosin and lung inflammation was semi-quantitatively scored between 0 and 3 (0: no inflammation, 1: low, 2: medium, 3: high). Control mice exhibited significantly higher histopathology scores than extract treated mice.…”

Section: In Vivo Experiments With S Lappa Extractsmentioning

confidence: 99%

“…Only one of the 10 animals in the treated group given S. lappa extract showed this condition (Figure 3B). [34][35][36][37] One out of 10 animals in the group 1 that was treated with S. lappa extract developed disease at 9 dpi, while nine animals (90%) were completely healthy and survived until the end of the experiment. Gross pathology showed that most of the lungs in the control group had pneumonic and hyperemic areas, while the lungs in the treated group were clean and showed no signs of infection.…”

Section: In Vivo Experiments With S Lappa Extractsmentioning

confidence: 99%

Investigation of the Antiviral Effects of Saussurea lappa Root Extract against SARS‐CoV‐2 Virus

Ulbegi Polat,

Serhatli,

Taş Ekiz

et al. 2023

Molecular Nutrition Food Res

Self Cite

View full text Add to dashboard Cite

Scope The purpose of this study was to look into the antiviral activity of a plant extract derived from the roots of the Saussurea lappa as a food supplement against SARS‐CoV‐2 infection. Methods and results Vero E6 cells are employed in the study to test the neutralizing effect of Saussurea lappa extract against the SARS‐CoV‐2 virus. For anti‐viral activity detection, a sensitive real‐time cell analyzer (xCELLigence RTCA) with a high repetition rate is used. A challenge experiment in mice is planned as a result of the in vitro analysis. A challenge test against SARS‐CoV‐2 is performed with 10 adult female K18‐hACE2 transgenic mice in each group for this purpose. The mice in the S. lappa Group are gavaged 2 days before the virus is administered intranasally (i.n.). The control group received PBS instead of the extract. SARS‐CoV‐2 virus is administered i.n. under anesthesia for the first 3 days of the experiment, and S. lappa extract was administered by gavage in the afternoon. On the 10th day, mice in the S. lappa group survived the study, whereas animals in the control group grew ill and/or died. In this study, the extract protects the mice against the SARS‐CoV‐2 virus in 90% of the cases. Conclusions This study demonstrates that the Saussurea plant has antiviral effects against SARS‐CoV‐2 in vitro and in animal models.

show abstract

Protein Scaffold‐Based Multimerization of Soluble ACE2 Efficiently Blocks SARS‐CoV‐2 Infection In Vitro and In Vivo

Cited by 13 publications

References 81 publications

Process development for an effective COVID-19 vaccine candidate harboring recombinant SARS-CoV-2 delta plus receptor binding domain produced by Pichia pastoris

Process development for an effective COVID-19 vaccine candidate harboring recombinant SARS-CoV-2 delta plus receptor binding domain produced by Pichia pastoris

S Protein, ACE2 and Host Cell Proteases in SARS-CoV-2 Cell Entry and Infectivity; Is Soluble ACE2 a Two Blade Sword? A Narrative Review

Investigation of the Antiviral Effects of Saussurea lappa Root Extract against SARS‐CoV‐2 Virus

Contact Info

Product

Resources

About