Rbd pseudotyped lentivirus

Manufactured by VectorBuilder

Sourced in United States

RBD-pseudotyped lentivirus is a laboratory tool used to study SARS-CoV-2 receptor binding domain (RBD) interactions. It is a lentiviral vector that has been engineered to display the SARS-CoV-2 RBD on its surface, allowing for the investigation of RBD binding and entry mechanisms.

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Lab products found in correlation

Polybrene, by VectorBuilder (2 mentions) Human ace2 mcherry construct, by VectorBuilder (2 mentions) Sp8 confocal microscope, by Leica (2 mentions) Sars cov 2 rbd protein, by Sino Biological (1 mentions) Ace2 sars cov 2 spike inhibitor screening assay kit, by BPS Biosciences (1 mentions) Methyl thiazolyl tetrazolium (mtt), by Thermo Fisher Scientific (1 mentions)

3 protocols using rbd pseudotyped lentivirus

Screening Assay for ACE2-SARS-CoV-2 Spike Inhibitors

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Corilagin (purity > 99%) was purchased from Chenguang Biological Technology Co., Ltd (Baoji, China) and was also verified by NMR and UPLC-UV (Fig. S1-3). Its chemical structure was shown in the Fig. 1. DMSO was supported by Sigma-Aldrich (St. Louis, MO, USA). MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumromide) powder was obtained from Thermo Fisher (Waltham, MA, USA). FluorSave reagent was purchased from Calbiochem (San Diego, CA, USA). The ACE2: SARS-CoV-2 Spike Inhibitor Screening Assay Kit (Cat: #79936) was purchased from BPS Biosciences (San Diego, CA, USA). Purified ACE2-His tag protein and SARS-CoV-2 RBD protein were supported by Sino Biological (Beijing, China). ACE2-EGFP plasmid, ACE2-mCherry construct and RBD-pseudotyped lentivirus were purchased from Vectorbuilder (Chicago, IL, USA).Fig. 1

Chemical structure of corilagin.

Fig 1

Yang L.J., Chen R.H., Hamdoun S., Coghi P., Ng J.P., Zhang D.W., Guo X., Xia C., Law B.Y, & Wong V.K. (2021). Corilagin prevents SARS-CoV-2 infection by targeting RBD-ACE2 binding. Phytomedicine, 87, 153591.

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ACE2-Mediated SARS-CoV-2 Infection Assay

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HEK 293 cells were seeded in 100 mm culture dish and cultured overnight. The cells were transfected with human ACE2/mCherry construct (Vectorbuilder Inc.) for 24 h. After that, 1×10⁵ transfected cells were seeded on the cover glasses of 24-well plate. The next day, PGG, RBD-pseudotyped lentivirus (Vectorbuilder Inc.) and polybrene (Vectorbuilder Inc.) were premixed in blank medium, and then added into the cells. After 12h, the medium was replaced with fresh FBS-medium and continually incubated for 48 h. The cells were then washed 3 times with PBST and then fixed with 4% PFA. The cover glasses were mounted with FluorSave reagent (Calbiochem, United States). The cells were imaged by Leica SP8 confocal microscope. For semi-quantitative determination, cells were cultured as described before. Briefly, a standard calibration curve of viral infection using 46.6, 37.3, 23.3, 4.66, 0 ×10⁶ TU of RBD-pseudotyped lentivirus were established. 3 points of cells fluorescence intensity for every titer of viral infection were calculated by Image J. Calibration curve was established for the evaluation of virus inhibition. Quantification bar chart represents the data from 5 independent experiments.
Viral infection formula: y = 286200x - 98,383, R² = 0.9729.

Chen R.H., Yang L.J., Hamdoun S., Chung S.K., Lam C.W., Zhang K.X., Guo X., Xia C., Law B.Y, & Wong V.K. (2021). 1,2,3,4,6-Pentagalloyl Glucose, a RBD-ACE2 Binding Inhibitor to Prevent SARS-CoV-2 Infection. Frontiers in Pharmacology, 12, 634176.

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Corilagin Inhibits SARS-CoV-2 Infection

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HEK293 cells were transfected with human ACE2-mCherry construct (Vectorbuilder) for 24 h. After that, 1 × 10⁵ transfected cells were seeded overnight on a coverslip inside a 24-well plate. Corilagin, RBD-pseudotyped lentivirus (Vectorbuilder) and polybrene (Vectorbuilder) were premixed in medium before being added into the cells. After 12 h, the cells were replaced with normal medium and incubated for a further 48 h. The cells were then washed with PBST and fixed with 4% PFA. The coverslips with cells were mounted with FluorSave reagent (Calbiochem). Cells were visualized by Leica SP8 confocal microscope and then subjected to semi-quantitative determination as described before. In addition, a standard calibration curve on viral infection using 46.6, 37.3, 23.3, 4.66, 0 × 10⁶ TU of RBD-pseudotyped lentivirus were established for the evaluation of virus inhibition. Fluorescence intensity of cells for every titer of viral infection were collected and then calculated by Image J. Quantification bar chart represented the data from 3 independent experiments.
Viral infection formula: y = 286200x - 98383, R² = 0.9729

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