COVID 19 Pseudoviral Particles, Indian B.1.617 Coronavirus Pseudovirus | COVID-19 pseudovirus

SARS-CoV-2 Pseudoviral Particles, Indian B.1.617 Variant

Cat. Num. AAA434306

• Synonyms: COVID 19 Pseudoviral Particles; Indian B.1.617 Coronavirus; SARS-CoV-2 Pseudoviral Particles; Indian B.1.617 Variant; 2019 Novel Coronavirus; Coronavirus; CoV; COVID-19 virus; HCoV-2; Human Coronavirus 2019; SARS2; SARS-CoV-2; Severe acute respiratory syndrome coronavirus 2; Pseudoviral Particles; COVID-19 pseudovirus

• Form/Format: 10 ml (2 tubes, 5 mL/tube), for 2 multi-well plates

• Application Notes: Our Pseudovirus Particles generate robust chemiluminescent signals in cell assays when coupled with our firefly luciferase assay kit (Catalog# MBS434279), useful for 1) screening potential inhibitor to block SARS-CoV-2 entry and viral protein translation; 2) measuring the activity of and screening for neutralizing antibody against SARS-CoV-2-IN.; ; ASSAY PROTOCOL I (Fig. 2 & 3); Note: requires a luciferase assay reagent. (Catalog# MBS434279); Cell Infection:; 1. Count HEK293-ACE2 cells (Catalog# MBS434276) to be infected and seed ~20K cells per well into 96-well plates (50 ul per well) DMEM with 10% FC II Serum (no antibiotics) or 5K cells per well into 384-well plates (15 ul per well).; 2. Culture cells overnight to make sure the cells stably adhere to the plates.; 3. On the 2nd day, remove media, add 50 ul SARS-CoV-2-IN pseudoviral particles into each well (12.5 ul for 384-well plate). Spin at 700 rpm for 15 min at 4°C.; Note: thaw the pseudoviral particles immediately before use (take about 1 hour to thaw in ice) and use it within 2 hours.; 4. Incubate for 2 hrs at 37°C.; 5. Add 50 ul DMEM with 10% FC into each well (12.5 ul for 384-well plates).; 6. Incubate for 48 hrs at 37 °C.; Measurement of Luciferase Activity in Infected cells; 1. Remove supernatant.; 2. Add 100 ul luciferase assay reagent (20 ul for 384-well plates).; 3. Read in a luminescence plate reader and record the data.; ; ASSAY PROTOCOL II (Fig. 4); Note: requires a luciferase assay reagent. (Catalog# MBS434279); Cell Infection:; 1. Count HEK293-ACE2 cells (Catalog # MBS434276) to be infected and seed ~7.5K cells per well into 384-well plates (15 ul per well) in DMEM with 10% FC II Serum (no antibiotics).; 2. On the 2nd day, dilute the antibodies (5X of the final concentrations) in DMEM with 10% FC II Serum (no antibiotics).; Note: for most antibodies, we recommend testing 11 different doses (1:3 serial dilutions), starting from 30 ug/ml or 10 ug/ml (final concentration in the well).; 3. Take 5 ul of the antibody solution and add into a compound plate containing 12.5 ul of SARS-CoV-2-PP pINer well (i.e. SARS-CoV-2-614G PP, Catalog # MBS434278).; 4. Incubate at 37°C for 1 hr.; 5. Take a cell plate out from the incubator, remove the medium, add 17.5 ul of SARS-CoV-2-PP and antibody mix into each well of the cell plate. Centrifuge at 700 rpm for 15 min at 4°C.; 6. Add additional 7.5 ul of DMEM with 10% FC into each well.; 7. Incubate at 37°C for 42 hr.; Measurement of Luciferase Activity in Infected cells; 1. Remove supernatant.; 2. Add 20 ul luciferase assay reagent into each well.; 3. Read in a luminescence plate reader and record the data.
• Features: Robust: Excellent signal to noise (basal) ratio; Easy to use: Amenable to HTS format (96-well, 384-well and 1536-well format)
• Shipping Note: MBS434306 is available for shipment in the United States, Canada and European countries. Please inquire for shipment to other countries.
• Dry Ice Shipment: Extra charge fee may add to your shipping cost as dry ice is required to ship this product.
• Preparation and Storage: Ships on dry ice. Store at -70°C.

Figure

(Figure 1. Illustration of the replication-deficient MLV particle pseudotyped with SARS-CoV-2 Spike protein)

Testing Data

(Figure 2. Pseudoviral Particle (PP) Infection AssaysSARS-CoV-2-IN variant pseudoviral particles on HEK293-ACE2 cells in 384-well formatLegends: SARS-CoV-2-India-PP: SARS-CoV-2 Indian B.1.617 variant MLV Pseudovirus Particles (MBS434306)(-): MLV control (w/o envelope spike protein) (MBS434280))

Testing Data

(Figure 3. SARS-CoV-2 Viral Infection Inhibiting Test by Neutralization Antibodies.HEK293-ACE2 cells incubated with SARS-CoV-2 India Variant Pseudoviral Particles (MBS434306) under various amount of neutralizing antibody.Legend: SARS-C0V2-India-PP, SARS-CoV-2 India variant Pseudoviral Particles (MBS434287)“Control”, neutralizing antibody used in this viral infection inhibition assay, MBS434300.)

Testing Data

(Figure 4. Antibody (Ab) Neutralization Assays with SARS-CoV-2-614G-PP)

Related Product Information for COVID-19 pseudovirus

SARS-CoV-2 Pseudoviral Particles, replication-deficient MLV pseudotyped with the SARS-CoV-2 spike protein (the Indian B.1.617 variant).

The SARS-CoV-2 Pseudoviral Particles are replication-deficient Maloneymurine leukemia virus (MLV) pseudotyped with the SARS-CoV-2 spike protein of the Indian B.1.617 variant (GISAID sequence accession number EPI_ISL_1547802).

It has been known that SARS coronavirus 2 (SARS-CoV-2) uses human ACE2 as entry receptor and human proteases as entry activators. The virus surface spike protein (S) mediates SARS-CoV-2 entry into cells. To fulfill its function, SARS-CoV-2 spike binds to its human ACE2 (hACE2) receptor through its receptor-binding domain (RBD) and is proteolytically activated by human proteases. Our SARS-CoV-2-IN Pseudoviral Particles are replication-deficient MLV pseudotyped with the SARS-CoV-2 spike protein of the Indian variant B.1.617, also known as G/452R.V3, (GISAID sequence accession number EPI_ISL_1547802). They also contain the ORF for firefly luciferase as a reporter. They establish a pseudovirus entry assay for SARS-CoV-2 as the spike protein mediated cell entry can be conveniently measured via luciferase reporter activity. This pseudovirus assay isolates the SARS-CoV-2 viral entry from other steps of the viral infection cycle.