COVID-19 Omicron Variant (B.1.1.529) Coronavirus Pseudovirus | COVID-19 pseudovirus

SARS-CoV-2 Pseudoviral Particles, Omicron Variant (B.1.1.529)

Cat. Num. AAA434315

• Synonyms: COVID-19 Omicron Variant (B.1.1.529) Coronavirus; SARS-CoV-2 Pseudoviral Particles; Omicron Variant (B.1.1.529); 2019 Novel Coronavirus; Coronavirus; CoV; COVID-19 virus; HCoV-2; Human Coronavirus 2019; SARS2; SARS-CoV-2; Severe acute respiratory syndrome coronavirus 2; COVID-19 pseudovirus

• Application Notes: Our Pseudovirus Particles (PP) 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-Omicron.; ; 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-Omicron 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 quickly in the room temperature water (< 30 minutes, do not shake) and use right away. Discard the unused portion (do not re-freeze or leave it on ice for later use).; 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 42 hrs at 37°C.; Measurement of Luciferase Activity in Infected cells; 1. Do not remove supernatant. Add 100 ul luciferase assay reagent (25 ul for 384-well plates) directly into each well.; 2. 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)
• Contents: 10 ml (2 tubes, 5 mL/tube), for 2 multi-well plates; PP per mL > 1.0E+07
• Dry Ice Shipment: Extra charge fee may add to your shipping cost as dry ice is required to ship this product.
• Preparation and Storage: Upon receipt, store at -70°C right away. Thaw* before immediate use.; *Note: read the instruction for thawing in the following protocol carefully. Do not aliquot and refreeze.; ; Shelf Life: Six months from the date of receipt when stored 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 assays (SARS-CoV-2-Omicron variant pseudoviral particles on HEK293-ACE2 cells)Legends: SARS-CoV-2 Omicron variant MLV pseudovirus particles (MBS434315) (-): MLV control (w/o envelope spike protein) (Catalog # MBS434280))

Testing Data

(Figure 3. Antibody neutralization assays using Omicron pseudoviral particle (PP) infection method. Each antibody dose-response curve represents the relative infectivity under serial dilutions of antibody. At each Ab concentration, HEK293-ACE2 cells were co-incubated with SARS-CoV-2 Omicron pseudoviral particles in the presence of the antibody. Among the 3 antibodies tested, Ab1 and Ab2 showed good neutralization capacity towards Omicron spike.)

Testing Data

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

Related Product Information for COVID-19 pseudovirus

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-Omicron Pseudoviral Particles are replication-deficient MLV pseudotyped with the SARS-CoV-2 spike protein (Accession ID: EPI_ISL_6699757) of the Omicron variant, originally known as lineage B.1.1.529. They also contain the ORF for firefly luciferase as a reporter. They establish a pseudovirus cell entry assay mediated by the SARS-CoV-2 spike protein that 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.