|Product Description||SARS-CoV-2 Spike (WT) Fluc Pseudovirus|
|Envelope protein||SARS-CoV-2 Spike Protein (WT)|
|Buffer||DMEM medium +10% FBS|
|Titer (TCID50）||>1.5×10^4 TCID50/mL|
Pseudovirus Neutralization Assay
a. The day before the assay begins, seed 2× 104 / well HEK293/Human ACE2 Overexpression Stable Cells (Acrobiosystems Cat # CHEK-ATP042) with DMEM medium +10% FBS in a 96-well plate (Plate A). Incubate Plate A in a 5% (vol/vol) CO2, 37 °C incubator overnight.
b. Dilute your samples with DMEM medium +10% FBS and mix well. Add 75 μL diluted samples per well of a new 96-well plate (Plate B), then add 25 μL pseudovirus suspension per well to reach a final volume of 100 μL per well. Incubate Plate B in a 5% (vol/vol) CO2, 37 °C incubator for 60 min.
c. Discard the medium in Plate A. Immediately transfer the sample-pseudovirus mixtures in Plate B into the wells of Plate A. Incubate Plate A in a 5% (vol/vol) CO2, 37 °C incubator for 48 h.
d. Take out the plate containing cells and balance it to room temperature. Add 100 μL Bright-Glo™ Luciferase Assay reagent (Promega Cat # E2620) and mix for 2 min using a 96-well plate shaker (avoid light).
e. Transfer the liquid into a 96-well white flat bottom plate (Thermo Cat # 136101) and read the RLU of the plate with a luminescence meter.
SARS-CoV-2 Spike (WT) Fluc Pseudovirus is derived from pseudotyped HIV-1 virus expressing SARS-CoV-2 spike protein. It contains firefly luciferase reporter gene for chemiluminescence detection. The pseudovirus can effectively infect human ACE2 overexpressing cells and can be used in screening viral entry inhibitors or neutralizing antibodies, characterizing the neutralizing antibodies in volunteers inoculated with candidate vaccine or convalescent patients.
a. Though pseudovirus particles has no pathogenicity and cannot replicate, the assays should be carried out carefully in a P2 or higher-level laboratory with a biosafety cabinet.
b. Serum samples from animals or humans should be inactivated in a water bath at 56 °C for 30 min before being tested.
c. Please avoid freezing and thawing, which would influence the titer of the pseudovirus.
d. The product is for Research Use Only.