Spike Variants (SARS-CoV-2) Pseudotyped Lentivirus Pack (Luciferase Reporter)
The pandemic coronavirus disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). As the first step of the viral replication, the virus attaches to the host cell surface before entering the cell. The viral Spike protein recognizes and attaches to the Angiotensin-Converting Enzyme 2 (ACE2) receptor found on the surface of type I and II pneumocytes, endothelial cells, and ciliated bronchial epithelial cells. Drugs targeting the interaction between the Spike protein of SARS-CoV-2 and human ACE2 may offer protection against the viral infection. Numerous SARS-CoV-2 variants have been identified so far. These variants contain a number of mutations that may increase morbidity and mortality and allow the virus to spread more easily and quickly than the original strain.
BPS Bioscience has launched a series of Spike Variants (SARS-CoV-2) Pseudotyped Lentivirus (Luc reporter). The Spike (SARS-CoV-2) Pseudotyped Lentiviruses were produced with SARS-CoV-2 Spike Variant (see below for mutation details) as the envelope glycoproteins instead of the commonly used VSV-G. These pseudovirions contain the firefly luciferase gene driven by a CMV promoter (Figure 1), therefore, the spike-mediated cell entry can be measured via luciferase activity. The Spike Variants (SARS-CoV-2) Pseudotyped Lentivirus Pack (Luciferase Reporter) contains a collection of 12 Spike variants (SARS-CoV-2) Pseudotyped lentivirus (Luc reporter). It is a great tool to screen for variant-specific antibodies or to test the binding or efficacy of drug candidates against the different Spike variants. The Spike (SARS-CoV-2) pseudotyped lentiviruses can be used to measure the activity of neutralizing antibody against SARS-CoV-2 infection in a Biosafety Level 2 facility.
Spike interacts with host cells by binding to membrane receptor ACE2 (angiotensin converting enzyme 2). Based on experiments performed by scientists at BPS Bioscience, we know that the wild-type SARS-CoV-2 spike pseudotyped lentiviruses transduce the following cells with great efficiency: ACE2-HEK293 cells (BPS Bioscience, #79951), ACE2-CHO cells (BPS Bioscience, #79959), ACE2-HeLa cells (BPS Bioscience, #79958). They also efficiently transduce TMPRSS2-Vero E6 cells (BPS Bioscience, #78081), which express high endogenous levels of ACE2 and were stably transfected with human serine protease TMPRSS2 required for the priming of Spike and fusion of the virion with the plasma membrane. By contrast, it has been shown by others that SARS-CoV-2 spike pseudotyped lentiviruses do not transduce parental Calu3 and Vero E6 cells very well [Neerukonda et al. 2021, PlosOne PMID: 33690649; Tandon et al. 2020, Scientific Reports PMID: 33154514; Condor Capcha et al. 2021, Front. Cardiovasc. Med. PMID: 33521067; Pisil et al. 2021, Pathogens PMID: 33540924].
SARS-CoV-2 variant pseudoviruses have been validated using ACE2-HEK293 cells but have not been tested in other cells.
As recommended in our protocol, 5 µl of virus/well in a 96-well plate provides a sufficient signal-to-noise ratio to perform inhibition studies. The amount of virus added to the cells can also be scaled down according to the user’s need.
The pack provides one vial of each of the following:
| SARS-CoV-2 Variant | BPS CAT# | Sequence details |
| WT Spike Wuhan-Hu-1 | 79942-1 | Genbank Accession #QHD43416.1 |
| D614G | 78028-1 | D614G |
| B.1.1.7 (Alpha variant, UK) | 78112-1 | Deletions of H69, V70, and Y144; N501Y, A570D, D614G, P681H, T716I, S982A, D1118H |
| B.1.351 (Beta variant, SA) | 78142-1 | L18F, D80A, D215G, R246I, K417N, E484K, N501Y, D614G, A701V |
| P.1 (Gamma variant, Brazil) | 78144-1 | L18F, T20N, P26S, D138Y, R190S, K417T, E484K, N501Y, D614G, H655Y, T1027I |
| B.1.429 (Epsilon variant) | 78172-1 | S13I, W152C, L452R, D614G |
| B.1.617 | 78204-1 | L452R, E484Q, D614G, P681R |
| B.1.617.1 (Kappa variant) | 78205-1 | G142D, E154K, L452R, E484Q, D614G, P681R, Q1071H |
| B.1.618 | 78206-1 | Y145del, H146del, E484K, D614G |
| B.1.617.2 (Delta variant) | 78215-1 | T19R, G142D, 156/157del, R158G, L452R, T478K, D614G, P681R, D950N |
| B.1.617.2.1 (Delta plus) | 78218-1 | T19R, G142D, 156/157del, R158G, K417N, L452R, T478K, D614G, P681R, D950N |
| B.1.621 (Mu variant) | 78618-1 | T95I, Y144S, Y145N, R346K, E484K, N501Y, D614G, P681H, D950N |
Figure 1. Schematic of the Luciferase Reporter in Spike (SARS-CoV-2) Pseudotyped Lentiviruses
| Name | Ordering Information |
| Thaw Medium 1 or HEK293 Growth Medium | BPS Bioscience, #60187 |
| ACE2-HEK293 Recombinant Cell Line | BPS Bioscience, #79951 |
| Spike S1 Neutralizing Antibody (Clone C-A11) (SARS-CoV-2) | BPS Bioscience, #101024 |
| 96-well tissue culture treated, white clear-bottom assay plate | Corning, #3610 |
| ONE-Step™ luciferase assay system | BPS Bioscience, #60690 |
The lentiviruses were produced from HEK293T cells. Supplied in medium containing 90% DMEM + 10% FBS.
Thaw Medium 1 (BPS Bioscience, #60187): MEM medium supplemented with 10% FBS, 1% non-essential amino acids, 1 mM Na pyruvate, 1% Penicillin/Streptomycin.
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