SARS-CoV-2 pseudovirus

COVID-19, a novel pathogenic coronavirus (2019-nCoV or SARS-CoV2) that causes the COVID-19 pandemic respiratory illness and spread globally rapidly. Due to its high pathogenicity and infectivity, live SARS-CoV-2 should be handled under biosafety level 3 (BSL-3) conditions, which has seriously hindered the development of COVID-19 vaccines and drugs. Combining the genomics and the biological characteristics of SARS - CoV- 2, BrainVTA has developed SARS-CoV-2 pseudovirus production system, from which the SARS-CoV-2 pseudotyped virus can be handled in biosafety level 2 (BSL-2).
SARS-CoV-2(2019nCoV) Pseudovirus (PSV) Based Research Field

-Neutralization Assay
In the pseudovirus based neutralization assay, the inhibition of viral entry into cells by Neutralize Antibody is correlated to the decreased levels of luciferase or fluorescence (GFP and mcherry) signals in the ACE2 expression cells (HEK293T-hACE2 and BHK-hACE2).This method is simplicity, higher sensitivity and accuracy, suitability for high-throughput experiments. Also, no live virus is used during the test. Therefore, this method could be used as an alternative for safely conducting serologic studies in a rapid response.

To evaluate the effectiveness of therapeutic monoclonal neutralizing antibodies.
To analyze and evaluate the neutralizing antibody levels of COVID-19 vaccines during the development phase and after future marketing.
The dynamic change of neutralizing antibody level in COVID-19 convalescent patients.

-To construct a study model simulating SARS-COV-2 infected cells/animals.

COVID-19 (SARS-CoV-2) Pseudovirus

Cat. No.	Product Name	Description
SARS-CoV-2-V03001	PV-SARS-CoV-2-S-VSV- △G-EGFP	VSV Backbone, SARS-CoV-2 Spike Protein (WT)-EGFP
SARS-CoV-2-V03005	PV-SARS-CoV-2-S-VSV- △G-Luciferase	VSV Backbone, SARS-CoV-2 Spike Protein (WT)-Luciferase
SARS-CoV-2-LV-0046	PV-SARS-CoV-2-S-LV-CMV-mcherry	HIV-1 Backbone,SARS-CoV-2 Spike Protein (WT)-mcherry
SARS-CoV-2-LV-0052	PV-SARS-CoV-2-S-LV-CMV-EGFP	HIV-1 Backbone, SARS-CoV-2 Spike Protein (WT)-EGFP
SARS-CoV-2-LV-0588	PV-SARS-CoV-2-S-LV-CMV-Luciferase	HIV-1 Backbone, SARS-CoV-2 Spike Protein (WT)-Luciferase
SARS-CoV-2-V03002	PV-SARS-CoV-2-S-(D614G)-VSV-△G-mcherry	VSV Backbone, SARS-CoV-2 Spike (D614G)-mcherry
SARS-CoV-2-K-LV-0588	PV-SARS-CoV-2-S-(D614G)-rLV-CMV-Luciferase	HIV-1 Backbone, SARS-CoV-2 Spike (D614G)-Luciferase
SARS-CoV-2-A-V03002	PV-SARS-CoV-2-S (B1.1.7) -VSV-△G-mCherry	VSV Backbone, SARS-CoV-2 Spike Protein (UK B.1.1.7;： N501YA570D，P681H，T716I，S982A，D1118H)-mCherry
SARS-CoV-2-K-LV-0588	PV-SARS-CoV-2-S(B.1.1.7)-rLV-CMV-Luciferase	HIV-1 Backbone, SARS-CoV-2 Spike Protein (UK B.1.1.7;： N501YA570D，P681H，T716I，S982A，D1118H)-Luciferase
SARS-CoV-2-B-V03002	PV-SARS-CoV-2-S(SA-B.1.351) -VSV-△G-mCherry	VSV Backbone, SARS-CoV-2 Spike Protein (South African variant Δ3 B.1.351, 20H/501Y.V2, Beta; L18F, D80A, D215G, ΔL242/A243/L244, R246I, K417N, N501Y, E484K, D614G, A701V)-mCherry
SARS-CoV-2-B-LV-0588	PV-SARS-CoV-2-S(SA-B.1.351)-LV-CMV-Luciferase	HIV Backbone, SARS-CoV-2 Spike Protein( South African variant Δ3 B.1.351, 20H/501Y.V2, Beta; L18F, D80A, D215G, ΔL242/A243/L244, R246I, K417N, N501Y, E484K, D614G, A701V)-Luciferase
V04006	PV-SARS-CoV-2-S(501Y.V2)-VSV-△G-mCherry	VSV Backbone, SARS-CoV-2 Spike ( South African variant B.1.351, 20C/501Y.V2; L18F, D80A, D215G, L242H, R246I, K417N, N501Y, E484K, D614G, A701V)-mCherry
SARS-CoV-2-K-LV-0588	PV-SARS-CoV-2-S(501Y.V2)-rLV-CMV-Luciferase	HIV Backbone, SARS-CoV-2 Spike ( South African variant B.1.351, 20C/501Y.V2; L18F, D80A, D215G, L242H, R246I, K417N, N501Y, E484K, D614G, A701V)-Luciferase
V04007	PV-SARS-CoV-2-S(B1.1.617)-VSV-△G-mCherry	VSV Backbone, SARS-CoV-2 Spike ( Indian variant B.1.617, RBD mutations only; L452R, E484Q, D614G)-mCherry
SARS-CoV-2-K-LV-0588	PV-SARS-CoV-2-S(B1.1.617)-rLV-CMV-Luciferase	HIV Backbone, SARS-CoV-2 Spike ( Indian variant B.1.617, RBD mutations only; L452R, E484Q, D614G)-Luciferase
SARS-CoV-2-D-LV-0588	PV-SARS-CoV-2-S(IN-B.1.617.2)-LV-CMV-Luciferase	HIV Backbone, SARS-CoV-2 Spike ( Indian variant B.1.617.2; T19R, G142D, del156/157, R158G, L452R, T478K, D614G, P681R, D950N)-Luciferase
SARS-CoV-2-K-LV-0588	PV-SARS-CoV-2-S(IN-B.1.617.1)-LV-CMV-Luciferase	HIV Backbone, SARS-CoV-2 Spike (Indian variant B.1.617.1; G142D, E154K, L452R, E484Q, D614G, P681R, Q1071H)-Luciferase
SARS-CoV-2-G-LV-0588	PV-SARS-CoV-2-S(BZ-P.1)-LV-CMV-Luciferase	HIV Backbone, SARS-CoV-2 Spike (Brazilian variant： L18F，T20N，P26S，D138Y，R190S，K417T，E484K， N501Y，D614G，H655Y，T1027I)-Luciferase
SARS-CoV-2-G-LV-0588	PV-SARS-CoV-2-S(B.1.1.529)-LV-CMV-Luciferase	HIVBackbone, SARS-CoV-2 Spike(A67V, Δ69-70, T95I, G142D, Δ143-145, Δ211 212, ins214EPE, G339D, S371L, S373P, S375F, K417N, N440K, G446S, S477N, T478K, E484A, Q493R, G496S, Q498R, N501Y, Y505H, T547K, D614G, H655Y, N679K, P681H, N764K, D796Y, N856K, Q954H, N969K, L981F)

Recombinant viral express SARS-COV-2-S, ACE2 and TMPRSS2 protein

Vector	Cat.	Vector Name	Biosafety Level
Lentiviral vector	LV-0537	rLV-CMV-SARS-CoV-2-S-2A-mCherry-WPRE	BSL-2
	LV-0538	rLV-Ef1a-ACE2-2A-EGFP-WPRE
	LV-0539	rLV-Ef1a-ACE2-2A-EGFP--CMV-Puro-WPRE
	LV-0558	rLV-CMV-TMPRSS2-2A-mCherry-WPRE
	LV-0559	rLV-CMV-ACE2-2A-TMPRSS2-2A-EGFP-WPRE
AAV vector	PT-2701	rAAV-CMV-TMPRSS2-2A-BFP-WPRE
	PT-2702	rAAV-CMV-SARS-CoV-2-S-2A-mCherry-WPRE
	PT-2703	rAAV-Ef1a-ACE2-2A-EGFP-WPRE

Product Features

● PV-SARS-CoV-2-S-VSV-△G-EGFP/mCherry
1) As the genome of the pseudovirus can be replicate and the fluorescence signal will be enlarged, so pseudovirus in VSV has a high sensitivity.
2) Add PV-SARS-CoV-2-S-VSV-△G-EGFP/mCherry can only simulate the single infection process of the COVID-19, and if you want to simulate the infection continuously, the S protein needs to be compensated by infecting LV/AAV that express it.
3) It can be handled in biosafety level 2 (BSL-2) and the fluorescence expression time is 7-8h.
● PV-SARS-CoV-2-S-LV-CMV-EGFP/mCherry
1) Suitable for difficult to infect cells or tissues, such as stem cells, neurons, primary cells, etc.
2) Can only be used to simulate a single infection of COVID-19.
●Recombinant lentivirus express SARS-COV-2-S, ACE2 and TMPRSS2 protein

1) Suitable for difficult to infect cells or tissues, such as stem cells, neurons, primary cells, etc.
2) Suitable for the establishment of cell model.
3) Suitable for the establishment of effector cells.
●Recombinant AAV express SARS-COV-2-S, ACE2 and TMPRSS2 protein
1) Immunogenicity is low and suitable for long-term expression.
2) Suitable for the establishment of animal models.

References
1. Jean K. Millet1, Tiffany Tang3,et al. Production of Pseudotyped Particles to Study Highly Pathogenic Coronaviruses in a Biosafety Level 2 Setting. J Vis Exp, doi:10.3791/59010 (2019).
2. Nie J , Li Q , Wu J , et al. Establishment and validation of a pseudovirus neutralization assay for SARS-CoV-2[J]. Emerging Microbes & Infections.2020.9(1):680-686.
3. Thomas T . SARS-CoV-2 receptor ACE2 expression in the human heart: cause of a post-pandemic wave of heart failure [J]. European Heart Journal.2020.9(1):45.
4. Wrapp , Wang NS, Corbett KS et al.Cryo-EM structure of the 2019-nCoV spike in the prefusion conformation[J].Science.2020. 367:1260–1263

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