Rapid Alpha-Pseudoviruses for Coronavirus Research

$400.00 – $6,800.00

NEW! SARS-CoV-2 EG.5.1 and BA.2.86 variant particles are now available!

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Betacoronavirus Species SARS-CoV-2, SARS-CoV, MERS-CoV
Variant Wild Type, XBB.1.9.1, XBB.1.16, D614G Single Mutation, XBB.1.5, XBB.1, BQ.1.1, BQ.1, BA.4.6, BA.2.75, BA.2.12.1, BA.4 and BA.5, AY4/BA.1 "Deltacron", Alpha (B.1.1.7), BA.2, Beta (B.1.351), Delta (B.1.617.2), Delta Plus (AY.2), Delta Plus (AY4.2), Epsilon (B.1.427/429), Epsilon (B.1.429), Eta (B.1.525), Gamma (P.1), Kappa (B.1.617.1), Lambda (C.37), Mu (B.1.621), Omicron (B.1.1.529), (B.1.1.207), (B.1.1.298), (B.1.2), (B.1.258), (B.1.494)
Reporter Gene Firefly Luciferase, GFP, RFP
Concentration 1x Not Concentrated, 10x, 60x
Size 5 x 200 µL ~20 wells/96 well plate, 10 x 200 µL ~40 wells/96 well plate, 25 x 200 µL ~100 wells/96 well plate, 50 x 200 µL ~200 wells/96 well plate, 100 x 200 µL ~400 wells/96 well plate, 5 x 100 µL (Concentrated Only)
SKU: HaCoV2(Luc) Categories: , Tags: , , , , , , , , , , , , , , , ,

Product Description

Rapid alpha-pseudovirus for SARS-CoV-2, SARS-CoV and MERS-CoV

 
Applications:
  • Screening and quantification of anti-coronavirus-neutralizing antibodies
  • Anti-coronavirus drug screening
  • Quantification of viral mutants’ infectivity
  • Identification of host co-factors and restriction factors of coronaviruses
Major advances:
  • Faster speed – As fast as 6 hours for the detection of reporter expression for Ha-CoV-2 versus 2-3 days for S protein pseudotyped lentivirus.
  • More robust reporter signal – Advantage of the rapid and robust gene expression capacity of alphavirus vector for reporter expression.
  • High fidelity in particle structure – Contains all 4 SARS-CoV-2 structural proteins (S, M, E, and N), but no structural proteins from other viruses. In contrast, lenti- and VSV- pseudoviruses contain only the S protein from SARAS-CoV-2, and multiple structural proteins from other viruses, such as HIV-1 Gag and Pol.
  • Strong correlation (coefficient value r2 = 0.87) with wild-type SARS-CoV-2 in neutralizing antibody assays (Hetrick et al., 2020).

Virongy offers pre-assembled Ha-CoV-2 and other coronaviral alpha-pseudovirus particles.  We also help customers to perform antiviral drug screening and quantification of neutralizing antibodies. For more information, please contact us by email: info@virongy.com

Description:

The hybrid alpha-pseudovirus for SARS-CoV-2 (Ha-CoV-2) is a newly developed SARS-CoV-2 virus-like particle (VLP) that encapsulates an alphavirus-derived RNA genome for rapid report expression (luciferase or GFP) in target cells (Hetrick et al., 2022). Different from commonly used S protein pseudotyped lenti- or vesicular stomatitis virus (VSV)-pseudoviruses, Ha-CoV-2 is assembled with all 4 structural proteins (S, M, N, and E) of SARS-CoV-2, and contains a reporter genome derived from an alphavirus-based vector for rapid (6 hours) and robust expression of reporter genes. The alphavirus vector does not contain any of the structural proteins from the authentic virus and the alpha-pseudovirus particles are single-cycle.

Ha-CoV-2 represents a major technological advancement in the development of SARS-CoV-2 pseudoviruses and serves as a platform for rapid and robust quantification of neutralizing antibodies, viral mutants, and antiviral drugs (Dabbagh et al., 2021; He et al., 2021). Virongy has now expanded its library of rapid alpha-pseudoviruses to other viruses within the coronavirus family.

Fig. 1. Schematic of the assembly of Ha-CoV-2 particles.

Fig. 2. Comparison of Ha-CoV-2 with S protein pseudotyped lentivirus in a time course of infection and reporter expression.

HEK293T(ACE2/TMPRESS2) cells were used as the target cell.

Fig. 3. Neutralizing antibody activity measured with Ha-CoV-2(Luc).

50 μl of Ha-CoV-2(Luc) (Cat# HaCoV2Luc-01) was incubated with serially diluted anti-serum for 1 hour at 37°C. The virus-antibody complex was used to infect 4×105 HEK293T(ACE2/TMPRSS2) target cells in a 96-well plate. Luciferase assay was performed at 12 hours post-infection. Coronavirus alpha-pseudoviruses, Coronavirus alpha-pseudoviruses, Coronavirus alpha-pseudoviruses

Cornavirus alpha-pseudoviruses

Example of product Certificate of Analysis

Ha-CoV-2 is intended for Research Use Only and is not for diagnostic or therapeutic purposes or uses in humans or animals.

SARS-CoV-2 Variants Available

SARS-CoV-2 Spikes Available Spike Mutations Full Protein Sequences
D614G Single AA Mutant Spike D614G
Alpha (B.1.1.7)

Spike A570D, Spike D614G, Spike D1118H, Spike H69del, Spike N501Y, Spike P681H, Spike S982A, Spike T716I, Spike V70del, Spike Y145del
(B.1.1.207)

Spike D614G, Spike E484K, Spike P681H
(B.1.1.298)

Spike D614G, Spike H69del, Spike I692V, Spike M1229I, Spike V70del, Spike Y453F
(B.1.2)

Spike D614G, Spike E484K, Spike G446V, Spike Y453F
(B.1.258)
Spike D614G, Spike H69del, Spike N439K, Spike V70del
Beta (B.1.351) Spike A243del, Spike A701V, Spike D80A, Spike D215G, Spike D614G, Spike E484K, Spike K417N, Spike L242del, Spike L244del, Spike N501Y
Epsilon (B.1.429)
Spike A222V, Spike D614G, Spike L452R, Spike S13I, Spike W152C
(B.1.494)
Spike A262S, Spike D614G, Spike D796Y, Spike H49Y, Spike L452R, Spike N501Y, Spike P681R, Spike Q613H
Gamma (P.1)Spike D138Y, Spike D614G, Spike E484K, Spike H655Y, Spike K417T, Spike L18F, Spike N501Y, Spike P26S, Spike R190S, Spike T20N, Spike T1027I, Spike V1176F
Kappa (B.1.617.1) Spike E154K, Spike L452R, Spike E484Q, Spike D614G, Spike P681R, Spike Q1071H
Eta (B.1.525)
Spike A67V, Spike D614G, Spike E484K, Spike F888L, Spike H69del, Spike Q52R, Spike Q677H, Spike V70del, Spike Y144del
Epsilon (B.1.427/429)
Spike D614G, Spike E484K, Spike L452R, Spike S13I, Spike W152C
Delta (B.1.617.2)
Spike T19R, Spike del157/158, Spike L452R, Spike T478K, Spike D614G, Spike P681R, Spike D950N
Delta Plus (AY.2)
Spike T19R, Spike V70F, Spiked del157/158, Spike A222V, Spike K417N, Spike L452R, Spike T478K, Spike D614G, Spike P681R, Spike D950N
Lambda (C.37)Spike G75V, Spike T76I, Spike R246N, Spike del247/253, Spike L452Q, Spike F490S, Spike D614G, Spike T859N
Mu (B.1.621)Spike T95I, Spike Y144S, Spike Y145N, Spike R346K, Spike E484K, Spike N501Y, Spike D614G, Spike P681H, Spike D950N
Delta Plus (AY4.2)Spike T19R, Spike T95I, Spike G142D, Spike Y145H, Spike E156G, Spike del157/158, Spike A222V, Spike L452R, Spike T478K, Spike D614G, Spike P681R, Spike D950N
Omicron (B.1.1.529)Spike A67V, Spike H69del,Spike V70del, Spike T95I, Spike G142D, Spike V143del, Spike Y144del, Spike Y145del, Spike N211del, Spike L212I, Spike ins214EPE, Spike G339D, Spike S371L, Spike S373P, Spike S375F, Spike K417N, Spike N440K, Spike G446S, Spike S477N, Spike T478K, Spike E484A, Spike Q493R, Spike G496S, Spike Q498R, Spike N501Y, Spike Y505H, Spike T547K, Spike D614G, Spike H655Y, Spike N679K, Spike P681H, Spike N764K, Spike D796Y, Spike N856K, Spike Q954H, Spike N969K, Spike L981F
BA.2 Spike T19I, Spike L24del, Spike P25del, Spike P26del, Spike A27S, Spike G142D, Spike V213G, Spike G339D, Spike S371F, Spike S373P, Spike S375F, Spike T376A, Spike D405N, Spike R408S, Spike K417N, Spike N440K, Spike S477N, Spike T478K, Spike E484A, Spike Q493R, Spike Q498R, Spike N501Y, Spike Y505H, Spike D614G, Spike H655Y, Spike N679K, Spike P681H, Spike N764K, Spike D796Y, Spike Q954H, Spike N969K
AY.4/BA.1Spike T19R, Spike A27S, Spike T95I, Spike G142D, Spike E156G, Spike F157del, Spike R158del, Spike ins214EPE, Spike N211del, Spike L212I, Spike G339D, Spike S371L, Spike S373P, Spike S375F Spike K417N, Spike N440K, Spike G446S, Spike S477N, Spike T478K, Spike E484A, Spike Q493R, Spike G496S, Spike Q498R, Spike N501Y, Spike Y505H, Spike T547K, Spike D614G, Spike H655Y, Spike N679K, Spike P681H, Spike N764K, Spike D796Y, Spike N856K, Spike Q954H, Spike L981F, Spike N969K
BA.4 and BA.5 (Share the Same Spike Mutations) Spike T19I, Spike L24S, Spike del25/27, Spike del69/70, Spike G142D, Spike V213G, Spike G339D,Spike S371F, Spike S373P, Spike S375F, Spike T376A, Spike D405N, Spike R408S, Spike K417N, Spike N440K, Spike L452R, Spike S477N, Spike T478K, Spike E484A, Spike F486V, Spike Q498R, Spike N501Y, Spike Y505H, Spike D614G, Spike H655Y, Spike N679K, Spike P681H, Spike N764K, Spike D796Y, Spike Q954H, Spike N969K
BA.2.12.1Spike T19I, Spike L24del, Spike P25del, Spike P26del, Spike A27S, Spike G142D, Spike V213G, Spike G339D, Spike S371F, Spike S373P, Spike S375F, Spike T376A, Spike D405N, Spike R408S, Spike K417N, Spike N440K, Spike L452Q, Spike S477N, Spike T478K, Spike E484A, Spike Q493R, Spike Q498R, Spike N501Y, Spike Y505H, Spike D614G, Spike H655Y, Spike N679K, Spike P681H, Spike S704L, Spike N764K, Spike D796Y, Spike Q954H, Spike N969K
BA.2.75Spike T19I, Spike K147E, Spike W152R, Spike F157L, Spike I210V, Spike V213G, Spike G257S, Spike G339H, Spike S371F, Spike S373P, Spike S375F, Spike T376A, Spike D405N, Spike R408S, Spike K417N, Spike N440K, Spike G446S, Spike N460K, Spike S477N, Spike T478K, Spike E484A, Spike Q498R, Spike N501Y, Spike Y505H, Spike D614G, Spike H655Y, Spike N679K, Spike P681H, Spike N764K, Spike D796Y, Spike Q954H, Spike N969K
BA.4.6Spike T19I, Spike L24S, Spike del25/27, Spike del69/70, Spike G142D, Spike V213G, Spike G339D, Spike R346T, Spike S371F, Spike S373P, Spike S375F, Spike T376A, Spike D405N, Spike R408S, Spike K417N, Spike N440K, Spike L452R, Spike S477N, Spike T478K, Spike E484A, Spike F486V, Spike Q498R, Spike N501Y, Spike Y505H, Spike D614G, Spike H655Y, Spike N658S, Spike N679K, Spike P681H, Spike N764K, Spike D796Y, Spike Q954H, Spike N969K
BQ.1Spike T19I, Spike L24S, Spike del25/27, Spike del69/70, Spike G142D, Spike V213G, Spike G339D, Spike S371F, Spike S373P,Spike S375F, Spike T376A, Spike D405N, Spike R408S, Spike K417N, Spike N440K, Spike K444T, Spike L452R, Spike N460K, Spike S477N, Spike T478K, Spike E484A, Spike F486V, Spike Q498R, Spike N501Y, Spike Y505H, Spike D614G, Spike H655Y, Spike N679K, Spike P681H, Spike N764K, Spike D796Y, Spike Q954H, Spike N969K
BQ.1.1Spike T19I, Spike L24S, Spike del25/27, Spike del69/70, Spike G142D, Spike V213G, Spike G339D, Spike R346T, Spike S371F, Spike S373P,Spike S375F, Spike T376A, Spike D405N, Spike R408S, Spike K417N, Spike N440K, Spike K444T, Spike L452R, Spike N460K, Spike S477N, Spike T478K, Spike E484A, Spike F486V, Spike Q498R, Spike N501Y, Spike Y505H, Spike D614G, Spike H655Y, Spike N679K, Spike P681H, Spike N764K, Spike D796Y, Spike Q954H, Spike N969K
XBB.1Spike T19I, Spike L24S, Spike del25/27, Spike V83A, Spike G142D,Spike del144/144, Spike H146Q, Spike Q183E, Spike V213E, Spike G252V, Spike G339H, Spike R346T, Spike L368I, Spike S371F, Spike S373P, Spike S375F, Spike T376A, Spike D405N, Spike R408S, Spike K417N,Spike N440K, Spike V445P, Spike G446S, Spike N460K, Spike S477N, Spike T478K, Spike E484A, Spike F486S, Spike F490S, Spike Q498R, Spike N501Y, Spike Y505H, Spike D614G, Spike H655Y, Spike N679K, Spike P681H, Spike N764K, Spike D796Y, Spike Q954H, Spike N969K
XBB.1.5 Spike T19I, Spike L24S, Spike del25/27, Spike V83A, Spike G142D,Spike del144/144, Spike H146Q, Spike Q183E, Spike V213E, Spike G252V, Spike G339H, Spike R346T, Spike L368I, Spike S371F, Spike S373P, Spike S375F, Spike T376A, Spike D405N, Spike R408S, Spike K417N,Spike N440K, Spike V445P, Spike G446S, Spike N460K, Spike S477N, Spike T478K, Spike E484A, Spike F486P, Spike F490S, Spike Q498R, Spike N501Y, Spike Y505H, Spike D614G, Spike H655Y, Spike N679K, Spike P681H, Spike N764K, Spike D796Y, Spike Q954H, Spike N969K
XBB.1.16Spike T19I, Spike L24S, Spike 25/27del, Spike V83A, Spike G142D, Spike Del144/144, Spike H146Q, SpikeE180V, Spike Q183E, Spike V213E, Spike G252V, Spike G339H, Spike R346T, Spike L368I, Spike S371F, Spike S373P, Spike S375F, Spike T376A, Spike D405N, Spike R408S, Spike K417N, Spike N440K, Spike V445P, Spike G446S,Spike N460K, Spike S477N, Spike T478R, Spike E484A, Spike F486P, Spike F490S, Spike Q498R, Spike N501Y, Spike Y505H, Spike D614G, Spike H655Y, Spike N679K, Spike P681H, Spike N764K, Spike D796Y, Spike Q954H, Spike N969K
XBB.1.9.1Spike T19I, Spike L24S, Spike Del25/27, Spike V83A, Spike G142D, Spike Del144/144, Spike H146Q, Spike Q183E, Spike V213E, Spike G339H, Spike R346T, Spike L368I, Spike S371F, Spike S373P, Spike S375F, Spike T376A, Spike D405N, Spike R408S, Spike K417N, Spike N440K, Spike V445P, Spike G446S, Spike N460K, Spike S477N, Spike T478K, Spike E484A, Spike F486P, Spike F490S, Spike Q498R, Spike N501Y, Spike Y505H, Spike D614G, Spike H655Y, Spike N679K, Spike P681H, Spike N764K, Spike D796Y, Spike Q954H, Spike N969K
BA.2.86
EG.5.1
FL.1.5.1
HV.1

Documentation

Alpha Pseudovirus – MSDS
Alpha Pseudovirus – Handbook

Suggested protocol

(For the infection of target cells in a 12-well plate. For more protocol details in other formats, such as in 96-well plate, please contact info@virongy.com)

Count target cells, g. HEK293T(ACE2/TMPRSS2), to be infected, and seed 1 x 105 cells per well in 0.5 ml culture medium in each well of a 12-well plate. Culture cells overnight. Note: Cell viability should be ≥ 80%.
Before infection, remove the medium, and add 300 μl fresh medium.
Add 100 μl Ha-CoV-2 virus, and infect for 2-6 hours. *
Wash cells by adding 1 ml fresh medium.
After washing, add 1 ml fresh complete medium (DMEM + 10% FBS).
Culture cells for 12-18 hours to read signals (e.g. luciferase assay).

* if performing neutralizing antibody assays, incubate the virus with antibodies at 37°C for 1 hour, then add the virus-antibody complex to cells for infection.

References

Dabbagh, D., He, S., Hetrick, B., Chilin, L., Andalibi, A., and Wu, Y. (2021). Identification of the SHREK family of proteins as broad-spectrum host antiviral factors. bioRxiv, doi: https://doi.org/10.1101/2021.02.02.429469

He, S., Waheed, A.A., Hetrick, B., Dabbagh, D., Akhrymuk, I.V., Kehn-Hall, K., Freed, E.O., and Wu, Y. (2021). PSGL-1 Inhibits the Incorporation of SARS-CoV and SARS-CoV-2 Spike Glycoproteins into Pseudovirions and Impairs Pseudovirus Attachment and Infectivity. Viruses 13, 46.

Hetrick, B., Chilin, L. D., He, S., Dabbagh, D., Alem, F., Narayanan, A., … & Wu, Y. (2022). Development of a hybrid alphavirus-SARS-CoV-2 pseudovirion for rapid quantification of neutralization antibodies and antiviral drugs. Cell Reports Methods, 100181.

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