SARS-CoV-2 Pseudovirus Infection Enhancer (CoV-2-PIE)

A cutting-edge technology to facilitate your COVID-19 research 

CoV-2-PIE is a viral infection enhancer designed specifically to facilitate coronavirus and pseudovirus infection of target cells. CoV-2-PIE is formulated based on our current technology of Infectin-2TM, which is designed to penetrate the cortical actin barrier, thereby greatly enhancing productive viral infection. CoV-2-PIE can be used to facilitate the infection of a variety of host cells by coronaviruses and pseudoviruses. CoV-2-PIE can enhance viral infection rates by 5 to 20 fold. Virongy developed CoV-2-PIE based on the scientific theory that the actin cytoskeleton is a natural barrier for viral entry and post-entry intracellular migration (Yoder et al., Cell, 2008, 134:782). CoV-2-PIE is formulated to mainly enhance viral infection of adherent target cells, which are frequently also limited by poor absorption of virion particles to target cells. CoV-2-PIE combines our proprietary technology of InfectinTM with additional technologies that promote viral absorption onto target cells.

Important

  • Once thawed, CoV-2-PIE should be stored at 4oC, and is stable for 3 months. Do not re-freeze and do not leave CoV-2-PIE at room temperature.
  • CoV-2-PIE viral infection enhancer works with most cell lines to enhance viral infection. On average, CoV-2-PIE enhances productive viral infection by 3- to 20-fold*.
  • CoV-2 PIE is formulated as 10 X concentrated.

 (*The degree of enhancement is affected by the types of viruses and cells.)

Applications

  • Enhancing coronavirus and pseudovirus transduction of target cells
  • Enhancing infection of other enveloped viruses
  • Facilitating recovery of infectious viruses from cell or tissue cultures
  • Facilitating anti-viral drug and neutralizing antibody screening efficiency

                                                   Table 1.  10 X  concentrated CoV-2-PIE

Protocol

Example  – CoV-2-PIE enhances SARS-CoV-2 lenti-pseudovirus transduction of Vero E6 cells:

  1. Count Vero E6 cells to be infected and seed ~1 x 105 cells per well into 12-well plates (0.5 ml per well). Culture cells until cells stably adhere to the plates (4–12 hours). Note: Cell viability should be 80%.
  2. Before infection, wash cells with 2 ml medium, and leave 250 μl medium in each well.Pre-treat cells by adding 25 μl of CoV-2-PIE (10 X) so that the CoV-2-PIE concentration is 1 X. Mix and incubate for 30–60 minutes.
  3. Add virus to the cells and mix. Note volume of virus used.
  4. Add CoV-2-PIE (10 X) in an amount equal to 1/10 of the virus volume used, g., if 100 μl of virus is used, add 10 μl of CoV-2-PIE. Incubate at 37°C for 4-6 hours.
  5. Add 2 ml fresh media to wash cells.
  6. After washing, add 2 ml fresh complete medium.
  7. Culture infected cells for 2-3 days to signal detection.

Example of results from our customers:

CoV-2-PIE enhances SARS-CoV-2-S(Luc) lenti-pseudovirus transduction of Vero E6 cells:

Vero E6 cells were transduced with SARS-CoV-2-S(Luc) lenti-pseudovirus (with a luciferase reporter) in the presence or absence of CoV-2-PIE. Reporter expression was quantified at 3 days post transduction (luciferase assay).

Selected publication from our users

DeMarino, C., Cowen, M., Pleet, M.L., Pinto, D.O., Khatkar, P., Erickson, J., Docken, S.S., Russell, N., Reichmuth, B., Phan, T., et al. (2020). Differences in Transcriptional Dynamics Between T-cells and Macrophages as Determined by a Three-State Mathematical Model. Sci Rep 10, 2227.

Fu, Y., He, S., Waheed, A.A., Dabbagh, D., Zhou, Z., Trinite, B., Wang, Z., Yu, J., Wang, D., Li, F., et al. (2020). PSGL-1 restricts HIV-1 infectivity by blocking virus particle attachment to target cells. Proc Natl Acad Sci U S A.

He, S., Hetrick, B., Dabbagh, D., Akhrymuk, I.V., Kehn-Hall, K., Freed, E.O., and Wu, Y. (2020). PSGL-1 blocks SARS-CoV-2 S protein-mediated virus attachment and infection of target cells. bioRxiv. 

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