Certificate of Analysis

1. Product Information

Product name:                                        Ha-CoV-2(Luc)

Product No:                                             HaCoV2Luc-01, HaCoV2Luc-10C (10 x concentrated)

Product Batch No:                                  HaCoV2Luc-01-Batch#020721

HaCoV2Luc-10C-Batch#020721

Product Manufacture Date:                    Feb. 07, 2021

 

Manufacturer details:

Production Technician Name:               D. Yu

Company Address:                                 Virongy Biosciences 11225 Assett Loop STE 111, Manassas, VA 20109

Company Phone Number:                     (703) 257 5500

Email address:                                          info@virongy.com

 

 

2. Quality Control Procedure

Ha-CoV-2 infection Protocol

  • Seed 4 x 104 HEK293T(ACE2/TMPRESS2) cells or HEK293T cells into each well of 96-well plates in a total volume of 100 ul cell culture medium (DMEM + 10% FBS + Pen/Strep). Culture overnight at 37°C.
  • Remove medium from each well, add 50 ul of Ha-CoV-2(Luc) or 50 ul of concentrated Ha-CoV-2(Luc). Infect for 2 hours.
  • Remove virus, add 100 ul freh medium (DMEM + 10% FBS + Pen/Strep). Culture overnight at 37°C for 12-18 hours.
  • Remove medium, wash once with PBS, lyse cells with 100 ul of 1 x Cell lysis buffer (Promega).
  • Add 100 ul of Luciferease Assay Buffer into each well of 98-well Luciferease Assay Plate. Mix with 30 ul cell lysates.
  • Read luciferease activity within 10-15 minutes on a GloMax Discover Microplate Reader.

 

3. Luciferase Results

Ha-CoV-2 + HEK293T(ACE2/TMPRESS2) cells:                  40,480.00 ± 4,925.76 (from 1.3 x 104 infected cells).

10 x Ha-CoV-2 + HEK293T(ACE2/TMPRESS2) cells:          94,366.67 ± 7,902.74 (from 1.3 x 104 infected cells).

HEK293T(ACE2/TMPRESS2) cells only:                               42.67 ± 15.28 (from 1.3 x 104 un-infected cells).

 

Ha-CoV-2 + HEK293T cells:                                                   120.00 ± 2.00 (from 1.3 x 104 infected cells).

HEK293T cells only:                                                               42.7 ± 15.27 (from 1.3 x 104  un-infected cells).

 

Date of Luciferease Assay:  Feb 12, 2021