COVID Target Stable Cell Lines

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COVID Target Stable Cell Lines

Spike protein of SARS-CoV-2 (previously named 2019-nCoV) is composed of a S1 domain and S2 domain. The S1 domain contains a receptor-binding domain (RBD) that can specifically bind to angiotensin-converting enzyme 2 (ACE2), the receptor on target cells. Cell entry of coronavirus depends on the spike (S) protein binding to a cellular receptor and priming by a cellular protease. Studies demonstrate that angiotensin-converting enzyme 2 (ACE2) serves as the cell surface receptor to bind S protein in SARS-CoV-2 and facilitates entry of these coronaviruses into the cell.
GenScript has developed single clone-derived stable cell lines with high expression of Spike protein of SARS-CoV-2 or human ACE2 (hACE2) to assist and accelerate COVID-19 research.

: Validated Expression

: Single Clone

: High Signal-to-Background Ratio

: >15 Passages Validated

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Application

Cell-based binding assay with HEK293/ACE2 stable cell lines

Figure 1: Cell-based binding assay of SARS-CoV-2 spike protein RBD with HEK293/ACE2 cells.

HEK293/ACE2 cells were incubated with SARS-CoV-2 Spike protein (RBD, His Tag) (Cat. No. Z03479, GenScript) in serial dilution. Then the cells were stained with THE™ His Tag Antibody [iFluor 488], mAb, Mouse (Cat. No. A01800, GenScript) and analyzed with flow cytometer. FACS analysis shows the EC50 of spike protein RBD binding with ACE2 on HEK293 cells is 1.071 ug/ml.
Neutralization assay with HEK293/ACE2 stable cell lines

Figure 2: Neutralization assay of SARS-CoV-2 spike protein RBD with HEK293/ACE2 stable cell lines.

FACS analysis histogram shows ACE2 expression in HEK293/ACE2 stable cell line (Cat. No. M00770) compared with negative control. HEK293/ACE2 cells are plated at 3E5+ cells /well in 100ul PBS in 96-well plate, incubated with 10 µg/ml S-RBD-His (GenScript, Cat. No. Z03479) on ice for 1 hour, or left untreated. Both groups are then washed with PBS, incubated with Anti-His 2nd antibody (GenScript, Cat. No. A01800) at 1:200 on ice for 1 hour, washed with PBS, and then analyzed on flow cytometer.