GenScript COVID Target Stable Cell Lines 제품을 소개합니다.

SARS-CoV-2(2019-nCoV)의 Spike protein은 S1 도메인과 S2 도메인으로 구성되어 있습니다. S1 도메인에는 수용체 결합 도메인(RBD)이 포함되어 있으며, 이는 표적 세포 표면의 수용체인 angiotensin-converting enzyme 2 (ACE2)에 특이적으로 결합할 수 있습니다. 코로나바이러스의 세포 침입은 Spike(S) 단백질이 세포 표면 수용체에 결합하고, 이후 세포 내 프로테아제에 의해 활성화(priming)되는 과정에 의존합니다. 연구에 따르면 ACE2는 SARS-CoV-2에서 S 단백질과 결합하는 세포 표면 수용체로 작용하며, 바이러스의 세포 내 유입을 촉진합니다. 

GenScript는 COVID-19 연구를 지원하고 가속화하기 위해, SARS-CoV-2 Spike protein 또는 인간 ACE2(hACE2)를 고발현하는 단일 클론 유래 안정 세포주(stable cell line)를 개발하였습니다.

[Applications]

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.