101Bio品牌国内代理商---默瑞生物

101Bio针对细胞培养上清，血浆/血清，干细胞培养上清，尿液及其他体液的外泌体提取试剂盒、3D细胞培养基凝胶等。

We help our customers to assemble SARS-CoV and SARS-CoV-2 S protein pseudotyped lentiviruses. These particles carry reporters that can be used for antiviral drug screening or the quantification of neutralizing antibodies. Use reporter construct from customer, and we can assemble particles within 1-2 weeks. 可用于抗病毒药物筛选或中和抗体的定量。

We also offer 6 pre-assembled, concentrated (60 x concentrated) particles for your initial testing (see table below). Our proprietary SARS-CoV-2 Pseudovirus Infection Enhancer (Cat#: CoV2-1ml) can greatly promote productive viral infection. This enhancer is used to facilitate the infection of a variety of host cells and enhance viral infection rates by 5 to 20 folds（假病毒感染增强剂用于多种宿主细胞的感染，并将病毒感染率提高5-20倍）.

1) The SARS-CoV-2 S protein pseudotyped lentiviral particles naturally have much lower infectivity than SARS-CoV S protein pseudotyped particles. The GFP reporter virus is designed for use in microscope imaging studies of GFP positive cells.

SARS-CoV-2 S 蛋白假型慢病毒比 SARS-CoV S 蛋白假型慢病毒拥有更低的传染性，GFP报告病毒用于GFP阳性细胞的显微成像研究。

2) Use our enhancer(Cov2-1ml ) for Luc virus to get robust signal. This enhancer works perfect for the SARS-CoV-2 S protein pseudotyped Luc (luciferase) reporter virus so it can effectively infect cells that naturally express ACE2 (such as Vero E6 cells). There is no need to over-express ACE2 in target cells by transient DNA transfection (see Fig. 1).

使用Luc病毒增强剂（Cov2-1ml）获得稳健的信号。这种增强剂对SARS-CoV-2s蛋白假型Luc（luciferase荧光素酶）报告病毒非常有效，因此它能有效感染自然表达ACE2的细胞（如Vero E6细胞）。瞬时DNA转染不需要在靶细胞中过度表达ACE2

3) The Luc reporter is expressed from the HIV-1 LTR promoter, which is driven by co-expression of the Tat protein that is present only in infected cells. The Luc reporter system is more robust than common promoters used for reporter expression.

       Luc报告者是从HIV-1ltr启动子中表达的，该启动子由仅存在于受感染细胞中的Tat蛋白共同表达驱动。Luc报告程序系统比用于报告程序表达式的普通启动程序更健壮。

Lenti-pseudovirus infection of Vero E6 cells (applicable to SARS-CoV S, SARS-CoV-2 S, and VSV-G pseudotyped viruses)

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.

3) Pre-treat cells by adding 25 μl of pseudovirus infection enhancer (Cat# CoV2-1ml) (10 X) so that the

enhancer concentration is 1 X. Mix and incubate for 30–60 minutes.

4) Add virus to the cells and mix. Note volume of virus used.

5) Add infection enhancer (10 X) in an amount equal to 1/10 of the virus volume used, e.g., if 100

μl of virus is used, add 10 μl of infection enhancer. Incubate at 37°C for 4-6 hours.

6) Add 2 ml fresh media to wash cells.

7) After washing, add 2 ml fresh complete medium.

8) Culture infected cells for 2-3 days to signal detection.

SARS-CoV-2-S(Luc) pseudovirus infection of Vero E6 cells (with pseudovirus infection enhancer ):

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

60 X concentrated, SARS-CoV and SAR-CoV-2 pseudoviral particles

SARS-CoV-2 S protein pseudotyped lentiviral particle transduction of Vero E6 cells (Left):

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

SARS-CoV S protein pseudotyped lentiviral particle transduction of Vero cells (Right):

Vero cells were transduced with SARS-CoV-S(GFP) lenti-pseudovirus (with a GFP reporter) in the presence of CoV-2-PIE. Reporter expression was quantified at 2 days post transduction (GFP flow cytometry).

SARS-CoVandSARS-CoV-2pseudovirus transduction of target cells for viral entry and functional studies.

Anti-SARS-CoV and SARS-CoV-2 drug screening

Anti-SARS-CoVandSARS-CoV-2 neutralizing antibody screening SARS-CoV and CoV-2 pseudoviruses are intended for Research Use Only.

HIV Rev-dependent Reporter Cells (Cat.# HRC-1) and HIV infection enhancer (H901-1)

Real 3D Cell Culture Gel Col-Tgel (Cat.# P720)

Lentivirus / Retrovirus  10X Titer-Up Reagent  (Cat.# P906 / P909)95% pure Exosome Isolation kits 

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