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Anti-SARS-CoV-2 Spike RBD Neutralizing Antibody, Human IgG1 (AS35)

产品描述(Description)

Anti-SARS-CoV-2 RBD Neutralizing Antibody, Human IgG1 (SAD-S35) is isolated from a SARS-CoV-2 infected patient and is recombinantly produced from human 293 cells (HEK293). This antibody recognizes the SARS-CoV-2 Spike Protein RBD domain and inhibits the interaction between SARS-CoV-2 RBD and ACE2 with an IC50 of 1.47 μg/mL using SARS-CoV-2 Inhibitor Screening Kit (Cat. No. EP-105).Pseudovirus assay shows that this antibody has potent neutralizing activity against pseudovirus bearing SARS-CoV-2 Spike protein.

亚型(Isotype)

Human IgG1/kappa

特异性(Specificity)

This product is a specific antibody against SARS-CoV-2 Spike protein RBD domain. No cross-reactivity is detected with Spike protein RBD domain of other coronaviruses, including SARS-CoV, MERS-CoV, HCoV-229E, HCoV-NL63, HCoV-OC43 and HCoV-HKU1.

纯度(Purity)

>95% as determined by SDS-PAGE.

内毒素(Endotoxin)

制剂(Formulation)

Lyophilized from 0.22 μm filtered solution in PBS, pH7.4 . Normally trehalose is added as protectant before lyophilization.

重构方法(Reconstitution)

Please see Certificate of Analysis for specific instructions.

For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.

存储(Storage)

For long term storage, the product should be stored at lyophilized state at -20°C or lower.

Please avoid repeated freeze-thaw cycles.

This product is stable after storage at:

  1. -20 to -70°C for 12 months in lyophilized state from date of receipt;
  2. -70°C for 3 months under sterile conditions after reconstitution.
 

电泳(SDS-PAGE)

Anti-SARS-CoV-2 RBD Neutralizing Antibody, Human IgG1 (Cat. No. SAD-S35) SDS-PAGE gel

Anti-SARS-CoV-2 RBD Neutralizing Antibody, Human IgG1 on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.

 

活性(Bioactivity)-ELISA

Biotinylated Human ELISA

Serial dilutions of Anti-SARS-CoV-2 RBD Neutralizing Antibody, Human IgG1 (Cat.No. SAD-S35) was detected by SARS-CoV-2 Inhibitor screening Kit (Cat.No. EP-105) with a half maximal inhibitory concentration (IC50) of 1.472 μg/mL (QC tested).

Protocol

Biotinylated Human  ELISA

Immobilized SARS-CoV-2 S1 protein, His Tag (Cat. No. S1N-C52H4) at 2 μg/mL (100 μL/well) can bind Anti-SARS-CoV-2 RBD Neutralizing Antibody, Human IgG1 (Cat. No. SAD-S35) with a linear range of 0.2-1.95 ng/mL (QC tested).

Protocol

Biotinylated Human ELISA

Serial dilutions of Anti-SARS-CoV-2 RBD Neutralizing Antibody, Human IgG1 (Cat.No. SAD-S35) was detected by Anti-SARS-CoV-2 Neutralizing Antibody Titer Serologic Assay Kit (Cat.No. TAS-K022) with a half maximal inhibitory concentration (IC50) of 0.6265 μg/mL (Routinely tested).

Protocol

Biotinylated Human ELISA

Serial dilutions of Anti-SARS-CoV-2 Spike RBD Neutralizing Antibody (Cat. No. SAD-S35) was incubated with SARS-CoV-2 pseudotyped virus at 37 ℃ for 1 hour. Afterward, Huh-7 cells were added, followed by incubation at 37 ℃ for 24h. Chemiluminescence detection was performed and the virus neutralization titers (IC50) is 2.181 nM using the Reed-Muench method.

Protocol

Biotinylated Human ELISA

Anti-SARS-CoV-2 Spike RBD Neutralizing antibody (Cat.No. SAD-S35) neutralizes SARS-CoV-2 Spike RBD by inhibiting RBD:ACE2 interaction. The ACE2-coated plate is incubated with the wild type (WT) RBD or B.1.1.7, B.1.351, P.1, B.1.617.1 mutant and treated with the antibody at increasing concentration. Percent inhibition is calculated based on the OD value.

Protocol

活性(Bioactivity)-SPR

Biotinylated Human SPR

Anti-SARS-CoV-2 RBD Neutralizing Antibody, Human IgG1 (Cat. No. SAD-S35) captured on CM5 chip via Anti-human IgG Fc antibodies surface can bind SARS-CoV-2 S protein RBD (A475V), His Tag (Cat. No. SPD-C52Hd) with an affinity constant of 87.7 nM as determined in a SPR assay (Biacore 8K) (Routinely tested).

Protocol

 

活性(Bioactivity)-FACS

Biotinylated Human FACS

FACS analysis shows that the binding of SARS-CoV-2 S protein RBD, Mouse IgG2a Fc Tag (Cat. No. SPD-C5259) to Vero E6 cells surface ACE2 was inhibited by increasing concentration of Anti-SARS-CoV-2 Neutralizing Antibody, Human IgG1 (Cat. No. SAD-S35). The concentration of SARS-CoV-2 S protein RBD used is 5μg/ml. The IC50 is 10.33 μg/ml (Routinely tested).

Protocol

Biotinylated Human FACS

FACS analysis shows that the binding of SARS-CoV-2 S1 protein, Mouse IgG2a Fc Tag (Cat. No. S1N-C5257) to Vero E6 cells surface ACE2 was inhibited by increasing concentration of Anti-SARS-CoV-2 RBD Neutralizing Antibody, Human IgG1 (Cat. No. SAD-S35). The concentration of SARS-CoV-2 S1 protein used is 3ug/ml. The IC50 is 1.352 μg/ml (Routinely tested).

Protocol

 

文献引用(Citations)

背景(Background)

It's been reported that Coronavirus can infect the human respiratory epithelial cells through interaction with the human ACE2 receptor. The spike protein is a large type I transmembrane protein containing two subunits, S1 and S2. S1 mainly contains a receptor binding domain (RBD), which is responsible for recognizing the cell surface receptor. S2 contains basic elements needed for the membrane fusion.The S protein plays key parts in the induction of neutralizing-antibody and T-cell responses, as well as protective immunity.

 

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