SARS-CoV-2 (COVID-19) Spike Neutralization Antibody [A10]

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ProSci
Product Code: PSI-SD9853
Product Group: Primary Antibodies
Supplier: ProSci
CodeSizePrice
PSI-SD9853-0.02mg0.02mg£166.00
Quantity:
PSI-SD9853-0.1mg0.1mg£531.00
Quantity:
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Overview

Host Type: Llama
Antibody Isotype: sdAb
Antibody Clonality: Recombinant Antibody
Antibody Clone: A10
Regulatory Status: RUO
Target Species: SARS-CoV-2
Applications:
  • Enzyme-Linked Immunosorbent Assay (ELISA)
  • Neutralisation
Shipping:
Dry Ice
Storage:
SARS-CoV-2 (COVID-19) Spike Neutralization Antibody should be stored in working aliquots at -20˚C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.

Images

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<strong>Figure 1 Neutralization Assay of SARS-CoV-2 Pseudovirus by SARS-CoV-2 Spike Antibody </strong><br> Neutralization antibody: Anti-SARS-CoV-2 Spike S1 antibody, <strong>SD9853</strong>, dilution: 0.1 ? 200 ng/mL. Pseudovirus: SARS-CoV-2 Spike Pseudovirus, <strong>95-200</strong>. Cells: ACE2-overexpressing 293T cell. Diluted antibody was incubated with Spike pseudovirus at 37 ˚C for 1 hr. ACE2-overexpressing 293T cells were added in each well and incubated at 37 ˚C for 48 hrs. Luminescence reporter reagent was added in each well and the final results were read with the luminescence plate reader. Percent inhibition is calculated based on the RLU value.
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<strong>Figure 2 ELISA Validation with RBDs of SARS-CoV-2 Variants</strong><br> Antibodies: SARS-CoV-2 (COVID-19) Spike RBD Antibody, SD9853. A direct ELISA was performed using RBD protein of SARS-CoV-2 variants (wild-type, alpha, beta, gamma and Delta) as coating antigens at 1 μg/mL and the anti-SARS-CoV-2 (COVID-19) RBD antibody (SD9853) as the capture antibody, followed by anti-cMyc-tag antibody (PM-7669) at 1 μg/mL. Secondary: Goat anti-mouse IgG HRP conjugate at 1:5000 dilution. SD9853 binds to RBDs of wild-type and alpha but not to the beta, gamma and Delta variants.
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<strong>Figure 3 ACE2-RBD binding inhibitory ELISA </strong><br> Antibodies: SARS-CoV-2 (COVID-19) Spike RBD Antibodies, SD9853 (A10) and SD9855 (E10). ACE2-RBD binding inhibitory ELISA was performed using RBD protein of SARS-CoV-2 variants (wild-type and alpha) as coating antigens at 1 μg/mL and the anti-SARS-CoV-2 (COVID-19) RBD antibody (SD9853 and SD9855) as the capture antibody, followed by incubation with 20 ng/mL human ACE2-Fc. Bound h-ACE2-Fc was detected with a goat anti-human IgG-HRP conjugate (1:20,000 dilution) using the TMB chromogenic substrate system. Both SD9853 (A10) and SD9855 (E10) exhibited a dose dependent inhibitory effect on ACE2 binding to RBDs of wild-type and alpha strains, and the combination of the two (A10+E10) showed a significantly synergistic effect.
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<strong>Figure 4 Pseudovirus neutralization assay</strong><br> Antibodies: SARS-CoV-2 (COVID-19) Spike RBD Antibodies, SD9853 (A10) and SD9855 (E10). Luciferase reporter virus-like particles and 293T-hsACE2 cells were used for the assay. After 72 hrs incubation infectivity was measured by luciferase expression using Promega Renilla-Glo Luciferase Assay System. Infectivity was measured as RLUs and no sdAb control was set to 100% infectivity. Neutralization activity of SD9853 (A10) and SD9855 (E10) was measured over a serial dilution series to determine the half-maximal inhibitory concentration (IC50). Both SD9853 (A10) and SD9855 (E10) exhibited a dose dependent neutralizing effect on both wild-type pseudoviruses, and the combination of the two showed a significantly synergistic effect.
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<strong>Figure 5 Pseudovirus neutralization assay</strong><br> Antibodies: SARS-CoV-2 (COVID-19) Spike RBD Antibodies, SD9853 (A10) and SD9855 (E10). Luciferase reporter virus-like particles and 293T-hsACE2 cells were used for the assay. After 72 hrs incubation infectivity was measured by luciferase expression using Promega Renilla-Glo Luciferase Assay System. Infectivity was measured as RLUs and no sdAb control was set to 100% infectivity. Neutralization activity of SD9853 (A10) and SD9855 (E10) was measured over a serial dilution series to determine the half-maximal inhibitory concentration (IC50). Both SD9853 (A10) and SD9855 (E10) exhibited a dose dependent neutralizing effect on both alpha pseudoviruses, and the combination of the two showed a significantly synergistic effect.
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<strong>Figure 1 Neutralization Assay of SARS-CoV-2 Pseudovirus by SARS-CoV-2 Spike Antibody </strong><br> Neutralization antibody: Anti-SARS-CoV-2 Spike S1 antibody, <strong>SD9853</strong>, dilution: 0.1 ? 200 ng/mL. Pseudovirus: SARS-CoV-2 Spike Pseudovirus, <strong>95-200</strong>. Cells: ACE2-overexpressing 293T cell. Diluted antibody was incubated with Spike pseudovirus at 37 ˚C for 1 hr. ACE2-overexpressing 293T cells were added in each well and incubated at 37 ˚C for 48 hrs. Luminescence reporter reagent was added in each well and the final results were read with the luminescence plate reader. Percent inhibition is calculated based on the RLU value.
<strong>Figure 2 ELISA Validation with RBDs of SARS-CoV-2 Variants</strong><br> Antibodies: SARS-CoV-2 (COVID-19) Spike RBD Antibody, SD9853. A direct ELISA was performed using RBD protein of SARS-CoV-2 variants (wild-type, alpha, beta, gamma and Delta) as coating antigens at 1 μg/mL and the anti-SARS-CoV-2 (COVID-19) RBD antibody (SD9853) as the capture antibody, followed by anti-cMyc-tag antibody (PM-7669) at 1 μg/mL. Secondary: Goat anti-mouse IgG HRP conjugate at 1:5000 dilution. SD9853 binds to RBDs of wild-type and alpha but not to the beta, gamma and Delta variants.
<strong>Figure 3 ACE2-RBD binding inhibitory ELISA </strong><br> Antibodies: SARS-CoV-2 (COVID-19) Spike RBD Antibodies, SD9853 (A10) and SD9855 (E10). ACE2-RBD binding inhibitory ELISA was performed using RBD protein of SARS-CoV-2 variants (wild-type and alpha) as coating antigens at 1 μg/mL and the anti-SARS-CoV-2 (COVID-19) RBD antibody (SD9853 and SD9855) as the capture antibody, followed by incubation with 20 ng/mL human ACE2-Fc. Bound h-ACE2-Fc was detected with a goat anti-human IgG-HRP conjugate (1:20,000 dilution) using the TMB chromogenic substrate system. Both SD9853 (A10) and SD9855 (E10) exhibited a dose dependent inhibitory effect on ACE2 binding to RBDs of wild-type and alpha strains, and the combination of the two (A10+E10) showed a significantly synergistic effect.
<strong>Figure 4 Pseudovirus neutralization assay</strong><br> Antibodies: SARS-CoV-2 (COVID-19) Spike RBD Antibodies, SD9853 (A10) and SD9855 (E10). Luciferase reporter virus-like particles and 293T-hsACE2 cells were used for the assay. After 72 hrs incubation infectivity was measured by luciferase expression using Promega Renilla-Glo Luciferase Assay System. Infectivity was measured as RLUs and no sdAb control was set to 100% infectivity. Neutralization activity of SD9853 (A10) and SD9855 (E10) was measured over a serial dilution series to determine the half-maximal inhibitory concentration (IC50). Both SD9853 (A10) and SD9855 (E10) exhibited a dose dependent neutralizing effect on both wild-type pseudoviruses, and the combination of the two showed a significantly synergistic effect.
<strong>Figure 5 Pseudovirus neutralization assay</strong><br> Antibodies: SARS-CoV-2 (COVID-19) Spike RBD Antibodies, SD9853 (A10) and SD9855 (E10). Luciferase reporter virus-like particles and 293T-hsACE2 cells were used for the assay. After 72 hrs incubation infectivity was measured by luciferase expression using Promega Renilla-Glo Luciferase Assay System. Infectivity was measured as RLUs and no sdAb control was set to 100% infectivity. Neutralization activity of SD9853 (A10) and SD9855 (E10) was measured over a serial dilution series to determine the half-maximal inhibitory concentration (IC50). Both SD9853 (A10) and SD9855 (E10) exhibited a dose dependent neutralizing effect on both alpha pseudoviruses, and the combination of the two showed a significantly synergistic effect.

Documents

Further Information

Additional Names:
SARS-CoV-2 (COVID-19) Spike S1 Antibody: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Surface Glycoprotein, Spike protein
Background:
Coronavirus disease 2019 (COVID-19), formerly known as 2019-nCoV acute respiratory disease, is an infectious disease caused by SARS-CoV-2, a virus closely related to the SARS virus (1). The disease is the cause of the 2019?20 coronavirus outbreak (2). The structure of 2019-nCoV consists of the following: a Spike protein (S), hemagglutinin-esterease dimer (HE), a membrane glycoprotein (M), an envelope protein (E) a nucleoclapid protein (N) and RNA. Coronavirus invades cells through Spike (S) glycoproteins, a class I fusion protein. It is the major viral surface protein that coronavirus uses to bind to the human cell surface receptor. It also mediates the fusion of host and viral cell membrane, allowing the virus to enter human cells and begin infection (3). The spike protein is the major target for neutralizing antibodies and vaccine development (4). The protein modeling suggests that there is strong interaction between Spike protein receptor-binding domain and its host receptor angiotensin-converting enzyme 2 (ACE2), which regulate both the cross-species and human-to-human transmissions of COVID-19 (5). The recent study has shown that the SARS-CoV-2 spike protein binds ACE2 with higher affinity than SARS-CoV spike protein (6).
Background References:
  • Gorbalenya. bioRxiv: 2020.
  • Hui et al. Int J Infect Dis. 2020;91:264-266.
  • Belouzard et al. Viruses. 2012;4(6):1011-33.
  • Lee et al. J Virol. 2006;80(8):4079-87.
  • Wan et al. J Virol. 2020.
  • Wrapp et al. Science. 2020.
Buffer:
SARS-CoV-2 (COVID-19) Spike Neutralization Antibody is supplied in PBS.
Concentration:
batch dependent
Conjugate:
Unconjugated
Immunogen:
SARS-CoV-2 S protein RBD containing C-terminal His Tag. The protein was expressed in human 293 cells (HEK293). It contains amino acids Arg 319 - Lys 537.
NCBI Gene ID #:
43740568
NCBI Official Name:
surface glycoprotein
NCBI Official Symbol:
S
NCBI Organism:
SARS-CoV-2
Physical State:
Liquid
Protein Accession #:
QHD43419
Protein GI Number:
1791269090
Purification:
SARS-CoV-2 (COVID-19) Spike Neutralization Antibody is affinity chromatography purified via Nickel column. Antibody is supplied as a His-tagged purified protein.??It also contains a myc-tag for detection.
Research Area:
Infectious Disease,COVID-19
Swissprot #:
P0DTC2
User NOte:
Optimal dilutions for each application to be determined by the researcher.