SARS-CoV-2 (COVID-19) Spike Neutralization Single Domain Antibody [E10]

• Catalog Number: orb1274179
• Category: Antibodies
• Description: SARS-CoV-2 (COVID-19) Spike Neutralization Single Domain Antibody [E10]
• Species/Host: Camelus
• Clonality: Recombinant
• Clone Number: E10
• Tested applications: ELISA, NeA
• Reactivity: Virus
• Isotype: sdAb
• 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.
• Concentration: batch dependent
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• Target: S
• UniProt ID: P0DTC2
• NCBI: QHD43419
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: SARS-CoV-2 (COVID-19) Spike Neutralization Antibody is supplied in PBS.
• Alternative names: SARS-CoV-2 (COVID-19) Spike S1 Antibody: Severe ac
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Pseudovirus neutralization assay. Antibodies: SARS-CoV-2 (COVID-19) Spike RBD Antibodies, orb1274179 (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 orb1274179 (E10) was measured over a serial dilution to determine the half-maximal inhibitory concentration (IC50). orb1274179 (E10) exhibited a dose dependent neutralizing effect on all the variant pseudoviruses tested.

ACE2-RBD binding inhibitory ELISA. Antibodies: SARS-CoV-2 (COVID-19) Spike RBD Antibodies, orb1274179. ACE2-RBD binding inhibitory ELISA was performed using RBD protein of SARS-CoV-2 variants (wild-type, alpha, beta, epsilon, gamma and kappa) as coating antigens at 1 μg/mL and the anti-SARS-CoV-2 (COVID-19) RBD antibody orb1274179 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 (1:20000 dilution) using the TMB chromogenic substrate system. orb1274179 exhibited a dose dependent inhibitory effect on ACE2 binding to RBDs of all the variants tested.

ELISA Validation with RBDs of SARS-CoV-2 Variants. Antibodies: SARS-CoV-2 (COVID-19) Spike RBD Antibody, orb1274179. 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 (orb1274180) as the capture antibody, followed by anti-cMyc-tag antibody (orb1239964) at 1 μg/mL. Secondary: Goat anti-mouse IgG HRP conjugate at 1:5000 dilution. orb1274180 binds to RBDs of all the variants tested.

Pseudovirus neutralization assay. Antibodies: SARS-CoV-2 (COVID-19) Spike RBD Antibodies, orb1274180 (A10) and orb1274179 (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 orb1274180 (A10) and orb1274179 (E10) was measured over a serial dilution series to determine the half-maximal inhibitory concentration (IC50). Both orb1274180 (A10) and orb1274179 (E10) exhibited a dose dependent neutralizing effect on wild-type pseudoviruses, and the combination of the two showed a significantly synergistic effect.

Pseudovirus neutralization assay. Antibodies: SARS-CoV-2 (COVID-19) Spike RBD Antibodies, orb1274180 (A10) and orb1274179 (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 orb1274180 (A10) and orb1274179 (E10) was measured over a serial dilution series to determine the half-maximal inhibitory concentration (IC50). Both orb1274180 (A10) and orb1274179 (E10) exhibited a dose dependent neutralizing effect on alpha pseudoviruses, and the combination of the two showed a significantly synergistic effect.

ACE2-RBD binding inhibitory ELISA. Antibodies: SARS-CoV-2 (COVID-19) Spike RBD Antibodies, orb1274180 (A10) and orb1274179 (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 (orb1274180 and orb1274179) 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 (1:20000 dilution) using the TMB chromogenic substrate system. Both orb1274180 (A10) and orb1274179 (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.