Activity Assay- ELISA The Binding Activity of SARS-CoV-2 Spike RBD Nanobody with SARS-CoV-2-S1-RBD Activity: Measured by its binding ability in a functional ELISA. Immobilized SARS-CoV-2-S1-RBD (CSB-YP3324GMY1) at 2 µg/ml can bind SARS-CoV-2 Spike RBD Nanobody, the EC50 is 0.8674 ng/ml.
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GICA In the Colloidal Gold Immunochromatography Assay detection system, the background of antibody (CSB-RA33245A2GMY) is clean, the detection limit can be as low as 25ng/ml (1.75ng/0.07ml), and the sensitivity is very good.
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SARS-CoV-2 Spike RBD Nanobody (CSB-RA33245A2GMY) competed with ACE2-HRP conjugate (CSB-MP866317HU) for binding to SARS-CoV-2-S1-RBD (CSB-YP3324GMY1). The binding signal of SARS-CoV-2-S1-RBD and ACE2-HRP conjugate was gradually reduced as the SARS-CoV-2 Spike RBD Nanobody concentrations increased. It indicated that this SARS-CoV-2 Spike RBD Nanobody effectively inhibited the SARS-CoV-2-S1-RBD/ACE2 binding. And the IC50 of this SARS-CoV-2 Spike RBD Nanobody is 1.296 nM.
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SARS-CoV-2 Spike RBD Nanobody (CSB-RA33245A2GMY) competitively prevented SARS-CoV-2-S1-RBD (CSB-YP3324GMY1) from binding to ACE2-HRP conjugate (CSB-MP866317HU). The inhibition efficacy of the SARS-CoV-2-S1-RBD/ACE2 binding was positively proportionally to the SARS-CoV-2 Spike RBD Nanobody concentrations. It showed that this SARS-CoV-2 Spike RBD Nanobody effectively inhibited the SARS-CoV-2-S1-RBD/ACE2 binding. And the IC50 of this SARS-CoV-2 Spike RBD Nanobody is 0.1074 µg/ml.
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SARS-CoV-2 Spike protein RBD His/Sumostar Tag (CSB-YP3324GMY1) captured on COOH chip binding to the SARS-CoV-2 Spike RBD Nanobody (CSB-RA33245A2GMY), increases the local refractive index (RI), leading to a red shift of the LSPR peak position. The higher concentrations of SARS-CoV-2 Spike RBD Nanobody, the larger the wavelength shift. The detected affinity constant of SARS-CoV-2 Spike protein RBD/SARS-CoV-2 Spike RBD Nanobody binding is 28.2nM.
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ELISA: Immobilize various types of SARS proteins at concentration of 2µg/ml on solid substrate, then react with SARS-CoV-2 Spike RBD Nanobody at concentration of 100µg/ml, 10µg/ml and 1µg/ml. It shows the SARS-CoV-2 Spike RBD Nanobody (CSB-RA33245A2GMY) is specific for SARS-CoV-2-S1-RBD protein, without any cross-reactivity with MERS-CoV, SARS-CoV, HCoV-OC43 or HCoV-229E.