Multiplex Assay:Article Title: SARS-CoV-2-specific antibody detection for sero-epidemiology: a multiplex analysis approach accounting for accurate seroprevalence.
Article Snippet: Assay ProcedureThe steps in assay validation were similar to recently developed bead-based multiplex immunoassays for CMV, EBV, and RSV, with minor modifications as described below [16, 17]. .. For the multiplex bead-based immune assay the following antigens obtained from Sino Biological were used: SARS-CoV-2 monomeric spike S1 (40591-V08H), RBD (40592-V08B), and nucleoprotein (N) (40588-V08B). ..
Article Title: SARS-CoV-2–Specific Antibody Detection for Seroepidemiology: A Multiplex Analysis Approach Accounting for Accurate Seroprevalence
Article Snippet: Assay Procedure The steps in assay validation were similar to recently developed bead-based multiplex immunoassays for CMV, EBV, and RSV, with minor modifications as described below [ , ]. .. For the multiplex bead-based immune assay the following antigens obtained from Sino Biological were used: SARS-CoV-2 monomeric spike S1 (40591-V08H), RBD (40592-V08B), and nucleoprotein (N) (40588-V08B). ..
Enzyme-linked Immunosorbent Assay:Article Title: Intradermal-delivered DNA vaccine provides anamnestic protection in a rhesus macaque SARS-CoV-2 challenge model
Article Snippet: Ninety-six well immunosorbent plates (NUNC) were coated with 1ug/mL recombinant SARS-CoV-2 S1+S2 ECD protein (Sino Biological 40589-V08B1), S1 protein (Sino Biological 40591-V08H), S2 protein (Sino Biological 40590-V08B), or receptor-binding domain (RBD) protein (Sino Biological 40595-V05H) in PBS overnight at 4°C. .. ELISA plates were also coated with 1 μg/mL recombinant SARS-CoV S1 protein (Sino Biological 40150-V08B1) and RBD protein (Sino Biological 40592-V08B) or MERS-CoV spike (Sino Biological 40069-V08B). ..
Article Title: Disease severity dictates SARS-CoV-2-specific neutralizing antibody responses in COVID-19
Article Snippet: .. Enzyme-linked immunosorbent assay As previously described, 50 ng of SARS-CoV-2 S1 protein (Sino Biological, 40591-V08H) or SARS-CoV-2 RBD protein (Sino Biological, 40592-V08B) or SARS-CoV-2 S2 protein (Sino Biological, 40590-V08B) in 100 μl phosphate-buffered saline (PBS) per well was coated on ELISA plates (Costar, 42592) overnight at 4 °C. .. The ELISA plates were blocked for 1 h with 100 μl blocking buffer (5% fetal bovine serum (FBS) and 0.1% Tween 20 in PBS) and then incubated with diluted patient or healthy control sera in 100 μl blocking buffer for 1 h. After washing with PBST buffer (0.1% Tween 20 in PBS), the ELISA plates were incubated with anti-human IgG horseradish peroxidase (HRP) antibody (Bioss Biotech, 0297D) for 45 min, followed by PBST washing and addition of 3,3′,5,5′-tetramethylbenzidine (TMB) buffer (Beyotime, P0209).
Article Title: Identification of IgG antibody response to SARS-CoV-2 spike protein and its receptor binding domain does not predict rapid recovery from COVID-19
Article Snippet: .. Reagents Recombinant protein Vendor Catalog SARS-CoV-2 (2019-nCoV) Spike RBD-His Recombinant Protein Sino Biological 40592-V08H Antibodies Vendor Catalog ELISA Dilution WB Dilution Strep TagII antibody Sigma 71590-3 1:500 1:1000 His Tag antibody Cell Signaling 2365S n/a 1:1000 SARS-CoV-2 (2019-nCoV) Spike RBD Antibody Sino Biological 40592-T62 n/a 1:1000 Goat anti-Human IgG (Fc specific)-Peroxidase Sigma-Aldrich A0170 1:10,000 n/a Goat anti-Mouse IgG (Fc specific)-Peroxidase Sigma-Aldrich A0168 1:10,000 n/a Goat anti-Rabbit IgG (whole molecule)–Peroxidase Sigma-Aldrich A0545 1:5,000 n/a Goat anti-mouse IgG (H) HRP Promega W4021 n/a 1:2000 Donkey anti-rabbit IgG (H) HRP Abcam ab16284 n/a 1:2000 Reagents Vendor Catalog Sodium carbonate (Na2CO3) Fisher Scientific S495-500 Sodium bicarbonate (NaHCO3) Sigma-Aldrich S6014-25G Hydrochoric acid (HCl) Fisher Scientific A144-212 Sodium hydroxide (NaOH) Fisher Scientific S318 Sulfuric acid (H2SO4) Fluka 35276 Tris base Fisher Scientific BP152 Sodium chloride (NaCl) Fisher Scientific BP358-1 Tween 20 Fisher Scientific BP337-500 3,3′,5,5′-Tetramethylbenzidine reagent (TMB) KPL 50-76-06 Bolt™ 4 to 12%, Bis-Tris, 1.0 mm, Mini Protein Gel, 10-well Invitrogen NW04120BOX Bolt™ 10%, Bis-Tris, 1.0 mm, Mini Protein Gel, 12-well Invitrogen NW00102BOX 20X Bolt™ MES SDS Running Buffer Invitrogen B0002 4x Laemmli Sample Buffer Bio-Rad 1610747 Trans-Blot Turbo RTA Mini 0.2 µm PVDF Transfer Kit Bio-Rad 1704272 West-Q Pico ECL Solution GenDEPOT W3652-020 Amersham Hyperfilm ECL cytiva (formerly GE Healthcare) 28906835 SimplyBlue SafeStrain Invitrogen LC6060 EDTA-free protease inhibitor cocktail Roche 04-693-159-001 Precison Plus Protein Dual Color Standards Bio-Rad 161-0374 Protein Purification Reagents Vendor Catalog Qiagen Ni-NTA Superflow 5 mL Qiagen 30761 StrepTrap HP 1 mL cytiva (formerly GE Healthcare) 28907546 Superdex S200 Increase 10/300 GL cytiva (formerly GE Healthcare) 28990944 Amicon® Ultra-4 Centrifugal Filter Unit MilliporeSigma UFC803024 Biotin Sigma-Aldrich B4501. ..
Recombinant:Article Title: Intradermal-delivered DNA vaccine provides anamnestic protection in a rhesus macaque SARS-CoV-2 challenge model
Article Snippet: Ninety-six well immunosorbent plates (NUNC) were coated with 1ug/mL recombinant SARS-CoV-2 S1+S2 ECD protein (Sino Biological 40589-V08B1), S1 protein (Sino Biological 40591-V08H), S2 protein (Sino Biological 40590-V08B), or receptor-binding domain (RBD) protein (Sino Biological 40595-V05H) in PBS overnight at 4°C. .. ELISA plates were also coated with 1 μg/mL recombinant SARS-CoV S1 protein (Sino Biological 40150-V08B1) and RBD protein (Sino Biological 40592-V08B) or MERS-CoV spike (Sino Biological 40069-V08B). ..
Article Title: Rapid SARS-CoV-2 Detection Using Electrochemical Immunosensor
Article Snippet: The spike protein detection antibody (#40591-MM43, Sino Biological) was biotinylated using a 40:1 molar ratio of biotin (#A39259, Thermo Fisher Scientific) to antibody. .. Recombinant Spike subunit 1 protein was applied as a calibrator (#40592-V08B, Sino Biological). .. 22E6 PFU/mL recombinant SARS CoV-2 virus (Linköping University, Linköping, Sweden) was diluted in PBS with 1% BSA, followed by a two-fold dilution in lysis buffer (100 mM Tris-HCl, 800 mM NaCl, pH 9.0, 1% Triton x-100, 1% BSA).
Article Title: Identification of IgG antibody response to SARS-CoV-2 spike protein and its receptor binding domain does not predict rapid recovery from COVID-19
Article Snippet: .. Reagents Recombinant protein Vendor Catalog SARS-CoV-2 (2019-nCoV) Spike RBD-His Recombinant Protein Sino Biological 40592-V08H Antibodies Vendor Catalog ELISA Dilution WB Dilution Strep TagII antibody Sigma 71590-3 1:500 1:1000 His Tag antibody Cell Signaling 2365S n/a 1:1000 SARS-CoV-2 (2019-nCoV) Spike RBD Antibody Sino Biological 40592-T62 n/a 1:1000 Goat anti-Human IgG (Fc specific)-Peroxidase Sigma-Aldrich A0170 1:10,000 n/a Goat anti-Mouse IgG (Fc specific)-Peroxidase Sigma-Aldrich A0168 1:10,000 n/a Goat anti-Rabbit IgG (whole molecule)–Peroxidase Sigma-Aldrich A0545 1:5,000 n/a Goat anti-mouse IgG (H) HRP Promega W4021 n/a 1:2000 Donkey anti-rabbit IgG (H) HRP Abcam ab16284 n/a 1:2000 Reagents Vendor Catalog Sodium carbonate (Na2CO3) Fisher Scientific S495-500 Sodium bicarbonate (NaHCO3) Sigma-Aldrich S6014-25G Hydrochoric acid (HCl) Fisher Scientific A144-212 Sodium hydroxide (NaOH) Fisher Scientific S318 Sulfuric acid (H2SO4) Fluka 35276 Tris base Fisher Scientific BP152 Sodium chloride (NaCl) Fisher Scientific BP358-1 Tween 20 Fisher Scientific BP337-500 3,3′,5,5′-Tetramethylbenzidine reagent (TMB) KPL 50-76-06 Bolt™ 4 to 12%, Bis-Tris, 1.0 mm, Mini Protein Gel, 10-well Invitrogen NW04120BOX Bolt™ 10%, Bis-Tris, 1.0 mm, Mini Protein Gel, 12-well Invitrogen NW00102BOX 20X Bolt™ MES SDS Running Buffer Invitrogen B0002 4x Laemmli Sample Buffer Bio-Rad 1610747 Trans-Blot Turbo RTA Mini 0.2 µm PVDF Transfer Kit Bio-Rad 1704272 West-Q Pico ECL Solution GenDEPOT W3652-020 Amersham Hyperfilm ECL cytiva (formerly GE Healthcare) 28906835 SimplyBlue SafeStrain Invitrogen LC6060 EDTA-free protease inhibitor cocktail Roche 04-693-159-001 Precison Plus Protein Dual Color Standards Bio-Rad 161-0374 Protein Purification Reagents Vendor Catalog Qiagen Ni-NTA Superflow 5 mL Qiagen 30761 StrepTrap HP 1 mL cytiva (formerly GE Healthcare) 28907546 Superdex S200 Increase 10/300 GL cytiva (formerly GE Healthcare) 28990944 Amicon® Ultra-4 Centrifugal Filter Unit MilliporeSigma UFC803024 Biotin Sigma-Aldrich B4501. ..
Western Blot:Article Title: Identification of IgG antibody response to SARS-CoV-2 spike protein and its receptor binding domain does not predict rapid recovery from COVID-19
Article Snippet: .. Reagents Recombinant protein Vendor Catalog SARS-CoV-2 (2019-nCoV) Spike RBD-His Recombinant Protein Sino Biological 40592-V08H Antibodies Vendor Catalog ELISA Dilution WB Dilution Strep TagII antibody Sigma 71590-3 1:500 1:1000 His Tag antibody Cell Signaling 2365S n/a 1:1000 SARS-CoV-2 (2019-nCoV) Spike RBD Antibody Sino Biological 40592-T62 n/a 1:1000 Goat anti-Human IgG (Fc specific)-Peroxidase Sigma-Aldrich A0170 1:10,000 n/a Goat anti-Mouse IgG (Fc specific)-Peroxidase Sigma-Aldrich A0168 1:10,000 n/a Goat anti-Rabbit IgG (whole molecule)–Peroxidase Sigma-Aldrich A0545 1:5,000 n/a Goat anti-mouse IgG (H) HRP Promega W4021 n/a 1:2000 Donkey anti-rabbit IgG (H) HRP Abcam ab16284 n/a 1:2000 Reagents Vendor Catalog Sodium carbonate (Na2CO3) Fisher Scientific S495-500 Sodium bicarbonate (NaHCO3) Sigma-Aldrich S6014-25G Hydrochoric acid (HCl) Fisher Scientific A144-212 Sodium hydroxide (NaOH) Fisher Scientific S318 Sulfuric acid (H2SO4) Fluka 35276 Tris base Fisher Scientific BP152 Sodium chloride (NaCl) Fisher Scientific BP358-1 Tween 20 Fisher Scientific BP337-500 3,3′,5,5′-Tetramethylbenzidine reagent (TMB) KPL 50-76-06 Bolt™ 4 to 12%, Bis-Tris, 1.0 mm, Mini Protein Gel, 10-well Invitrogen NW04120BOX Bolt™ 10%, Bis-Tris, 1.0 mm, Mini Protein Gel, 12-well Invitrogen NW00102BOX 20X Bolt™ MES SDS Running Buffer Invitrogen B0002 4x Laemmli Sample Buffer Bio-Rad 1610747 Trans-Blot Turbo RTA Mini 0.2 µm PVDF Transfer Kit Bio-Rad 1704272 West-Q Pico ECL Solution GenDEPOT W3652-020 Amersham Hyperfilm ECL cytiva (formerly GE Healthcare) 28906835 SimplyBlue SafeStrain Invitrogen LC6060 EDTA-free protease inhibitor cocktail Roche 04-693-159-001 Precison Plus Protein Dual Color Standards Bio-Rad 161-0374 Protein Purification Reagents Vendor Catalog Qiagen Ni-NTA Superflow 5 mL Qiagen 30761 StrepTrap HP 1 mL cytiva (formerly GE Healthcare) 28907546 Superdex S200 Increase 10/300 GL cytiva (formerly GE Healthcare) 28990944 Amicon® Ultra-4 Centrifugal Filter Unit MilliporeSigma UFC803024 Biotin Sigma-Aldrich B4501. ..
Protease Inhibitor:Article Title: Identification of IgG antibody response to SARS-CoV-2 spike protein and its receptor binding domain does not predict rapid recovery from COVID-19
Article Snippet: .. Reagents Recombinant protein Vendor Catalog SARS-CoV-2 (2019-nCoV) Spike RBD-His Recombinant Protein Sino Biological 40592-V08H Antibodies Vendor Catalog ELISA Dilution WB Dilution Strep TagII antibody Sigma 71590-3 1:500 1:1000 His Tag antibody Cell Signaling 2365S n/a 1:1000 SARS-CoV-2 (2019-nCoV) Spike RBD Antibody Sino Biological 40592-T62 n/a 1:1000 Goat anti-Human IgG (Fc specific)-Peroxidase Sigma-Aldrich A0170 1:10,000 n/a Goat anti-Mouse IgG (Fc specific)-Peroxidase Sigma-Aldrich A0168 1:10,000 n/a Goat anti-Rabbit IgG (whole molecule)–Peroxidase Sigma-Aldrich A0545 1:5,000 n/a Goat anti-mouse IgG (H) HRP Promega W4021 n/a 1:2000 Donkey anti-rabbit IgG (H) HRP Abcam ab16284 n/a 1:2000 Reagents Vendor Catalog Sodium carbonate (Na2CO3) Fisher Scientific S495-500 Sodium bicarbonate (NaHCO3) Sigma-Aldrich S6014-25G Hydrochoric acid (HCl) Fisher Scientific A144-212 Sodium hydroxide (NaOH) Fisher Scientific S318 Sulfuric acid (H2SO4) Fluka 35276 Tris base Fisher Scientific BP152 Sodium chloride (NaCl) Fisher Scientific BP358-1 Tween 20 Fisher Scientific BP337-500 3,3′,5,5′-Tetramethylbenzidine reagent (TMB) KPL 50-76-06 Bolt™ 4 to 12%, Bis-Tris, 1.0 mm, Mini Protein Gel, 10-well Invitrogen NW04120BOX Bolt™ 10%, Bis-Tris, 1.0 mm, Mini Protein Gel, 12-well Invitrogen NW00102BOX 20X Bolt™ MES SDS Running Buffer Invitrogen B0002 4x Laemmli Sample Buffer Bio-Rad 1610747 Trans-Blot Turbo RTA Mini 0.2 µm PVDF Transfer Kit Bio-Rad 1704272 West-Q Pico ECL Solution GenDEPOT W3652-020 Amersham Hyperfilm ECL cytiva (formerly GE Healthcare) 28906835 SimplyBlue SafeStrain Invitrogen LC6060 EDTA-free protease inhibitor cocktail Roche 04-693-159-001 Precison Plus Protein Dual Color Standards Bio-Rad 161-0374 Protein Purification Reagents Vendor Catalog Qiagen Ni-NTA Superflow 5 mL Qiagen 30761 StrepTrap HP 1 mL cytiva (formerly GE Healthcare) 28907546 Superdex S200 Increase 10/300 GL cytiva (formerly GE Healthcare) 28990944 Amicon® Ultra-4 Centrifugal Filter Unit MilliporeSigma UFC803024 Biotin Sigma-Aldrich B4501. ..
Protein Purification:Article Title: Identification of IgG antibody response to SARS-CoV-2 spike protein and its receptor binding domain does not predict rapid recovery from COVID-19
Article Snippet: .. Reagents Recombinant protein Vendor Catalog SARS-CoV-2 (2019-nCoV) Spike RBD-His Recombinant Protein Sino Biological 40592-V08H Antibodies Vendor Catalog ELISA Dilution WB Dilution Strep TagII antibody Sigma 71590-3 1:500 1:1000 His Tag antibody Cell Signaling 2365S n/a 1:1000 SARS-CoV-2 (2019-nCoV) Spike RBD Antibody Sino Biological 40592-T62 n/a 1:1000 Goat anti-Human IgG (Fc specific)-Peroxidase Sigma-Aldrich A0170 1:10,000 n/a Goat anti-Mouse IgG (Fc specific)-Peroxidase Sigma-Aldrich A0168 1:10,000 n/a Goat anti-Rabbit IgG (whole molecule)–Peroxidase Sigma-Aldrich A0545 1:5,000 n/a Goat anti-mouse IgG (H) HRP Promega W4021 n/a 1:2000 Donkey anti-rabbit IgG (H) HRP Abcam ab16284 n/a 1:2000 Reagents Vendor Catalog Sodium carbonate (Na2CO3) Fisher Scientific S495-500 Sodium bicarbonate (NaHCO3) Sigma-Aldrich S6014-25G Hydrochoric acid (HCl) Fisher Scientific A144-212 Sodium hydroxide (NaOH) Fisher Scientific S318 Sulfuric acid (H2SO4) Fluka 35276 Tris base Fisher Scientific BP152 Sodium chloride (NaCl) Fisher Scientific BP358-1 Tween 20 Fisher Scientific BP337-500 3,3′,5,5′-Tetramethylbenzidine reagent (TMB) KPL 50-76-06 Bolt™ 4 to 12%, Bis-Tris, 1.0 mm, Mini Protein Gel, 10-well Invitrogen NW04120BOX Bolt™ 10%, Bis-Tris, 1.0 mm, Mini Protein Gel, 12-well Invitrogen NW00102BOX 20X Bolt™ MES SDS Running Buffer Invitrogen B0002 4x Laemmli Sample Buffer Bio-Rad 1610747 Trans-Blot Turbo RTA Mini 0.2 µm PVDF Transfer Kit Bio-Rad 1704272 West-Q Pico ECL Solution GenDEPOT W3652-020 Amersham Hyperfilm ECL cytiva (formerly GE Healthcare) 28906835 SimplyBlue SafeStrain Invitrogen LC6060 EDTA-free protease inhibitor cocktail Roche 04-693-159-001 Precison Plus Protein Dual Color Standards Bio-Rad 161-0374 Protein Purification Reagents Vendor Catalog Qiagen Ni-NTA Superflow 5 mL Qiagen 30761 StrepTrap HP 1 mL cytiva (formerly GE Healthcare) 28907546 Superdex S200 Increase 10/300 GL cytiva (formerly GE Healthcare) 28990944 Amicon® Ultra-4 Centrifugal Filter Unit MilliporeSigma UFC803024 Biotin Sigma-Aldrich B4501. ..
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