Blocking Assay:Article Title: CD98hc is a target for brain delivery of biotherapeutics
Article Snippet: The total test article concentrations in cynomolgus monkey serum and brain lysate samples were quantified using a generic anti-human IgG sandwich-format electrochemiluminescence immunoassay (ECLIA) on a Meso Scale Discovery (MSD) platform. .. Briefly, 1% casein-based PBS blocking buffer (Thermo Scientific, 37528) was added to an MSD GOLD 96-well small-spot streptavidin-coated microtiter plate (Meso Scale Discovery, L45SA) and incubated for approximately 1 h. Following the plate blocking and wash steps, biotinylated goat anti-human IgG (SouthernBiotech 2049-08) at a working concentration of 0.5 μg/mL was added to coat the assay plate and allowed to incubate for 1–2 h. Subsequently, test samples were diluted (MRD of 1:100 in 0.5% casein-based PBS assay buffer) and added to the assay plate. .. Following the 1–2 h incubation in the capture step, a pre-adsorbed secondary ruthenylated (SULFO-TAG) goat anti-human IgG antibody (Meso Scale Discovery, R32AJ) at a working solution of 0.5 μg/mL was added to the assay plate and incubated for approximately 1 h. An assay read buffer (1X MSD Read Buffer T, R92TC) was then added to generate the electrochemiluminescence (ECL) assay signal, expressed in ECL units (ECLU).
Article Title: Dual targeting of transferrin receptor and CD98hc enhances brain exposure of large molecules.
Article Snippet: .. Quantification of huIgG concentration in brain lysate by MSD The total test article concentrations in brain lysate samples (Figures 2B, 2D, and 2F) were quantified using a generic anti-human IgG sandwich-format electrochemiluminescence immunoassay (ECLIA) on a Meso Scale Discovery (MSD) platform, as described previously.17 Briefly, 1% casein-based PBS blocking buffer (ThermoFisher, 37528) was added to an MSD GOLD 96-well small-spot streptavidin-coated microtiter plate (Meso Scale Discovery, L45SA) and incubated for approximately 1 h. Following the plate blocking and wash steps, biotinylated goat anti-human IgG (SouthernBiotech 2049-08) at a working concentration of 0.5 μg/mL was added to coat the assay plate and allowed to incubate for 1–2 h. Subsequently, test samples were diluted (MRD of 1:100 in 0.5% casein-based PBS assay buffer) and added to the assay plate. .. Following the 1–2 h incubation in the capture step, a pre-adsorbed secondary ruthenylated (SULFO-TAG) goat anti-human IgG antibody (Meso Scale Discovery, R32AJ) at a working solution of 0.5 μg/mL was added to the assay plate and incubated for approximately 1 h. An assay read buffer (1× MSD Read Buffer T, R92TC) was then added to generate the electrochemiluminescence (ECL) assay signal, expressed in ECL units (ECLU).
Article Title: Anti-TREM2 antibodies and methods of use thereof
Article Snippet: For anti-TREM2 antibody PK analysis, the total antibody concentrations in monkey plasma were quantified using a generic anti-human IgG sandwich electrochemiluminescence immunoassay (ECLIA) on a Meso Scale Discovery (MSD) platform. .. Briefly, 1% casein-based PBS blocking buffer (Thermo Scientific, MA) was added to an MSD GOLD 96-well small-spot streptavidin-coated microtiter plate (Meso Scale Discovery, MD) and incubated for approximately 1 hr. .. Following the plate blocking and wash steps, a biotinylated anti-human IgG goat antibody (SouthernBiotech, AL) at a working solution of 0.5 μg/mL was added to the assay plate and allowed to incubate for 1-2 hrs.
Article Title: Dual targeting of transferrin receptor and CD98hc enhances brain exposure of large molecules
Article Snippet: Similar to a protocol described previously , the total test article concentrations in brain lysate samples ( , d, f ) were quantified using a generic anti-human IgG sandwich-format electrochemiluminescence immunoassay (ECLIA) on a Meso Scale Discovery (MSD) platform. .. Briefly, 1% casein-based PBS blocking buffer (Thermo Scientific, 37528) was added to an MSD GOLD 96-well small-spot streptavidin-coated microtiter plate (Meso Scale Discovery, L45SA) and incubated for approximately 1 hour. .. Following the plate blocking and wash steps, biotinylated goat anti-human IgG (SouthernBiotech 2049-08) at a working concentration of 0.5 μg/mL was added to coat the assay plate and allowed to incubate for 1-2 hours.
Incubation:Article Title: CD98hc is a target for brain delivery of biotherapeutics
Article Snippet: The total test article concentrations in cynomolgus monkey serum and brain lysate samples were quantified using a generic anti-human IgG sandwich-format electrochemiluminescence immunoassay (ECLIA) on a Meso Scale Discovery (MSD) platform. .. Briefly, 1% casein-based PBS blocking buffer (Thermo Scientific, 37528) was added to an MSD GOLD 96-well small-spot streptavidin-coated microtiter plate (Meso Scale Discovery, L45SA) and incubated for approximately 1 h. Following the plate blocking and wash steps, biotinylated goat anti-human IgG (SouthernBiotech 2049-08) at a working concentration of 0.5 μg/mL was added to coat the assay plate and allowed to incubate for 1–2 h. Subsequently, test samples were diluted (MRD of 1:100 in 0.5% casein-based PBS assay buffer) and added to the assay plate. .. Following the 1–2 h incubation in the capture step, a pre-adsorbed secondary ruthenylated (SULFO-TAG) goat anti-human IgG antibody (Meso Scale Discovery, R32AJ) at a working solution of 0.5 μg/mL was added to the assay plate and incubated for approximately 1 h. An assay read buffer (1X MSD Read Buffer T, R92TC) was then added to generate the electrochemiluminescence (ECL) assay signal, expressed in ECL units (ECLU).
Article Title: Dual targeting of transferrin receptor and CD98hc enhances brain exposure of large molecules.
Article Snippet: .. Quantification of huIgG concentration in brain lysate by MSD The total test article concentrations in brain lysate samples (Figures 2B, 2D, and 2F) were quantified using a generic anti-human IgG sandwich-format electrochemiluminescence immunoassay (ECLIA) on a Meso Scale Discovery (MSD) platform, as described previously.17 Briefly, 1% casein-based PBS blocking buffer (ThermoFisher, 37528) was added to an MSD GOLD 96-well small-spot streptavidin-coated microtiter plate (Meso Scale Discovery, L45SA) and incubated for approximately 1 h. Following the plate blocking and wash steps, biotinylated goat anti-human IgG (SouthernBiotech 2049-08) at a working concentration of 0.5 μg/mL was added to coat the assay plate and allowed to incubate for 1–2 h. Subsequently, test samples were diluted (MRD of 1:100 in 0.5% casein-based PBS assay buffer) and added to the assay plate. .. Following the 1–2 h incubation in the capture step, a pre-adsorbed secondary ruthenylated (SULFO-TAG) goat anti-human IgG antibody (Meso Scale Discovery, R32AJ) at a working solution of 0.5 μg/mL was added to the assay plate and incubated for approximately 1 h. An assay read buffer (1× MSD Read Buffer T, R92TC) was then added to generate the electrochemiluminescence (ECL) assay signal, expressed in ECL units (ECLU).
Article Title: Anti-TREM2 antibodies and methods of use thereof
Article Snippet: For anti-TREM2 antibody PK analysis, the total antibody concentrations in monkey plasma were quantified using a generic anti-human IgG sandwich electrochemiluminescence immunoassay (ECLIA) on a Meso Scale Discovery (MSD) platform. .. Briefly, 1% casein-based PBS blocking buffer (Thermo Scientific, MA) was added to an MSD GOLD 96-well small-spot streptavidin-coated microtiter plate (Meso Scale Discovery, MD) and incubated for approximately 1 hr. .. Following the plate blocking and wash steps, a biotinylated anti-human IgG goat antibody (SouthernBiotech, AL) at a working solution of 0.5 μg/mL was added to the assay plate and allowed to incubate for 1-2 hrs.
Article Title: Dual targeting of transferrin receptor and CD98hc enhances brain exposure of large molecules
Article Snippet: Similar to a protocol described previously , the total test article concentrations in brain lysate samples ( , d, f ) were quantified using a generic anti-human IgG sandwich-format electrochemiluminescence immunoassay (ECLIA) on a Meso Scale Discovery (MSD) platform. .. Briefly, 1% casein-based PBS blocking buffer (Thermo Scientific, 37528) was added to an MSD GOLD 96-well small-spot streptavidin-coated microtiter plate (Meso Scale Discovery, L45SA) and incubated for approximately 1 hour. .. Following the plate blocking and wash steps, biotinylated goat anti-human IgG (SouthernBiotech 2049-08) at a working concentration of 0.5 μg/mL was added to coat the assay plate and allowed to incubate for 1-2 hours.
Concentration Assay:Article Title: CD98hc is a target for brain delivery of biotherapeutics
Article Snippet: The total test article concentrations in cynomolgus monkey serum and brain lysate samples were quantified using a generic anti-human IgG sandwich-format electrochemiluminescence immunoassay (ECLIA) on a Meso Scale Discovery (MSD) platform. .. Briefly, 1% casein-based PBS blocking buffer (Thermo Scientific, 37528) was added to an MSD GOLD 96-well small-spot streptavidin-coated microtiter plate (Meso Scale Discovery, L45SA) and incubated for approximately 1 h. Following the plate blocking and wash steps, biotinylated goat anti-human IgG (SouthernBiotech 2049-08) at a working concentration of 0.5 μg/mL was added to coat the assay plate and allowed to incubate for 1–2 h. Subsequently, test samples were diluted (MRD of 1:100 in 0.5% casein-based PBS assay buffer) and added to the assay plate. .. Following the 1–2 h incubation in the capture step, a pre-adsorbed secondary ruthenylated (SULFO-TAG) goat anti-human IgG antibody (Meso Scale Discovery, R32AJ) at a working solution of 0.5 μg/mL was added to the assay plate and incubated for approximately 1 h. An assay read buffer (1X MSD Read Buffer T, R92TC) was then added to generate the electrochemiluminescence (ECL) assay signal, expressed in ECL units (ECLU).
Article Title: Dual targeting of transferrin receptor and CD98hc enhances brain exposure of large molecules.
Article Snippet: .. Quantification of huIgG concentration in brain lysate by MSD The total test article concentrations in brain lysate samples (Figures 2B, 2D, and 2F) were quantified using a generic anti-human IgG sandwich-format electrochemiluminescence immunoassay (ECLIA) on a Meso Scale Discovery (MSD) platform, as described previously.17 Briefly, 1% casein-based PBS blocking buffer (ThermoFisher, 37528) was added to an MSD GOLD 96-well small-spot streptavidin-coated microtiter plate (Meso Scale Discovery, L45SA) and incubated for approximately 1 h. Following the plate blocking and wash steps, biotinylated goat anti-human IgG (SouthernBiotech 2049-08) at a working concentration of 0.5 μg/mL was added to coat the assay plate and allowed to incubate for 1–2 h. Subsequently, test samples were diluted (MRD of 1:100 in 0.5% casein-based PBS assay buffer) and added to the assay plate. .. Following the 1–2 h incubation in the capture step, a pre-adsorbed secondary ruthenylated (SULFO-TAG) goat anti-human IgG antibody (Meso Scale Discovery, R32AJ) at a working solution of 0.5 μg/mL was added to the assay plate and incubated for approximately 1 h. An assay read buffer (1× MSD Read Buffer T, R92TC) was then added to generate the electrochemiluminescence (ECL) assay signal, expressed in ECL units (ECLU).
Electrochemiluminescence:Article Title: Dual targeting of transferrin receptor and CD98hc enhances brain exposure of large molecules.
Article Snippet: .. Quantification of huIgG concentration in brain lysate by MSD The total test article concentrations in brain lysate samples (Figures 2B, 2D, and 2F) were quantified using a generic anti-human IgG sandwich-format electrochemiluminescence immunoassay (ECLIA) on a Meso Scale Discovery (MSD) platform, as described previously.17 Briefly, 1% casein-based PBS blocking buffer (ThermoFisher, 37528) was added to an MSD GOLD 96-well small-spot streptavidin-coated microtiter plate (Meso Scale Discovery, L45SA) and incubated for approximately 1 h. Following the plate blocking and wash steps, biotinylated goat anti-human IgG (SouthernBiotech 2049-08) at a working concentration of 0.5 μg/mL was added to coat the assay plate and allowed to incubate for 1–2 h. Subsequently, test samples were diluted (MRD of 1:100 in 0.5% casein-based PBS assay buffer) and added to the assay plate. .. Following the 1–2 h incubation in the capture step, a pre-adsorbed secondary ruthenylated (SULFO-TAG) goat anti-human IgG antibody (Meso Scale Discovery, R32AJ) at a working solution of 0.5 μg/mL was added to the assay plate and incubated for approximately 1 h. An assay read buffer (1× MSD Read Buffer T, R92TC) was then added to generate the electrochemiluminescence (ECL) assay signal, expressed in ECL units (ECLU).
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