Adalimumab ADA were measured using MSD GOLD 96-well Streptavidin SECTOR Plates (L15SA) and a Meso Scale Discovery (MSD) MESO® QuickPlex SQ 120 Instrument. The MSD technology is based on electrochemiluminescence in a bridging format. Adalimumab (Humira®, Abbvie) was independently labeled with either EZ-Link® Sulfo-NHS-LC-Biotin (Thermo scientific 21327) or MSD SULFO-TAG NHS-Ester (Meso Scale Discovery, R91AO) according to the supplier's instructions. Plasma samples diluted at 1/10 (Minimum required dilution) from patients were incubated overnight with biotin-labeled adalimumab and then transferred on GOLD 96-well Streptavidin SECTOR Plates (L15SA) blocked prior with 150 μl of casein in PBS. The plate was then read on a MSD Sector Imager. The negative control was composed of a pool of plasma from 10 negative healthy donors supplied by the French Blood Bank (EFS, Rungis, France). The positive control was a pool of four monoclonal anti-Adalimumab antibodies produced and purified from a human B cell hybridoma by the Immune Regulation Laboratory Institute for Research in Biomedicine (IRB, Bellinzona) in the frame of the ABIRISK program. The sensitivity of the assay was determined by testing eight serial dilutions of the positive control spanning the cut point in three independent assays. The sensitivity was 18 ng/ml for anti-Adalimumab.
Meso quickplex sq 120 instrument
Manufactured by MSD
Sourced in United States
The MESO QuickPlex SQ 120 is an instrument designed for quantitative protein analysis. It utilizes electrochemiluminescence technology to detect and measure multiple analytes in a single sample. The core function of this instrument is to provide accurate and reliable protein quantification for research and diagnostic applications.
Automatically generated - may contain errors
Lab products found in correlation
Sector imager, by MSD (1 mentions)
Ez link sulfo nhs lc biotin, by Thermo Fisher Scientific (1 mentions)
Discovery workbench software, by MSD (1 mentions)
Spd c82e9, by ACROBiosystems (1 mentions)
Ez link sulfo nhs lc biotin kit, by Thermo Fisher Scientific (1 mentions)
Msd gold sulfo tag nhs ester conjugation pack, by MSD (1 mentions)
Contour next, by Ascensia Diabetes Care (1 mentions)
Discovery workbench 4, by MSD (1 mentions)
Contour xt, by Bayer (1 mentions)
5 protocols using meso quickplex sq 120 instrument
1
Detection of Anti-Adalimumab Antibodies
2
Multiplex Immunoassay Quantification of Plasma Biomarkers
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Plasma concentrations of ARG1, CCL20, CD163, CORIN, CXCL9 and PCSK9 were quantified using customized multiplex immunoassay kits from MSD (Meso Scale Discovery, Rockville, MD). CD163 (MSD, FCorreccted21J4) and PCSK9 (MSD, F21ABA) were multiplexed on a 2-assay plate (MSD, K15227N) according to the manufacturer’s instructions with samples diluted 1:20 in assay diluent and run in duplicate. ARG1 (MSD, F21Q1), CCL20 (MSD, B21UZ), CORIN (MSD, F210B) and CXCL9 (MSD, F210I) were multiplexed on a 4-assay plate (MSD, K15229N) according to the manufacturer’s instructions with samples diluted 1:2 in assay diluent and run in duplicate. Two control plasma samples were included on each plate to account for interplate variation. All plates were analyzed on the Meso Quick Plex SQ120 Instrument (MSD). Data were generated on the Methodological Mind software (version 1.0.36) and analyzed using Discovery Workbench software (version 4.0, MSD).
3
Mucosal IgA Response to SARS-CoV-2 Variants
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Vaccine-induced mucosal IgA specific to SARS-CoV-2 variant spike and RBD were measured with the Mesoscale Discovery (MSD) 4-spot U-PLEX Development Pack (MSD cat#K15229N). Spots were linked with anti-Syrian hamster IgA antibody (Brookwood Biomedical cat#sab3001a) biotinylated with the EZ-Link Sulfo-NHS-LC-Biotin kit (Thermo Fisher Scientific cat#A39257) and either biotinylated Wuhan, delta, or omicron variant SARS-CoV-2 spike trimer and RBD proteins (ACROBiosystems cat#SPD-C82E9, SPN-C82Ec, SPD-C82Ed, SPN-C82Ee, SPD-C82E4). Plates were coated, blocked, washed, and incubated with sample and detection antibody according to the manufacturer’s recommendations. Nasal samples were diluted at 1:15 and oral samples were diluted at 1:5 in Diluent-100 (MSD cat#R50AA). The anti-Syrian hamster IgA antibody was sulfo-tagged with the MSD GOLD SULFO-TAG NHS-Ester Conjugation Pack (MSD cat#R31AA) and diluted with Diluent-100 to 2 µg/mL for the nasal samples and 1 µg/mL for the oral samples. Plates were read on a Meso QuickPlex SQ 120 instrument (MSD). Samples were reported as relative light units (RLUs). Due to the variability in mucosal sampling, samples were normalized by total IgA and expressed as fold change was reported as vaccinated over unvaccinated sample. Data analysis was performed in GraphPad Prism (Version 9.4.1).
4
Quantifying Anti-RBD IgG Antibodies
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Anti-RBD IgG was measured in the EU-COVAT-1 AGED cohort using the commercially available MSD V-PLEX SARS-CoV-2 Panel 2 Kit (MSD, Rockville, MD, USA), as per the manufacturer’s instructions. Briefly, plates were blocked with MSD Blocker A for 30 min then washed with MSD wash buffer. Plasma samples, standards and control samples were diluted in MSD diluent 100, added to the plate and incubated for 2 h. Plates were then washed with MSD wash buffer and the detection antibody diluted in MSD diluent 100 to a concentration of 1 μg/mL was added to the plate and incubated for 1 h. Plates were washed with MSD wash buffer, MSD GOLD Read Buffer B was added, and plates were analysed with a MESO QuickPlex SQ 120 instrument (MSD, Rockville, MD, USA). Results were normalised to the WHO international standard using the conversion factor provided by the manufacturer.
5
Plasma Cytokine Profiling for Inflammation
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Glucose was measured from plasma samples using Bayer’s ContourXT with Contour next sensors for blood glucose determination (Ascensia Diabetes Care DeutschlandAG, Leverkusen, Germany). The concentration of glucose was given in mmol/l.
To assess early inflammatory processes, the plasma cytokines interleukin-6 (IL-6), interleukin 1β (lL-1β), tumor necrosis factor α (TNFα), and the signal molecule vascular endothelial growth factor A (VEGF-A) were measured using electrochemiluminescence-based assays from Meso Scale Diagnostics (MSD, Rockville, MD, USA). Therefore, a U-PLEX assay was used according to the manufacturer’s recommendations and all samples were analyzed in duplicates. Analyses were done using the MESO QuickPlex SQ 120 instrument (MSD) and DISCOVERY WORKBENCH® 4.0 software (MSD). For the purposes of statistical analyses, any value that was below the lowest limit of detection (LLOD) for the assay was considered negative and assigned a value of 0 pg/ml.
To assess early inflammatory processes, the plasma cytokines interleukin-6 (IL-6), interleukin 1β (lL-1β), tumor necrosis factor α (TNFα), and the signal molecule vascular endothelial growth factor A (VEGF-A) were measured using electrochemiluminescence-based assays from Meso Scale Diagnostics (MSD, Rockville, MD, USA). Therefore, a U-PLEX assay was used according to the manufacturer’s recommendations and all samples were analyzed in duplicates. Analyses were done using the MESO QuickPlex SQ 120 instrument (MSD) and DISCOVERY WORKBENCH® 4.0 software (MSD). For the purposes of statistical analyses, any value that was below the lowest limit of detection (LLOD) for the assay was considered negative and assigned a value of 0 pg/ml.
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