The Simoa HD-1 Analyzer: A Novel Fully Automated Digital Immunoassay Analyzer with Single-Molecule Sensitivity and Multiplexing

Wilson, David H.; Rissin, David M.; Kan, Cheuk W.; Fournier, David R.; Piech, Tomasz; Campbell, Todd; Meyer, Raymond E.; Fishburn, Matthew W.; Cabrera, Carlos; Patel, Purvish P.; Frew, Erica; Chen, Yao; Chang, Lei; Ferrell, Evan P.; Einem, Volker von; McGuigan, William M.; Reinhardt, Marcus; Sayer, Heiko; Vielsack, Claus; Duffy, David C.

doi:10.1177/2211068215589580

Cited by 313 publications

(230 citation statements)

References 38 publications

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“…To measure tau in mouse plasma we utilized a Si ngle mo lecule a rray – (Simoa) assay, a sensitive digital ELISA platform (51). Homebrew assays specific for total tau were developed according to the manufacturer’s recommendations (Quanterix Corp).…”

Section: Methodsmentioning

confidence: 99%

Anti-tau antibody administration increases plasma tau in transgenic mice and patients with tauopathy

et al. 2017

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Tauopathies are a group of disorders in which the cytosolic protein tau aggregates and accumulates in cells within the brain, resulting in neurodegeneration. A promising treatment being explored for tauopathies is passive immunization with anti-tau antibodies. We previously found that administration of an anti-tau antibody to human tau transgenic mice increased the concentration of plasma tau. We further explored the effects of administering an anti-tau antibody on plasma tau. After peripheral administration of an anti-tau antibody to human patients with tauopathy and to mice expressing human tau in the central nervous system, there was a dose-dependent increase in plasma tau. In mouse plasma, we found that tau had a short half-life of 8 min that increased to more than 3 hours after administration of anti-tau antibody. As tau transgenic mice accumulated insoluble tau in the brain, brain soluble and interstitial fluid tau decreased. Administration of anti-tau antibody to tau transgenic mice that had decreased brain soluble tau and interstitial fluid tau resulted in an increase in plasma tau, but this increase was less than that observed in tau transgenic mice without these brain changes. Tau transgenic mice subjected to acute neuronal injury using 3-nitropropionic acid showed increased interstitial fluid tau and plasma tau. These data suggest that peripheral administration of an anti-tau antibody results in increased plasma tau, which correlates with the concentration of extracellular and soluble tau in the brain.

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Section: Methodsmentioning

confidence: 99%

Anti-tau antibody administration increases plasma tau in transgenic mice and patients with tauopathy

et al. 2017

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“…The prepared capture beads and biotinylated detection antibody detailed above were used to develop a digital ELISA for Aβ 1–42 measurement using the Simoa technology on a fully automated HD-1 Analyzer as described elsewhere [36, 39–41]. During the first step, 100 μl of calibrators or diluted samples were mixed with the capture beads (3 × 10 6 /ml) and biotinylated detection antibody (0.1 μg/ml) for 35 minutes at room temperature, then washed three times with wash buffer containing 5× PBS and 0.1% Tween 20.…”

Section: Methodsmentioning

confidence: 99%

“…Following a second wash, a solution of enzyme substrate, resorufin β- d -galactopyranoside, was added, and the capture beads were resuspended and then loaded into Simoa arrays, each containing 216,000 femtoliter-sized wells for detection. The assay was quantified by measurement of signals from bound analyte targets on the beads in units of average enzymes per bead (AEB) as previously described [36, 37, 41]. …”

Section: Methodsmentioning

confidence: 99%

“…Beads are subsequently loaded and sealed into an array of femtoliter-sized wells for digital measurement of signals. This ability to trap and detect single protein molecules provides unprecedented sensitivity compared with standard ELISA assays, where signal measurement usually occurs within the reacting mixture in comparatively large volumes [38–41]. The goals of this study were also to develop a digital ELISA using the same monoclonal antibodies used in previous clinical trials sponsored by Eli Lilly and Company (Indianapolis, IN, USA), evaluate the analytical performance, and demonstrate its ability to quantify Aβ 1–42 in samples from subjects treated with a previously characterized Aβ-lowering agent using the fully automated Simoa HD-1 Analyzer (Quanterix, Lexington, MA, USA).…”

Section: Introductionmentioning

confidence: 99%

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A digital enzyme-linked immunosorbent assay for ultrasensitive measurement of amyloid-β 1–42 peptide in human plasma with utility for studies of Alzheimer’s disease therapeutics

et al. 2016

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BackgroundAmyloid-β 1–42 peptide (Aβ1–42) is associated with plaque formation in the brain of patients with Alzheimer’s disease (AD). Pharmacodynamic studies of AD therapeutics that lower the concentrations of Aβ1–42 in peripheral blood require highly sensitive assays for its measurement. A digital enzyme-linked immunosorbent assay (ELISA) using single molecule array (Simoa) technology has been developed that provides improved sensitivity compared with conventional ELISA methods using the same antibody reagents.MethodsA sensitive digital ELISA for measurement of Aβ1–42 using antibodies 3D6 and 21F12 was developed. Assay performance was evaluated by repeated testing of pooled human plasma and buffer diluent quality control samples to determine relative accuracy, intra- and inter-assay precision, limit of detection (LOD), lower limit of quantification (LLOQ), dilutional linearity, and spike recovery. The optimized assay was used to quantify Aβ1–42 in clinical samples from patients treated with the β-site amyloid precursor protein cleaving enzyme 1 inhibitor LY2886721.ResultsThe prototype assay measured Aβ1–42 with an LOD of 0.3 pg/ml and an LLOQ of 2.8 pg/ml in plasma, calibrated using an Aβ1–42 peptide standard from Fujirebio. Assay precision was acceptable with intra- and inter-assay coefficients of variation both being ≤10%. Dilutional linearity was demonstrated in sample diluent and immunodepleted human plasma. Analyte spike recovery ranged from 51% to 93% with a mean of 80%. This assay was able to quantify Aβ1–42 in all of the 84 clinical samples tested. A rapid reduction in levels of Aβ1–42 was detected within 1 h after drug treatment, and a dose-dependent decrease of Aβ1–42 levels was also observed over the time course of sample collection.ConclusionsThis digital ELISA has potential utility in clinical applications for quantification of Aβ1–42 in plasma where high sensitivity and precision are required.

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“…However, relevant biomarkers are often present in very low abundance and conventional immunoassays lack in sensitivity [26]. Similarly to TILDA and iCARED, Simoa digital ELISA from Quanterix Corporation represents a novel promising assay for ultrasensitive detection of proteins at femtogram per milliliter levels [26–28]. This represents a >1:200-fold sensitivity improvement compared with classical ELISA [26].…”

Section: The Next-generation Assaysmentioning

confidence: 99%

HIV-1 Latent Reservoir: Size Matters

et al. 2016

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More than 35 million people remain infected with HIV-1. Upon antiretroviral therapy cessation, HIV-1-positive individuals systematically fail to achieve sustained virological remission, revealing the presence of a reservoir. This reservoir takes into account anatomical sanctuaries where HIV-1 continues to replicate, and latently infected cells also known as the latent reservoir (LR). A better understanding of the nature and features of the LR and its quantification are crucial to evaluate the efficiency of therapeutic strategies aiming at purging HIV-1. Culture- and PCR-based assays have already been implemented to measure the LR, and new assays are continuously being developed. In this review, we will discuss these methods highlighting the difficulties to accurately measure the LR, one main obstacle in curing HIV-1.

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The Simoa HD-1 Analyzer: A Novel Fully Automated Digital Immunoassay Analyzer with Single-Molecule Sensitivity and Multiplexing

Cited by 313 publications

References 38 publications

Anti-tau antibody administration increases plasma tau in transgenic mice and patients with tauopathy

Anti-tau antibody administration increases plasma tau in transgenic mice and patients with tauopathy

A digital enzyme-linked immunosorbent assay for ultrasensitive measurement of amyloid-β 1–42 peptide in human plasma with utility for studies of Alzheimer’s disease therapeutics

HIV-1 Latent Reservoir: Size Matters

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