Emerging Technologies for Biotherapeutic Bioanalysis from a High-Throughput and Multiplexing Perspective: Insights from an AAPS Emerging Technology Action Program Committee

Purushothama, Shobha; Dysinger, Mark; Chen, Yao; Österlund, Karolina; Mora, Johanna; Chunyk, Allison G.; Peloquin, Russ

doi:10.4155/bio-2017-0196

Cited by 12 publications

(4 citation statements)

References 9 publications

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“…Despite the challenge of selecting visits/samples from a prospectively assessed cohort in which patients were not followed in lockstep nor flare rates predicted to have an equal number of preflare and pre-nonflare visits, we confirmed that immune changes occurred prior to clinical disease flare, whether assessed en masse or on an individual patient level, and this information was harnessed to inform a refined FRI. The microfluidic, automated Ella immunoassay platform, designed for commercial use with minimal batch effects between users, sites, and lots (11,(22)(23)(24), allowed for design of the refined FRI-11 for clinical application.…”

Section: Discussionmentioning

confidence: 99%

A Flare Risk Index Informed by Select Immune Mediators in Systemic Lupus Erythematosus

Munroe

Blankenship

DeFreese³

et al. 2023

Arthritis & Rheumatology

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Objective. Systemic lupus erythematosus (SLE) is marked by immune dysregulation linked to varied clinical disease activity. Using a unique longitudinal cohort of SLE patients, this study sought to identify optimal immune mediators informing an empirically refined flare risk index (FRI) reflecting altered immunity prior to clinical disease flare.Methods. Thirty-seven SLE-associated plasma mediators were evaluated by microfluidic immunoassay in 46 samples obtained in SLE patients with an imminent clinical disease flare (preflare) and 53 samples obtained in SLE patients without a flare over a corresponding period (pre-nonflare). SLE patients were selected from a unique longitudinal cohort of 106 patients with classified SLE (meeting the American College of Rheumatology 1997 revised criteria for SLE or the Systemic Lupus International Collaborating Clinics 2012 revised criteria for SLE). Autoantibody specificities, hybrid SLE Disease Activity Index (hSLEDAI) scores, clinical features, and medication usage were also compared at preflare (mean ± SD 111 ± 47 days prior to flare) versus pre-nonflare (99 ± 21 days prior to nonflare) time points. Variable importance was determined by random forest analysis with logistic regression subsequently applied to determine the optimal number and type of analytes informing a refined FRI.Results. Preflare versus pre-nonflare differences were not associated with demographics, autoantibody specificities, hSLEDAI scores, clinical features, nor medication usage. Forward selection and backward elimination of mediators ranked by variable importance resulted in 17 plasma mediator candidates differentiating preflare from pre-nonflare visits. A final combination of 11 mediators best informed a newly refined FRI, which achieved a maximum sensitivity of 97% and maximum specificity of 98% after applying decision curve analysis to define low, medium, and high FRI scores.Conclusion. We verified altered immune mediators associated with imminent disease flare, and a subset of these mediators improved the FRI to identify SLE patients at risk of imminent flare. This molecularly informed, proactive management approach could be critical in prospective clinical trials and the clinical management of lupus.

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Section: Discussionmentioning

confidence: 99%

A Flare Risk Index Informed by Select Immune Mediators in Systemic Lupus Erythematosus

Munroe

Blankenship

DeFreese³

et al. 2023

Arthritis & Rheumatology

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“…The sensitivity of the available Ella™ assays is in the lower pg/mL range with a required sample volume between 2.5 and 50 µL. 16,17 While a vast literature is available on the use of SiMoA™ for the detection of plasma NfL levels, the assay based on Ella™ has been introduced more recently and its performance in comparison with the one observed on SiMoA™ needs to be understood. In a rst study, Ella™ and SiMoA™ were used to detect blood levels of NfL in serum samples of patients with multiple sclerosis.…”

Section: Introductionmentioning

confidence: 99%

“…12− 15 Ella™ is a micro uidic cartridge-based immunoassay platform from ProteinSimple (part of Bio-Techne), which is widely used to quantify soluble biomarkers. 16,17 Different cartridge formats allow measuring 72 samples in one assay. Measures are performed in triplicates in three glass nanoreactors (GNRs) for each sample using a uorescent substrate, which helps to achieve a wide dynamic measuring range.…”

Section: Introductionmentioning

confidence: 99%

Neurofilament-light chain quantification by SimoaTM and EllaTM in plasma from patients with dementia: a comparative study

Truffi

Ricciardi

Ramusino

et al. 2022

Preprint

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Neurofilament light chains (NfL) are neuron-specific cytoskeletal proteins whose plasmatic concentrations have been explored as a clinically useful marker in several types of dementia. Plasma concentrations of NfL are extremely low, and just two assays are commercially available for their study: one based on the SiMoA™ technology and one based on Ella™. We thus studied plasma levels of NfL with both platforms to check the correlation between them and to assess their potential in the diagnosis of dementia. Plasma NfL levels were measured on 50 subjects: 18 healthy controls, 20 Alzheimer’s disease, and 12 frontotemporal dementia patients. Ella™ returned plasmatic NfL levels significantly higher than SiMoA™, however the results were strongly correlated (r = 0.94), and a proportional coefficient of 0.58 between the two assays was calculated. Both assays detected higher plasma NfL levels in patients with dementia than in the control group (p < 0.0001) and allowed their discrimination with excellent diagnostic performance (AUC > 0.95). No difference was found between Alzheimer’s and Frontotemporal dementia either using SiMoA™ or Ella™. In conclusion, both the analytical platform resulted effective in analysing plasma levels of NfL. However, the correct interpretation of results requires the precise knowledge of the assay used.

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“…Array-based immunoassays were first described by Ekins et al and have been actively researched over the last 30 years. , In these immunoassays, capture antibodies are printed onto a planar surface and bind the target molecules in a sample that are subsequently labeled and detected. This type of immunoassay is amenable to multiplex protein detection by immobilizing capture antibodies specific to different proteins at predefined locations on a planar surface.…”

Section: Introductionmentioning

confidence: 99%

Customizable Multiplex Antibody Array Immunoassays with Attomolar Sensitivities

Tobos

Sheehan²,

Duffy

et al. 2020

Anal. Chem.

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We have developed a customizable contact printed multiplex immunoassay capable of simultaneously measuring up to five analytes with attomolar sensitivities. This enzyme-linked immunosorbent assay (ELISA) was based on spotting different antibodies in a circular pattern at the bottom of a microtiter plate well. Unlike traditional antibody printing for ELISA that prints a capture antibody specific to a target of interest, in this ELISA we printed unique “anchor” antibodies at the well surface, each having a high affinity for a specific peptide target. By coupling each peptide to a unique assay capture antibody, this array of anchor antibodies enabled a customizable contact printed multiplex immunoassay workflow. As a proof of concept, we developed a 5-plex assay measuring interleukin 5 (IL-5), interleukin 6 (IL-6), interleukin 10 (IL-10), interleukin 22 (IL-22), and tumor necrosis factor alpha (TNF-α). Measurements of these five analytes in serum and plasma correlated well between the method utilizing the anchor antibodies and peptides and the traditional capture antibody printing approach, with r 2 values of 0.99, 0.93, 0.99, 0.96, and 0.75 for IL-5, IL-6, IL-10, IL-22, and TNFα, respectively. This approach makes customizable multiplex ultrasensitive ELISA available to laboratories without access to the precision printing instrumentation and will be useful for antibody screening, custom assay development, biomarker detection, and protein profiling for diagnostic applications.

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Emerging Technologies for Biotherapeutic Bioanalysis from a High-Throughput and Multiplexing Perspective: Insights from an AAPS Emerging Technology Action Program Committee

Cited by 12 publications

References 9 publications

A Flare Risk Index Informed by Select Immune Mediators in Systemic Lupus Erythematosus

A Flare Risk Index Informed by Select Immune Mediators in Systemic Lupus Erythematosus

Neurofilament-light chain quantification by SimoaTM and EllaTM in plasma from patients with dementia: a comparative study

Customizable Multiplex Antibody Array Immunoassays with Attomolar Sensitivities

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