Customizable Multiplex Antibody Array Immunoassays with Attomolar Sensitivities

Tobos, Carmen; Sheehan, Antony J; Duffy, David C.; Rissin, David M.

doi:10.1021/acs.analchem.0c00631

Cited by 12 publications

(14 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The ability to simultaneously measure multiple analytes in a single sample can accelerate sample throughput and biomarker signature discovery and is especially critical in applications with limited sample volumes, such as neonatal saliva or fingerprick blood. Multiplexing in digital ELISA as well as in a recently developed ultrasensitive planar ELISA platform has shown great utility in various diagnostic applications, but the number of bead types that can be analyzed simultaneously in bead confinement methods is limited by the total number of compartments and low bead analysis efficiencies. − In contrast, the solution-based readout of MOSAIC enables, in principle, an unlimited number of bead types to be analyzed in one sample with readily tunable sensitivities and dynamic ranges via assay bead number for each analyte (Figure A). Furthermore, the versatility of flow cytometry enables a wide range of beads to be distinguished by fluorescence wavelengths and intensities, as well as bead sizes.…”

Section: Results and Discussionmentioning

confidence: 99%

High-Throughput, High-Multiplex Digital Protein Detection with Attomolar Sensitivity

Dougan

Walt

2022

ACS Nano

View full text Add to dashboard Cite

A major challenge in many clinical diagnostic applications is the measurement of low-abundance proteins and other biomolecules in biological fluids. Digital technologies such as the digital enzyme-linked immunosorbent assay (ELISA) have enabled 1000-fold increases in sensitivity over conventional protein detection methods. However, current digital ELISA technologies still possess insufficient sensitivities for many rare protein biomarkers and require specialized instrumentation or time-consuming workflows that have limited their widespread implementation. To address these challenges, we have developed a more sensitive and streamlined digital ELISA platform, Molecular On-bead Signal Amplification for Individual Counting (MOSAIC), which attains low attomolar limits of detection, with an order of magnitude enhancement in sensitivity over these other methods. MOSAIC uses a rapid, automatable flow cytometric readout that vastly increases throughput and is easily integrated into existing laboratory infrastructure. As MOSAIC provides high sampling efficiencies for rare target molecules, assay bead number can readily be tuned to enhance signal-to-background with high measurement precision. Furthermore, the solution-based signal readout of MOSAIC expands the number of analytes that can simultaneously be measured for higher-order multiplexing with femtomolar sensitivities or below, compared with microwell- or droplet-based digital methods. As a proof of principle, we apply MOSAIC toward improving the detectability of low-abundance cytokines in saliva and ultrasensitive multiplexed measurements of eight protein analytes in plasma and saliva. The attomolar sensitivity, high throughput, and broad multiplexing abilities of MOSAIC provide highly accessible and versatile ultrasensitive capabilities that can potentially accelerate protein biomarker discovery and diagnostic testing for diverse disease applications.

show abstract

Section: Results and Discussionmentioning

confidence: 99%

High-Throughput, High-Multiplex Digital Protein Detection with Attomolar Sensitivity

Dougan

Walt

2022

ACS Nano

View full text Add to dashboard Cite

show abstract

“…However, it is rarely applied in food safety assays due to the rigid requirement of dedicated signal-collecting instrumentation that suitable for 384-well or 813-spot microtiter plates. Tobos et al (2020) sought to develop a simple, customized multiplex ELISA array without the need for complex and expensive equipment by spotting five kinds of anchor antibodies at the bottom of a single microtiter plate well. This customizable contact printed 5-plex ELISA can simultaneously detect 5 proteins with attomolar sensitivity.…”

Section: Array Of Microtiter Platementioning

confidence: 99%

“…Tobos et al. (2020) sought to develop a simple, customized multiplex ELISA array without the need for complex and expensive equipment by spotting five kinds of anchor antibodies at the bottom of a single microtiter plate well. This customizable contact printed 5‐plex ELISA can simultaneously detect 5 proteins with attomolar sensitivity.…”

Section: Strategies and Applications Of Mobas In Food Safety Analysismentioning

confidence: 99%

Multiplex optical bioassays for food safety analysis: Toward on‐site detection

Guan

Jin

et al. 2022

Comp Rev Food Sci Food Safe

View full text Add to dashboard Cite

Food safety analysis plays a significant role in controlling food contamination and supervision. In recent years, multiplex optical bioassays (MOBAs) have been widely applied to analyze multiple hazards due to their efficiency and low cost. However, due to the challenges such as multiplexing capacity, poor sensitivity, and bulky instrumentation, the further application of traditional MOBAs in food screening has been limited. In this review, effective strategies regarding food safety MOBAs are summarized, such as spatial‐resolution modes performed in multi‐T lines/dots strips or arrays of strip/microplate/microfluidic chip/SPR chip and signal‐resolution modes employing distinguishable colorimetric/luminescence/fluorescence/surface plasma resonance/surface‐enhanced Raman spectrum as signal tags. Following this, new trends on how to design engineered sensor architecture and exploit distinguishable signal reporters, how to improve both multiplexing capacity and sensitivity, and how to integrate these formats into smartphones so as to be mobile are summarized systematically. Typically, in the case of enhancing multiplexing capacity and detection throughput, microfluidic array chips with multichannel architecture would be a favorable approach to overcome the spatial and physical limitations of immunochromatographic assay (ICA) test strips. Moreover, noble metal nanoparticles and single‐excitation, multiple‐emission luminescence nanomaterials hold great potential in developing ultrasensitive MOBAs. Finally, the exploitation of innovative multiplexing strategy hybridized with powerful and widely available smartphones opens new perspectives to MOBAs. In future, the MOBAs should be more sensitive, have higher multiplexing capacity, and easier instrumentation.

show abstract

“…As with ADCs, total antibody can be quantified with LBA or hybrid LBA-LC-MS/ MS methods. Historically, higher sensitivity can be achieved with an LBA approach, whereas the LC-MS platform can offer the flexibility of a generic capture approach [71,86]. This is contingent upon assay requirements and available reagents.…”

Section: Antibody-oligonucleotide Conjugatesmentioning

confidence: 99%

Bioanalytical Methods and Strategic Perspectives Addressing the Rising Complexity of Novel Bioconjugates and Delivery Routes for Biotherapeutics

Yuan

Huang

et al. 2022

BioDrugs

View full text Add to dashboard Cite

In recent years, an increase in the discovery and development of biotherapeutics employing new modalities, such as bioconjugates or novel routes of delivery, has created bioanalytical challenges. The inherent complexity of conjugated molecular structures means that quantification of the bioconjugate and its multiple components is critical for preclinical/clinical studies to inform drug discovery and development. Moreover, bioconjugates involve additional multifactorial complexity because of the potential for in vivo catabolism and biotransformation, which may require thorough investigations in multiple biological matrices. Furthermore, excipients that enhance absorption are frequently evaluated and employed for the development of oral and inhaled biotherapeutics. Risk-benefit assessments are required for novel or existing excipients that utilize dosages above previously approved levels. Bioanalytical methods that can measure both excipients and potential drug metabolites in biological matrices are highly relevant to these emerging bioanalysis challenges. We discuss the bioanalytical strategies for analyzing bioconjugates such as antibody-drug conjugates and antibody-oligonucleotide conjugates and review recent advances in bioanalytical methods for the quantification and characterization of novel bioconjugates. We also discuss bioanalytical considerations for both biotherapeutics and excipients through novel administration routes and review analyses in various biological matrices, from the extensively studied serum or plasma to tissue biopsy in the context of preclinical and clinical studies from both technical and regulatory perspectives.Ruipeng Mu and Jiaqi Yuan have contributed equally to this work.

show abstract

Customizable Multiplex Antibody Array Immunoassays with Attomolar Sensitivities

Cited by 12 publications

References 14 publications

High-Throughput, High-Multiplex Digital Protein Detection with Attomolar Sensitivity

High-Throughput, High-Multiplex Digital Protein Detection with Attomolar Sensitivity

Multiplex optical bioassays for food safety analysis: Toward on‐site detection

Bioanalytical Methods and Strategic Perspectives Addressing the Rising Complexity of Novel Bioconjugates and Delivery Routes for Biotherapeutics

Contact Info

Product

Resources

About