Composable Microfluidic Plates (cPlate): A Simple and Scalable Fluid Manipulation System for Multiplexed Enzyme-Linked Immunosorbent Assay (ELISA)

He, Ziyi; Huffman, Justin L.; Curtin, Kathrine; Garner, Krista L; Bowdridge, Elizabeth C.; Li, Xiaojun; Nurkiewicz, Timothy R.

doi:10.1021/acs.analchem.0c03651

Cited by 27 publications

(19 citation statements)

References 55 publications

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“…The ELISA chip generated aliquots with various volumes and timed the assay steps both within the CV of ≤ 3.5%, rivaling the common pipetting robots utilized in laboratory ELISA and other automated microfluidic ELISA. 29,30 The ELISA chip could operate with 0.05% Tween 20 commonly used in assays. The LOD of the ELISA chip for the SARS-CoV-2 N protein in 4×-diluted saliva was 91 pg/mL, in line with classical microplate ELISA and outperforming conventional lateral flow assays by ~25×.…”

Section: Discussionmentioning

confidence: 99%

3D-printed capillaric ELISA-on-a-chip with aliquoting

Parandakh

Ymbern

Jogia

et al. 2022

Preprint

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Sandwich immunoassays such as the enzyme-linked immunosorbent assay (ELISA) have been miniaturized and performed in a lab-on-a-chip format, but the execution of the multiple assay steps typically requires a computer or complex peripherals. Recently, an ELISA for detecting antibodies was encoded structurally in a chip thanks to the microfluidic chain reaction but the need for precise pipetting and intolerance to commonly used surfactant concentrations limited the potential for broader adoption. Here, we introduce the ELISA-on-a-chip with aliquoting functionality that obviates the need for precise pipetting, accommodates higher surfactant concentrations, includes barrier channels that delay the contact between solutions and prevent undesired mixing, and that executed a quantitative, high sensitivity assay for the SARS-CoV-2 nucleocapsid protein in 4×-diluted saliva. Upon loading the chip using disposable pipettes, capillary flow draws each reagent and the sample into a separate volumetric measuring reservoir for detection antibody (70 μL), enzyme conjugate (50 μL), substrate (80 μL), and sample (210 μL), and splits washing buffer into 4 different reservoirs of 40, 40, 60, and 20 μL. The excess volume is autonomously drained via a structurally encoded capillaric aliquoting circuit, creating aliquots with an accuracy of >93%. Next, the user click-connects the assay module, comprising a nitrocellulose membrane with immobilized capture antibodies and a capillary pump, to the chip which triggers the step-by-step, timed flow of all aliquoted solutions. A colored precipitate forming a line on a nitrocellulose strip serves as an assay readout, and upon digitization, yielded a binding curve with a limit of detection of 54 and 91 pg/mL for buffer and diluted saliva respectively, vastly outperforming rapid tests. The ELISA chip is 3D-printed, modular, adaptable to other targets and assays, and could be used to automate ELISA in the lab; or as a diagnostic test at the point of care with the convenience and form factor of rapid tests while preserving the protocol and performance of central laboratory ELISA.

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Section: Discussionmentioning

confidence: 99%

3D-printed capillaric ELISA-on-a-chip with aliquoting

Parandakh

Ymbern

Jogia

et al. 2022

Preprint

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“…[301][302][303][304] Compared with conventional macroscale vessels, microfluidic devices offer several advantages, including i) low cost and large-scale production with desired parameters, ii) portability, iii) low reagent consumption, iv) short assay time, v) easy processing of samples, vi) the development of multiple control units with the modulation of surface properties, and vii) high mixing efficiency. [305][306][307][308][309][310][311] Furthermore, microfluidic devices provide a high surface-tovolume ratio, leading to high mass and heat transfer rates. [312] To date, microfluidic devices have several applications in various research fields, including protein engineering, [313] immunoassays, [314] DNA research.…”

Section: An Integrative Platform: Microfluidicsmentioning

confidence: 99%

Carbon‐Based Nanomaterials and Sensing Tools for Wearable Health Monitoring Devices

Erdem

Derin

Shirejini

et al. 2021

Adv Materials Technologies

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In this manner, sensor technologies have garnered great attention in various fields, including biomedicine, [2][3][4][5] environmental monitoring, [6][7][8][9] smart devices, [10] wearable devices, [11] automobile manufacturing [12] since the semiconductor materials and circuits have been developed. In particular, biosensors are powerful and innovative analytical tools that incorporate biological receptors to recognize biological analytes through either physical or chemical transducers. Primarily, bio-receptors are responsible for identifying and capturing target analytes, and the transducer basically translates biological and chemical information into the detectable signals, which are eventually converted into the concentration of the analyte. [13,14] Considering gold standard methods, such as enzyme-linked immunosorbent assay (ELISA) [15] and polymerase chain reaction (PCR)-based strategies, [16] biosensors mostly hold crucial features, such as i) short assay time, [17] ii) affordable tools and reagents, [18] iii) portability, [19] and iv) facile use and minimum user interpretation. [20] Nowadays, the applications of biosensors have been leveraged by the advancements of portable and miniaturized platforms. In particular, over the past years, wearable health monitoring devices have notable impact on continuous and real-time monitoring of health parameters, thereby accelerating the deployment of biosensing strategies to daily lives. Besides, non-invasive and ease-of-collecting information supports the benefits of the wearable systems for enhancing the awareness of individuals and communities. [21][22][23] The special features of the mechanically flexible and stable wearable sensors include remarkable means, such as portability, comfortability, light-weight, non-invasive, and reliable performance. To put it simply, wearable sensors are readily attached to skin or organ surfaces through an adhesive tape [24] or microneedles, [25] and because of such easy integrations, several researchers have focused on developing wearable sensors for real-time health monitoring. A wearable sensor is basically composed of some vital elements, including a flexible base material attached to the skin or an organ, a signal transfer electrode, and a biorecognition element. Recently, researchers have concentrated on creating integrated sensors that are able to measure various parameters simultaneously, such as pressure, temperature,The healthcare system has a drastic paradigm shift from centralized care to home-based and self-monitoring strategies; aiming to reach more individuals, minimize workload in hospitals, and reduce healthcare-associated expenses. Particularly, wearable technologies are garnering considerable interest by tracking physiological parameters through motion and activities, and monitoring biochemical markers from sweat, saliva, and tears. Through their integrations with sensors, microfluidics, and wireless communication systems, they allow physicians, family members, or individuals to monitor multiple parameters withou...

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“…He et al recently developed miniaturized 96-well plates that require only 5 μL of the sample for each well and minimal equipment (a pipette and a magnet). 16 ELISA using streptavidin beads in femtoliter-sized wells improves the limit of protein detection to sub-attomolar concentrations, but analysis times are still 2 h. 17 …”

Section: Introductionmentioning

confidence: 99%

Quantitation of Trastuzumab and an Antibody to SARS-CoV-2 in Minutes Using Affinity Membranes in 96-Well Plates

et al. 2022

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Quantitation of therapeutic monoclonal antibodies (mAbs) in human serum could ensure that patients have adequate levels of mAbs for effective treatment. This research describes the use of affinity, glass-fiber membranes in a 96-well-plate format for rapid (<5 min) quantitation of the therapeutic mAb trastuzumab and a mAb against the SARS-CoV-2 spike protein. Adsorption of a poly(acrylic acid)-containing film in membrane pores and activation of the −COOH groups in the film enable covalent-linking of affinity peptides or proteins to the membrane. Passage of mAb-containing serum through the affinity membrane results in mAb capture within 1 min. Subsequent rinsing, binding of a secondary antibody conjugated to a fluorophore, and a second rinse yield mAb-concentration-dependent fluorescence intensities in the wells. Calibration curves established from analyses on different days have low variability and allow determination of mAb levels in separately prepared samples with an average error <10%, although errors in single-replicate measurements may reach 40%. The assays can occur in diluted serum with physiologically relevant mAb concentrations, as well as in undiluted serum. Thus, the combination of 96-well plates containing affinity membranes, a microplate reader, and a simple vacuum manifold affords convenient mAb quantitation in <5 min.

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Composable Microfluidic Plates (cPlate): A Simple and Scalable Fluid Manipulation System for Multiplexed Enzyme-Linked Immunosorbent Assay (ELISA)

Cited by 27 publications

References 55 publications

3D-printed capillaric ELISA-on-a-chip with aliquoting

3D-printed capillaric ELISA-on-a-chip with aliquoting

Carbon‐Based Nanomaterials and Sensing Tools for Wearable Health Monitoring Devices

Quantitation of Trastuzumab and an Antibody to SARS-CoV-2 in Minutes Using Affinity Membranes in 96-Well Plates

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