Immunoassays in the Clinical Chemistry Laboratory

Plaut, David

doi:10.1093/labmed/30.11.728

Cited by 2 publications

(1 citation statement)

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“…Immunoassays are important for the detection of proteins to enable disease identification and monitor treatment. Since its introduction in the 1950s, immunoassays have been widely used in many areas including medical diagnostics and drug monitoring. , While there are many immunoassay variations, they can be generally grouped into two categories: bulk immunoassays − and digital immunoassays. , They both share steps to form immunocomplexes involving a target molecule and a reporter that results in a measurable signal.…”

Section: Introductionmentioning

confidence: 99%

Wash-Free, Digital Immunoassay in Polydisperse Droplets

et al. 2020

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Immunoassays are important for the detection of proteins to enable disease identification and monitor treatment, but many immunoassays suffer from sensitivity limitations. The development of digital assays has enabled highly sensitive biomarker detection and quantification, but the necessary devices typically require precisely controlled volumes to reduce biases in concentration estimates from compartment size variation. These constraints have led to systems that are often expensive, cumbersome, and challenging to operate, confining many digital assays to centralized laboratories. To overcome these limitations, we have developed a simplified digital immunoassay performed in polydisperse droplets that are prepared without any specialized equipment. This polydisperse digital droplet immunoassay (ddIA) uses proximity ligation to remove the need for wash steps and simplifies the system to a single reagent addition step. Using interleukin-8 (IL-8) as an example analyte, we demonstrated the concept with samples in buffer and diluted whole blood with limits of detection of 0.793 pM and 1.54 pM, respectively. The development of a one-pot, washless assay greatly improves usability compared to traditional immunoassays or digital-based systems that rely heavily on wash steps and can be run with common and readily available laboratory equipment such as a heater and simple fluorescent microscope. We also developed a stochastic model with physically meaningful parameters that can be utilized to optimize the assay and enable quantification without standard curves, after initial characterization of the parameters. Our polydisperse ddIA assay serves as an example of sensitive, lower-cost, and simpler immunoassays suitable for both laboratory and point-of-care applications.

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