Multiplex microarray ELISA versus classical ELISA, a comparison study of pollutant sensing for environmental analysis

Abstract

The present study describes the development, optimization and performance comparison of three ELISAs and one multiplex immunoassay in a microarray format. The developed systems were dedicated to the detection of three different classes of pollutants (pesticide, explosive and toxin) in water. The characteristics and performances of these two types of assays were evaluated and compared, in order to verify that multiplex immunoassays can replace ELISA for multiple analyte sensing. 2,4-Dichlorophenoxyacetic acid, 2,4,6-trinitrotoluene and okadaic acid were chosen as model targets and were immobilized in classical microtiter plate wells or arrayed at the surface of a microarray integrated within a classical 96-well plate. Once optimized, the classical ELISAs and microarray-based ELISA performances were evaluated and compared in terms of limit of detection, IC50, linearity range and reproducibility. Classical ELISAs provided quite good sensitivity (limit of detection down to 10 μg L(-1)), but the multiplex immunoassay was proven to be more sensitive (limit of detection down to 0.01 μg L(-1)), more reproducible and an advantageous tool in terms of cost and time expenses. This multiplex tool was then used for the successful detection of the three target molecules in spiked water samples and achieved very promising recovery rates.

Time-resolved digital immunoassay for rapid and sensitive quantitation of procalcitonin with plasmonic imaging

“…Recent technological advances have made it possible to detect single molecules, which have been used to measure the binding of a single biomarker molecule to a capture antibody with improved detection limit. This single-molecule approach has been referred to as digital immunoassay [10][11][12][13] .…”

“…Several digital immunoassay methods have been developed or proposed. One is to replace the large reaction well in the traditional ELISA with an array of microwells [10][11][12][13] . The volume of the microwells is small, such that each microwell has either no biomarker present or a single biomarker molecule that binds to a capture antibody conjugated with a magnetic bead in the microwell.…”

A High-Throughput, Precipitating Colorimetric Sandwich ELISA Microarray for Shiga Toxins

“…Though similar to a recently described high-throughput detection platform [11,12], the colorimetric enzyme-linked immunosorbent assay (ELISA) microarray presented herein was developed using intact, 96-well, polystyrene microtiter plates. Employed as a toxin typing array, this technique is not only rapid, but also has exhibited sufficient sensitivity to detect Stx at the low nanogram per milliliter limit.…”

Exploring Marine Resources for Bioactive Compounds

“…The most common are based on antibodies and enzymes. For example, Marquette et al developed an immunosensor for okadaic acid integrated in a fiber optic-based chemiluminescence flow injection system [95][96][97].…”