title: High-throughput multiplexed competitive immunoassay for pollutants sensing in water.
abstract: The present study described the development and evaluation of a new fully automated multiplex competitive immunoassay enabling the simultaneous detection of five water pollutants (okadaic acid (OA), 2-chloro-4-ethylamino-6-isopropylamino-1,3,5-triazine (atrazine), 2.4-dichlorophenoxyacetic acid (2,4-D), 2,4,6-trinitrotoluene (TNT), and 1,3,5-trinitroperhydro-1,3,5-triazine (RDX)). The technology is taking advantage of an optical-clear pressure-sensitive adhesive on which biomolecules can be immobilized and that can be integrated within a classical 96-well format. The optimization of the microarray composition and cross-reaction was performed using an original approach where probe molecules (haptens) were conjugated to different carriers such as protein (bovine serum albumin or ovalbumin), amino-functionalized latex beads, or dextran polymer and arrayed at the surface of the adhesive. A total of 17 different probes were then arrayed together with controls on the adhesive surface and screened toward their specific reactivity and cross-reactivity. Once optimized, the complete setup was used for the detection of the five target molecules (less than 3 h for 96 samples). Limits of detection of 0.02, 0.01, 0.01, 100, and 0.02 μg L(-1) were found for OA, atrazine, 2,4-D, TNT, and RDX, respectively. The proof of concept of the multiplex competitive detection (semiquantitative or qualitative) of the five pollutants was also demonstrated on 16 spiked samples.
authors.affiliation: Equipe Génie Enzymatique, Membranes Biomimétiques et Assemblages Supramoléculaires, Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, Université Lyon 1-CNRS ICBMS, France.
authors.forename: Cloé
authors.initials: C
authors.lastname: Desmet
authors.forename: Loic J
authors.initials: LJ
authors.lastname: Blum
authors.forename: Christophe A
authors.initials: CA
authors.lastname: Marquette
mesh_terms: D015084:2,4-Dichlorophenoxyacetic Acid / Q000032:analysis; D000818:Animals; D001280:Atrazine / Q000032:analysis; D001331:Automation; D002417:Cattle; D006241:Haptens / Q000737:chemistry; D007118:Immunoassay / Q000379:methods*; D046228:Microarray Analysis; D019319:Okadaic Acid / Q000032:analysis; D010047:Ovalbumin / Q000737:chemistry; D011108:Polymers / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry; D014227:Triazines / Q000032:analysis; D014303:Trinitrotoluene / Q000032:analysis; D014874:Water Pollutants, Chemical / Q000032:analysis*
chemical_list: D006241:Haptens; D011108:Polymers; D014227:Triazines; D014874:Water Pollutants, Chemical; D014303:Trinitrotoluene; D019319:Okadaic Acid; D015084:2,4-Dichlorophenoxyacetic Acid; D012710:Serum Albumin, Bovine; D010047:Ovalbumin; D001280:Atrazine