Kinetic analyses and performance of a colloidal magnetic nanoparticle based immunoassay dedicated to allergy diagnosis |
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Authors: | Bruno Teste Frédéric Kanoufi Stéphanie Descroix Pascal Poncet Thomas Georgelin Jean-Michel Siaugue Jan Petr Anne Varenne Marie-Claire Hennion |
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Institution: | 1.Physicochimie des Electrolytes, Collo?des et Sciences Analytiques (PECSA) UMR 7195 CNRS-ESPCI-UPMC,Paris Cedex 05,France;2.CNRS-UPMC-ESPCI,Paris Cedex 05,France;3.CNRS-ENSCP-UPMC, Ecole Nationale Supérieure de Chimie de Paris, Chimie Paris Tech,Paris Cedex 05,France;4.Institut Pasteur,Paris,France |
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Abstract: | In this paper, we demonstrate the possibility to use magnetic nanoparticles as immunosupports for allergy diagnosis. Most
immunoassays used for immunosupports and clinical diagnosis are based on a heterogeneous solid-phase system and suffer from
mass-transfer limitation. The nanoparticles’ colloidal behavior and magnetic properties bring the advantages of homogeneous
immunoassay, i.e., species diffusion, and of heterogeneous immunoassay, i.e., easy separation of the immunocomplex and free
forms, as well as analyte preconcentration. We thus developed a colloidal, non-competitive, indirect immunoassay using magnetic
core–shell nanoparticles (MCSNP) as immunosupports. The feasibility of such an immunoassay was first demonstrated with a model
antibody and described by comparing the immunocapture kinetics using macro (standard microtiter plate), micro (microparticles)
and nanosupports (MCSNP). The influence of the nanosupport properties (surface chemistry, antigen density) and of the medium
(ionic strength, counter ion nature) on the immunocapture efficiency and specificity was then investigated. The performances
of this original MCSNP-based immunoassay were compared with a gold standard enzyme-linked immunosorbent assay (ELISA) using
a microtiter plate. The capture rate of target IgG was accelerated 200-fold and a tenfold lower limit of detection was achieved.
Finally, the MCSNP-based immunoassay was successfully applied to the detection of specific IgE from milk-allergic patient’s
sera with a lower LOD and a good agreement (CV < 6%) with the microtiter plate, confirming the great potential of this analytical
platform in the field of immunodiagnosis. |
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