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Marquette C.A.,CNRS Institute of Molecular and Supramolecular Chemistry and Biochemistry | Corgier B.P.,AXO Science | Blum L.J.,CNRS Institute of Molecular and Supramolecular Chemistry and Biochemistry
Bioanalysis | Year: 2012

The present review reports on the lastest developments in multiplex immunoassays. The selected examples are classified through their detection strategy (fluorescence, chemiluminescence, colorimetry or labeless) and their assay format (standard microtiter plate, polymeric membranes and glass slides). Finally, the degree of integration in a complete system, incorporating fluid handling and detection was also taken into account. © 2012 Future Science Ltd.

Mandon C.A.,CNRS Institute of Molecular and Supramolecular Chemistry and Biochemistry | Berthuy O.I.,CNRS Institute of Molecular and Supramolecular Chemistry and Biochemistry | Corgier B.P.,AXO Science | Le Goff G.C.,CNRS Institute of Molecular and Supramolecular Chemistry and Biochemistry | And 4 more authors.
Biosensors and Bioelectronics | Year: 2013

The present report describes the integration and application possibilities of a new microarray concept based on adhesive surface. The method was shown to enable the straightforward production of 384 and 1536-well plates modified with 100 and 25 spots per well, respectively. Such in-well densities were only possible thanks to the fabrication process which implies first the deposition of the microarray on a flat adhesive surface and then its assembly with bottomless 384 or 1536-well plates.The concept was also confronted to various applications such as oligonucleotide detection, localised cell culture onto spotted adhesion proteins and immobilisation of peptide or active antibodies for immunoassays. In the particular case of immunotesting, the study focused on liver diseases diagnosis and more particularly on the detection of either one liver cancer marker, the alpha-fetoprotein, or the detection of Hepatitis C Virus infection. In every cases, interesting performances were obtained directly in crude patient serum, proof of the robust and generic aspect of the platform. © 2012 Elsevier B.V..

Berthuy O.I.,CNRS Institute of Molecular and Supramolecular Chemistry and Biochemistry | Mandon C.A.,AXO Science | Corgier B.P.,AXO Science | Octobre G.G.,CNRS Institute of Molecular and Supramolecular Chemistry and Biochemistry | And 4 more authors.
Journal of Materials Science | Year: 2014

In this study, we develop a new concept for multiplexed and localized cell co-culture. This cell chip consists of a polystyrene spin-coated solid support bearing gold-bottomed microwells. The cell-chip support is fabricated as follows: (i) electrosputtering of a thin layer of gold (40 nm) onto a polycarbonate substrate, (ii) spin coating of a polystyrene thin film (500 ± 50 nm) over the gold layer, followed by (iii) polystyrene etching through the spotting of toluene nanovolume (300-900 pL). In each gold-bottomed microwell, a small population of adherent cells (approx. 100 cells) can be cultured. In this miniaturized system, different cell lines can be co-cultured on a 1-cm2 surface, opening the way to multiplexed cell-chip development. In order to keep the cells in a properly hydrated environment and to physically retain them before they adhere, a biocompatible alginate polymer was used during the robotized micropipetting. This approach allows for the encapsulation of the cell in a very small volume (50 nL), directly in the microwells. After 24 h of culture, the cells adhered on the gold bottom of the microwells, and the alginate matrix was removed by addition of calcium-free culture medium. Graphical Abstract: Multiplex culture of cells was obtained using in situ produced microwells and encapsulated cells. The microwells are produced by organic solvent etching (nanovolume spotting) of a spin-coated polystyrene thin film, and the living multiple cell line deposition is obtained using on-site encapsulation in an alginate bead. [Figure not available: see fulltext.] © 2014 Springer Science+Business Media New York.

Harwanegg C.,Thermo Fisher Scientific | Marquette C.A.,AXO Science | Corgier B.P.,AXO Science | Soh W.T.,Chulalongkorn University | And 3 more authors.
Allergy: European Journal of Allergy and Clinical Immunology | Year: 2016

Background Understanding patterns of IgE sensitization in Dermatophagoides-allergic patients living in various geographical areas is necessary to design a product suitable for worldwide allergen immunotherapy (AIT). Methods Using a HIFI Allergy customized microarray assay, IgEs specific for 12 purified allergens from Dermatophagoides pteronyssinus or D. farinae were assessed in sera from 1302 house dust mite (HDM)-allergic patients living in various areas. Comprehensive mass spectrometric (MS) analyses were conducted to characterize HDM extracts, as well as purified bodies and feces. Results Patterns of IgE reactivity to HDM allergens are comparable in all cohorts of patients analyzed, encompassing adults and 5- to 17-year-old children, as well as American, Canadian, European, and Japanese patients. Overall, >70% and >80% of HDM-allergic patients are sensitized to group 1 and group 2 allergens, respectively, from D. pteronyssinus and/or D. farinae species. Furthermore, 20-47% of patients also have IgEs to allergens from groups 4, 5, 7, 13, 15, 21, and 23. All patients have IgEs to allergens present in mite bodies and feces. MS-based analyses confirmed the presence of mite allergens recorded by IUIS in D. pteronyssinus and D. farinae extracts, with groups 2, 8, 10, 11, 14, and 20 prominent in bodies and groups 1, 6, 18, and 23 well represented in feces. Conclusions Mite-specific AIT should rely upon a mixture of D. pteronyssinus and D. farinae extracts, manufactured from both feces and bodies. Such a combination is appropriate to treat children and adult Dermatophagoides-allergic patients from Asia, Europe, and North America. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

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