title: Development of miniaturized immunoassay: influence of surface chemistry and comparison with enzyme-linked immunosorbent assay and Western blot.
abstract: Protein microarray technology provides a useful approach for the simultaneous serodetection of various antibodies in low sample volumes. To implement functional protein microarrays, appropriate surface chemistry must be designed so that both the protein structure and the biological activity can be retained. In the current study, two surface chemistries for protein microarrays and immunofluorescent assays were developed. Glass slides were functionalized with N-hydroxysuccinimide (NHS) ester via a monofunctional silane or maleic anhydride-alt-methyl vinyl ether (MAMVE) copolymer to allow covalent grafting of histone proteins. Analytical performance of these microarrays was then evaluated for the detection of anti-histone autoantibodies present in the sera of patients suffering from a systemic autoimmune disease, namely systemic lupus erythematosus (SLE), and the results were compared with those of the classical enzyme-linked immunosorbent assay (ELISA) and Western blot. The detection limit of our MAMVE copolymer microarrays was 50-fold lower than that of the classical ELISA. Furthermore, 100-fold less volume of biological samples was required with these miniaturized immunoassays.
authors.affiliation: Institut des Nanotechnologies de Lyon-INL, UMR CNRS 5270, Université de Lyon, 69134 Ecully, France.
authors.forename: Graziella
authors.initials: G
authors.lastname: El Khoury
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Laurenceau
authors.forename: Yann
authors.initials: Y
authors.lastname: Chevolot
authors.forename: Yves
authors.initials: Y
authors.lastname: Mérieux
authors.forename: Agnès
authors.initials: A
authors.lastname: Desbos
authors.forename: Nicole
authors.initials: N
authors.lastname: Fabien
authors.forename: Dominique
authors.initials: D
authors.lastname: Rigal
authors.forename: Eliane
authors.initials: E
authors.lastname: Souteyrand
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Cloarec
mesh_terms: D001323:Autoantibodies / Q000097:blood; D015153:Blotting, Western / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006657:Histones / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D007118:Immunoassay / Q000379:methods*; D008180:Lupus Erythematosus, Systemic / Q000378:metabolism; D008904:Miniaturization; D040081:Protein Array Analysis / Q000379:methods*; D011713:Pyran Copolymer / Q000737:chemistry; D012821:Silanes / Q000737:chemistry
chemical_list: D001323:Autoantibodies; D006657:Histones; D055767:Immobilized Proteins; D012821:Silanes; D011713:Pyran Copolymer