Development of miniaturized immunoassay: influence of surface chemistry and comparison with enzyme-linked immunosorbent assay and Western blot.
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.
El Khoury
Graziella
G
Institut des Nanotechnologies de Lyon-INL, UMR CNRS 5270, Université de Lyon, 69134 Ecully, France.
Laurenceau
Emmanuelle
E
Chevolot
Yann
Y
Mérieux
Yves
Y
Desbos
Agnès
A
Fabien
Nicole
N
Rigal
Dominique
D
Souteyrand
Eliane
E
Cloarec
Jean-Pierre
JP
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
D001323:Autoantibodies; D006657:Histones; D055767:Immobilized Proteins; D012821:Silanes; D011713:Pyran Copolymer