title: Amination of polystyrene microwells: application to the covalent grafting of DNA probes for hybridization assays.
abstract: Amine functionalization of polystyrene microwells for covalent binding of DNA is described. Polystyrene support was first carboxylated by permanganate oxidation in diluted sulfuric acid. These functions were activated with water-soluble carbodiimide and grafted with N-methyl-1,3-propane diamine to introduce a free secondary amino group on the support. The samples were characterized by X-ray photoelectron spectroscopy and radiochemical assay. The conditions for covalent linkage of secondary amine on polystyrene microwells were optimized. Functionalized supports were used for covalent binding of a DNA capture probe for the detection of human cytomegalovirus in a sandwich hybridization assay. Sensitivity of the assay compared very well with a commercially available surface, Covalink-NH, microwell plate obtained by electromagnetic irradiation.
authors.affiliation: Laboratoire de Biochimie Cellulaire, Facultés Universitaires Notre-Dame de la Paix, rue de Bruxelles 61, Namur, 5000, Belgium.
authors.forename: N
authors.initials: N
authors.lastname: Zammatteo
authors.forename: C
authors.initials: C
authors.lastname: Girardeaux
authors.forename: D
authors.initials: D
authors.lastname: Delforge
authors.forename: J J
authors.initials: JJ
authors.lastname: Pireaux
authors.forename: J
authors.initials: J
authors.lastname: Remacle
mesh_terms: D000588:Amines / Q000737:chemistry*; D003587:Cytomegalovirus / Q000737:chemistry; D004247:DNA / Q000032:analysis*; D015342:DNA Probes / Q000737:chemistry*; D004279:DNA, Viral / Q000032:analysis; D017895:Manganese Compounds / Q000737:chemistry; D009693:Nucleic Acid Hybridization; D010087:Oxides / Q000737:chemistry; D011137:Polystyrenes / Q000737:chemistry*
chemical_list: D000588:Amines; D015342:DNA Probes; D004279:DNA, Viral; D017895:Manganese Compounds; D010087:Oxides; D011137:Polystyrenes; C048856:permanganic acid; D004247:DNA