title: A miniaturized heterogeneous fluorescence immunoassay on gold-coated nano-titer plates.
abstract: A miniaturized heterogeneous fluorescence immunoassay utilizing radiationless energy transfer to gold surfaces for fluorescence quenching is described. The phase-separation fluorescence immunoassay (PSFIA), a competitive heterogeneous assay, is carried out in the nanowells of a gold-coated nano-titer plate (NTP). Small analytes such as atrazine and histamine can be detected in nanoliter volumes with low limits of detection, maintaining a high degree of parallelization and miniaturization. The assay is characterized and optimized with respect to gold-layer thickness, surface concentration of immobilized analyte derivative, and antibody concentration. Various immobilization strategies are discussed.
authors.affiliation: Institute of Physical and Theoretical Chemistry, University of Tübingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Seidel
authors.forename: Daniela M
authors.initials: DM
authors.lastname: Dankbar
authors.forename: Günter
authors.initials: G
authors.lastname: Gauglitz
mesh_terms: D031541:Fluorescence Resonance Energy Transfer; D006046:Gold / Q000737:chemistry*; D007118:Immunoassay; D036103:Nanotechnology / Q000295:instrumentation* / Q000379:methods; D013050:Spectrometry, Fluorescence / Q000379:methods; D013499:Surface Properties
chemical_list: D006046:Gold