title: Bioconjugated fluorescent zeolite L nanocrystals as labels in protein microarrays.
abstract: Zeolite L nanocrystals, as inorganic host material containing hydrophobic fluorophore N,N'-bis(2,6-dimethylphenyl)perylene-3,4,9,10-tetracarboxylic diimide in the unidirectional channels, are developed as new labels for biosensor systems. The external surface of the particles is modified with carboxylic acid groups for conjugation to primary amines of biomolecules such as antibodies. Anti-digoxigenin (anti-DIG) is selected to be immobilized on zeolite L via N-hydroxysulfosuccinimide ester linker. Together with DIG, it serves as a good universal binding pair for diverse analyte detection owing to the high binding affinity and low background noise. The conjugates are characterized by the dynamic light scattering technique for their hydrodynamic diameters and by enzyme-linked immunosorbent assay for antigen-antibody binding behavior. The characterizations prove that anti-DIG antibodies are successfully immobilized on zeolite L with their binding activities maintained. The microarray fluorescent sandwich immunoassay based on such nanocrystalline labels shows high sensitivity in a thyroid-stimulating hormone assay with the lower detection limit down to the femtomolar range. These new fluorescent labels possess great potential for in vitro diagnostics applications.
authors.affiliation: Physikalisches Institut and Center for Nanotechnology, CeNTech, Universität Münster, Heisenbergstr. 11, 48149 Muenster, Germany.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Li
authors.forename: Gianluigi
authors.initials: G
authors.lastname: Luppi
authors.forename: Albert
authors.initials: A
authors.lastname: Geiger
authors.forename: Hans-Peter
authors.initials: HP
authors.lastname: Josel
authors.forename: Luisa
authors.initials: L
authors.lastname: De Cola
mesh_terms: D004076:Digoxigenin / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005022:Ethyldimethylaminopropyl Carbodiimide / Q000737:chemistry; D005453:Fluorescence; D007118:Immunoassay; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D040081:Protein Array Analysis / Q000379:methods*; D013194:Staining and Labeling; D013972:Thyrotropin / Q000032:analysis; D017641:Zeolites / Q000378:metabolism*
chemical_list: D017641:Zeolites; D013972:Thyrotropin; D004076:Digoxigenin; D005022:Ethyldimethylaminopropyl Carbodiimide