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  • Bioconjugated fluorescent zeolite L nanocrystals as labels in protein microarrays.

Bioconjugated fluorescent zeolite L nanocrystals as labels in protein microarrays.

Small (Weinheim an der Bergstrasse, Germany) (2011-10-01)
Zhen Li, Gianluigi Luppi, Albert Geiger, Hans-Peter Josel, Luisa De Cola
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide, ≥97.0% (T)
Sigma-Aldrich
StreptaWell (transparent 96-well), pkg of 5 plates (11664778001 [12 x 8 well strips and frame]), pkg of 15 plates (11734776001 [96 wells])
Supelco
Digoxigenin, analytical standard
Digoxigenin, European Pharmacopoeia (EP) Reference Standard