ENSAM: Europium Nanoparticles for Signal Enhancement of Antibody Microarrays on Nanoporous Silicon.
Författare
Summary, in English
To improve the sensitivity of antibody microarray assays, we developed ENSAM ( Europium Nanoparticles for Signal enhancement of Antibody Microarrays). ENSAM is based on two nanomaterials. The first is polystyrene nanoparticles incorporated with europium chelate (beta-diketone) and coated with streptavidin. The multiple fluorophores incorporated into each nanoparticle should increase signal obtained from a single binding event. The second nanomaterial is array surfaces of nanoporous silicon, which creates high capacity for antibody adsorption. Two antibody microarray assays were compared: ENSAM and use of streptavidin labeled with a nine-dentate europium chelate. Analyzing biotinylated prostate-specific antigen (PSA) spiked into human female serum, ENSAM yielded a 10-fold signal enhancement compared to the streptavidin-europium chelate. Similarly, we observed around 1 order of magnitude greater sensitivity for the ENSAM assay (limit of detection </= 0.14 ng/mL, dynamic range > 10 (5)) compared to the streptavidin-europium chelate assay (limit of detection </= 0.7 ng/mL, dynamic range > 10 (4)). Analysis of a titration series showed strong linearity of ENSAM ( R (2) = 0.99 by linear regression). This work demonstrates the novel utility of nanoparticles with time-resolved fluorescence for signal enhancement of antibody microarrays, requiring as low as 100-200 zmol biotinylated PSA per microarray spot. In addition, proof of principle was shown for analyzing PSA in plasma obtained from patients undergoing clinical PSA-testing.
Avdelning/ar
Publiceringsår
2008
Språk
Engelska
Sidor
1308-1314
Publikation/Tidskrift/Serie
Journal of Proteome Research
Volym
7
Länkar
Dokumenttyp
Artikel i tidskrift
Förlag
The American Chemical Society (ACS)
Ämne
- Nano Technology
Status
Published
Forskningsgrupp
- Clinical Chemistry, Malmö
- Urological cancer, Malmö
ISBN/ISSN/Övrigt
- ISSN: 1535-3893