ENSAM: Europium Nanoparticles for Signal Enhancement of Antibody Microarrays on Nanoporous Silicon.
Forskningsoutput: Tidskriftsbidrag › Artikel i vetenskaplig tidskrift
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.
|Enheter & grupper|
Ämnesklassifikation (UKÄ) – OBLIGATORISK
|Tidskrift||Journal of Proteome Research|
|Status||Published - 2008|
|Peer review utförd||Ja|
2010, Department of Laboratory Medicine, Lund University. 158 s.
Forskningsoutput: Avhandling › Doktorsavhandling (sammanläggning)