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ENSAM: Europium Nanoparticles for Signal Enhancement of Antibody Microarrays on Nanoporous Silicon.

Järås, Kerstin LU ; Tajudin, Asilah; Ressine, Anton LU ; Soukka, Tero; Marko-Varga, György LU ; Bjartell, Anders LU ; Malm, Johan LU ; Laurell, Thomas LU and Lilja, Hans LU (2008) In Journal of Proteome Research 7. p.1308-1314
Abstract
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... (More)
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. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Proteome Research
volume
7
pages
1308 - 1314
publisher
The American Chemical Society
external identifiers
  • pmid:18257515
  • wos:000253825100044
  • scopus:44449165077
ISSN
1535-3893
DOI
10.1021/pr700591j
language
English
LU publication?
yes
id
7aa9dfcc-424a-43a7-af66-fde689fb1786 (old id 1042152)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/18257515?dopt=Abstract
date added to LUP
2008-03-06 11:58:46
date last changed
2017-09-10 04:46:14
@article{7aa9dfcc-424a-43a7-af66-fde689fb1786,
  abstract     = {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 &lt;/= 0.14 ng/mL, dynamic range &gt; 10 (5)) compared to the streptavidin-europium chelate assay (limit of detection &lt;/= 0.7 ng/mL, dynamic range &gt; 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.},
  author       = {Järås, Kerstin and Tajudin, Asilah and Ressine, Anton and Soukka, Tero and Marko-Varga, György and Bjartell, Anders and Malm, Johan and Laurell, Thomas and Lilja, Hans},
  issn         = {1535-3893},
  language     = {eng},
  pages        = {1308--1314},
  publisher    = {The American Chemical Society},
  series       = {Journal of Proteome Research},
  title        = {ENSAM: Europium Nanoparticles for Signal Enhancement of Antibody Microarrays on Nanoporous Silicon.},
  url          = {http://dx.doi.org/10.1021/pr700591j},
  volume       = {7},
  year         = {2008},
}