ENSAM: Europium Nanoparticles for Signal Enhancement of Antibody Microarrays on Nanoporous Silicon.
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1042152
- author
- 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
- organization
- publishing date
- 2008
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of Proteome Research
- volume
- 7
- pages
- 1308 - 1314
- publisher
- The American Chemical Society (ACS)
- external identifiers
-
- pmid:18257515
- wos:000253825100044
- scopus:44449165077
- pmid:18257515
- ISSN
- 1535-3893
- DOI
- 10.1021/pr700591j
- language
- English
- LU publication?
- yes
- additional info
- The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Analytical Chemistry (S/LTH) (011001004), Clinical Chemistry, Malmö (013016000), Biomedical Engineering (011200011), Division of Urological Cancers (013243420)
- 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
- 2016-04-04 08:36:57
- date last changed
- 2022-03-23 02:42:23
@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 </= 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.}}, 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 (ACS)}}, 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}}, doi = {{10.1021/pr700591j}}, volume = {{7}}, year = {{2008}}, }