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Aptamer microarray mediated capture and mass spectrometry identification of biomarker in serum samples.

Ji-Young, Ahn; Lee, Sang Wook LU ; Kang, Hye Suk; Jo, Minjoung; Lee, Dong-Ki; Laurell, Thomas LU and Kim, Soyoun (2010) In Journal of Proteome Research 9(11). p.5568-5573
Abstract
Sensitive detection of molecular biomarkers in clinical samples is crucially important in disease diagnostics. This paper reports the developement of an aptamer microarray platform combined with sol-gel technology to identify low-abundance targets in complex serum samples. Because of the nanoporous structure of the sol-gel, a high capacity to immobilize the affinity specific aptamers is accomplished which allows binding and detection of target molecules with high sensitivity. The captured protein is digested in situ and the obtained digest was analyzed by ESI-MS without any interference from the affinity probe. TBP (TATA Box Protein) and its specific aptamers were chosen as a model system. A proof of concept with protein concentrations... (More)
Sensitive detection of molecular biomarkers in clinical samples is crucially important in disease diagnostics. This paper reports the developement of an aptamer microarray platform combined with sol-gel technology to identify low-abundance targets in complex serum samples. Because of the nanoporous structure of the sol-gel, a high capacity to immobilize the affinity specific aptamers is accomplished which allows binding and detection of target molecules with high sensitivity. The captured protein is digested in situ and the obtained digest was analyzed by ESI-MS without any interference from the affinity probe. TBP (TATA Box Protein) and its specific aptamers were chosen as a model system. A proof of concept with protein concentrations ranging between nanomolar to micromolar is reported, showing a good linearity up to 400 nM when characterized in an aptamer sandwich assay. Moreover, as low as 0.001% of target protein present in total serum proteins could be identified without any pretreatment step using ESI MS/MS mass spectrometry. We believe this novel strategy could become an efficient method for aptamer-based biomarker detection linked directly to mass spectrometry readout. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
sol-gel immobilization, aptamer microarray, sandwich assay, biomarkers, ESI MS/MS mass spectrometry
in
Journal of Proteome Research
volume
9
issue
11
pages
5568 - 5573
publisher
The American Chemical Society
external identifiers
  • pmid:20806970
  • wos:000283810500006
  • scopus:78149380835
ISSN
1535-3893
DOI
10.1021/pr100300t
language
English
LU publication?
yes
id
ea7a50e6-006f-4694-a7f7-824cee5b83bb (old id 1688687)
date added to LUP
2010-10-12 13:45:15
date last changed
2018-11-21 19:43:44
@article{ea7a50e6-006f-4694-a7f7-824cee5b83bb,
  abstract     = {Sensitive detection of molecular biomarkers in clinical samples is crucially important in disease diagnostics. This paper reports the developement of an aptamer microarray platform combined with sol-gel technology to identify low-abundance targets in complex serum samples. Because of the nanoporous structure of the sol-gel, a high capacity to immobilize the affinity specific aptamers is accomplished which allows binding and detection of target molecules with high sensitivity. The captured protein is digested in situ and the obtained digest was analyzed by ESI-MS without any interference from the affinity probe. TBP (TATA Box Protein) and its specific aptamers were chosen as a model system. A proof of concept with protein concentrations ranging between nanomolar to micromolar is reported, showing a good linearity up to 400 nM when characterized in an aptamer sandwich assay. Moreover, as low as 0.001% of target protein present in total serum proteins could be identified without any pretreatment step using ESI MS/MS mass spectrometry. We believe this novel strategy could become an efficient method for aptamer-based biomarker detection linked directly to mass spectrometry readout.},
  author       = {Ji-Young, Ahn and Lee, Sang Wook and Kang, Hye Suk and Jo, Minjoung and Lee, Dong-Ki and Laurell, Thomas and Kim, Soyoun},
  issn         = {1535-3893},
  keyword      = {sol-gel immobilization,aptamer microarray,sandwich assay,biomarkers,ESI MS/MS mass spectrometry},
  language     = {eng},
  number       = {11},
  pages        = {5568--5573},
  publisher    = {The American Chemical Society},
  series       = {Journal of Proteome Research},
  title        = {Aptamer microarray mediated capture and mass spectrometry identification of biomarker in serum samples.},
  url          = {http://dx.doi.org/10.1021/pr100300t},
  volume       = {9},
  year         = {2010},
}