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Aptamer/ISET-MS: A New Affinity-Based MALDI Technique for Improved Detection of Biomarkers

Lee, Sujin LU ; Adler, Belinda LU ; Ekström, Simon LU ; Rezeli, Melinda LU ; Végvári, Ákos LU ; Park, Jeewoong LU ; Malm, Johan LU and Laurell, Thomas LU (2014) In Analytical Chemistry 86(15). p.7627-7634
Abstract
With the rapid progress in the development of new clinical biomarkers there is an unmet need of fast and sensitive multiplex analysis methods for disease specific protein monitoring. Immunoaffinity extraction integrated with matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis offers a route to rapid and sensitive protein analysis and potentially multiplex biomarker analysis. In this study, the previously reported integrated selective enrichment target (ISET)-MALDI-MS analysis was implemented with ssDNA aptamer functionalized microbeads to address the specific capturing of thrombin in complex samples. The main objective for using an aptamer as the capturing ligand was to avoid the inherently high background... (More)
With the rapid progress in the development of new clinical biomarkers there is an unmet need of fast and sensitive multiplex analysis methods for disease specific protein monitoring. Immunoaffinity extraction integrated with matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis offers a route to rapid and sensitive protein analysis and potentially multiplex biomarker analysis. In this study, the previously reported integrated selective enrichment target (ISET)-MALDI-MS analysis was implemented with ssDNA aptamer functionalized microbeads to address the specific capturing of thrombin in complex samples. The main objective for using an aptamer as the capturing ligand was to avoid the inherently high background components, which are produced during the digestion step following the target extraction when antibodies are used. By applying a thrombin specific aptamer linked to ISET-MALDI-MS detection, a proof of concept of antibody fragment background reduction in the ISET-MALDI-MS readout is presented. Detection sensitivity was significantly increased compared to the corresponding system based on antibody-specific binding as the aptamer ligand does not induce any interfering background residues from the antibodies. The limit of detection for thrombin was 10 fmol in buffer using the aptamer/ISET-MALDI-MS configuration as confirmed by MS/MS fragmentation. The aptamer/ISET-MALDI-MS platform also displayed a limit of detection of 10 fmol for thrombin in five different human serum samples (1/10 diluted), demonstrating the applicability of the aptamer/ISET-MALDI-MS analysis in clinical samples. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Analytical Chemistry
volume
86
issue
15
pages
7627 - 7634
publisher
The American Chemical Society
external identifiers
  • pmid:25001319
  • wos:000340081100068
  • scopus:84905695347
ISSN
1520-6882
DOI
10.1021/ac501488b
language
English
LU publication?
yes
id
363e82fb-6516-4a7e-9e1a-e46cb9e8e3c8 (old id 4438527)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/25001319
date added to LUP
2014-08-14 10:19:51
date last changed
2017-01-01 03:52:26
@article{363e82fb-6516-4a7e-9e1a-e46cb9e8e3c8,
  abstract     = {With the rapid progress in the development of new clinical biomarkers there is an unmet need of fast and sensitive multiplex analysis methods for disease specific protein monitoring. Immunoaffinity extraction integrated with matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis offers a route to rapid and sensitive protein analysis and potentially multiplex biomarker analysis. In this study, the previously reported integrated selective enrichment target (ISET)-MALDI-MS analysis was implemented with ssDNA aptamer functionalized microbeads to address the specific capturing of thrombin in complex samples. The main objective for using an aptamer as the capturing ligand was to avoid the inherently high background components, which are produced during the digestion step following the target extraction when antibodies are used. By applying a thrombin specific aptamer linked to ISET-MALDI-MS detection, a proof of concept of antibody fragment background reduction in the ISET-MALDI-MS readout is presented. Detection sensitivity was significantly increased compared to the corresponding system based on antibody-specific binding as the aptamer ligand does not induce any interfering background residues from the antibodies. The limit of detection for thrombin was 10 fmol in buffer using the aptamer/ISET-MALDI-MS configuration as confirmed by MS/MS fragmentation. The aptamer/ISET-MALDI-MS platform also displayed a limit of detection of 10 fmol for thrombin in five different human serum samples (1/10 diluted), demonstrating the applicability of the aptamer/ISET-MALDI-MS analysis in clinical samples.},
  author       = {Lee, Sujin and Adler, Belinda and Ekström, Simon and Rezeli, Melinda and Végvári, Ákos and Park, Jeewoong and Malm, Johan and Laurell, Thomas},
  issn         = {1520-6882},
  language     = {eng},
  number       = {15},
  pages        = {7627--7634},
  publisher    = {The American Chemical Society},
  series       = {Analytical Chemistry},
  title        = {Aptamer/ISET-MS: A New Affinity-Based MALDI Technique for Improved Detection of Biomarkers},
  url          = {http://dx.doi.org/10.1021/ac501488b},
  volume       = {86},
  year         = {2014},
}