Direct access to aptamer-protein complexes via MALDI-MS

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Author(s) Chen, Fan, Gülbakan, Basri, Zenobi, Renato
Publication Type Journal Items, Publication Status: Published
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Title Direct access to aptamer-protein complexes via MALDI-MS
Author(s) Chen, Fan
Gülbakan, Basri
Zenobi, Renato
Journal or Series Title Chemical science
Volume Number 4
Issue Number 10
Start Page 4071
End Page 4078
ISSN 2041-6520
2041-6539
Publisher Royal Society of Chemistry
Publication Place Cambridge
Publication Date 2013-10-01
Abstract We report on the direct detection of protein–aptamer complexes by matrix-assisted laser desorption ionization (MALDI) mass spectrometry (MS). By using optimized conditions, we were able to observe the complexes of thrombin and two different thrombin binding aptamers (TBAs) directly. We also detected the complex of PDGF-AB/BB with the specific PDGF binding aptamer (Apt-35) in a 1 : 2 stoichiometry. Detection of the complex between lysozyme and its corresponding aptamer further confirmed the capability of MALDI-MS for studying such systems. All these analyses could be performed with very low sample concentrations (1 pmol) and volumes (1–10 mL). Well-designed control experiments confirmed that the complex observation is due to specific non-covalent interactions, rather than non-specific clusters formed in the MALDI plume. The stronger thrombin–TBA29 complex showed a larger signal at the m/z of the intact complex than the weaker thrombin–TBA15 complex; the complex signal of Apt-35 and PDGF-BB was stronger in MALDI compared with that of PDGF-AB and PDGF-AA. These observations indicate that the noncovalent interaction strength in solution is reflected in the MALDI mass spectra.
DOI 10.1039/c3sc51410b
Additional Notes Received 21 May 2013, Accepted 29 July 2013, Published online 31 July 2013
Document Type Article
Publication Status Published
Language English
NEBIS System Number 006104946
Source Database ID SCOPUS-84883301904
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@article{Chn2013,
  author = "Chen, Fan and G{\"{u}}lbakan, Basri and Zenobi, Renato",
  title = "{D}irect access to aptamer-protein complexes via {M}{A}{L}{D}{I}-{M}{S}",
  journal = "Chemical science",
  year = 2013,
  volume = "4",
  number = "10",
  pages = "4071--4078",
  month = oct,
}


E-Citations record created: Tue, 10 Sep 2013, 07:00:54 CET