Use of DNA-aptamers for enrichment of low abundant proteins in cellular extracts for quntitative detection by selected reaction monitoring

   
Ptitsyn K.G.1, Novikova S.E.1, Kiseleva Y.Y.2, Moysa A.A.1, Kurbatov L.K.1, Farafonova T.E.1, Radko S.P.1 , Zgoda V.G.1, Archakov A.I.1

1. Institute of Biomedical Chemistry, Moscow, Russia
2. Russian Scientific Center of Roentgenoradiology, Moscow, Russia
Section: Experimental Study
DOI: 10.18097/PBMC20186401005      PubMed Id: 29460828
Year: 2018  Volume: 64  Issue: 1  Pages: 5-9
The relationship between the amount of a target protein in a complex biological sample and its amount measured by selected reaction monitoring (SRM) mass spectrometry upon the affinity enrichment of target protein with aptamers immobilized on a solid phase was studied. Human thrombin added in known concentrations to cellular extracts derived from bacterial cells was used as model target protein. It has been demonstrated that the affinity enrichment of thrombin in cellular extracts by means of the thrombin-binding aptamer immobilized on the surface of magnetic microbeads results in an approximately 10-fold increase of the concentration of target protein and a 100-fold decrease of the low limit of a target protein concentration range where its quantitative detection by SRM is possible without an interference from other peptides present in a tryptic digest.
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Keywords: aptamers, thrombin, enrichment, quantitative detection, selected reaction monitoring
Citation:

Ptitsyn, K. G., Novikova, S. E., Kiseleva, Y. Y., Moysa, A. A., Kurbatov, L. K., Farafonova, T. E., Radko, S. P., Zgoda, V. G., Archakov, A. I. (2018). Use of DNA-aptamers for enrichment of low abundant proteins in cellular extracts for quntitative detection by selected reaction monitoring. Biomeditsinskaya khimiya, 64(1), 5-9.
This paper is also available as the English translation: 10.1134/S1990750818020105
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