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Use of DNA-Aptamers for Enrichment of Low Abundant Proteins in Cellular Extracts for Quantitative Detection by Selected Reaction Monitoring

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Abstract

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 the target protein with aptamers immobilized on a solid phase has been investigated. Human thrombin added in known concentrations to cellular extracts derived from bacterial cells was used as a model target protein. The affinity enrichment of thrombin in cellular extracts by means of the thrombin-binding aptamer immobilized on the surface of magnetic microbeads resulted in an approximately 10-fold increase of the concentration of the target protein and a 100-fold decrease of the low limit of a target protein concentration range where its quantitative detection by SRM was possible without interference from other peptides present in the tryptic digest.

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Authors and Affiliations

  1. Institute of Biomedical Chemistry, ul. Pogodinskaya 10, Moscow, 119121, Russia

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

  2. Russian Scientific Center of Roentgenoradiology, Moscow, Russia

    Y. Y. Kiseleva

Authors
  1. K. G. Ptitsyn
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  2. S. E. Novikova
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  3. Y. Y. Kiseleva
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  4. A. A. Moysa
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  5. L. K. Kurbatov
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  6. T. E. Farafonova
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  7. S. P. Radko
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  8. V. G. Zgoda
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  9. A. I. Archakov
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Corresponding author

Correspondence to S. P. Radko.

Additional information

Original Russian Text © K.G. Ptitsyn, S.E. Novikova, Y.Y. Kiseleva, A.A. Moysa, L.K. Kurbatov, T.E. Farafonova, S.P. Radko, V.G. Zgoda, A.I. Archakov, 2018, published in Biomeditsinskaya Khimiya.

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Ptitsyn, K.G., Novikova, S.E., Kiseleva, Y.Y. et al. Use of DNA-Aptamers for Enrichment of Low Abundant Proteins in Cellular Extracts for Quantitative Detection by Selected Reaction Monitoring. Biochem. Moscow Suppl. Ser. B 12, 176–180 (2018). https://doi.org/10.1134/S1990750818020105

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  • DOI: https://doi.org/10.1134/S1990750818020105

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