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Optimization of serum proteome profiling of healthy humans

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Abstract

Proteomic research is currently used worldwide for search biomarkers of different types of cancer. However, the assessment of the changes in proteomic profiles that are not associated with the progress of pathology is also an important task of clinical proteomics. This problem can be solved by developing a reproducible and exact method for profiling serum from healthy humans. The technologies of prefractionation of human serum samples by magnetic beads MB WCX and MB IMAC Cu using ClinProtrobot (Bruker Daltonics) and microchromatographic zip-tips prior to direct mass-spectrometric profiling were tested. Quality control was performed for all of the obtained spectra, with elimination of drastically different samples from the total data set, and the variation coefficient was calculated for each sample preparation technology. The study has demonstrated that the most reproducible method of serum prefractionation is the pretreatment of samples with magnetic beads MB WCX, which show the lowest number of ineffectual spectra and low variation coefficient (CV = 0.19) as compared with MB IMAC Cu (CV = 0.26) and zip-tip microcolumns (CV = 0.40).

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Authors and Affiliations

  1. Institute for Medical and Biological Problems, Russian Academy of Sciences, Moscow, 123007, Russia

    N. A. Pakharukova, L. Kh. Pastushkova, O. P. Trifonova & I. M. Larina

  2. Orekhovich Research Institute of Biomedical Chemistry, Russian Academy of Medical Sciences, Moscow, 109028, Russia

    M. A. Pyatnitsky, M. A. Vlasova, S. A. Moshkovsky & E. N. Nikolayev

  3. Bruker Daltonics Gmbh, Bremen, Germany

    I. P. Nikitin

Authors
  1. N. A. Pakharukova
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  2. L. Kh. Pastushkova
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  3. O. P. Trifonova
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  4. M. A. Pyatnitsky
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  5. M. A. Vlasova
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  6. I. P. Nikitin
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  7. S. A. Moshkovsky
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  8. E. N. Nikolayev
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  9. I. M. Larina
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Additional information

Original Russian Text © N.A. Pakharukova, L.Kh. Pastushkova, O.P. Trifonova, M.A. Pyatnitsky, M.A. Vlasova, I.P. Nikitin, S.A. Moshkovsky, E.N. Nikolayev, I.M. Larina, 2009, published in Fiziologiya Cheloveka, 2009, Vol. 35, No. 3, pp. 101–107.

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Pakharukova, N.A., Pastushkova, L.K., Trifonova, O.P. et al. Optimization of serum proteome profiling of healthy humans. Hum Physiol 35, 350–356 (2009). https://doi.org/10.1134/S0362119709030116

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  • DOI: https://doi.org/10.1134/S0362119709030116

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