Mass-spectrometric MRM analysis of FDA-verified proteins in the blood plasma of healthy volunteers

Novikova S.E.1 , Farafonova T.E.1, Tikhonova O.V.1, Shushkova N.A.1, Pyatnitsky M.A.1, Zgoda V.G.1, Ponomarenko E.A.1, Lisitsa A.V.1, Grigoryev A.I.2, Tutelyan V.A.3, Archakov A.I.1

1. Institute of Biomedical Chemistry, Moscow, Russia
2. Institute of Biomedical Problems, Russian Academy of Sciences, Moscow, Russia
3. Institute of Nutrition, Moscow, Russia
Section: Experimental Study
DOI: 10.18097/PBMC20206604294      PubMed Id: 32893820
Year: 2020  Volume: 66  Issue: 4  Pages: 294-316
The proteomic composition of a biological sample serves as the most important feature of a biological object, and it allows discriminating normal and pathological conditions. Targeted mass spectrometric analysis, namely, multiple reaction monitoring (MRM) using synthetic isotopically-labeled internal standard (SIS), is the main alternative to the ELISA method for the analysis of diagnostically significant proteins. Based on the MRM results, a prototype test system has been developed; it employs the targeted mass spectrometric method for multiplex, quantitative analysis of FDA-verified proteins in whole blood plasma. Using this approach, it was possible to measure the content of 42 proteins in 31 samples in a concentration range spanning five orders of magnitude. The interindividual variability for 30 of the 42 registered proteins was less than 40%. The largest scatter was observed for haptoglobin (68%), immunoglobulin heavy constant delta IGHD (90%), angiotensin (72%), sex hormone-binding globulin SHBG (100%) and lipoprotein-(a) (136%). The obtained results on the concentration of proteins correlate with published data (Hortin et al., 2008, Clinical Chemistry, 54, 1608) with R2=0.84. The developed prototype test system based on targeted mass spectrometric analysis of proteins can be considered as an alternative to methods using monoclonal antibodies.
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Keywords: SRM analysis, isotope-labeled internal peptide standards, FDA-verified proteins
Supplementary materials:

Novikova, S. E., Farafonova, T. E., Tikhonova, O. V., Shushkova, N. A., Pyatnitsky, M. A., Zgoda, V. G., Ponomarenko, E. A., Lisitsa, A. V., Grigoryev, A. I., Tutelyan, V. A., Archakov, A. I. (2020). Mass-spectrometric MRM analysis of FDA-verified proteins in the blood plasma of healthy volunteers. Biomeditsinskaya khimiya, 66(4), 294-316.
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