Mass spectrometric detection of siRNA in plasma samples for doping control purposes

Analytical and Bioanalytical Chemistry
Maxie KohlerMario Thevis

Abstract

Small interfering ribonucleic acid (siRNA) molecules can effect the expression of any gene by inducing the degradation of mRNA. Therefore, these molecules can be of interest for illicit performance enhancement in sports by affecting different metabolic pathways. An example of an efficient performance-enhancing gene knockdown is the myostatin gene that regulates muscle growth. This study was carried out to provide a tool for the mass spectrometric detection of modified and unmodified siRNA from plasma samples. The oligonucleotides are purified by centrifugal filtration and the use of an miRNA purification kit, followed by flow-injection analysis using an Exactive mass spectrometer to yield the accurate masses of the sense and antisense strands. Although chromatography and sensitive mass spectrometric analysis of oligonucleotides are still challenging, a method was developed and validated that has adequate sensitivity (limit of detection 0.25-1 nmol mL(-1)) and performance (precision 11-21%, recovery 23-67%) for typical antisense oligonucleotides currently used in clinical studies.

References

Jan 1, 1995·Rapid Communications in Mass Spectrometry : RCM·M Greig, R H Griffey
Apr 16, 2003·European Journal of Biochemistry·Jens Kurreck
Jun 25, 2004·The New England Journal of Medicine·Markus SchuelkeSe-Jin Lee
Jul 12, 2005·Journal of Biotechnology·Jannel AcostaMario Pablo Estrada
Aug 5, 2006·Mass Spectrometry Reviews·Mario Thevis, Wilhelm Schänzer
Nov 17, 2006·Nature Chemical Biology·David BumcrotDinah W Y Sah
Dec 22, 2006·Nature Reviews. Molecular Cell Biology·Tariq M Rana
Feb 1, 2007·Analytical and Bioanalytical Chemistry·Mario Thevis, Wilhelm Schänzer
Dec 7, 2007·The Journal of Clinical Investigation·David R Corey
Apr 26, 2008·Human Gene Therapy·Dirk Haussecker
May 13, 2008·Rapid Communications in Mass Spectrometry : RCM·Yan ZouJerome Hochman
Jul 30, 2008·Proceedings of the National Academy of Sciences of the United States of America·Patrick S MitchellMuneesh Tewari
Oct 25, 2008·Analytical Chemistry·Teng-Yi HuangScott A McLuckey
Nov 26, 2008·Oligonucleotides·Mark A Behlke
Jan 21, 2009·Angewandte Chemie·Jens Kurreck
Jan 31, 2009·Nature Reviews. Drug Discovery·Kathryn A WhiteheadDaniel G Anderson
Feb 28, 2009·Proceedings of the National Academy of Sciences of the United States of America·Kai WangDavid J Galas
Apr 7, 2009·Analytical Biochemistry·Sean M McCarthyJohn Gebler
Apr 29, 2009·Molecular Pharmaceutics·R JulianoX Ming
Apr 30, 2009·Molecular Therapy : the Journal of the American Society of Gene Therapy·Shan GaoJørgen Kjems
Mar 11, 2010·Methods : a Companion to Methods in Enzymology·Ditte AndreasenPeter Mouritzen

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Citations

Mar 23, 2011·Analytical and Bioanalytical Chemistry·Mario ThevisWilhelm Schänzer
Jul 6, 2013·Analytical and Bioanalytical Chemistry·Andreas ThomasMario Thevis
Aug 9, 2013·Analytical and Bioanalytical Chemistry·Mario ThevisWilhelm Schänzer
Jun 8, 2014·Journal of Pharmaceutical and Biomedical Analysis·Mario Thevis, Wilhelm Schänzer
Apr 18, 2012·Drug Testing and Analysis·Elmo W I NeubergerPerikles Simon
Oct 28, 2011·Drug Testing and Analysis·Maxie KohlerMario Thevis
Jan 28, 2012·Journal of Mass Spectrometry : JMS·Kai SchuhmannAndrej Shevchenko
Aug 6, 2013·Drug Testing and Analysis·Andreas ThomasMario Thevis
Oct 28, 2011·Drug Testing and Analysis·Elmo W I NeubergerPerikles Simon
Jul 28, 2015·Mass Spectrometry Reviews·Wilhelm Schänzer, Mario Thevis
Jan 22, 2011·Drug Testing and Analysis·Mario ThevisWilhelm Schänzer

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