NMR Separation of Intra- and Extracellular Compounds Based on Intermolecular Coherences
Identifieur interne : 000266 ( Pmc/Corpus ); précédent : 000265; suivant : 000267NMR Separation of Intra- and Extracellular Compounds Based on Intermolecular Coherences
Auteurs : Verena Hoerr ; Armin Purea ; Cornelius FaberSource :
- Biophysical Journal [ 0006-3495 ] ; 2010.
Abstract
NMR spectroscopy is a powerful tool for detection and characterization of chemical compounds in biological systems. Its application in pharmaceutical studies in cell cultures, however, has been hampered by the enormous technical challenges in separating intra- from extracellular amounts of one substance. We introduce a novel approach to separate intra- from extracellular NMR signal based on the detection of intermolecular zero-quantum coherences in presence of a chemical shift agent. In a sample of large cells in culture, the investigation of cellular uptake of pharmacological substances becomes feasible. The addition of 10 mM Tm-DOTP to a suspension of 100
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DOI: 10.1016/j.bpj.2010.06.068
PubMed: 20923669
PubMed Central: 3042554
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PMC:3042554Le document en format XML
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<author><name sortKey="Hoerr, Verena" sort="Hoerr, Verena" uniqKey="Hoerr V" first="Verena" last="Hoerr">Verena Hoerr</name>
<affiliation><nlm:aff id="aff1">Department of Experimental Physics 5, University of Würzburg, Würzburg, Germany</nlm:aff>
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<affiliation><nlm:aff id="aff2">Department of Clinical Radiology, University Hospital Münster, Münster, Germany</nlm:aff>
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<author><name sortKey="Purea, Armin" sort="Purea, Armin" uniqKey="Purea A" first="Armin" last="Purea">Armin Purea</name>
<affiliation><nlm:aff id="aff1">Department of Experimental Physics 5, University of Würzburg, Würzburg, Germany</nlm:aff>
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<affiliation><nlm:aff id="aff3">RAPID Biomedical, Rimpar, Germany</nlm:aff>
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<author><name sortKey="Faber, Cornelius" sort="Faber, Cornelius" uniqKey="Faber C" first="Cornelius" last="Faber">Cornelius Faber</name>
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<author><name sortKey="Purea, Armin" sort="Purea, Armin" uniqKey="Purea A" first="Armin" last="Purea">Armin Purea</name>
<affiliation><nlm:aff id="aff1">Department of Experimental Physics 5, University of Würzburg, Würzburg, Germany</nlm:aff>
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<author><name sortKey="Faber, Cornelius" sort="Faber, Cornelius" uniqKey="Faber C" first="Cornelius" last="Faber">Cornelius Faber</name>
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<series><title level="j">Biophysical Journal</title>
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<front><div type="abstract" xml:lang="en"><p>NMR spectroscopy is a powerful tool for detection and characterization of chemical compounds in biological systems. Its application in pharmaceutical studies in cell cultures, however, has been hampered by the enormous technical challenges in separating intra- from extracellular amounts of one substance. We introduce a novel approach to separate intra- from extracellular NMR signal based on the detection of intermolecular zero-quantum coherences in presence of a chemical shift agent. In a sample of large cells in culture, the investigation of cellular uptake of pharmacological substances becomes feasible. The addition of 10 mM Tm-DOTP to a suspension of 100 <italic>Xenopus laevis</italic>
oocytes resulted in sufficient separation of resonance frequencies between intra- and extracellular water. Upon selective excitation of either intra- or extracellular water signal, only intra- or extracellular components were observed, respectively. The presented localization technique provides intrinsic averaging over a large number of cells, resulting in a significant signal gain. The method works on standard NMR spectrometers, which are available at most scientific research institutions today. On a high-resolution NMR system with a cryoprobe, a 20-fold sensitivity gain was observed as compared to conventionally localized NMR spectroscopy of a single <italic>X. laevis</italic>
oocyte on dedicated NMR microscopes.</p>
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<front><journal-meta><journal-id journal-id-type="nlm-ta">Biophys J</journal-id>
<journal-title-group><journal-title>Biophysical Journal</journal-title>
</journal-title-group>
<issn pub-type="ppub">0006-3495</issn>
<issn pub-type="epub">1542-0086</issn>
<publisher><publisher-name>The Biophysical Society</publisher-name>
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<article-meta><article-id pub-id-type="pmid">20923669</article-id>
<article-id pub-id-type="pmc">3042554</article-id>
<article-id pub-id-type="publisher-id">BPJ1956</article-id>
<article-id pub-id-type="doi">10.1016/j.bpj.2010.06.068</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Spectroscopy, Imaging, and Other Techniques</subject>
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</article-categories>
<title-group><article-title>NMR Separation of Intra- and Extracellular Compounds Based on Intermolecular Coherences</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Hoerr</surname>
<given-names>Verena</given-names>
</name>
<xref rid="aff1" ref-type="aff">†</xref>
<xref rid="aff2" ref-type="aff">‡</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Purea</surname>
<given-names>Armin</given-names>
</name>
<xref rid="aff1" ref-type="aff">†</xref>
<xref rid="aff3" ref-type="aff">§</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Faber</surname>
<given-names>Cornelius</given-names>
</name>
<email>faberc@uni-muenster.de</email>
<xref rid="aff1" ref-type="aff">†</xref>
<xref rid="aff2" ref-type="aff">‡</xref>
<xref rid="cor1" ref-type="corresp">∗</xref>
</contrib>
</contrib-group>
<aff id="aff1"><label>†</label>
Department of Experimental Physics 5, University of Würzburg, Würzburg, Germany</aff>
<aff id="aff2"><label>‡</label>
Department of Clinical Radiology, University Hospital Münster, Münster, Germany</aff>
<aff id="aff3"><label>§</label>
RAPID Biomedical, Rimpar, Germany</aff>
<author-notes><corresp id="cor1"><label>∗</label>
Corresponding author <email>faberc@uni-muenster.de</email>
</corresp>
</author-notes>
<pub-date pub-type="ppub"><day>06</day>
<month>10</month>
<year>2010</year>
</pub-date>
<volume>99</volume>
<issue>7</issue>
<fpage>2336</fpage>
<lpage>2343</lpage>
<history><date date-type="received"><day>1</day>
<month>4</month>
<year>2010</year>
</date>
<date date-type="accepted"><day>28</day>
<month>6</month>
<year>2010</year>
</date>
</history>
<permissions><copyright-statement>© 2010 by the Biophysical Society.</copyright-statement>
<copyright-year>2010</copyright-year>
<copyright-holder>Biophysical Society</copyright-holder>
</permissions>
<abstract><p>NMR spectroscopy is a powerful tool for detection and characterization of chemical compounds in biological systems. Its application in pharmaceutical studies in cell cultures, however, has been hampered by the enormous technical challenges in separating intra- from extracellular amounts of one substance. We introduce a novel approach to separate intra- from extracellular NMR signal based on the detection of intermolecular zero-quantum coherences in presence of a chemical shift agent. In a sample of large cells in culture, the investigation of cellular uptake of pharmacological substances becomes feasible. The addition of 10 mM Tm-DOTP to a suspension of 100 <italic>Xenopus laevis</italic>
oocytes resulted in sufficient separation of resonance frequencies between intra- and extracellular water. Upon selective excitation of either intra- or extracellular water signal, only intra- or extracellular components were observed, respectively. The presented localization technique provides intrinsic averaging over a large number of cells, resulting in a significant signal gain. The method works on standard NMR spectrometers, which are available at most scientific research institutions today. On a high-resolution NMR system with a cryoprobe, a 20-fold sensitivity gain was observed as compared to conventionally localized NMR spectroscopy of a single <italic>X. laevis</italic>
oocyte on dedicated NMR microscopes.</p>
</abstract>
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</front>
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