Recent advances in the field of in-cell NMR spectroscopy have made it possible to study proteins in the context of bacterial or mammalian cell extracts or even entire cells. As most mammalian cells are part of a multi-cellular complex, there is a need to develop novel NMR approaches enabling the study of proteins within the complexity of a 3D cellular environment. Here we investigate the use of the hanging drop method to grow spheroids which are homogenous in size and shape as a model system to study solid tumors using solid-state NMR (ssNMR) spectroscopy. We find that these spheroids are stable under magic-angle-spinning conditions and show a clear change in metabolic profile as compared to single cell preparations. Finally, we utilize dynamic nuclear polarization (DNP)-supported ssNMR measurements to show that low concentrations of labelled nanobodies targeting EGFR (7D12) can be detected inside the spheroids. These findings suggest that solid-state NMR can be used to directly examine proteins or other biomolecules in a 3D cellular microenvironment with potential applications in pharmacological research.
[NMR paper] (1) H-Detected Solid-State NMR Studies of Water-Inaccessible Proteins In Vitro and In Situ.
(1) H-Detected Solid-State NMR Studies of Water-Inaccessible Proteins In Vitro and In Situ.
Related Articles (1) H-Detected Solid-State NMR Studies of Water-Inaccessible Proteins In Vitro and In Situ.
Angew Chem Int Ed Engl. 2016 Sep 27;
Authors: Medeiros-Silva J, Mance D, Daniëls M, Jekhmane S, Houben K, Baldus M, Weingarth M
Abstract
(1) H detection can significantly improve solid-state NMR spectral sensitivity and thereby allows studying more complex proteins. However, the common prerequisite for (1) H detection is the...
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09-28-2016 06:22 PM
[NMR paper] Abnormal lipid metabolism in collagen-induced arthritis rat model: in vitro, high resolution NMR spectroscopy based analysis.
Abnormal lipid metabolism in collagen-induced arthritis rat model: in vitro, high resolution NMR spectroscopy based analysis.
Related Articles Abnormal lipid metabolism in collagen-induced arthritis rat model: in vitro, high resolution NMR spectroscopy based analysis.
Indian J Exp Biol. 2014 Jul;52(7):673-82
Authors: Srivastava NK, Sharma S, Purusottam RN, Sinha N, Singh R, Sharma D
Abstract
Collagen-induced arthritis (CIA) was induced in female Wistar rats by intradermal injection of porcine immunization grade native collagen...
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08-06-2014 08:39 PM
Press Release: NIH Funds Development of Integrated THz System, Making DNP-NMR Spectroscopy More Affordable
From The DNP-NMR Blog:
Press Release: NIH Funds Development of Integrated THz System, Making DNP-NMR Spectroscopy More Affordable
Press Release
NIH Funds Development of Integrated THz System, Making DNP-NMR Spectroscopy More Affordable
Bridge12’s SBIR Fast-Track Grant Will Eliminate Need for Second Superconducting Magnet, Greatly Reducing Cost for Dynamic Nuclear Polarization
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11-21-2013 01:14 AM
[NMR paper] KML001 Displays Vascular Disrupting Properties and Irinotecan Combined Antitumor Activities in a Murine Tumor Model.
KML001 Displays Vascular Disrupting Properties and Irinotecan Combined Antitumor Activities in a Murine Tumor Model.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www.plosone.org-images-pone_120x30.png http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www.pubmedcentral.nih.gov-corehtml-pmc-pmcgifs-pubmed-pmc.gif Related Articles KML001 Displays Vascular Disrupting Properties and Irinotecan Combined Antitumor Activities in a Murine Tumor Model.
PLoS One. 2013;8(1):e53900
Authors: Moon CH, Lee SJ, Lee HY, Lee JC, Cha H, Cho WJ, Park JW, Park HJ,...
Development of 43Ca NMR Solid State NMR Spectroscopy as a Probe of Local Structure in Inorganic and Molecular Materials
Development of 43Ca NMR Solid State NMR Spectroscopy as a Probe of Local Structure in Inorganic and Molecular Materials
Publication year: 2012
Source:Progress in Nuclear Magnetic Resonance Spectroscopy</br>
Danielle Laurencin, Mark E. Smith</br>
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http://origin-ars.els-cdn.com/content/image/1-s2.0-S0079656512000684-fx1.jpg Graphical abstract Highlights
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05-17-2012 11:37 PM
Solid-state (55)Mn NMR spectroscopy of bis(?-oxo)dimanganese(IV) [Mn(2)O(2)(salpn)(2)], a model for the oxygen evolving complex in photosystem II.
Solid-state (55)Mn NMR spectroscopy of bis(?-oxo)dimanganese(IV) , a model for the oxygen evolving complex in photosystem II.
Solid-state (55)Mn NMR spectroscopy of bis(?-oxo)dimanganese(IV) , a model for the oxygen evolving complex in photosystem II.
J Am Chem Soc. 2010 Dec 1;132(47):16727-9
Authors: Ellis PD, Sears JA, Yang P, Dupuis M, Boron TT, Pecoraro VL, Stich TA, Britt RD, Lipton AS
We have examined the antiferromagneticly coupled bis(?-oxo)dimanganese(IV) complex (1) with (55)Mn solid-state NMR at cryogenic temperatures and...
Development of in vitro-grown spheroids as a 3D tumor model system for solid-state NMR spectroscopy
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