Name: preparation, purification, and characterization of lanthanide complexes for use as contrast agents for magnetic resonance imaging
Uploaded: 2011-07-21T13:00:00
Description: Watch this Scientific Journal Video about Preparation, Purification, and Characterization of Lanthanide Complexes for Use as Contrast Agents for Magnetic Resonance Imaging at JoVE.com

Agradecemos fundos de inicialização de Wayne State University (MJA), uma bolsa da Fundação Americana para Aging Research (SMV), e um Caminho para a Independência Award Transição de Carreira (R00EB007129) do Instituto Nacional de Imagem Biomédica e Bioengenharia do Instituto Nacional da Saúde (MJA).

1. Metalation usando LnCl três sais <ol><li> Dissolver o ligante na água para produzir uma solução de 30-265 mM. O ligante 2 - (4-isothiocyanatobenzyl)-ácido dietileno pentaacetic (p-SCN Bn-DTPA) foi usado neste vídeo a uma concentração de 73 mM. </li><li> Ajustar o pH da solução do ligante para entre 5,5 e 7,0 pela adição de 1 M NH 4 OH. Neste vídeo, 0,2 mL de NH M 1 4 OH solução foi usado. </li><li> Dissolver 02/01 equivalentes de LnCl 3 em água pa...

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	<li>Barge, A., Cravotto, G., Gianolio, E., Fedeli, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+to+determine+free+Gd+and+free+ligand+in+solution+of+Gd+chelates.+A+technical+note.">How to determine free Gd and free ligand in solution of Gd chelates. A technical note.</a> Contrast Med. Mol. Imaging. 1, 184-188 (2006).</li><li>Nagaraja, T. N., Croxen, R. L., Panda, S., Knight, R. A., Keenan, K. A., Brown, S. L., Fenstermacher, J. D., Ewing, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+arsenazo+III+in+the+preparation+and+characterization+of+an+albumin-linked,+gadolinium-based+macromolecular+magnetic+resonance+contrast+agent.">Application of arsenazo III in the preparation and characterization of an albumin-linked, gadolinium-based macromolecular magnetic resonance contrast agent.</a> J. Neurosci. Methods. 157, 238-245 (2006).</li><li>Supkowski, R. M., Horrocks, W. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=On+the+determination+of+the+number+of+water+molecules,+q,+coordinated+to+europium(III)+ions+in+solution+from+luminescence+decay+lifetimes.">On the determination of the number of water molecules, q, coordinated to europium(III) ions in solution from luminescence decay lifetimes.</a> Inorg. Chim. Acta. 340, 44-48 (2002).</li><li>Menjoge, A. R., Kannan, R. M., Tomalia, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dendrimer-based+drug+and+imaging+conjugates:+design+considerations+for+nanomedical+applications.">Dendrimer-based drug and imaging conjugates: design considerations for nanomedical applications.</a> Drug Discovery Today. 15, 171-185 (2010).</li><li>Que, E. L., Chang, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Responsive+magnetic+resonance+imaging+contrast+agents+as+chemical+sensors+for+metals+in+biology+and+medicine.">Responsive magnetic resonance imaging contrast agents as chemical sensors for metals in biology and medicine.</a> Chem. Soc. Rev. 39, 51-60 (2010).</li><li>Uppal, R., Caravan, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeted+probes+for+cardiovascular+MR+imaging.">Targeted probes for cardiovascular MR imaging.</a> Future Med. Chem. 2, 451-470 (2010).</li><li>Major, J. L., Meade, T. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bioresponsive,+cell-penetrating,+and+multimeric+MR+contrast+agents.">Bioresponsive, cell-penetrating, and multimeric MR contrast agents.</a> Acc. Chem. Res. 42, 893-903 (2009).</li><li>Datta, A., Raymond, K. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gd-hydroxypyridinone+(HOPO)-based+high-relaxivity+magnetic+resonance+imaging+(MRI)+contrast+agents.">Gd-hydroxypyridinone (HOPO)-based high-relaxivity magnetic resonance imaging (MRI) contrast agents.</a> Acc. Chem. Res. 42, 938-947 (2009).</li><li>Le&#243;n-Rodr&#237;guez, L. M. D., Lubag, A. J. M., Malloy, C. R., Martinez, G. V., Gillies, R. J., Sherry, A. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Responsive+MRI+agents+for+sensing+metabolism+in+vivo.">Responsive MRI agents for sensing metabolism in vivo.</a> Acc. Chem. Res. 42, 948-957 (2009).</li><li>Castelli, D. D., Gianolio, E., Crich, S. G., Terreno, E., Aime, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metal+containing+nanosized+systems+for+MR-molecular+imaging+applications.">Metal containing nanosized systems for MR-molecular imaging applications.</a> Coord. Chem. Rev. 252, 2424-2443 (2008).</li><li>Caravan, P., Ellison, J. J., McMurry, T. J., Lauffer, R. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gadolinium(III)+chelates+as+MRI+contrast+agents:+structure,+dynamics,+and+applications.">Gadolinium(III) chelates as MRI contrast agents: structure, dynamics, and applications.</a> Chem. Rev. 99, 2293-2352 (1999).</li><li>Lauffer, R. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Paramagnetic+metal+complexes+as+water+proton+relaxation+agents+for+NMR+imaging:+theory+and+design.">Paramagnetic metal complexes as water proton relaxation agents for NMR imaging: theory and design.</a> Chem. Rev. 87, 901-927 (1987).</li><li>Yoo, B., Pagel, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+overview+of+responsive+MRI+contrast+agents+for+molecular+imaging.">An overview of responsive MRI contrast agents for molecular imaging.</a> Front. Biosci. 13, 1733-1752 (2008).</li><li>Pandya, S., Yu, J., Parker, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Engineering+emissive+europium+and+terbium+complexes+for+molecular+imaging+and+sensing.">Engineering emissive europium and terbium complexes for molecular imaging and sensing.</a> Dalton Trans. 23, 2757-2766 (2006).</li><li>Nwe, K., Xu, H., Regino, C. A. S., Bernardo, M., Ileva, L., Riffle, L., Wong, K. J., Brechbiel, M. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+approach+in+the+preparation+of+dendrimer-based+bifunctional+diethylenetriaminepentaacetic+acid+MR+contrast+agent+derivatives.">A new approach in the preparation of dendrimer-based bifunctional diethylenetriaminepentaacetic acid MR contrast agent derivatives.</a> Bioconjugate Chem. 20, 1412-1418 (2009).</li><li>Nwe, K., Bernardo, M., Regino, C. A. S., Williams, M., Brechbiel, M. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+MRI+properties+between+derivatized+DTPA+and+DOTA+gadolinium-dendrimer+conjugates.">Comparison of MRI properties between derivatized DTPA and DOTA gadolinium-dendrimer conjugates.</a> Bioorg. Med. Chem. 18, 5925-5931 (2010).</li><li>Caravan, P., Das, B., Deng, Q., Dumas, S., Jacques, V., Koerner, S. K., Kolodziej, A., Looby, R. J., Sun, W. -. C., Zhang, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+lysine+walk+to+high+relaxivity+collagen-targeted+MRI+contrast+agents.">A lysine walk to high relaxivity collagen-targeted MRI contrast agents.</a> Chem. Commun. , 430-432 (2009).</li><li>Le&#243;n-Rodr&#237;guez, L. M. D., Kovacs, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+synthesis+and+chelation+chemistry+of+DOTA-peptide+conjugates.">The synthesis and chelation chemistry of DOTA-peptide conjugates.</a> Bioconjugate Chem. 19, 391-402 (2008).</li><li>Boswell, C. A., Eck, P. K., Regino, C. A. S., Bernardo, M., Wong, K. J., Milenic, D. E., Choyke, P. L., Brechbiel, M. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis,+characterization,+and+biological+evaluation+of+integrin+&#945;V&#946;3-targeted+PAMAM+dendrimers.">Synthesis, characterization, and biological evaluation of integrin &#945;V&#946;3-targeted PAMAM dendrimers.</a> Mol. Pharm. 5, 527-539 (2008).</li></ol>

Dado o número crescente de publicações que incluem agentes de contraste à base de lantanídeos 14/04, é importante que seja tomado cuidado na preparação, purificação e caracterização de produtos para garantir resultados reprodutíveis e comparáveis. Estes complexos são muitas vezes considerados desafiadores para purificar e caracterizar moléculas orgânicas em relação ao devido à sua natureza paramagnética ea sensibilidade de qualquer grupo funcional que pode ser usado para bioconjugation. N...

<table border="1"><tbody><tr><th> Reagentes e Equipamentos </th><th> Companhia </th><th> Número de catálogo </th></tr><tr><td> EuCl 3 • 6H 2 O </td><td> Sigma-Aldrich </td><td> 203254-5G </td></tr><tr><td> p SCN-Bn-DTPA </td><td> Macrocyclics </td><td> B-305 </td></tr><tr><td> hidróxido de amônio </td><td> EMD </td><td> AX1303-3 </td></tr><tr><td> Spectra / Por Biotech Celulose Ester (CE) Membrana de Diálise - 500 D MWCO </td><td> Fisher Scientific </td><td> 68-671-24 </td></tr><tr><td> Millipore IC Millex-LG Unidades Filtro </td><td> Fisher Scientific </td><td> SLLG C13 NL </td></tr><tr><td> xilenol laranja tetrasodium sal </td><td> Alfa Aesar </td><td> 41379 </td></tr><tr><td> ácido acético </td><td> Fluka </td><td> 49199 </td></tr><tr><td> D 2 O </td><td> Cambridge Isotope Laboratories, Inc. </td><td> DLM-4-25 </td></tr><tr><td> depurador de água </td><td> ELGA </td><td> PURELAB Ultra </td></tr><tr><td> cromatografia líquida de alta eficiência e espectrometria de massas </td><td> Shimadzu </td><td> LCMS-2010EV </td></tr><tr><td> analisador de tempo de relaxamento </td><td> Bruker </td><td> mq60 minispec </td></tr><tr><td> Espectrofotômetro UV-vis </td><td> Fisher Scientific </td><td> 20-624-00092 </td></tr><tr><td> liofilizador </td><td> Fisher Scientific </td><td> 10-030-133 </td></tr><tr><td> medidor de pH </td><td> Hanna Instruments </td><td> HI 221 </td></tr><tr><td> espectrofluorímetro </td><td> HORIBA Jobin Yvon </td><td> Fluoromax-4 </td></tr><tr><td> Versão Calculadora Peso Molecular 6,46 por Matthew Monroe, baixado 17 out, 2009 </td><td> <a href="http://ncrr.pnl.gov/software/">http://ncrr.pnl.gov/software/</a> </td><td> Calculadora de Peso Molecular </td></tr></tbody></table>

preparation, purification, and characterization of lanthanide complexes for use as contrast agents for magnetic resonance imaging

Polyaminopolycarboxylate baseado ligantes são comumente usados ​​para quelar íons lantanídeos, e os complexos resultantes são úteis como agentes de contraste para ressonância magnética (MRI). Muitos ligantes disponíveis comercialmente são especialmente úteis porque contêm grupos funcionais que permitem rápido, de alta pureza e alto rendimento conjugação de macromoléculas e biomoléculas através de amina-reativo ésteres ativados e grupos isotiocianato ou tiol-reativa Maleimidas. Enquanto metalation desses ligantes é considerada o conhecimento comum no campo da química bioconjugation, diferenças sutis em procedimentos metalation devem ser levados em consideração na seleção de materiais metálicos de partida. Além disso, várias opções para a purificação e caracterização de existir, e seleção do procedimento mais eficaz depende em parte da seleção de matérias-primas. Essas diferenças sutis são muitas vezes negligenciados nos protocolos publicados. Aqui, nosso objetivo é demonstrar os métodos comuns para metalation, purificação e caracterização de complexos de lantanídeos que podem ser usados ​​como agentes de contraste para ressonância magnética (Figura 1). Esperamos que esta publicação irá permitir que os cientistas biomédicos para incorporar as reações de complexação lantanídeos em seu repertório de reações comumente utilizados, facilitando a seleção de matérias-primas e métodos de purificação.

Watch this Scientific Journal Video about Preparation, Purification, and Characterization of Lanthanide Complexes for Use as Contrast Agents for Magnetic Resonance Imaging at JoVE.com

Preparation, Purification, and Characterization of Lanthanide Complexes for Use as Contrast Agents for Magnetic Resonance Imaging

Demonstramos a metalation, purificação e caracterização de complexos de lantanídeos. Os complexos descrito aqui pode ser conjugado com macromoléculas para habilitar o rastreamento destas moléculas usando ressonância magnética.

Purificação de preparação, e Caracterização de Complexos de lantanídeos para uso como agentes de contraste para Imagem por Ressonância Magnética

Polyaminopolycarboxylate-based ligands are commonly used to chelate lanthanide ions, and the resulting complexes are 
useful as contrast agents for ...

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