Name: synthesis of aptamer-pei-g-peg modified gold nanoparticles loaded with doxorubicin for targeted drug delivery
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Description: Watch this Scientific Journal Video about Synthesis of Aptamer-PEI-g-PEG Modified Gold Nanoparticles Loaded with Doxorubicin for Targeted Drug Delivery at JoVE.com

Denne forskning blev finansieret af National Natural Science Foundation of China (31700840); Henanprovinsens vigtigste videnskabelige forskningsprojekt (18B430013, 18A150049). Denne forskning blev støttet af Nanhu Scholars Program for Young Scholars of XYNU. Forfatterne vil gerne takke bachelorstuderende Zebo Qu fra College of Life Sciences i XYNU for hans nyttige værker. Forfatterne vil gerne anerkende Analysis &Testing Center af XYNU for brugen af deres udstyr.

ADVARSEL: Sørg for at konsultere alle relevante datablade om materialesikkerhed (MSDS), før du bruger alle kemikalier. Flere af de kemikalier, der anvendes til fremstilling af copolymer og nanopartikler, er akut giftige. Nanopartikler har også potentielle farer. Sørg for at bruge alle relevante sikkerhedspraksisser og personlige værnemidler, herunder handsker, laboratoriekittel, hætter, bukser i fuld længde og nærtåede sko.
1. Syntese af dobbelt-carboxyl polyethylen glycol (CT-PEG)<sup ...

<ol>
	<li>Abad, J. M., Bravo, I., Pariente, F., Lorenzo, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multi-tasking+base+ligand:+a+new+concept+of+AuNPs+synthesis.">Multi-tasking base ligand: a new concept of AuNPs synthesis.</a> Analytical and Bioanalytical Chemistry. 408 (9), 2329-2338 (2016).</li><li>Siegel, R. L., Miller, K. D., Jemal, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cancer+statistics,+2019.">Cancer statistics, 2019.</a> CA-A Cancer Journal for Clinicians. 69 (1), 7-34 (2019).</li><li>Jang, B., Kwon, H., Katila, P., Lee, S. 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S., Murphy, 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=Recent+Progress+in+Cancer+Thermal+Therapy+Using+Gold+Nanoparticles.">Recent Progress in Cancer Thermal Therapy Using Gold Nanoparticles.</a> The Journal of Physical Chemistry C. 120 (9), 4691-4716 (2016).</li><li>Riley, R. S., Day, E. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gold+nanoparticle-mediated+photothermal+therapy:+applications+and+opportunities+for+multimodal+cancer+treatment.">Gold nanoparticle-mediated photothermal therapy: applications and opportunities for multimodal cancer treatment.</a> WIREs Nanomedicine and Nanobiotechnology. 9 (4), 1449 (2017).</li><li>Fratoddi, I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Highly+Hydrophilic+Gold+Nanoparticles+as+Carrier+for+Anticancer+Copper(I)+Complexes:+Loading+and+Release+Studies+for+Biomedical+Applications.">Highly Hydrophilic Gold Nanoparticles as Carrier for Anticancer Copper(I) Complexes: Loading and Release Studies for Biomedical Applications.</a> Nanomaterials (Basel). 9 (5), 772 (2019).</li><li>Lee, S. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drug-loaded+gold+plasmonic+nanoparticles+for+treatment+of+multidrug+resistance+in+cancer.">Drug-loaded gold plasmonic nanoparticles for treatment of multidrug resistance in cancer.</a> Biomaterials. 35 (7), 2272-2282 (2014).</li><li>Dreaden, E. C., Alkilany, A. M., Huang, X., Murphy, C. J., El-Sayed, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+golden+age:+gold+nanoparticles+for+biomedicine.">The golden age: gold nanoparticles for biomedicine.</a> Chemical Society Reviews. 41 (7), 2740-2779 (2012).</li><li>Wei, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anticancer+drug+nanomicelles+formed+by+self-assembling+amphiphilic+dendrimer+to+combat+cancer+drug+resistance.">Anticancer drug nanomicelles formed by self-assembling amphiphilic dendrimer to combat cancer drug resistance.</a> Proceedings of the National Academy of Sciences of the United States of America. 112 (10), 2978-2983 (2015).</li><li>Galluzzi, L., Buque, A., Kepp, O., Zitvogel, L., Kroemer, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunological+Effects+of+Conventional+Chemotherapy+and+Targeted+Anticancer+Agents.">Immunological Effects of Conventional Chemotherapy and Targeted Anticancer Agents.</a> Cancer Cell. 28 (6), 690-714 (2015).</li><li>Muddineti, O. S., Ghosh, B., Biswas, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+trends+in+using+polymer+coated+gold+nanoparticles+for+cancer+therapy.">Current trends in using polymer coated gold nanoparticles for cancer therapy.</a> International Journal of Pharmaceutics. 484 (1-2), 252-267 (2015).</li><li>Hu, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methyl+Orange+removal+by+a+novel+PEI-AuNPs-hemin+nanocomposite.">Methyl Orange removal by a novel PEI-AuNPs-hemin nanocomposite.</a> Journal of Environmental Sciences. 53, 278-283 (2017).</li><li>Gu, F. X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeted+nanoparticles+for+cancer+therapy.">Targeted nanoparticles for cancer therapy.</a> Nano Today. 2 (3), 14-21 (2007).</li><li>Srinivasarao, M., Galliford, C. V., Low, P. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Principles+in+the+design+of+ligand-targeted+cancer+therapeutics+and+imaging+agents.">Principles in the design of ligand-targeted cancer therapeutics and imaging agents.</a> Nature Reviews Drug Discovery. 14 (3), 203-219 (2015).</li><li>Liu, Z., Shi, Y., Chen, Z., Duan, L., Wang, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+progress+towards+the+use+of+aptamers+in+targeted+cancer+therapy.">Current progress towards the use of aptamers in targeted cancer therapy.</a> Chinese Science Bulletin (Chinese Version). 59 (14), 1267 (2014).</li><li>Ghosh, P., Han, G., De, M., Kim, C. K., Rotello, V. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gold+nanoparticles+in+delivery+applications.">Gold nanoparticles in delivery applications.</a> Advanced Drug Delivery Reviews. 60 (11), 1307-1315 (2008).</li><li>Vandghanooni, S., Eskandani, M., Barar, J., Omidi, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antisense+LNA-loaded+nanoparticles+of+star-shaped+glucose-core+PCL-PEG+copolymer+for+enhanced+inhibition+of+oncomiR-214+and+nucleolin-mediated+therapy+of+cisplatin-resistant+ovarian+cancer+cells.">Antisense LNA-loaded nanoparticles of star-shaped glucose-core PCL-PEG copolymer for enhanced inhibition of oncomiR-214 and nucleolin-mediated therapy of cisplatin-resistant ovarian cancer cells.</a> International Journal of Pharmaceutics. 573, 118729 (2020).</li><li>Andghanooni, S., Eskandani, M., Barar, J., Omidi, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=AS1411+aptamer-decorated+cisplatin-loaded+poly(lactic-co-glycolic+acid)+nanoparticles+for+targeted+therapy+of+miR-21-inhibited+ovarian+cancer+cells.">AS1411 aptamer-decorated cisplatin-loaded poly(lactic-co-glycolic acid) nanoparticles for targeted therapy of miR-21-inhibited ovarian cancer cells.</a> Nanomedicine. 13 (21), 2729-2758 (2018).</li><li>Palmieri, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+anti-nucleolin+recombinant+immunoagent+for+cancer+therapy.">Human anti-nucleolin recombinant immunoagent for cancer therapy.</a> Proceedings of the National Academy of Sciences of the United States of America. 112 (30), 9418-9423 (2015).</li><li>Pichiorri, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+NCL+targeting+affects+breast+cancer+aggressiveness+through+miRNA+regulation.">In vivo NCL targeting affects breast cancer aggressiveness through miRNA regulation.</a> Journal of Experimental Medicine. 210 (5), 951-968 (2013).</li><li>Hou, S., McCauley, L. K., Ma, P. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+and+erosion+properties+of+PEG-containing+polyanhydrides.">Synthesis and erosion properties of PEG-containing polyanhydrides.</a> Macromolecular Bioscience. 7 (5), 620-628 (2007).</li><li>Nie, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Injectable+Vaginal+Hydrogels+as+a+Multi-Drug+Carrier+for+Contraception.">Injectable Vaginal Hydrogels as a Multi-Drug Carrier for Contraception.</a> Applied Sciences. 9 (8), 1638 (2019).</li><li>Zou, P., Suo, J., Nie, L., Feng, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Temperature-responsive+biodegradable+star-shaped+block+copolymers+for+vaginal+gels.">Temperature-responsive biodegradable star-shaped block copolymers for vaginal gels.</a> Journal of Materials Chemistry. 22 (13), 6316-6326 (2012).</li><li>Etrych, T., &#352;ubr, V., Laga, R., &#344;&#237;hov&#225;, B., Ulbrich, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Polymer+conjugates+of+doxorubicin+bound+through+an+amide+and+hydrazone+bond:+Impact+of+the+carrier+structure+onto+synergistic+action+in+the+treatment+of+solid+tumours.">Polymer conjugates of doxorubicin bound through an amide and hydrazone bond: Impact of the carrier structure onto synergistic action in the treatment of solid tumours.</a> European Journal of Pharmaceutical Sciences. 58, 1-12 (2014).</li><li>Safari, F., Tamaddon, A. M., Zarghami, N., Abolmali, S., Akbarzadeh, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Polyelectrolyte+complexes+of+hTERT+siRNA+and+polyethyleneimine:+Effect+of+degree+of+PEG+grafting+on+biological+and+cellular+activity.">Polyelectrolyte complexes of hTERT siRNA and polyethyleneimine: Effect of degree of PEG grafting on biological and cellular activity.</a> Artificial Cells, Nanomedicine, and Biotechnology. 44 (6), 1561-1568 (2016).</li></ol>

1H NMR-spektret (figur 2) bekræfter den vellykkede syntese af CT-PEG-copolymer og PEI-g-PEG-copolymer. De molekylære vægte af PEG og PEI var henholdsvis 1.000 og 1.200. Derudover blev EDC/NHS-katalysatorsystemet brugt til at syntetisere PEI-g-PEG-copolymer via amide reaktioner. Det skal bemærkes, at hvis peg's og PEI's molekylvægte ændres for syntetisering af PEI-g-PEG-copolymer, skal reaktionstiden og katalysatorsystemet revurderes. Reaktionstilstanden for PEI-g-PEG-copolym...

At være den største folkesundhed problem på verdensplan, kræft er bredt karakteriseret som havende en lav helbredelse, høj gentagelsesrate, og højdødelighed 1,2. Nuværende konventionelle anti-cancer metoder omfatter kirurgi, kemoterapi, ogstrålebehandling 3, blandt hvilke kemoterapi er den primære behandling for kræftpatienter i klinikken4. Klinisk anvendte kræftlægemidler omfatter hovedsageligt paclitaxel (PTX)5 og doxorubicin (DOX)6,7. DOX, et antineoplastisk lægemiddel, er stort set blevet anvendt i klinisk kemoterapi på grund af fordelene ved kræft cytotoksicitet og hæmning af kræftcellespredning8,9. DOX forårsager dog kardiotoksicitet10,11, og DOX's korte halveringstid begrænser dens anvendelse i klinikken12. Derfor er nedbrydelige lægemiddelbærere nødvendige for at indlæse DOX og diskret frigive på en kontrolleret måde til et målrettet område.
Nanopartikler er blevet udbredt i målrettede narkotikaleveringssystemer og har flere fordele i kræftbehandling (dvs. betydeligt overflade-til-volumen-forhold, lille størrelse, evne til at indkapsle forskellige lægemidler og tunabel overfladekemi osv.) 13,14,15. Især guld nanopartikler (AuNPs) har været meget udbredt i biologiske og biomedicinske applikationer, såsom fototermisk kræftbehandling16,17. De unikke egenskaber ved AuNPs, såsom let syntese og generel overfladefunktionalisering, har fremragende udsigter inden for det kliniske område af kræftbehandling18. Også, AuNPs er blevet brugt til at identificere narkotika levering strategier, diagnosticere tumorer, og overvinde resistens i mange undersøgelser19,20.
Ikke desto mindre skal auNPs skræddersys yderligere for at overvinde resistens over for lægemidler via høj lokal frigivelse ved tumorlæsioner gennem øget gennemtrængning og fastholdelse (EPR), såsom målretnings- og tilgængelighedsegenskaber. Polymerfunktionaliserede godkendelsesprogrammer har udvist unikke fordele, såsom forbedret vandopløselighed af hydrofobe anticancermedicin og forlænget cirkulationstid21,22. Forskellige biokompatible polymerer er blevet brugt til AuNP-belægninger, såsom polyethylenglycol (PEG), polyethylenetimine (PEI), hyaluronsyre, heparin og xanthangummi. Så stabiliteten, samt nyttelasten, af AuNPs er forbedret godt23. Specifikt er PEI en stærkt forgrenet polymer, der består af mange gentagne enheder af primære, sekundære og tertiære aminer24. PEI har fremragende opløselighed, lav viskositet og en høj grad af funktionalitet, som er velegnet til belægning på AuNPs.
På den anden side skal kræftmedicin leveres til kræftceller direkte med forbedret belastningseffektivitet og med lavere toksicitet til behandling af primære og avancerede metastatiske tumorer25. Målrettede ligands har et stort potentiale for anti-cancer lægemiddel målrettet levering systemer26. Dens selektivitet for målmolekylebinding giver anti-cancer lægemiddel rettet mod specificitet og øger lægemiddelberigelsen i syge væv27. Flere ligands omfatter antistoffer, polypeptider og små molekyler. Sammenlignet med andre ligands, nukleinsyre aptamers kan syntetiseres in vitro og er nemme at ændre. AS1411 er en uændret 26 bp phosphodiester oligonukleotid , der danner en stabil dimerisk G-tetramer struktur til specifikt at binde sig til en overekspresseret mål nuklear protein receptor på kræftceller28,29,30. AS1411 hæmmer spredningen af mange kræftceller, men påvirker ikke væksten af raske celler31,32. Som et resultat, AS1411 er blevet brugt til at fremstille en ideel målrettet lægemiddellevering system.
I denne undersøgelse syntetiseres en PEI-g-PEG copolymer via en midtreaktion, hvorefter PEI-g-PEG copolymer coatede guld nanopartikler (PEI-g-PEG@AuNPs) fremstilles. Derudover er DOX og AS1411 fortløbende knyttet til de forberedte PEI-g-PEG@AuNPs, som vist i figur 1. Denne detaljerede protokol er beregnet til at hjælpe forskere med at undgå mange af de almindelige faldgruber forbundet med fremstillingen af nye PEI-g-PEG@AuNPs fyldt med DOX og AS1411.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>4-Dimethylaminopyridine</td>
			<td>Macklin</td>
			<td>D807273</td>
			<td></td>
		</tr>
		<tr>
			<td>A549 cell</td>
			<td></td>
			<td>ATCC CCL-185TM</td>
			<td></td>
		</tr>
		<tr>
			<td>AS1411</td>
			<td>BBI Life Sciences Corporation</td>
			<td></td>
			<td>5&#39;-d (TTTGGTGGTGGTGGTTGTGGTGGTGGTGG) FL-AS1411 (fluorophore-labeled AS1411)</td>
		</tr>
		<tr>
			<td>Anhydrous Tetrahydrofuran (THF)</td>
			<td>SinoPharm Chemical Reagent Co., Ltd</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cell counting kit-8 (CCK-8)</td>
			<td>Sigma Aldrich</td>
			<td>96992-500TESTS-F</td>
			<td></td>
		</tr>
		<tr>
			<td>Dichloromethane</td>
			<td>Traditional Chinese medicine</td>
			<td>80047318</td>
			<td></td>
		</tr>
		<tr>
			<td>Diethyl ether (Et2O)</td>
			<td>SinoPharm Chemical Reagent Co., Ltd</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Dimethyl sulfoxide</td>
			<td>Macklin</td>
			<td>D806645</td>
			<td></td>
		</tr>
		<tr>
			<td>Dulbecco&#39;s modified Eagle&#39;s medium (DMEM)</td>
			<td>Sigma Aldrich</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Doxorubicin hydrochloride</td>
			<td>Rhawn</td>
			<td>R017518</td>
			<td></td>
		</tr>
		<tr>
			<td>Ether absolute</td>
			<td>Traditional Chinese medicine</td>
			<td>80059618</td>
			<td></td>
		</tr>
		<tr>
			<td>Field Emission Transmission Electron Microscope</td>
			<td>FEI Company</td>
			<td>Tecnai G2 F 20</td>
			<td></td>
		</tr>
		<tr>
			<td>Gold(III) chloride trihydrate</td>
			<td>Rhawn</td>
			<td>R016035</td>
			<td></td>
		</tr>
		<tr>
			<td>Laser Particle-size Instrument</td>
			<td>Malvern Instruments Ltd</td>
			<td>ZetasizerNanoZS/Masterszer3000E</td>
			<td></td>
		</tr>
		<tr>
			<td>Microplate Reader</td>
			<td>Molecular Devices</td>
			<td>SpectraMax 190</td>
			<td></td>
		</tr>
		<tr>
			<td>N-(3-Dimethylaminopropyl)-N&#39;-ethylcarbodiimide hydrochloride</td>
			<td>Macklin</td>
			<td>N808856</td>
			<td></td>
		</tr>
		<tr>
			<td>N-Hydroxysuccinimide</td>
			<td>Macklin</td>
			<td>H6231</td>
			<td></td>
		</tr>
		<tr>
			<td>NMR software</td>
			<td>Delta 5.2.1</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Nuclear Magnetic Resonance Spectrometer</td>
			<td>JEOL</td>
			<td>JNM-ECZ600R/S3</td>
			<td></td>
		</tr>
		<tr>
			<td>Origin 8.5</td>
			<td>OriginLab</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Penicillin</td>
			<td>Sigma Aldrich</td>
			<td>V900929-100ML</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate-buffered saline</td>
			<td>Sigma Aldrich</td>
			<td>P4417-100TAB</td>
			<td></td>
		</tr>
		<tr>
			<td>Poly(ethylene glycol)</td>
			<td>Sigma Aldrich</td>
			<td>81188</td>
			<td>BioUltra, average Mn ~ 1000</td>
		</tr>
		<tr>
			<td>Poly (ethyleneimine) solution</td>
			<td>Sigma Aldrich</td>
			<td>482595</td>
			<td>average Mn ~ 1200, 50 wt.% in H2O</td>
		</tr>
		<tr>
			<td>Sodium borohydride, powder</td>
			<td>Acros</td>
			<td>C18930</td>
			<td></td>
		</tr>
		<tr>
			<td>Streptomycin</td>
			<td>Sigma Aldrich</td>
			<td>85886-10ML</td>
			<td></td>
		</tr>
		<tr>
			<td>Succinic anhydride</td>
			<td>Traditional Chinese medicine</td>
			<td>30171826</td>
			<td></td>
		</tr>
		<tr>
			<td>Tetrahydrofuran</td>
			<td>Traditional Chinese medicine</td>
			<td>40058161</td>
			<td></td>
		</tr>
		<tr>
			<td>Triethylamine</td>
			<td>Traditional Chinese medicine</td>
			<td>80134318</td>
			<td></td>
		</tr>
		<tr>
			<td>UV/VIS/NIR Spectrometer</td>
			<td>Lambda950</td>
			<td>Lambda950</td>
			<td></td>
		</tr>
		<tr>
			<td>X-ray Photoelectron Spectrometer</td>
			<td>Thermo&#8194;Fisher&#8194;Scientific</td>
			<td>K-ALPHA 0.5EV</td>
			<td></td>
		</tr>
	</tbody>
</table>

synthesis of aptamer-pei-g-peg modified gold nanoparticles loaded with doxorubicin for targeted drug delivery

På grund af resistens over for lægemidler og toksicitet i raske celler, brug af doxorubicin (DOX) har været begrænset i klinisk kræftbehandling. Denne protokol beskriver udformningen af poly (ethylenimine) podet med polyethylen glycol (PEI-g-PEG) copolymer funktionaliserede guld nanopartikler (AuNPs) med lastet aptamer (AS1411) og DOX gennem amide reaktioner. AS1411 er specifikt bundet med målrettede nukleolinreceptorer på kræftceller, så DOX målretter kræftceller i stedet for raske celler. For det første podes PEG til forgrenet PEI for at opnå en PEI-g-PEG-copolymer, hvilket bekræftes af 1H NMR-analyse. Dernæst syntetiseres PEI-g-PEG copolymer coated gold nanopartikler (PEI-g-PEG@AuNPs), og DOX og AS1411 er kovalent bundet til AuNPs gradvist via midt reaktioner. Diameteren af den forberedte AS1411-g-DOX-g-PEI-g-PEG@AuNPs er ~39,9 nm, med et zetapotentiale på -29,3 mV, hvilket indikerer, at nanopartiklerne er stabile i vand- og cellemedie. Celle cytotoksicitet assays viser, at den nydesignede DOX indlæst AuNPs er i stand til at dræbe kræftceller (A549). Denne syntese viser den delikate arrangement af PEI-g-PEG copolymers, aptamers, og DOX på AuNPs, der opnås ved sekventielle midt reaktioner. Sådanne aptamer-PEI-g-PEG funktionaliserede aunps giver en lovende platform for målrettet lægemiddellevering i kræftbehandling.

1 1 1 1 H NMR-spektroskopi blev anvendt til at bekræfte den vellykkede syntese af CT-PEG polymer og PEI-g-PEG copolymerer (figur 2). Figur 2a viser, at methylenprotonsignalet ved δ = 3,61 ppm og carboxylprotonsignalet ved δ = 2,57 ppm bekræfter den vellykkede syntese af CT-PEG polymerer. Figur 2b viser, at methylenprotonsignalet for PEG ved δ = 2,6 ppm og protonsignalet for PEI ved δ</...

Watch this Scientific Journal Video about Synthesis of Aptamer-PEI-g-PEG Modified Gold Nanoparticles Loaded with Doxorubicin for Targeted Drug Delivery at JoVE.com

Synthesis of Aptamer-PEI-g-PEG Modified Gold Nanoparticles Loaded with Doxorubicin for Targeted Drug Delivery

I denne protokol syntetiseres doxorubicin-loaded AS1411-g-PEI-g-PEG modificerede guld nanopartikler via tre-trins midt reaktioner. Derefter er doxorubicin lastet og leveret til at målrette kræftceller til kræftbehandling.

Syntese af Aptamer-PEI-g-PEG Modificeret Guld Nanopartikler Fyldt med Doxorubicin for målrettet drug delivery

Due to drug resistance and toxicity in healthy cells, use of doxorubicin (DOX) has been limited in clinical cancer therapy. This protocol describes the ...

Research

JoVE Journal

Chemistry

Template Directed Synthesis of Plasmonic Gold Nanotubes with Tunable IR Absorbance

Skabelon Instrueret Syntese af plasmoniske Guld Nanorør med Justerbar IR Absorbans

A nearly parallel array of pores can be produced by anodizing aluminum foils in acidic environments1, 2. Applications of anodic aluminum oxide (AAO) ...

Preparation and Characterization of SDF-1&#945;-Chitosan-Dextran Sulfate Nanoparticles

Preparation and Characterization of SDF-1ÃƒÅ½Ã‚Â±-Chitosan-Dextran Sulfate Nanoparticles

Fremstilling og karakterisering af SDF-1α-Chitosan-dextransulfat Nanopartikler

Chitosan (CS) and dextran sulfate (DS) are charged polysaccharides (glycans), which form polyelectrolyte complex-based nanoparticles when mixed under ...

A Simple Method for the Size Controlled Synthesis of Stable Oligomeric Clusters of Gold Nanoparticles under Ambient Conditions

En simpel metode til størrelse Kontrolleret Syntese af stabile Oligomere Klynger af guld nanopartikler under omgivende betingelser

Reducing dilute aqueous HAuCl4 with sodium thiocyanate (NaSCN) under alkaline conditions produces 2 to 3 nm diameter nanoparticles. Stable grape-like ...

Protocol for the Synthesis of Ortho-trifluoromethoxylated Aniline Derivatives

Protocol for the Synthesis of Ortho-trifluoromethoxylated Aniline Derivatives

Protokol til syntese af<em&gt; Ortho</em&gt; -trifluoromethoxylated anilinderivater

Molecules bearing trifluoromethoxy (OCF3) group often show desired pharmacological and biological properties. However, facile synthesis of ...

Hydroquinone Based Synthesis of Gold Nanorods

Hydroquinon Based Syntese af guld nanorods

Gold nanorods are an important kind of nanoparticles characterized by peculiar plasmonic properties. Despite their widespread use in nanotechnology, the ...

Chemical Precipitation Method for the Synthesis of Nb2O5 Modified Bulk Nickel Catalysts with High Specific Surface Area

Chemical Precipitation Method for the Synthesis of Nb2O5 Modified Bulk Nickel Catalysts with High Specific Surface Area

Kemisk udfældning metode til syntese af Nb2O5 ændret Bulk nikkel katalysatorer med høje specifikke overfladeareal

We demonstrate a method for the synthesis of NixNb1-xO catalysts with sponge-like and fold-like nanostructures. By varying the Nb:Ni ratio, a series of ...

Rapid Nanoprobe Signal Enhancement by In Situ Gold Nanoparticle Synthesis

Rapid Nanoprobe Signal Enhancement by In Situ Gold Nanoparticle Synthesis

Hurtig Nanoprobe Signal forstærkning af In Situ guld nanopartikel syntese

The use of nanoprobes such as gold, silver, silica or iron-oxide nanoparticles as detection reagents in bioanalytical assays can enable high sensitivity ...

Synthesis and Characterization of 1,2-Dithiolane Modified Self-Assembling Peptides

Syntese og karakterisering af 1,2-Dithiolane ændret selvsamlende peptider

This report focuses on the synthesis of an N-terminus 1,2-dithiolane modified self-assembling peptide and the characterization of the resulting ...

Molten-Salt Synthesis of Complex Metal Oxide Nanoparticles

Smeltet Salt syntese af komplekse metaloxid nanopartikler

The development of feasible synthesis methods is critical for the successful exploration of novel properties and potential applications of nanomaterials. ...

Methionine Functionalized Biocompatible Block Copolymers for Targeted Plasmid DNA Delivery

Methionin Funktionaliserede biokompatible blok copolymere til målrettet plasmid DNA levering

Reversible addition-fragmentation chain transfer (RAFT) polymerization integrates the advantages of radical polymerization and living polymerization. This ...