Questa ricerca è stata finanziata dalla National Natural Science Foundation of China (31700840); il Progetto chiave di ricerca scientifica della provincia di Henan (18B430013, 18A150049). Questa ricerca è stata supportata dal Nanhu Scholars Program for Young Scholars of XYNU. Gli autori vorrebbero ringraziare lo studente di laurea Zebo Qu del College of Life Sciences di XYNU per le sue utili opere. Gli autori vorrebbero riconoscere il Centro analisi & test di XYNU per l'uso delle loro apparecchiature.

ATTENZIONE: Assicurarsi di consultare tutte le schede di dati di sicurezza dei materiali pertinenti (MSDS) prima di utilizzare tutte le sostanze chimiche. Molte delle sostanze chimiche utilizzate per preparare copolimeri e nanoparticelle sono acutamente tossiche. Anche le nanoparticelle hanno potenziali pericoli. Assicurati di utilizzare tutte le pratiche di sicurezza appropriate e i dispositivi di protezione individuale, tra cui guanti, camice da laboratorio, cappucci, pantaloni a figura intera e scarpe a punta stretta.
1. Sintesi del polietilene glicole polietilenilene doppio-carbossile (CT-PEG)33
<ol><li>Aggiungere 1,46 g (14,6 mmol) di anidride succinica (SA) e 209 mg (1,71 mmol) di 4-dimetilaminopiridina (DMAP) in un pallone inferiore rotondo da 100 ml.</li>
	<li>Aggiungere 15 ml di tetraidrofurano anidrofurano (THF) al pallone utilizzato nel passaggio 1.1 e montare un tappo di vetro. Mantenere il pallone a 0 °C per 30 minuti.</li>
	<li>Aggiungere 4,28 g (4,28 mmol) di polietilene glicole (PEG) e 1,8 ml (12,8 mmol) di trietilammina (TEA) in un nuovo pallone.</li>
	<li>Aggiungere 15 ml di THF anidro al pallone utilizzato nel passaggio 1.3 e montare un tappo di vetro. Trasferire lentamente la soluzione nel pallone utilizzato nel passaggio 1.2, utilizzando una siringa in atmosfera di azoto.</li>
	<li>Mescolare la soluzione a 0 °C per 2 ore, quindi continuare la reazione a temperatura ambiente (RT) durante la notte.</li>
	<li>Utilizzando un evaporatore rotante (40 °C, 0,1 MPa), concentrare la soluzione di reazione e rimuovere il solvente THF.</li>
	<li>A RT sciogliere la soluzione di reazione dal passaggio 1.6 in 15 mL di 1.325 g/mL diclorometano (DCM), quindi aggiungere 15 mL di etere dietile freddo (Et2O) per ottenere il prodotto di precipitazione (polietilene glicole diacid). Rimuovere il solvente tramite carta da filtro. NOTA: La fase di precipitazione può essere ripetuta 3x.</li>
	<li>Asciugare i precipitati sotto vuoto a RT per 48 ore.</li>
</ol>2. Sintesi del copolimero PEI-g-PEG
<ol><li>Aggiungere 305,47 mg di CT-PEG dai gradini 1.8 e 5 mL di solfossido di dimetile (DMSO) a un pallone e mescolare a RT per assicurarsi che CT-PEG sia completamente sciolto in DMSO.</li>
	<li>Sciogliere 49,46 mg di cloridrato di 1-(3-dimetilaminopropil)-3-etilcarbodiimide (EDC) in 5 ml di DMSO, quindi aggiungere la soluzione al pallone utilizzato nel passaggio 2.1 e mescolare per 30 minuti a RT.</li>
	<li>Sciogliere 29,69 mg di N-idrossisuccinimide (NHS) in 5 ml di DMSO e aggiungere la soluzione al pallone utilizzato nel passaggio 2.1. Continuare a mescolare a RT per 3 h.</li>
	<li>Sciogliere 28,6 μL di polietileneimina (PEI) in 10 ml di DMSO e aggiungere la soluzione per goccia al pallone utilizzato nel passaggio 2.1. Mescolare per almeno 3 giorni.</li>
	<li>Trasferire la soluzione reagì dal passaggio 2.4 a un sacchetto di dialisi (1.000 tagli di peso molecolare [MWCO]). Posizionare il sacchetto di dialisi in un becher da 1 L con 500 mL di acqua ultrapura come dialisi. Cambiare l'acqua ultrapura ogni 12 ore per 3 giorni.</li>
	<li>Trasferire la soluzione nel passaggio 2.5 in un altro sacchetto di dialisi (10.000 MWCO). Posizionare il sacchetto di dialisi in un becher da 1 L con 500 mL di acqua ultrapura come dialisi. Cambiare l'acqua ultrapura ogni 12 ore per 3 giorni.</li>
	<li>Concentrare la soluzione dal passaggio 2.6 utilizzando un evaporatore rotante (40 °C, 0,1 MPa) e congelare il campione per ottenere la polvere PEI-g-PEG.</li>
</ol>3. Sintesi di PEI-g-PEG@AuNPs
<ol><li>Sciogliere 5 mg di PEI-g-PEG preparato (fase 2.7) in 5 ml di acqua ultrapura in un nuovo pallone e montare con un tappo di vetro.</li>
	<li>Aggiungere 100 mL di soluzione HAuCl4 da 0,3 mM al pallone e mescolare la soluzione per 3 ore a RT. NOTA: Il colore della soluzione deve cambiare immediatamente dal giallo all'arancione.</li>
	<li>Aggiungere 1 ml di soluzione NaBH4 mg/mL al pallone e mescolare la soluzione per 3 ore a RT. NOTA: La soluzione di reazione dovrebbe girare istantaneamente bordeaux.</li>
	<li>Dializzare il prodotto di reazione utilizzando un sacchetto di dialisi (1.000 MWCO) per 3 giorni come descritto al passaggio 2.5 per ottenere la soluzione PEI-g-PEG@AuNPs.</li>
</ol>4. Sintesi di DOX-g-PEI-g-PEG@AuNPs
<ol><li>Aggiungere 1 ml di soluzione DOX da 2,2 mg/mL e 20 ml di soluzione PEI-g-PEG@AuNPs in un nuovo pallone e montare con un tappo di vetro.</li>
	<li>Sciogliere 0,727 mg di EDC in 1 ml di acqua ultrapura e aggiungere la soluzione EDC al pallone utilizzato nel passaggio 4.1.</li>
	<li>Sciogliere 0,437 mg di NHS in 1 mL di acqua ultrapura. Aggiungere la soluzione NHS al pallone e mescolare a RT per 1 h.</li>
	<li>Dializzare il prodotto di reazione utilizzando un sacchetto di dialisi (1.000 MWCO) per 3 giorni come descritto al passaggio 2.5 per ottenere la soluzione DOX-g-PEI-g-PEG@AuNPs.</li>
</ol>5. Sintesi di AS1411-g-DOX-g-PEI-g-PEG@AuNPs
<ol><li>Aggiungere 20 mL di soluzione DOX-g-PEI-g-PEG@AuNPs e 4OD di AS1411 (OD = densità ottica; 1OD ≈ 33 μg) a un nuovo pallone.</li>
	<li>Sciogliere 28,76 mg di EDC in 1 ml di acqua ultrapura e aggiungere la soluzione EDC al pallone utilizzato nel passaggio 5.1.</li>
	<li>Sciogliere 17,27 mg di NHS in 1 mL di acqua ultrapura. Aggiungere la soluzione NHS al pallone utilizzato nel passaggio 5.1 e mescolare la reazione per 1 h a RT.</li>
	<li>Dializzare il prodotto di reazione utilizzando un sacchetto di dialisi (1.000 MWCO) per 3 giorni come descritto al passaggio 2.5 per ottenere AS1411-g-DOX-g-PEI-g-PEG@AuNPs.</li>
</ol>6. Caratterizzazione del campione
<ol><li>Sciogliere il polimero CT-PEG (fase 1.8) e il copolimero PEI-g-PEG (fase 2.7) rispettivamente in cloroformio-d nei tubi di risonanza magnetica nucleare (NMR). Analizzare i campioni utilizzando uno spettrometro NMR a 600 MHz dotato di un magnete superconduttore da 14,09 T e di una sonda ad alta risoluzione a banda larga a banda larga 5,0 mm 600 MHz per confermare la strutturachimica 34.</li>
	<li>Disperdere AuNPs, DOX e AS1411 e preparare PEI-g-PEG@AuNPs (passaggio 3.4), DOX-g-PEI-g-PEG@AuNPs (passaggio 4.4) e AS1411-g-DOX-g-PEI-g-PEG@AuNPs (passaggio 5.4), rispettivamente in acqua ultrapura. Quindi, trasferire alle cuvette e registrare gli spettri visibili agli ultravioletti (UV-vis) utilizzando uno spettrofotometro UV-vis.</li>
	<li>Attaccare un adesivo a doppia faccia (~2 mm x 2 mm) alla lamina di alluminio e immergere la soluzione del campione (PEI-g-PEG@AuNPs, DOX-g-PEI-g-PEG@AuNPs e AS1411-g-DOX-g-PEI-g-PEG@AuNPs) sull'intero nastro in modo uniforme. Analizzare i campioni utilizzando un analizzatore di spettroscopia fotoelettronica a raggi X.</li>
	<li>Disperdere le soluzioni PEI-g-PEG@AuNPs, DOX-g-PEI-g-PEG@AuNPs e AS1411-g-DOX-g-PEI-g-PEG@AuNPs, rispettivamente in acqua ultrapura. Quindi, trasferire alle cuvette e valutare la distribuzione delle dimensioni utilizzando lo scattering dinamico della luce.</li>
	<li>Disperdere le soluzioni PEI-g-PEG@AuNPs, DOX-g-PEI-g-PEG@AuNPs e AS1411-g-DOX-g-PEI-g-PEG@AuNPs in acqua ultrapura (una goccia di campione per 5 mL di acqua ultrapura per ogni campione). Sonicare per 2 h. Immergere la griglia di rame in soluzioni campione e asciugare sotto una lampada a infrarossi. Caratterizzare la morfologia usando un microscopio elettronico a trasmissione.</li>
	<li>Iniettare 1 mg di AS1411-g-DOX-g-PEI-g-PEG@AuNPs in una cassetta di dialisi MWCO da 20 kDa, quindi mettere in 80 mL di saline tamponate di fosfato (PBS) con albumina di siero bovino al 5% (BSA). Mescolare a 37 °C.</li>
	<li>Nei punti di tempo predeterminati, raccogliere aliquote 100 μL e sostituire con PBS fresco. Utilizzare uno spettrofotometro UV-vis per misurare l'intensità di fluorescenza DOX delle aliquote.</li>
</ol>7. Test CCK-8 delle nanoparticelle di AS1411-g-DOX-g-PEI-g-PEG@AuNPs
<ol><li>Coltivare cellule A549 nel mezzo dell'Aquila modificato (DMEM) di Dulbecco integrato con il 10% di siero bovino fetale, 100 U/mL di penicillina e 100 μg/mL di streptomicina in un'atmosfera umidificata del 95% di aria e del 5% di CO2 a 37 °C. Sostituire il mezzo di coltura ogni 2 giorni. Utilizzare cellule al passaggio 5 per la proliferazione cellulare e test di citotossicità per valutare quantitativamente la citotossicità delle nanoparticelle AS1411-g-DOX-g-PEI-g-PEG@AuNPs preparate.</li>
	<li>Aggiungere 100 μL di soluzione di nanoparticelle in ogni pozzo contenente 1 mL di mezzo cellulare. Dopo aver coltivato per 24 ore e 48 ore, rimuovere i mezzi di coltura dalle piastre di coltura cellulare, quindi aggiungere immediatamente 300 μL di mezzi di coltura freschi e 30 μL di soluzioni di kit di conteggio cellulare-8 (CCK-8) immediatamente ad ogni pozzo. Incubare per 4 ore in un incubatore di CO2 a 37 °C.</li>
	<li>Trasferire 200 μL di soluzioni di reazione dal passo 7.2 in una piastra di pozzo 96. Leggere la densità ottica (OD) di ogni pozzo a 570 nm con un lettore di micropiatte.</li>
	<li>Osservare la morfologia delle cellule a 24 ore e 48 ore al microscopio.</li>
</ol>

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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>

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Lo spettro 1H NMR (Figura 2) conferma la sintesi riuscita del copolimero CT-PEG e del copolimero PEI-g-PEG. I pesi molecolari di PEG e PEI erano rispettivamente 1.000 e 1.200. Inoltre, il sistema catalitico EDC/NHS è stato utilizzato per sintetizzare il copolimero PEI-g-PEG attraverso reazioni di ammide. Va notato che se i pesi molecolari di PEG e PEI sono cambiati per sintetizzare il copolimero PEI-g-PEG, allora il tempo di reazione e il sistema catalitico devono essere rivaluta...

Essendo il principale problema di salute pubblica in tutto il mondo, il cancro è ampiamente caratterizzato come avente un basso tasso di cura, un alto tasso di recidiva e un alto tassodi mortalità 1,2. Gli attuali metodi anti-cancro convenzionali includono chirurgia, chemioterapia e radioterapia3, tra cui la chemioterapia è il trattamento primario per i pazienti oncologici nella clinica4. I farmaci antitumorali usati clinicamente includono principalmente paclitaxel (PTX)5 e doxorubicina (DOX)6,7. Dox, un farmaco antineoplastico, è stato ampiamente applicato nella chemioterapia clinica, a causa dei vantaggi della citotossicità del cancro e dell'inibizione della proliferazione delle celluletumorali 8,9. Tuttavia, DOX causa cardiotossicità 10,11e la breve emiprotezione di DOX ne limita l'applicazione nella clinica12. Pertanto, i portatori di farmaci degradabili sono necessari per caricare DOX e rilasciare in modo sottoequente in modo controllato in un'area mirata.
Le nanoparticelle sono state ampiamente utilizzate in sistemi mirati di somministrazione di farmaci e hanno diversi vantaggi nel trattamento del cancro (ad esempio, rapporto superficie/volume bilmente ridotto, piccole dimensioni, capacità di incapsulare vari farmaci e chimica delle superfici tonnibili, ecc.) 13,14,15. In particolare, le nanoparticelle d'oro (AuNPs) sono state ampiamente utilizzate in applicazioni biologiche e biomediche, come la terapia del cancro fototermico16,17. Le proprietà uniche degli AuNP, come la facile sintesi e la funzionalizzazione generale della superficie, hanno ottime prospettive nel campo clinico della terapia del cancro18. Inoltre, gli AuNP sono stati utilizzati per identificare le strategie di somministrazione dei farmaci, diagnosticare i tumori e superare la resistenza inmolti studi 19,20.
Nonostante ciò, gli AuRP devono essere ulteriormente personalizzati per superare la resistenza ai farmaci attraverso un elevato rilascio locale di lesioni tumorali attraverso una maggiore permeazione e ritenzione (EPR), come le proprietà di targeting e accessibilità. Gli AuRP funzionalizzati polimerici hanno mostrato vantaggi unici, come una migliore solubilità in acqua dei farmaci anticancro idrofobici e un tempo di circolazioneprolungato 21,22. Vari polimeri biocompatibili sono stati utilizzati per rivestimenti AuNP, come polietilene glicole (PEG), polietileneimina (PEI), acido ialuronico, eparina e gomma di xantano. Quindi la stabilità, così come il carico utile, degli AuNP è migliorata ben23. In particolare, pei è un polimero altamente ramificato composto da molte unità ripetute di ammine primarie, secondarie e terziarie24. PEI ha un'eccellente solubilità, bassa viscosità e un alto grado di funzionalità, adatto per il rivestimento su AuNPs.
D'altra parte, i farmaci anti-cancro devono essere consegnati alle cellule tumorali direttamente con una migliore efficienza di carico e con una minore tossicità per il trattamento dei tumori metastatici primari eavanzati 25. I ligandi mirati hanno un grande potenziale per i sistemi di somministrazione mirati ai farmaci anti-cancro26. La sua selettività per il legame con molecole bersaglio conferisce un farmaco anti-cancro mirato alla specificità e aumenta l'arricchimento dei farmaci nei tessutimalati 27. Altri ligandi includono anticorpi, polipeptidi e piccole molecole. Rispetto ad altri ligandi, gli aptameri di acido nucleico possono essere sintetizzati in vitro e sono facili da modificare. AS1411 è un oligonucleotide fosfodiestere da 26 bp non modificato che forma una struttura G-tetramero dimerica stabile per legarsi specificamente a un recettore delle proteine nucleari bersaglio sovraespressosulle cellule tumorali 28,29,30. AS1411 inibisce la proliferazione di molte cellule tumorali ma non influisce sulla crescita delle cellulesane 31,32. Di conseguenza, AS1411 è stato utilizzato per fabbricare un sistema di somministrazione di farmaci mirato ideale.
In questo studio, un copolimero PEI-g-PEG viene sintetizzato attraverso una reazione ammide, quindi vengono fabbricate nanoparticelle d'oro rivestite di copolimero PEI-g-PEG (PEI-g-PEG@AuNPs). Inoltre, DOX e AS1411 sono collegati in sequenza al pei-g-PEG@AuNPs preparato, come illustrato nella figura 1. Questo protocollo dettagliato ha lo scopo di aiutare i ricercatori a evitare molte delle insidie comuni associate alla fabbricazione di nuovi PEI-g-PEG@AuNPs caricati con DOX e 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

A causa della resistenza ai farmaci e della tossicità nelle cellule sane, l'uso di doxorubicina (DOX) è stato limitato nella terapia clinica del cancro. Questo protocollo descrive la progettazione di poli(etilenimina) innestate con polietilene glicole (PEI-g-PEG) copolimero funzionalizzato nanoparticelle d'oro (AuNPs) con aptamero caricato (AS1411) e DOX attraverso reazioni ammide. AS1411 è specificamente legato con recettori mirati della nucleolina sulle cellule tumorali in modo che il DOX si ritrae contro le cellule tumorali anziché le cellule sane. In primo luogo, PEG viene carbossilato, quindi innestato in PEI ramificata per ottenere un copolimero PEI-g-PEG, che è confermato dall'analisi NMR 1H. Successivamente, vengono sintetizzati nanoparticelle d'oro rivestite di copolimero PEI-g-PEG@AuNPs) e DOX e AS1411 sono covalentemente legati agli AuNP gradualmente attraverso reazioni ammide. Il diametro dell'AS1411-g-DOX-g-PEI-g-PEG@AuNPs preparato è di ~39,9 nm, con un potenziale zeta di -29,3 mV, indicando che le nanoparticelle sono stabili in acqua e mezzo cellulare. I test di citotossicità cellulare mostrano che gli AuNP caricati da DOX di nuova progettatezza sono in grado di uccidere le cellule tumorali (A549). Questa sintesi dimostra la delicata disposizione di copolimeri, aptameri e DOX PEI-g-PEG sugli AuRP che si ottengono con reazioni sequenziali di ammide. Tali ADUL funzionalizzati aptamer-PEI-g-PEG forniscono una piattaforma promettente per la somministrazione mirata di farmaci in terapia oncologica.

1 ) La commissione per la La spettroscopia H NMR è stata utilizzata per confermare la sintesi riuscita del polimero CT-PEG e dei copolimeri PEI-g-PEG (Figura 2). La figura 2a mostra che il segnale protonico di metilene a δ = 3,61 ppm e il segnale protonico carbossile a δ = 2,57 ppm confermano la sintesi riuscita dei polimeri CT-PEG. La figura 2b mostra che il segnale protonico di metilene di PEG a...

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

In questo protocollo, le nanoparticelle d'oro modificate AS1411-g-PEI-g-PEG caricate con doxorubicina vengono sintetizzati tramite reazioni di ammide in tre step. Quindi, la doxorubicina viene caricata e consegnata alle cellule tumorali bersaglio per la terapia del cancro.

Sintesi di Nanoparticelle d'oro modificate Aptamer-PEI-g-PEG Caricate con doxorubicina per la consegna mirata di farmaci

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

synthesis-aptamer-pei-g-peg-modified-gold-nanoparticles-loaded-with

Research

JoVE Journal

Chemistry

5.8K Views.  Xinyang Normal University. In questo protocollo, le nanoparticelle d'oro modificate AS1411-g-PEI-g-PEG caricate con doxorubicina vengono sintetizzati tramite reazioni di ammide in tre step. Quindi, la doxorubicina viene caricata e consegnata alle cellule tumorali bersaglio per la terapia del cancro.

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

Template Directed Synthesis of Plasmonic Gold Nanotubes with Tunable IR Absorbance

A nearly parallel array of pores can be produced by anodizing aluminum foils in acidic environments1, 2. Applications of anodic aluminum oxide (AAO) membranes have been under development since the 1990's and have become a common method to template the synthesis of high aspect ratio nanostructures, mostly by electrochemical growth or pore-wetting. Recently, these membranes have become commercially available in a wide range of pore sizes and densities, leading to an extensive library of functional nanostructures being synthesized from AAO membranes. These include composite nanorods, nanowires and nanotubes made of metals, inorganic materials or polymers 3-10. Nanoporous membranes have been used to synthesize nanoparticle and nanotube arrays that perform well as refractive index sensors, plasmonic biosensors, or surface enhanced Raman spectroscopy (SERS) substrates 11-16, as well as a wide range of other fields such as photo-thermal heating 17, permselective transport 18, 19, catalysis 20, microfluidics 21, and electrochemical sensing 22, 23. Here, we report a novel procedure to prepare gold nanotubes in AAO membranes. Hollow nanostructures have potential application in plasmonic and SERS sensing, and we anticipate these gold nanotubes will allow for high sensitivity and strong plasmon signals, arising from decreased material dampening 15.

Solution-suspendable gold nanotubes with controlled dimensions can be synthesized by electrochemical deposition in porous anodic aluminum oxide (AAO) membranes using a hydrophobic polymer core. Gold nanotubes and nanotube arrays hold promise for applications in plasmonic biosensing, surface-enhanced Raman spectroscopy, photo-thermal heating, ionic and molecular transport, microfluidics, catalysis and electrochemical sensing.

Modello Regia Sintesi di nanotubi d&#39;oro plasmoniche con Tunable Assorbanza IR

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

Ricerca

Chimica

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

Chitosan (CS) and dextran sulfate (DS) are charged polysaccharides (glycans), which form polyelectrolyte complex-based nanoparticles when mixed under appropriate conditions. The glycan nanoparticles are useful carriers for protein factors, which facilitate the in vivo delivery of the proteins and sustain their retention in the targeted tissue. The glycan polyelectrolyte complexes are also ideal for protein delivery, as the incorporation is carried out in aqueous solution, which reduces the likelihood of inactivation of the proteins. Proteins with a heparin-binding site adhere to dextran sulfate readily, and are, in turn, stabilized by the binding. These particles are also less inflammatory and toxic when delivered in vivo. In the protocol described below, SDF-1&#945; (Stromal cell-derived factor-1&#945;), a stem cell homing factor, is first mixed and incubated with dextran sulfate. Chitosan is added to the mixture to form polyelectrolyte complexes, followed by zinc sulfate to stabilize the complexes with zinc bridges. The resultant SDF-1&#945;-DS-CS particles are measured for size (diameter) and surface charge (zeta potential). The amount of the incorporated SDF-1&#945; is determined, followed by measurements of its in vitro release rate and its chemotactic activity in a particle-bound form.

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

The objective of this protocol is to incorporate SDF-1&#945;, a stem cell homing factor, into dextran sulfate-chitosan nanoparticles. The resultant particles are measured for their size and zeta potential, as well as the content, activity, and in vitro release rate of SDF-1&#945; from the nanoparticles.

Preparazione e caratterizzazione di SDF-1α-Chitosan-Destrano solfato nanoparticelle

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

Reducing dilute aqueous HAuCl4 with sodium thiocyanate (NaSCN) under alkaline conditions produces 2 to 3 nm diameter nanoparticles. Stable grape-like oligomeric clusters of these yellow nanoparticles of narrow size distribution are synthesized under ambient conditions via two methods. The delay-time method controls the number of subunits in the oligoclusters by varying the time between the addition of HAuCl4 to alkaline solution and the subsequent addition of reducing agent, NaSCN. The yellow oligoclusters produced range in size from ~3 to ~25 nm. This size range can be further extended by an add-on method utilizing hydroxylated gold chloride (Na+[Au(OH4-x)Clx]-) to auto-catalytically increase the number of subunits in the as-synthesized oligocluster nanoparticles, providing a total range of 3 nm to 70 nm. The crude oligocluster preparations display narrow size distributions and do not require further fractionation for most purposes. The oligoclusters formed can be concentrated &#62;300 fold without aggregation and the crude reaction mixtures remain stable for weeks without further processing. Because these oligomeric clusters can be concentrated before derivatization they allow expensive derivatizing agents to be used economically. In addition, we present two models by which predictions of particle size can be made with great accuracy.

We describe a simple method for producing highly stable oligomeric clusters of gold nanoparticles via the reduction of chloroauric acid (HAuCl4) with sodium thiocyanate (NaSCN). The oligoclusters have a narrow size distribution and can be produced with a wide range of sizes and surface coats.

Un metodo semplice per la dimensione sintesi controllata di stabili oligomeriche Grappoli di oro nanoparticelle in condizioni ambientali

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

Hydroquinone Based Synthesis of Gold Nanorods

Gold nanorods are an important kind of nanoparticles characterized by peculiar plasmonic properties. Despite their widespread use in nanotechnology, the synthetic methods for the preparation of gold nanorods are still not fully optimized. In this paper we describe a new, highly efficient, two-step protocol based on the use of hydroquinone as a mild reducing agent. Our approach allows the preparation of nanorods with a good control of size and aspect ratio (AR) simply by varying the amount of hexadecyl trimethylammonium bromide (CTAB) and silver ions (Ag+) present in the "growth solution". By using this method, it is possible to markedly reduce the amount of CTAB, an expensive and cytotoxic reagent, necessary to obtain the elongated shape. Gold nanorods with an aspect ratio of about 3 can be obtained in the presence of just 50 mM of CTAB (versus 100 mM used in the standard protocol based on the use of ascorbic acid), while shorter gold nanorods are obtained using a concentration as low as 10 mM.

This paper describes a protocol for the synthesis of gold nanorods, based on the use of hydroquinone as reducing agent, plus the different mechanisms for controlling their size and aspect ratio.

Idrochinone Based Sintesi di oro 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

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 NixNb1-xO nanoparticles with different atomic compositions (x = 0.03, 0.08, 0.15, and 0.20) have been prepared by chemical precipitation. These NixNb1-xO catalysts are characterized by X-ray diffraction, X-ray photoelectron spectroscopy, and scanning electron microscopy. The study revealed the sponge-like and fold-like appearance of Ni0.97Nb0.03O and Ni0.92Nb0.08O on the NiO surface, and the larger surface area of these NixNb1-xO catalysts, compared with the bulk NiO. Maximum surface area of 173 m2/g can be obtained for Ni0.92Nb0.08O catalysts. In addition, the catalytic hydroconversion of lignin-derived compounds using the synthesized Ni0.92Nb0.08O catalysts have been investigated.

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

A protocol for the synthesis of sponge-like and fold-like Ni1-xNbxO nanoparticles by chemical precipitation is presented.

Modificare il metodo di precipitazione chimica per la sintesi di Nb2O5 catalizzatori di nichel Bulk con elevata superficie specifica

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

The use of nanoprobes such as gold, silver, silica or iron-oxide nanoparticles as detection reagents in bioanalytical assays can enable high sensitivity and convenient colorimetric readout. However, high densities of nanoparticles are typically needed for detection. The available synthesis-based enhancement protocols are either limited to gold and silver nanoparticles or rely on precise enzymatic control and optimization. Here, we present a protocol to enhance the colorimetric readout of gold, silver, silica, and iron oxide nanoprobes. It was observed that the colorimetric signal can be improved by up to a 10000-fold factor. The basis for such signal enhancement strategies is the chemical reduction of Au3+ to Au0. There are several chemical reactions that enable the reduction of Au3+ to Au0. In the protocol, Good&#39;s buffers and H2O2 are used and it is possible to favor the deposition of Au0 onto the surface of existing nanoprobes, in detriment of the formation of new gold nanoparticles. The protocol consists of the incubation of the microarray with a solution consisting of chloroauric acid and H2O2 in 2-(N-morpholino)ethanesulfonic acid pH 6 buffer following the nanoprobe-based detection assay. The enhancement solution can be applied to paper and glass-based sensors. Moreover, it can be used in commercially available immunoassays as demonstrated by the application of the method to a commercial allergen microarray. The signal development requires less than 5 min of incubation with the enhancement solution and the readout can be assessed by naked eye or low-end image acquisition devices such as a table-top scanner or a digital camera.

Rapid Nanoprobe Signal Enhancement by In Situ Gold Nanoparticle Synthesis

In this work, a protocol for signal enhancement of nanoprobe-based biosensing is presented. The protocol is based on the reduction of chloroauric acid onto the surface of existing nanoprobes that consist of gold, silver, silica or iron-oxide nanoparticles.

Segnale Nanosonda rapido aumento di sintesi di nanoparticelle di oro In Situ

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

Methionine Functionalized Biocompatible Block Copolymers for Targeted Plasmid DNA Delivery

Reversible addition-fragmentation chain transfer (RAFT) polymerization integrates the advantages of radical polymerization and living polymerization. This work presents the preparation of methionine functionalized biocompatible block copolymers via RAFT polymerization. Firstly, N,N-bis(2-hydroxyethyl)methacrylamide-b-N-(3-aminopropyl)methacrylamide (BNHEMA-b-APMA, BA) was synthesized via RAFT polymerization using 4,4&#8217;-azobis(4-cyanovaleric acid) (ACVA) as an initiating agent and 4-cyanopentanoic acid dithiobenzoate (CTP) as the chain transfer agent. Subsequently, N,N-bis(2-hydroxyethyl)methacrylamide-b-N-(3-guanidinopropyl)methacrylamide (methionine grafted BNHEMA-b-GPMA, mBG) was prepared by modifying amine groups in APMA with methionine and guanidine groups. Three kinds of block polymers, mBG1, mBG2, and mBG3, were synthesized for comparison. A ninhydrin reaction was used to quantify the APMA content; mBG1, mBG2, and mBG3 had 21%, 37%, and 52% of APMA, respectively. Gel permeation chromatography (GPC) results showed that BA copolymers possess molecular weights of 16,200 (BA1), 20,900(BA2), and 27,200(BA3) g/mol. The plasmid DNA (pDNA) complexing ability of the obtained block copolymer gene carriers was also investigated. The charge ratios (N/P) were 8, 16, and 4 when pDNA was complexed completely with mBG1, mBG2, mBG3, respectively. When the N/P ratio of mBG/pDNA polyplexes was higher than 1, the Zeta potential of mBG was positive. At an N/P ratio between 16 and 32, the average particle size of mBG/pDNA polyplexes was between 100-200 nm. Overall, this work illustrates a simple and convenient protocol for the block copolymer carrier synthesis.

This work presents the preparation of methionine functionalized biocompatible block copolymers (mBG) via the reversible addition-fragmentation chain transfer (RAFT) method. The plasmid DNA complexing ability of the obtained mBG and their transfection efficiency were also investigated. The RAFT method is very beneficial for polymerizing monomers containing special functional groups.

Meticolosina Sistemi colatori di blocchi biocompatibili per la consegna mirata del DNA plasmide

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

Synthesis and Characterization of Amphiphilic Gold Nanoparticles

Gold nanoparticles covered with a mixture of 1-octanethiol (OT) and 11-mercapto-1-undecane sulfonic acid (MUS) have been extensively studied because of their interactions with cell membranes, lipid bilayers, and viruses. The hydrophilic ligands make these particles colloidally stable in aqueous solutions and the combination with hydrophobic ligands creates an amphiphilic particle that can be loaded with hydrophobic drugs, fuse with the lipid membranes, and resist nonspecific protein adsorption. Many of these properties depend on nanoparticle size and the composition of the ligand shell. It is, therefore, crucial to have a reproducible synthetic method and reliable characterization techniques that allow the determination of nanoparticle properties and the ligand shell composition. Here, a one-phase chemical reduction, followed by a thorough purification to synthesize these nanoparticles with diameters below 5 nm, is presented. The ratio between the two ligands on the surface of the nanoparticle can be tuned through their stoichiometric ratio used during synthesis. We demonstrate how various routine techniques, such as transmission electron microscopy (TEM), nuclear magnetic resonance (NMR), thermogravimetric analysis (TGA), and ultraviolet-visible (UV-Vis) spectrometry, are combined to comprehensively characterize the physicochemical parameters of the nanoparticles.

Amphiphilic gold nanoparticles can be used in many biological applications. A protocol to synthesize gold nanoparticles coated by a binary mixture of ligands and a detailed characterization of these particles is presented.

Sintesi e caratterizzazione delle nanoparticelle d'oro dell'anfifile

Gold nanoparticles covered with a mixture of 1-octanethiol (OT) and 11-mercapto-1-undecane sulfonic acid (MUS) have been extensively studied because of ...

Gold Nanoparticle Modified Carbon Fiber Microelectrodes for Enhanced Neurochemical Detection

For over 30 years, carbon-fiber microelectrodes (CFMEs) have been the standard for neurotransmitter detection. Generally, carbon fibers are aspirated into glass capillaries, pulled to a fine taper, and then sealed using an epoxy to create electrode materials that are used for fast scan cyclic voltammetry testing. The use of bare CFMEs has several limitations, though. First and foremost, the carbon fiber contains mostly basal plane carbon, which has a relatively low surface area and yields lower sensitivities than other nanomaterials. Furthermore, the graphitic carbon is limited by its temporal resolution, and its relatively low conductivity. Lastly, neurochemicals and macromolecules have been known to foul at the surface of carbon electrodes where they form non-conductive polymers that block further neurotransmitter adsorption. For this study, we modify CFMEs with gold nanoparticles to enhance neurochemical testing with fast scan cyclic voltammetry. Au3+ was electrodeposited or dipcoated from a colloidal solution onto the surface of CFMEs. Since gold is a stable and relatively inert metal, it is an ideal electrode material for analytical measurements of neurochemicals. Gold nanoparticle modified (AuNP-CFMEs) had a stability to dopamine response for over 4 h. Moreover, AuNP-CFMEs exhibit an increased sensitivity (higher peak oxidative current of the cyclic voltammograms) and faster electron transfer kinetics (lower &#916;EP or peak separation) than bare unmodified CFMEs. The development of AuNP-CFMEs provides the creation of novel electrochemical sensors for detecting fast changes in dopamine concentration and other neurochemicals at lower limits of detection. This work has vast applications for the enhancement of neurochemical measurements. The generation of gold nanoparticle modified CFMEs will be vitally important for the development of novel electrode sensors to detect neurotransmitters in vivo in rodent and other models to study neurochemical effects of drug abuse, depression, stroke, ischemia, and other behavioral and disease states.

In this study, we modify carbon-fiber microelectrodes with gold nanoparticles to enhance the sensitivity of neurotransmitter detection.

Microelettrodi in fibra di carbonio modificati con nanoparticle d'oro per una migliore rilevazione neurochimica

For over 30 years, carbon-fiber microelectrodes (CFMEs) have been the standard for neurotransmitter detection. Generally, carbon fibers are aspirated into ...

Gold Nanoparticle Synthesis

Gold nanoparticles (Au nanoparticles) that are ~12 nm in diameter were synthesized by rapidly injecting a solution of 150 mg (0.15 mmol) of tetrachloroauric acid in 3.0 g (3.7 mmol, 3.6 mL) of oleylamine (technical grade) and 3.0 mL of toluene into a boiling solution of 5.1 g (6.4 mmol, 8.7 mL) of oleylamine in 147 mL of toluene. While boiling and mixing the reaction solution for 2 hours, the color of the reaction mixture changed from clear, to light yellow, to light pink, and then slowly to dark red. The heat was then turned off, and the solution was allowed to gradually cool down to room temperature for 1 hour. The gold nanoparticles were then collected and separated from the solution using a centrifuge and washed three times; by vortexing and dispersing the gold nanoparticles in 10 mL portions of toluene, and then precipitating the gold nanoparticles by adding 40 mL portions of methanol and spinning them in a centrifuge. The solution was then decanted to remove any remaining byproducts and unreacted starting materials. Drying the gold nanoparticles in a vacuum environment produced a solid black pellet; which could be stored for long periods of time (up to one year) for later use, and then redissolved in organic solvents such as toluene.

A protocol for synthesizing ~12 nm diameter gold nanoparticles (Au nanoparticles) in an organic solvent is presented. The gold nanoparticles are capped with oleylamine ligands to prevent agglomeration. The gold nanoparticles are soluble in organic solvents such as toluene.