Bu araştırma Çin Ulusal Doğa Bilimleri Vakfı (31700840) tarafından finanse edildi; Henan Eyaleti'nin Temel Bilimsel Araştırma Projesi (18B430013, 18A150049). Bu araştırma, XYNU'nun Genç Bilginleri için Nanhu Bursiyerleri Programı tarafından desteklendi. Yazarlar, XYNU'daki Yaşam Bilimleri Koleji'nden lisans öğrencisi Zebo Qu'ya yardımcı çalışmaları için teşekkür ediyor. Yazarlar, ekipmanlarının kullanımı için XYNU Analiz ve Test Merkezi'ni kabul etmek isterler.

DİkKAT: Tüm kimyasalları kullanmadan önce ilgili tüm malzeme güvenliği veri sayfalarına (MSDS) başvurduğunuzdan emin olun. Koalyolimer ve nanopartiküllerin hazırlanmasında kullanılan kimyasalların birçoğu akut olarak toksiktir. Nanopartiküllerin de potansiyel tehlikeleri vardır. Eldivenler, laboratuvar önlüğü, davlumbazlar, tam boy pantolonlar ve yakın kullanımlı ayakkabılar da dahil olmak üzere tüm uygun güvenlik uygulamalarını ve kişisel koruyucu ekipmanları kullandığınızdan emin olun.
1. Çift karboksil polietilen glikol sentezi (CT-PEG)33
<ol><li>100 mL yuvarlak alt şişeye 1,46 g (14,6 mmol) süksinik anhidrit (SA) ve 4-dimetilamipikopyridin (DMAP) 209 mg (1,71 mmol) ekleyin.</li>
	<li>1.1. adımda kullanılan mataraya 15 mL susuz tetrahidrofuran (THF) ekleyin ve bir cam durdurucu takın. Matarayı 0 °C'de 30 dakika tutun.</li>
	<li>Yeni bir şişeye 4,28 g (4,28 mmol) polietilen glikol (PEG) ve 1,8 mL (12,8 mmol) trietilen (TEA) ekleyin.</li>
	<li>1.3. adımda kullanılan mataraya 15 mL susuz THF ekleyin ve bir cam durdurucu takın. Çözeltiyi, azot atmosferi altında bir şırınna kullanarak 1.2 adımında kullanılan şişeye yavaşça aktarın.</li>
	<li>Çözeltiyi 0 °C'de 2 saat karıştırın, ardından reaksiyona oda sıcaklığında (RT) gece boyunca devam edin.</li>
	<li>Döner evaporatör (40 °C, 0,1 MPa) kullanarak reaksiyon çözeltisini konsantre edin ve THF çözücüyü çıkarın.</li>
	<li>RT'de, reaksiyon çözeltisini 1.325 g/mL dikloromethanenin (DCM) 15 mL'sinde 1.6 adımından çözün, ardından çökeltme ürününü (polietilen glikol diacid) elde etmek için 15 mL soğuk dietil eter (Et2O) ekleyin. Çözücüyü filtre kağıdı ile çıkarın. NOT: Yağış adımı 3x tekrarlanabilir.</li>
	<li>Çökelticileri RT'de vakum altında 48 saat kurutun.</li>
</ol>2. PEI-g-PEG koalolimer sentezi
<ol><li>1.8 ve 5 mL dimetil sülfit (DMSO) adımından bir şişeye 305.47 mg CT-PEG ekleyin ve CT-PEG'in DMSO'da tamamen çözüldüğünden emin olmak için RT'de karıştırın.</li>
	<li>49,46 mg 1-(3-dimetilaminopropil)-3-etilkarbodiimid hidroklorür (EDC) 5 mL DMSO çözün, sonra 2.1 adımda kullanılan şişeye çözelti ekleyin ve RT'de 30 dakika karıştırın.</li>
	<li>29,69 mg N-hidroksisuçinid (NHS) 5 mL DMSO'da çözün ve çözeltiyi adım 2.1'de kullanılan şişeye ekleyin. RT'de 3 saat karıştırmaya devam edin.</li>
	<li>28,6 μL polietileniminin (PEI) 10 mL DMSO'da çözün ve çözeltiyi adım 2,1'de kullanılan şişeye damla yönünde ekleyin. En az 3 gün karıştırın.</li>
	<li>Reaksiyona giren çözeltiyi adım 2.4'ten diyaliz torbasına (1.000 moleküler ağırlık kesme [MWCO]) aktarın. Diyaliz torbasını kadran olarak 500 mL ultra saf su ile 1 L beher içine yerleştirin. Ultra saf suyu 3 gün boyunca her 12 saat değiştirin.</li>
	<li>2.5. adımdaki çözeltiyi başka bir diyaliz torbasına (10.000 MWCO) aktarın. Diyaliz torbasını kadran olarak 500 mL ultra saf su ile 1 L beher içine yerleştirin. Ultra saf suyu 3 gün boyunca her 12 saat değiştirin.</li>
	<li>Çözeltiyi 2.6.</li>
</ol>3. PEI-g-PEG@AuNPs Sentezi
<ol><li>5 mg hazırlanmış PEI-g-PEG'i (adım 2.7) 5 mL ultra saf suda yeni bir şişede çözün ve bir cam durdurucuya takın.</li>
	<li>Mataraya 100 mL 0,3 mM HAuCl4 çözeltisi ekleyin ve çözümü RT'de 3 saat karıştırın. NOT: Çözeltinin rengi hemen sarıdan turuncuya değişmelidir.</li>
	<li>Şişeye 1 mL 1 mg/mL NaBH4 çözeltisi ekleyin ve rt'de çözeltiyi 3 saat karıştırın. NOT: Reaksiyon çözeltisi anında bordoya dönmelidir.</li>
	<li>PEI-g-PEG@AuNPs çözeltisini elde etmek için 2.5 adımında açıklandığı gibi 3 gün boyunca bir diyaliz torbası (1.000 MWCO) kullanarak reaksiyon ürününü dialyze edin.</li>
</ol>4. DOX-g-PEI-g-PEG@AuNPs sentezi
<ol><li>Yeni bir şişeye 1 mL 2,2 mg/mL DOX çözeltisi ve 20 mL PEI-g-PEG@AuNPs çözeltisi ekleyin ve cam bir durdurucuya takın.</li>
	<li>1 mL ultra saf suda 0.727 mg EDC çözün ve EDC çözeltisini adım 4.1'de kullanılan şişeye ekleyin.</li>
	<li>0.437 mg NHS'yi 1 mL ultra saf suda çözün. NHS çözeltisini şişeye ekleyin ve RT'de 1 saat karıştırın.</li>
	<li>DOX-g-PEI-g-PEG@AuNPs çözümünü elde etmek için 2.5.</li>
</ol>5. AS1411-g-DOX-g-PEI-g-PEG@AuNPs sentezi
<ol><li>Yeni bir şişeye 20 mL DOX-g-PEI-g-PEG@AuNPs çözeltisi ve 4OD AS1411 (OD = optik yoğunluk; 1OD ≈ 33 μg) ekleyin.</li>
	<li>28,76 mg EDC'yi 1 mL ultra saf suda çözün ve 5.1 adımında kullanılan şişeye EDC çözeltisini ekleyin.</li>
	<li>17,27 mg NHS'yi 1 mL ultra saf suda çözün. 5.1. adımda kullanılan mataraya NHS solüsyonünü ekleyin ve RT'de reaksiyonun 1 saat karıştırın.</li>
	<li>AS1411-g-DOX-g-PEI-g-PEG@AuNPs elde etmek için 2.5.</li>
</ol>6. Örnek karakterizasyon
<ol><li>CT-PEG polimerini (adım 1.8) ve PEI-g-PEG koalolymerini (adım 2.7) sırasıyla nükleer manyetik rezonans (NMR) tüplerinde kloroform-d olarak çözün. Kimyasal yapıyı doğrulamak için 14,09 T süper iletken mıknatıs ve 5,0 mm 600 MHz geniş bant Z-gradyan yüksek çözünürlüklü prob ile donatılmış 600 MHz NMR spektrometre kullanarak numuneleri analiz edin34.</li>
	<li>AuNP'leri, DOX'u ve AS1411'i dağıtıp ultra saf suda sırasıyla PEI-g-PEG@AuNPs (adım 3.4), DOX-g-PEI-g-PEG@AuNPs (adım 4.4) ve AS1411-g-DOX-g-PEI-g-PEG@AuNPs (adım 5.4) hazırlayın. Ardından, cuvettes aktarın ve bir UV-vis spektrofotometre kullanarak ultraviyole görünür (UV-vis) spektrumu kaydedin.</li>
	<li>Alüminyum folyoya çift taraflı bir yapıştırıcı (~2 mm x 2 mm) takın ve numune çözeltisini (PEI-g-PEG@AuNPs, DOX-g-PEI-g-PEG@AuNPs ve AS1411-g-DOX-g-PEI-g-PEG@AuNPs) tüm bant üzerine eşit şekilde batırın. X-ışını fotoelektron spektroskopisi analizörü kullanarak numuneleri analiz edin.</li>
	<li>ULTRA SAF SUDA PEI-g-PEG@AuNPs, DOX-g-PEI-g-PEG@AuNPs ve AS1411-g-DOX-g-PEI-g-PEG@AuNPs çözeltilerini sırasıyla dağıtin. Ardından, cuvettes aktarın ve dinamik ışık saçılım kullanarak boyut dağılımını değerlendirin.</li>
	<li>PEI-g-PEG@AuNPs, DOX-g-PEI-g-PEG@AuNPs ve AS1411-g-DOX-g-PEI-g-PEG@AuNPs çözeltilerini ultra saf suya dağıtın (her numune için 5 mL ultra saf su başına bir damla numune). 2 saat boyunca sonicate. Bakır ızgarayı numune çözeltilerine batırın ve kızılötesi lambanın altında kurutun. İletim elektron mikroskobu kullanarak morfolojiyi karakterize edin.</li>
	<li>20 kDa MWCO diyaliz kasetine 1 mg AS1411-g-DOX-g-PEI-g-PEG@AuNPs enjekte edin, ardından % 5 sığır serum albümini (BSA) ile 80 mL fosfat tamponlu salin (PBS) yerleştirin. 37 °C'de karıştırın.</li>
	<li>Önceden belirlenmiş zaman noktalarında, 100 μL aliquots toplayın ve taze PBS ile değiştirin. Aliquotların DOX floresan yoğunluğunu ölçmek için bir UV-vis spektrofotometre kullanın.</li>
</ol>7. AS1411-g-DOX-g-PEI-g-PEG@AuNPs nanopartiküllerinin CCK-8 tahlilleri
<ol><li>Dulbecco'nun modifiye Eagle's medium'unda (DMEM) %10 fetal sığır serumu, 100 U/mL penisilin ve 100 μg/mL streptomisin ile desteklenmiş A549 hücrelerini %95 hava ve %5 CO2 ile 37 °C'de nemlendirilmiş bir atmosfer altında büyütün. Kültür ortamını her 2 günde bir değiştirin. Hazırlanan AS1411-g-DOX-g-PEI-g-PEG@AuNPs nanopartiküllerinin sitotoksisitesini nicel olarak değerlendirmek için hücre çoğalması ve sitotoksiklik tahlilleri için 5.</li>
	<li>Her kuyuya 1 mL hücre ortamı içeren 100 μL nanopartikül çözeltisi ekleyin. 24 saat ve 48 saat kültleme yaptıktan sonra, kültür medyasını hücre kültürü plakalarından çıkarın, ardından her kuyuya hemen 300 μL taze kültür ortamı ve 30 μL hücre sayım kiti-8 (CCK-8) kiti çözümleri ekleyin. 37 °C'de bir CO 2 inkübatörde4 saat kuluçkaya yatırın.</li>
	<li>200 μL reaksiyon çözümlerini adım 7.2'den 96 kuyu plakasına aktarın. Mikro plaka okuyucu ile 570 nm'de her kuyunun optik yoğunluğunu (OD) okuyun.</li>
	<li>Mikroskop altında 24 saat ve 48 saat hücre morfolojisini gözlemleyin.</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>

Yazarların açıklayacak bir şeyi yok.

1H NMR spektrumu (Şekil 2) CT-PEG koalolimer ve PEI-g-PEG koalolimerinin başarılı sentezini doğrular. PEG ve PEI'nin moleküler ağırlıkları sırasıyla 1.000 ve 1.200'dir. Ek olarak, EDC/NHS katalitik sistemi, PEI-g-PEG koalolymer'ı amid reaksiyonları yoluyla sentezlemek için kullanılmıştır. PEI-g-PEG koalizatör sentezlemek için PEG ve PEI'nin moleküler ağırlıkları değiştiyse, reaksiyon süresinin ve katalitik sistemin yeniden değerlendirilmesi gerektiği...

Dünya çapında en büyük halk sağlığı sorunu olan kanser, düşük kür oranı, yüksek nüks oranı ve yüksek ölüm oranı1,2. Mevcut konvansiyonel kanser karşıtı yöntemler cerrahi, kemoterapi ve radyoterapi3, kemoterapinin klinikteki kanser hastaları için birincil tedavi olduğu4. Klinik olarak kullanılan antikanser ilaçlar esas olarak paclitaxel (PTX)5 ve doxorubicin (DOX)6,7içerir. Bir antineoplastik ilaç olan DOX, kanser sitotoksisitesi ve kanser hücre çoğalmasının inhibisyonu avantajları nedeniyle klinik kemoterapide yaygın olarak uygulanmıştır8,9. Bununla birlikte, DOX kardiyotoksiteye neden olur10,11ve DOX'un kısa yarı ömrü klinikte uygulanmasını kısıtlar12. Bu nedenle, DOX'i yüklemek ve kontrollü bir şekilde hedeflenen bir alana serbest bırakmak için bozulabilir ilaç taşıyıcılarına ihtiyaç vardır.
Nanopartiküller hedefli ilaç dağıtım sistemlerinde yaygın olarak kullanılmaktadır ve kanser tedavisinde çeşitli avantajlara sahiptir (yani, büyük yüzey-hacim oranı, küçük boyut, çeşitli ilaçları kapsülleme yeteneği ve ayarlanabilir yüzey kimyası vb.) 13,14,15. Özellikle, altın nanopartiküller (AuNP'ler) fototermal kanser tedavisi16,17gibi biyolojik ve biyomedikal uygulamalarda yaygın olarak kullanılmaktadır. AuNP'lerin facile sentezi ve genel yüzey fonksiyonelleştirme gibi benzersiz özellikleri, kanser tedavisinin klinik alanında mükemmel beklentilere sahiptir18. Ayrıca, AuNP'ler ilaç dağıtım stratejilerini tanımlamak, tümörleri teşhis etmek ve birçok çalışmada direnci yenmek için kullanılmıştır19,20.
Buna rağmen, AuNP'lerin, hedefleme ve erişilebilirlik özellikleri gibi gelişmiş geçirgenlik ve tutma (EPR) yoluyla tümör lezyonlarında yüksek lokal salınım yoluyla ilaç direncinin üstesinden gelmek için daha fazla uyarlanması gerekir. Polimer fonksiyonelleştirilmiş AuNP'ler, hidrofobik antikanser ilaçlarının iyileştirilmiş su çözünürlüğü ve uzun sirkülasyon süresi21,22gibi benzersiz avantajlar sergilemiştir. AuNP kaplamaları için polietilen glikol (PEG), polietilenimin (PEI), hyaluronik asit, heparin ve ksantan sakızı gibi çeşitli biyouyumlu polimerler kullanılmıştır. Daha sonra AuNP'lerin stabilitesi ve yükü iyileştirildi23. Özellikle, PEI, birincil, ikincil ve üçüncül aminlerin24'ünbirçok tekrarlayan biriminden oluşan yüksek dallı bir polimerdir. PEI mükemmel çözünürlüğe, düşük viskoziteye ve AuNP'lerde kaplama için uygun olan yüksek derecede işlevselliğe sahiptir.
Öte yandan, anti-kanser ilaçlarının doğrudan kanser hücrelerine daha iyi yükleme verimliliği ve primer ve ileri metastatik tümörlerin tedavisi için daha düşük toksisite ile teslim edilmesi gerekir25. Hedeflenen ligandlar anti-kanser ilaç hedefli doğum sistemleri için büyük bir potansiyele sahiptir26. Hedef molekül bağlama için seçiciliği, özgüllüğü hedefleyen kanser önleyici ilaçlar ve hastalıklı dokularda ilaç zenginleştirmeyi arttırır27. Daha fazla ligand antikorlar, polipeptitler ve küçük moleküller içerir. Diğer ligandlara kıyasla, nükleik asit aptamerleri in vitro olarak sentezlenebilir ve değiştirilmesi kolaydır. AS1411, özellikle kanser hücreleri 28,29,30üzerinde aşırı ifade edilmiş bir hedef nükleer protein reseptörüne bağlanmak için kararlı bir dimerik G-tetramer yapısı oluşturan değiştirilmemişbir 26bp fosfodiester oligonükleotiddir. AS1411 birçok kanser hücresinin çoğalmasını engeller, ancak sağlıklı hücrelerin büyümesini etkilemez31,32. Sonuç olarak, AS1411 ideal bir hedefli ilaç dağıtım sistemi imal etmek için kullanılmıştır.
Bu çalışmada, bir PEI-g-PEG koalolymer amid reaksiyonu ile sentezlenir, daha sonra PEI-g-PEG koalolymer kaplı altın nanopartiküller (PEI-g-PEG@AuNPs) imal edilir. Ayrıca, DOX ve AS1411, Şekil 1'degösterildiği gibi, hazırlanan PEI-g-PEG@AuNPs ardışık olarak bağlanır. Bu ayrıntılı protokol, araştırmacıların DOX ve AS1411 yüklü yeni PEI-g-PEG@AuNPs üretimiyle ilgili ortak tuzakların çoğundan kaçınmalarına yardımcı olmak için tasarlanmıştır.

<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

Sağlıklı hücrelerde ilaç direnci ve toksisite nedeniyle klinik kanser tedavisinde doksofobisin (DOX) kullanımı sınırlanmıştır. Bu protokol, polietilen glikol (PEI-g-PEG) koaloksimer fonksiyonelleştirilmiş altın nanopartiküller (AuNP'ler) ile aşılanmış poli (etileninimin) yüklü aptamer (AS1411) ve DOX ile amid reaksiyonları yoluyla tasarlanmasını açıklar. AS1411 özellikle kanser hücrelerindeki hedefli nükleolin reseptörleri ile bağlanır, böylece DOX sağlıklı hücreler yerine kanser hücrelerini hedefler. İlk olarak, PEG karboksilatlanır, daha sonra 1H NMR analizi ile onaylanan bir PEI-g-PEG koalizör elde etmek için dallı PEI'ye aşılanır. Daha sonra, PEI-g-PEG koalizör kaplı altın nanopartiküller (PEI-g-PEG@AuNPs) sentezlenir ve DOX ve AS1411, reaksiyonlar arasında yavaş yavaş AuNP'lere bağlanır. Hazırlanan AS1411-g-DOX-g-PEI-g-PEG@AuNPs çapı ~39,9 nm dir ve nanopartiküllerin su ve hücre ortamında stabil olduğunu gösteren -29,3 mV zeta potansiyeline sahiptir. Hücre sitotoksikliği tahlilleri, yeni tasarlanan DOX yüklü AuNP'lerin kanser hücrelerini (A549) öldürebildiğini göstermektedir. Bu sentez, PEI-g-PEG koalizörlerinin, aptamerlerinin ve DOX'un, sıralı amid reaksiyonları ile elde edilen AuNP'ler üzerindeki hassas düzenini göstermektedir. Bu tür aptamer-PEI-g-PEG fonksiyonelleştirilmiş AuNP'ler kanser tedavisinde hedefe yönelik ilaç teslimatı için umut verici bir platform sağlar.

1 CT-PEG polimer ve PEI-g-PEG koalolimerlerinin başarılı sentezini doğrulamak için H NMR spektroskopisi kullanılmıştır (Şekil 2). Şekil 2a, δ 'daki metilen proton sinyalinin = 3.61 ppm'de ve δ ' de karboksil proton sinyalinin = 2.57 ppm'de CT-PEG polimerlerinin başarılı sentezini doğruladığını göstermektedir. Şekil 2b, PEG'in δ =2.6 ppm'deki metilen proton sinyalinin ve <...

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

Bu protokolde, doxorubicin yüklü AS1411-g-PEI-g-PEG modifiye altın nanopartiküller üç aşamalı reaksiyonlar yoluyla sentezlenir. Daha sonra doksofobikin yüklenir ve kanser tedavisi için hedef kanser hücrelerine teslim edilir.

Hedeflenen İlaç Teslimatı için Doxorubicin Yüklü Aptamer-PEI-g-PEG Modifiye Altın Nanopartiküllerinin Sentezi

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 Görüntüleme.  Xinyang Normal University. Bu protokolde, doxorubicin yüklü AS1411-g-PEI-g-PEG modifiye altın nanopartiküller üç aşamalı reaksiyonlar yoluyla sentezlenir. Daha sonra doksofobikin yüklenir ve kanser tedavisi için hedef kanser hücrelerine teslim edilir.

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.

Şablon Ayarlanabilir IR Absorbans ile plasmonik Altın Nanotüpler Sentez Yönetmen

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

Kimya

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.

SDF-1α-Kitosan-Dekstran Sülfat nano partiküllerin hazırlanması ve Karakterizasyonu

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.

Ortam Koşullarında Altın Nanopartiküller Kararlı Oligomerik Kümeleri Boyut Kontrollü Sentezi için Basit Bir Yöntem

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.

Altın nanorods hidrokinon Esaslı Sentezi

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.

Nb2O5 sentezi için kimyasal yağış yöntemi toplu nikel katalizör yüksek belirli yüzey alanı ile modifiye

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.

Hızlı Nanoprobe sinyal geliştirme içinde in Situ altın nanoparçacık sentezi tarafından

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.

Hedeflenen Plasmid DNA teslim için metiyonin functionalized biyouyumlu blok kopolimerler

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.

Amphiphilik altın nanopartiküller sentezi ve karakterizasyonu

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.

Gelişmiş nörokimyasal algılama için altın Nanopmakale modifiye karbon fiber Mikroelektrotlar

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.