本研究由中国国家自然科学基金委员会（31700840）资助:河南省重点科研项目（18B430013，18A150049）。这项研究得到了南湖青年学者项目的支持。作者要感谢来自XYNU生命科学学院的本科生曲泽波的乐于助人的作品。作者希望确认XYNU的分析和测试中心使用他们的设备。

注意:在使用所有化学品之前，请务必查阅所有相关材料安全数据表 （MSDS）。用于制备共聚物和纳米粒子的几种化学物质具有剧毒性。纳米粒子也有潜在的危害。确保使用所有适当的安全做法和个人防护设备，包括手套、实验室外套、头罩、全长裤子和近身鞋。
1. 双碳水化合物聚乙二醇（CT-PEG）合成33
<ol><li>将 1 . 46 克（ 14 . 6 mmol ）的氢化物（ SA ）和 209 毫克（ 1 . 71 mmol ）的 4 二甲基甲基二胺（ DMAP ）添加到 100 mL 圆形底部烧瓶中。</li>
	<li>将 15 毫升的无水四氢氢素 （THF） 添加到步骤 1.1 中使用的烧瓶中，并安装玻璃塞。将烧瓶保持在 0 °C 30 分钟。</li>
	<li>将 4.28 克（4.28 mmol） 聚乙二醇 （PEG） 和 1.8 mL （12.8 mmol） 添加到新的烧瓶中。</li>
	<li>将 15 毫升的无水 THF 添加到步骤 1.3 中使用的烧瓶中，并安装玻璃塞。使用氮气下的注射器，将溶液缓慢地转移到步骤 1.2 中使用的烧瓶中。</li>
	<li>在 0 °C 搅拌溶液 2 小时，然后在室温 （RT） 下持续反应过夜。</li>
	<li>使用旋转蒸发器（40 °C，0.1 MPa），浓缩反应溶液并去除THF溶剂。</li>
	<li>在RT，溶解步骤1.6在1.325克/毫升二氯甲烷（DCM）的15毫升反应溶液，然后加入15毫升的冷二乙醚（Et2O）以获得降水产品（聚乙二醇二恶英）。通过滤纸去除溶剂。 注:降水步骤可重复 3 倍。</li>
	<li>在RT的真空下干燥沉淀物48小时。</li>
</ol>2. PEI-g-PEG共聚体的合成
<ol><li>将 305.47 毫克 CT-PEG 从步骤 1.8 和 5 mL 的二甲基硫化物 （DMSO） 添加到烧瓶中，并在 RT 搅拌，以确保 CT-PEG 在 DMSO 中完全溶解。</li>
	<li>在 5 mL DMSO 中溶解 49.46 毫克 1-（3-二甲基丙烯酸丙基）-3-乙基二氧化硫 （EDC），然后将溶液添加到步骤 2.1 中使用的烧瓶中，并在 RT 搅拌 30 分钟。</li>
	<li>在 5 mL DMSO 中溶解 29.69 毫克的 N-羟基硫酰胺 （NHS），并将溶液添加到步骤 2.1 中使用的烧瓶中。继续在RT搅拌3小时。</li>
	<li>在 DMSO 的 10 mL 中溶解 28.6 μL 聚乙烯 （PEI），并将溶液顺向添加到步骤 2.1 中使用的烧瓶中。至少搅拌3天。</li>
	<li>将反应溶液从步骤 2.4 转移到透析袋（1，000 分子重量截止 [MWCO]）。将透析袋放入 1 升烧嘴中，并放入 500 毫升超纯水作为透析剂。每12小时更换一次超纯水3天。</li>
	<li>将第 2.5 步的溶液转移到另一个透析袋 （10，000 MWCO）。将透析袋放入 1 升烧嘴中，并放入 500 毫升超纯水作为透析剂。每12小时更换一次超纯水3天。</li>
	<li>使用旋转蒸发器（40 °C，0.1 MPa）浓缩步骤2.6的溶液，并冷冻干燥样品以获得PEI-g-PEG粉末。</li>
</ol>3. PEI-g-PEG@AuNPs综合
<ol><li>将5毫克准备好的PEI-g-PEG（步骤2.7）溶解在5毫升的超纯水中，放入新的烧瓶中，并配有玻璃塞。</li>
	<li>将 100 mL 的 0.3 mM HAuCl4 溶液添加到烧瓶中，并在 RT 搅拌解决方案 3 小时。 注意:溶液的颜色应立即从黄色更改为橙色。</li>
	<li>在烧瓶中加入 1 毫克/mL NaBH4 溶液，在 RT 搅拌溶液 3 小时。 注意:反应解决方案应立即变成酒红色。</li>
	<li>使用透析袋 （1，000 MWCO） 对反应产品进行透析，为期 3 天，如步骤 2.5 所述，以获得 PEI-g-PEG@AuNPs 解决方案。</li>
</ol>4. 多克斯-格-爱德华-格-PEG@AuNPs综合
<ol><li>将 1 毫升 2.2 毫克/毫升 DOX 解决方案和 20 mL 的 PEI-g-PEG@AuNPs 溶液添加到新的烧瓶中，并配有玻璃塞子。</li>
	<li>在 1 mL 的超纯水中溶解 0.727 毫克 EDC，并将 EDC 解决方案添加到步骤 4.1 中使用的烧瓶中。</li>
	<li>在 1 mL 的超纯水中溶解 0.437 毫克 NHS。将 NHS 解决方案添加到烧瓶中，在 RT 搅拌 1 小时。</li>
	<li>通过使用透析袋（1，000 MWCO）对反应产品进行透析，为期3天，如第2.5步所述，以获得DOX-g-PEI-g-PEG@AuNPs解决方案。</li>
</ol>5. AS1411-g-DOX-g-PEI-g-PEG@AuNPs综合
<ol><li>将 20 mL 的 DOX-g-PEI-g-PEG@AuNPs 溶液和 4OD AS1411 （OD = 光学密度;1OD ≈ 33 μg） 添加到新的烧瓶中。</li>
	<li>在 1 mL 的超纯水中溶解 28.76 毫克 EDC，并将 EDC 解决方案添加到步骤 5.1 中使用的烧瓶中。</li>
	<li>在 1 mL 的超纯水中溶解 17.27 毫克 NHS。将 NHS 解决方案添加到步骤 5.1 中使用的烧瓶中，并在 RT 中搅拌反应 1 小时。</li>
	<li>通过使用透析袋（1，000 MWCO）对反应产品进行透析3天，如步骤2.5所述，获得AS1411-g-DOX-g-PEI-g-PEG@AuNPs。</li>
</ol>6. 样本特征
<ol><li>分别在核磁共振（NMR）管中溶解氯仿中的CT-PEG聚合物（步骤1.8）和PEI-g-PEG共聚物（步骤2.7）。使用配备 14.09 T 超导磁体和 5.0 mm 600 MHz 宽带 Z 梯度高分辨率探头的 600 MHz NMR 光谱仪分析样品，以确认化学结构34。</li>
	<li>分散 AuNP、DOX 和 AS1411，并在超纯水中分别准备 PEI-g-PEG@AuNPs（步骤 3.4）、DOX-g-PEI-g-PEG@AuNPs（第 4.4 步）和 AS1411-g-DOX-g-PEI-g-PEG@AuNPs（步骤 5.4）。然后，转移到紫外线，并使用紫外线-维斯光谱仪记录紫外线可见光谱（UV-vis）。</li>
	<li>在铝箔上附加双面胶粘剂（~2 mm x 2 mm），并将样品溶液（PEI-g-PEG@AuNPs、DOX-g-PEI-g-PEG@AuNPs和AS1411-g-DOX-g-PEI-g-PEG@AuNPs）均匀地浸入整个磁带上。使用 X 射线光电子光谱分析仪分析样品。</li>
	<li>分别在超纯水中分散 PEI-g-PEG@AuNPs、DOX-g-PEI-g-PEG@AuNPs和 AS1411-g-DOX-g-PEI-PEG@AuNPs解决方案。然后，转移到小面包，并使用动态光散射来评估大小分布。</li>
	<li>将 PEI-g-PEG@AuNPs、DOX-g-PEI-g-PEG@AuNPs和 AS1411-g-DOX-g-PEI-g-PEG@AuNPs溶液分散在超纯水中（每个样品每 5 mL 超纯水一滴样品）。声波2小时。将铜网格浸入样品溶液中，并在红外灯下干燥。使用传输电子显微镜描述形态。</li>
	<li>将 1 毫克 AS1411-g-DOX-g-PEI-g-PEG@AuNPs注入 20 kDa MWCO 透析盒中，然后放入 80 mL 磷酸盐缓冲盐水 （PBS） 中，加入 5% 牛血清白蛋白 （BSA）。在37°C搅拌。</li>
	<li>在预先确定的时间点，收集 100 μL 别名，代之以新鲜的 PBS。使用紫外线光谱仪测量音素的 DOX 荧光强度。</li>
</ol>7. CCK-8 对AS1411-g-DOX-g-PEI-g-PEG@AuNPs纳米粒子的检测
<ol><li>在杜尔贝科改良的鹰介质 （DMEM） 中生长 A549 细胞，在 95% 空气和 5% CO2 在 37 °C 的潮湿大气中补充 10% 胎儿牛血清、100 U/mL 青霉素和 100μg/mL 链霉素。 每 2 天更换一次文化介质。使用通道 5 中的细胞进行细胞增殖和细胞毒性检测，定量评估制备 AS1411-g-DOX-g-PEI-g-PEG@AuNPs纳米粒子的细胞毒性。</li>
	<li>将 100 μL 的纳米粒子溶液添加到每口井中，其中含有 1 mL 的细胞介质。培养 24 小时和 48 小时后，从细胞培养板中去除培养介质，然后立即向每口井添加 300 μL 的新鲜培养介质和 30 μL 的细胞计数套件-8 （CCK-8） 套件解决方案。在 37 °C 的 CO2 孵化器中孵化 4 小时。</li>
	<li>将 200 μL 的反应解决方案从步骤 7.2 转移到 96 井板中。用微板读取器阅读每口井的光学密度 （OD）在 570 nm。</li>
	<li>在显微镜下以24小时和48小时观察细胞的形态。</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>

作者没有什么可透露的。

1H NMR光谱（图2）确认CT-PEG共聚合体和PEI-g-PEG共聚合体的成功合成。PEG和PEI的分子量分别为1，000和1，200。此外，EDC/NHS催化系统还用于通过阿米德反应合成PEI-g-PEG共聚物。需要注意的是，如果PEG和PEI的分子量因合成PEI-g-PEG共聚物而发生变化，则需要重新评估反应时间和催化系统。此外，需要进一步调整AUNPs上PEI-g-PEG共聚合体涂层的反应条件，主要是因为PEG-g-PEI聚合体?...

癌症是世界性的主要公共卫生问题，其广泛特征是治愈率低、复发率高、死亡率高。目前传统的抗癌方法包括手术、化疗和放疗3种，其中化疗是4号诊所癌症患者的主要治疗方法。临床使用的抗癌药物主要包括帕利塔塞尔（PTX）5和多索鲁比辛（DOX）6，7。DOX，一种抗肿瘤药物，由于癌症细胞毒性和抑制癌细胞增殖8，9的优点，已广泛应用于临床化疗。然而，DOX导致心毒性10，11，和DOX的短半生限制其应用在诊所12。因此，需要降解的药物携带者以可控的方式将 DOX 加载并以子等方式释放到目标区域。
纳米粒子已广泛应用于靶向药物输送系统，在癌症治疗方面具有若干优势（即表面与体积比大、体积小、封装各种药物的能力、可调谐表面化学等）。13，14，15.特别是金纳米粒子（AuNPs）已广泛应用于生物和生物医学应用，如光热癌治疗16，17。AuNPs的独特特性，如简体合成和一般表面功能化，在癌症治疗18的临床领域具有良好的前景。此外，AuNP已经被用来识别药物输送策略，诊断肿瘤，并克服阻力在许多研究19，20。
尽管如此，AuNP 需要进一步定制，通过增强渗透和保留 （EPR）（如靶向和辅助功能）在肿瘤病变时高局部释放来克服耐药性。聚合物功能化 AuNP 具有独特的优势，如疏水抗癌药物水溶解性提高，循环时间延长21、22。各种生物相容聚合物已用于 AuNP 涂料，如聚乙二醇 （PEG）、聚乙烯胺 （PEI）、透明质酸、肝素和黄豆胶。然后，AuNP的稳定性和有效载荷得到很好的改善。具体来说，PEI是一种高度分支的聚合物，由许多重复单位的初级，二级和三级胺24组成。PEI 具有极好的溶解度、低粘度和高功能性，适合涂在 AuNPs 上。
另一方面，抗癌药物需要直接输送到癌细胞，提高负荷效率，降低毒性，治疗原发性肿瘤和晚期转移性肿瘤25。靶向配体具有巨大的潜力，抗癌药物靶向输送系统26。其靶分子结合的选择性赋予抗癌药物针对特异性，并增加药物在病组织中的丰富性27。更多的配体包括抗体、多肽和小分子。与其他配体相比，核酸贴合剂可以在体外合成，易于修改。AS1411是一种未经改性26b磷化物寡核苷酸，形成稳定的二元G-四聚氰胺结构，专门与癌细胞28、29、30上过度表达的目标核蛋白受体结合。AS1411抑制许多癌细胞的增殖，但不影响健康细胞31，32的生长。因此，AS1411 被用来制造理想的有针对性的药物输送系统。
在这项研究中，PEI-g-PEG共聚物通过阿米德反应合成，然后制造PEI-g-PEG共聚物涂层金纳米粒子（PEI-g-PEG@AuNPs）。此外，DOX 和 AS1411 与准备好的 PEI-g-PEG@AuNPs有顺序链接，如 图 1所示。此详细的协议旨在帮助研究人员避免与制造装有 DOX 和 AS1411 的新 PEI-g-PEG@AuNPs相关的许多常见陷阱。

<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

由于健康细胞的耐药性和毒性，多索鲁比辛（DOX）在临床癌症治疗中的使用受到限制。该协议描述了用聚乙二醇（PEI-g-PEG）二甘醇（PEI-g-PEG）共聚聚（乙二烯氨酸）嫁接的聚（乙二烯氨酸）与加载的阿普塔默（AS1411）和DOX通过阿米特反应进行功能化金纳米粒子（AuNPs）的设计。AS1411与癌细胞上的靶向核素受体特别结合，使DOX针对的是癌细胞而不是健康细胞。首先，PEG 是盒装的，然后嫁接到分支 PEI 以获得 PEI-g-PEG 聚合物，这一点由 1H NMR 分析确认。其次，PEI-g-PEG copolymer 涂层金纳米粒子 （PEI-g-PEG@AuNPs） 进行合成，DOX 和 AS1411 通过阿米德反应逐渐与 AuNP 粘合。制备的 AS1411-g-DOX-g-PEI-g-PEG@AuNPs直径为 39.9 nm，Zeta 电位为 -29.3 mV，表明纳米粒子在水和细胞介质中是稳定的。细胞毒性检测表明，新设计的DOX加载的AuNPs能够杀死癌细胞（A549）。这种合成展示了PEI-g-PEG共聚物、贴花和DOX在AuNP上的微妙排列，这些排列是通过连续的阿米德反应实现的。这种阿普塔默-PEI-g-PEG功能化的 AuNP 为癌症治疗中的靶向药物输送提供了一个很有前途的平台。

1H NMR光谱学用于确认CT-PEG聚合物和PEI-g-PEG共聚合物（图2）的成功合成。图2a显示，δ的甲基质子信号=3.61ppm，δ=2.57ppm的纸箱质子信号确认CT-PEG聚合物的成功合成。图2b显示PEG的乙烯质子信号在δ=2.6ppm，PEI的质子信号在δ=1.66ppm确认PEI-g-PEG共聚物的合成。 
紫外线光谱学的进行，...

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

在此协议中，多索鲁比辛加载 AS1411-g-PEI-g-PEG 改性金纳米粒子通过三步阿米内反应合成。然后，多索鲁比辛被加载并交付给靶向癌细胞进行癌症治疗。

合成阿普塔默-PEI-g-PEG改性金纳米粒子装载多索鲁比辛用于靶向药物输送

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 次观看.  Xinyang Normal University. 在此协议中，多索鲁比辛加载 AS1411-g-PEI-g-PEG 改性金纳米粒子通过三步阿米内反应合成。然后，多索鲁比辛被加载并交付给靶向癌细胞进行癌症治疗。

视频: 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.

的模板导演电浆黄金纳米管的合成与可调谐红外吸收

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

科研

化学

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 - 壳聚糖硫酸葡聚糖纳米粒子的制备与表征

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.

根据环境条件的金纳米粒子的稳定低聚簇的大小控制合成的一种简便方法

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.

金纳米棒基于对苯二酚合成

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.

化学沉淀法合成铌2O5改性高比表面积镍催化剂

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.

原位金纳米粒子合成快速 Nanoprobe 信号增强

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.

用于靶向质粒DNA输送的美聚氨酸功能化生物相容性块共聚物

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.

两栖金纳米粒子的合成与表征

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.

用于增强神经化学检测的金纳米粒子改性碳纤维微电极

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.