Name: combinatorial synthesis of and high-throughput protein release from polymer film and nanoparticle libraries
Uploaded: 2012-09-06T12:00:00
Description: Watch this Scientific Journal Video about Combinatorial Synthesis of and High-throughput Protein Release from Polymer Film and Nanoparticle Libraries at JoVE.com

作者承认的ONR的MURI奖（NN00014-06-1-1176）和美国陆军医学研究和装备司令部（批准号：W81XWH-10-1-0806）的资金支持。这种材料是根据工作支持的美国国家科学基金会根据格兰特第EEC 0552584号和0851519。

1。组合聚合物图书馆的合成高分子化学变化-请参阅图1为机器人设定<ol><li>各单体溶解在适当的溶剂中（浓度为25毫克/毫升），和加载每个成10cc的气密注射器。 </li><li>将附加抗溶剂的诱惑锁毛细管各注射器的端部。 </li><li>将注射泵的注射器（新时代可编程注射泵），并锁定到位。 </li><li>线性致动器（Zaber）的起始位置。 </li><li>使用环形支架夹的两个毛细管，定位结束进入首发瓶/?...

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	<li>Petersen, L. K., Sackett, C. K., Narasimhan, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel,+high-throughput+method+to+study+in+vitro+protein+release+from+polymer+nanospheres.">A novel, high-throughput method to study in vitro protein release from polymer nanospheres.</a> J. Comb. Chem. 12, 51-56 (2010).</li><li>Petersen, L. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activation+of+innate+immune+responses+in+a+pathogen-mimicking+manner+by+amphiphilic+polyanhydride+nanoparticle+adjuvants.">Activation of innate immune responses in a pathogen-mimicking manner by amphiphilic polyanhydride nanoparticle adjuvants.</a> Biomaterials. 32, 6815-6822 (2011).</li><li>Vogel, B. M., Cabral, J. T., Eidelman, N., Narasimhan, B., Mallapragada, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Parallel+synthesis+and+high-throughput+dissolution+testing+of+biodegradable+polyanhydride+copolymers.">Parallel synthesis and high-throughput dissolution testing of biodegradable polyanhydride copolymers.</a> J. Comb. Chem. 7, 921-928 (2005).</li><li>Petersen, L. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-throughput+evaluation+of+in+vivo+biodistribution+of+polyanhydride+nanoparticles.">High-throughput evaluation of in vivo biodistribution of polyanhydride nanoparticles.</a> Adv. Healthcare Mater. , (2012).</li><li>Petersen, L. K., Narasimhan, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Combinatorial+design+of+biomaterials+for+drug+delivery:+opportunities+and+challenges.">Combinatorial design of biomaterials for drug delivery: opportunities and challenges.</a> Expert Opin. Drug Deliv. 5, 837-846 (2008).</li><li>Petersen, L. K., Oh, J., Sakaguchi, D. S., Mallapragada, S. K., Narasimhan, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Amphiphilic+polyanhydride+films+promote+neural+stem+cell+adhesion+and+differentiation.">Amphiphilic polyanhydride films promote neural stem cell adhesion and differentiation.</a> Tissue Eng. 17, 2533-2541 (2011).</li><li>Petersen, L. K., Sackett, C. K., Narasimhan, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-throughput+analysis+of+protein+stability+in+polyanhydride+nanoparticles.">High-throughput analysis of protein stability in polyanhydride nanoparticles.</a> Acta Biomater. 6, 3873-3881 (2010).</li><li>Petersen, L. K., Xue, L., Wannemuehler, M. J., Rajan, K., Narasimhan, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+simultaneous+effect+of+polymer+chemistry+and+device+geometry+on+the+in+vitro+activation+of+murine+dendritic+cells.">The simultaneous effect of polymer chemistry and device geometry on the in vitro activation of murine dendritic cells.</a> Biomaterials. 30, 5131-5142 (2009).</li><li>Thorstenson, J. B., Petersen, L. K., Narasimhan, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Combinatorial/high-throughput+methods+for+the+determination+of+polyanhydride+phase+behavior.">Combinatorial/high-throughput methods for the determination of polyanhydride phase behavior.</a> J. Comb. Chem. 11, 820-828 (2009).</li><li>Xue, L., Petersen, L., Broderick, S., Narasimhan, B., Rajan, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identifying+factors+controlling+protein+release+from+combinatorial+biomaterial+libraries+via+hybrid+data+mining+methods.">Identifying factors controlling protein release from combinatorial biomaterial libraries via hybrid data mining methods.</a> ACS Comb. Sci. 13, 50-58 (2011).</li><li>Adler, A. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-throughput+cell-based+screening+of+biodegradable+polyanhydride+libraries.">High-throughput cell-based screening of biodegradable polyanhydride libraries.</a> Comb. Chem. High Through. Screen. 12, 634-645 (2009).</li><li>Determan, A. S., Trewyn, B. G., Lin, V. S., Nilsen-Hamilton, M., Narasimhan, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Encapsulation,+stabilization,+and+release+of+BSA-FITC+from+polyanhydride+microspheres.">Encapsulation, stabilization, and release of BSA-FITC from polyanhydride microspheres.</a> J. Control. Release. 100, 97-109 (2004).</li><li>Determan, A. S., Wilson, J. H., Kipper, M. J., Wannemuehler, M. J., Narasimhan, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protein+stability+in+the+presence+of+polymer+degradation+products:+consequences+for+controlled+release+formulations.">Protein stability in the presence of polymer degradation products: consequences for controlled release formulations.</a> Biomaterials. 27, 3312-3320 (2006).</li><li>Torres, M. P., Determan, A. S., Anderson, G. L., Mallapragada, S. K., Narasimhan, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Amphiphilic+polyanhydrides+for+protein+stabilization+and+release.">Amphiphilic polyanhydrides for protein stabilization and release.</a> Biomaterials. 28, 108-116 (2007).</li><li>Torres, M. P., Vogel, B. M., Narasimhan, B., Mallapragada, S. K. <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+characterization+of+novel+polyanhydrides+with+tailored+erosion+mechanisms.">Synthesis and characterization of novel polyanhydrides with tailored erosion mechanisms.</a> J. Biomed. Mater. Res. A. 76, 102-110 (2006).</li><li>Carrillo-Conde, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Encapsulation+into+amphiphilic+polyanhydride+microparticles+stabilizes+Yersinia+pestis+antigens.">Encapsulation into amphiphilic polyanhydride microparticles stabilizes Yersinia pestis antigens.</a> Acta Biomater. 6, 3110-3119 (2010).</li></ol>

知识库的必要的合成条件和所合成的聚合物的玻璃化转变温度（T 克 ）是必不可少的图书馆制造。如果的 T g是低于室温下，纳米粒子的制造工序中，可能需要进行在受控温度以下的环境中的 T g的聚合物。此外，应采取谨慎，以确保所有设备在接触高温和溶剂必须适当处理这些情况。此协议的一些参数可以调整，以适应不同的聚合物体系（ 即 ，温度，真空度，孵育...

<table border="1"><tbody><tr><td> 名称 </td><td> 公司 </td><td> 目录号 </td></tr><tr><td>电动XYZ阶段：了3倍T-LSM050A，每轴50 mm行程</td><td> Zaber技术</td><td> T-XYZ-LSM050A-KT04 </td></tr><tr><td> NE-1000单注射泵</td><td>新的时代水泵系统</td><td> NE-1000 </td></tr><tr><td>高硼硅* Vista *的无框可重复使用的玻璃文化管</td><td>康宁</td><td> 07-250-125 </td></tr><tr><td>玻璃比色皿</td><td>科学战略</td><td> G102 </td></tr><tr><td> LabVIEW的</td><td>美国国家仪器公司</td><td> 776671-35 </td></tr><tr><td>的SGE气密性注射器，露儿禄</td><td> Sigma Aldrich公司</td><td> 509507 </td></tr><tr><td> U96深孔板1.3 ML＆2.0毫升的</td><td>赛默飞世尔科技：Nunc公司</td><td> 278743 </td></tr><tr><td>井帽垫</td><td>赛默飞世尔科技：Nunc公司</td><td> 276000 </td></tr><tr><td>台风9400 </td><td> GE医疗集团</td><td> 63-0055-79 </td></tr><tr><td> Whatman公司级50圈90毫米</td><td> Whatman公司</td><td> 1450-090 </td></tr></tbody></table>

combinatorial synthesis of and high-throughput protein release from polymer film and nanoparticle libraries

聚酸酐的生物材料是一类具有优良的生物相容性和药物输送能力。虽然他们已经被广泛研究，与传统的单样本在每次一个合成技术，最近的高通量方法已经开发使聚酐1的合成和试验的大型图书馆。这将有利于更有效地优化和设计过程中这些生物材料，药物和疫苗交付应用。在这项工作中描述的方法的组合合成的可生物降解的聚酐膜和纳米颗粒的图书馆和从这些库的蛋白释放的高通量的检测。在这种机器人的操作方法（ 图1），线性致动器和注射器泵控制的LabVIEW，它使免提自动化协议，消除了用户错误。此外，这种方法可以快速制造微观尺度聚合物图书馆，红色ucing创造多元聚合物系统而导致的批量大小。这个组合的聚合物的合成方法便于用相当量的时间，将采取以往合成聚合物中的15种不同的聚合物的合成。此外，聚合物组合库，可以制作成空白或蛋白质加载的几何形状，包括在溶剂中和沉淀到非溶剂（纳米粒子），或通过真空干燥（薄膜）的薄膜或纳米颗粒溶解时的聚合物库。在加载到聚合物中的图书馆荧光染料共轭蛋白，蛋白质的释放动力学可以在高吞吐量的评估使用基于荧光的检测方法（ 图2和图3），如前面描述1。组合平台已验证常规方法2和聚酐膜和纳米库的特点<suP&gt; 1 H NMR和FTIR。蛋白释放动力学，稳定性和抗原性的图书馆已筛选， 体外细胞毒性，细胞因子的产生，表面标志物的表达，黏附，增殖和分化，以及体内生物分布和粘膜粘附1-11。开发本发明的组合的方法，使高吞吐量的聚合物合成和加载蛋白质纳米粒子的制造和膜的图书馆，可以反过来， 在体外和体内筛选的生物材料的性能的优化。

Watch this Scientific Journal Video about Combinatorial Synthesis of and High-throughput Protein Release from Polymer Film and Nanoparticle Libraries at JoVE.com

Combinatorial Synthesis of and High-throughput Protein Release from Polymer Film and Nanoparticle Libraries

该方法描述了组合化学合成的可生物降解的聚酸酐薄膜和纳米粒子库，，蛋白释放这些库和高通量检测。

组合合成和高通量的蛋白质释放的聚合物薄膜和纳米粒子库

Polyanhydrides are a class of biomaterials with excellent biocompatibility and drug delivery capabilities. While they have been studied extensively with ...

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