Functionalizing αvβ3- or α5β1-Selective Integrin Antagonists for Surface Coating: A Method To Discriminate Integrin Subtypes In Vitro
Functionalizing αvβ3- or α5β1-Selective Integrin Antagonists for Surface Coating: A Method To Discriminate Integrin Subtypes In Vitro
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DOI：
10.1002/anie.201206370
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发表时间：
2013-01-01
影响因子：
16.6
通讯作者：
Kessler, Horst
中科院分区：
文献类型：
--
作者：
Rechenmacher, Florian;Neubauer, Stefanie;Kessler, Horst
在一个典型的合成中，7 mg/mL聚苯乙烯(154)-嵌段聚（2-乙烯基吡啶）（33）（PS154-b-P2VP33，聚合物源）在室温下溶解在对二甲苯（Sigma-Aldrich, St. Louis， USA）中，搅拌2天。相对于P2VP单元数（NP2VP）计算无机前驱体的数量，并定义为加载参数(L)。L在所有实验中都保持不变，等于0.5（即每2个P2VP单位中有1个HAuCl4分子）。将四氯酸氢（III）三水合物（HAuCl4∙3H2O; Sigma-Aldrich）加入嵌段共聚物溶液中，在密封玻璃容器中搅拌2天。玻璃盖（卡尔罗斯，卡尔斯鲁厄，德国）在水虎鱼溶液中清洁至少5小时，用MilliQ水广泛冲洗，并在氮气流下干燥。以24mm /min的恒定速度将玻璃盖盖浸入先前制备的溶液中制备胶束单层膜。最后，为了去除有机模板并形成无机纳米颗粒，将浸涂玻片暴露于氧等离子体（150 W, 0.15 mbar, 45 min， PVA TEPLA 100等离子体系统）中。为了防止非特异性蛋白质吸附或细胞结合，金纳米颗粒周围或之间的底物区域用聚乙二醇（PEG，分子量2000）共价修饰然后将聚乙二醇功能化的底物置于100 μl的100 μM乙醇溶液中，用拟肽剂对金纳米颗粒进行功能化。然后用乙醇和水彻底冲洗底物，在水中孵育过夜，最后用氮气干燥。
In a typical synthesis, 7 mg/mL of polystyrene (154)-block-poly (2-vinylpyridine)(33)(PS154-b-P2VP33, Polymer Source) was dissolved at room temperature in p-xylene (Sigma-Aldrich, St. Louis, USA) and stirred for 2 days. The quantity of inorganic precursor was calculated relative to the number of P2VP units (NP2VP) and defined as the loading parameter (L). L was kept constant in all experiments, equal to 0.5 (ie, 1 molecule of HAuCl4 per 2 P2VP units). Hydrogen tetrachloroaurate (III) trihydrate (HAuCl4∙ 3H2O; Sigma-Aldrich) was added to the block copolymer solution and stirred for 2 days in a sealed glass vessel.Glass coverslips (Carl Roth, Karlsruhe, Germany) were cleaned in a piranha solution for at least 5 h and extensively rinsed with MilliQ water and dried under a stream of nitrogen. Micellar monolayers were prepared by dip-coating a glass coverslip into the previously prepared solutions with a constant velocity equal to 24 mm/min. Finally, to remove the organic template and form inorganic nanoparticles, the dip-coated glass slides were exposed to oxygen plasma (150 W, 0.15 mbar, 45 min, PVA TEPLA 100 Plasma System). To prevent nonspecific protein adsorption or cell binding, the substrate area surrounding or in between the gold nanoparticles was covalently modified with polyethylene glycol (PEG; molecular weight 2000).[1] The gold nanoparticles were then functionalized with peptidomimetics by incubating the PEG-functionalized substrates in 100 μl of a 100 μM ethanolic solution. The substrates were then thoroughly rinsed with ethanol and water, incubated overnight in water, and finally dried with nitrogen.