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用于高通量研究蛋白质-聚合物相互作用的聚合物微阵列表面等离子体共振成像

Surface plasmon resonance imaging of polymer microarrays to study protein-polymer interactions in high throughput.

作者信息

Hook Andrew L, Thissen Helmut, Voelcker Nicolas H

机构信息

School of Chemistry, Physics and Earth Sciences, Flinders University, Adelaide, 5001, South Australia, Australia.

出版信息

Langmuir. 2009 Aug 18;25(16):9173-81. doi: 10.1021/la900735n.

Abstract

Polymer microarrays provide a high-throughput format in which to assess biointerfacial interactions. This endeavor greatly assists with the development of advanced biomaterials. In order to increase the scope of this platform technology, the development of analytical tools that are compatible with the microarray format and are capable of analyzing biomolecular interactions in high throughput is needed. Here, we show that surface plasmon resonance imaging (SPRi) is such a tool. SPRi enables spatially resolved, surface sensitive, label free, real-time analysis of multiple surface-biomolecular interactions in parallel. In order to demonstrate this, we first printed phenylazide-modified polymers onto a slide coated with a low fouling base polymer. UV irradiation of the slide resulted in the cross-linking of the printed polymer spots to the surface. SPRi was then employed to study the adsorption and desorption of bovine serum albumin, collagen, and fibronectin to these adhesive microarray spots. The spots were also incubated with an adherent cell line, enabling insight into the underlying mechanisms of cell attachment to the polymers studied. For the system analyzed here, electrostatic interactions were shown to dominate cell attachment.

摘要

聚合物微阵列提供了一种高通量形式,可用于评估生物界面相互作用。这项工作极大地有助于先进生物材料的开发。为了扩大该平台技术的应用范围,需要开发与微阵列形式兼容且能够高通量分析生物分子相互作用的分析工具。在此,我们展示了表面等离子体共振成像(SPRi)就是这样一种工具。SPRi能够对多个表面生物分子相互作用进行空间分辨、表面敏感、无标记的实时并行分析。为了证明这一点,我们首先将叠氮苯基修饰的聚合物打印到涂有低污染基础聚合物的载玻片上。对载玻片进行紫外线照射导致打印的聚合物斑点与表面交联。然后采用SPRi研究牛血清白蛋白、胶原蛋白和纤连蛋白在这些粘性微阵列斑点上的吸附和解吸。这些斑点还与一种贴壁细胞系一起孵育,从而深入了解细胞附着在所研究聚合物上的潜在机制。对于此处分析的系统,静电相互作用被证明在细胞附着过程中起主导作用。

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