Wang Ruhung, Murali Vasanth S, Draper Rockford
Department of Biological Sciences, The University of Texas at Dallas, Richardson, TX, USA.
Department of Chemistry & Biochemistry, The University of Texas at Dallas, Richardson, TX, USA.
Methods Mol Biol. 2017;1530:147-164. doi: 10.1007/978-1-4939-6646-2_10.
Polyethylene glycol (PEG) and related polymers are often used in the solubilization and noncovalent functionalization of carbon nanomaterials by sonication. For example, carbon nanotubes are frequently sonicated with PEG-containing surfactants of the Pluronic series or phospholipid-PEG polymers to noncovalently functionalize the nanotubes. However, PEG is very sensitive to degradation upon sonication and the degradation products can be toxic to mammalian cells and to organisms such as zebrafish embryos. It is therefore useful to have a simple and inexpensive method to determine the extent of potential PEG sonolysis, as described in this chapter. Intact PEG polymers and degraded fragments are resolved on sodium dodecyl sulfate polyacrylamide gels by electrophoresis and visualized by staining with barium iodine (BaI). Digitized images of gels are acquired using a flatbed photo scanner and the intensities of BaI-stained PEG bands are quantified using ImageJ software. Degradation of PEG polymers after sonication is readily detected by the reduction of band intensities in gels compared to those of non-sonicated, intact PEG polymers. In addition, the approach can be used to rapidly screen various sonication conditions to identify those that might minimize PEG degradation to acceptable levels.
聚乙二醇(PEG)及相关聚合物常用于通过超声处理实现碳纳米材料的增溶和非共价功能化。例如,碳纳米管常与普朗尼克系列含PEG的表面活性剂或磷脂-PEG聚合物一起进行超声处理,以对纳米管进行非共价功能化。然而,PEG在超声处理时对降解非常敏感,其降解产物可能对哺乳动物细胞以及斑马鱼胚胎等生物体有毒性。因此,拥有一种简单且廉价的方法来确定潜在的PEG超声降解程度是很有用的,本章将对此进行描述。完整的PEG聚合物和降解片段通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳进行分离,并用碘化钡(BaI)染色进行可视化。使用平板照片扫描仪获取凝胶的数字化图像,并使用ImageJ软件对BaI染色的PEG条带强度进行定量。与未超声处理的完整PEG聚合物相比,通过凝胶中条带强度的降低可以很容易地检测到超声处理后PEG聚合物的降解。此外,该方法可用于快速筛选各种超声处理条件,以确定那些可能将PEG降解降至可接受水平的条件。