Gel Electrophoresis Method to Measure the Concentration of Single-Walled Carbon Nanotubes Extracted from Biological Tissue
Gel Electrophoresis Method to Measure the Concentration of Single-Walled Carbon Nanotubes Extracted from Biological Tissue
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DOI：
10.1021/ac802485n
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发表时间：
2009-04-15
影响因子：
7.4
通讯作者：
Draper, Rockford K.
中科院分区：
文献类型：
--
作者：
Wang, Ruhung;Mikoryak, Carole;Draper, Rockford K.
提出了一种快速、灵敏的检测生物样品中单壁碳纳米管（SWNT）的方法。该方法使用聚丙烯酰胺凝胶电泳 (PAGE)，然后对 SWNT 带进行定量。该方法使用分散在牛血清白蛋白（BSA）中的单壁碳纳米管来开发。当 BSA-SWNT 分散体进行十二烷基硫酸钠 (SDS)-PAGE 时，BSA 通过浓缩胶，进入分离胶，并按预期向阳极迁移。然而，单壁碳纳米管在加样孔和堆积凝胶之间的界面处聚集成尖锐的带状。这些条带的数字化图像的强度与加载到凝胶上的单壁碳纳米管的量成正比，检测限为 5 纳克单壁碳纳米管。为了测试该方法，培养中的正常大鼠肾 (NRK) 细胞在暴露于含有不同浓度 BSA-SWNT 的培养基中不同时间和温度后，可以吸收 SWNT。细胞裂解物样品的 SDS-PAGE 分析表明，暴露于含有 98 μg/mL SWNT 的培养基后，BSA-SWNT 通过液相内吞作用以 30 fg/天/细胞的速率进入 NRK 细胞。
A rapid and sensitive method to detect single-walled carbon nanotubes (SWNTs) in biological samples is presented. The method uses polyacrylamide gel electrophoresis (PAGE) followed by quantification of SWNT bands. SWNTs dispersed in bovine serum albumin (BSA) were used to develop the method. When BSA-SWNT dispersions were subjected to sodium dodecyl sulfate (SDS)-PAGE, BSA passed through the stacking gel, entered the resolving gel, and migrated toward the anode as expected. The SWNTs, however, accumulated in a sharp band at the interface between the loading well and the stacking gel. The intensities from digitized images of these bands were proportional to the amount of SWNTs loaded onto the gel with a detection limit of 5 ng of SWNTs. To test the method, normal rat kidney (NRK) cells in culture were allowed to take up SWNTs upon exposure to medium containing various concentrations of BSA-SWNTs for different times and temperatures. The SDS-PAGE analyses of cell lysate samples suggest that BSA-SWNTs enter NRK cells by fluid-phase endocytosis at a rate of 30 fg/day/cell upon exposure to medium containing 98 mu g/mL SWNTs.