Wang Ruhung, Mikoryak Carole, Chen Elena, Li Synyoung, Pantano Paul, Draper Rockford K
Department of Molecular and Cell Biology, The University of Texas at Dallas, Richardson, Texas 75080, USA.
Anal Chem. 2009 Apr 15;81(8):2944-52. doi: 10.1021/ac802485n.
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 microg/mL SWNTs.
本文介绍了一种快速灵敏的检测生物样品中单壁碳纳米管（SWNTs）的方法。该方法采用聚丙烯酰胺凝胶电泳（PAGE），随后对SWNT条带进行定量分析。使用分散在牛血清白蛋白（BSA）中的SWNTs来开发此方法。当BSA-SWNT分散液进行十二烷基硫酸钠（SDS）-PAGE时，BSA穿过堆积胶，进入分离胶，并如预期那样向阳极迁移。然而，SWNTs在加样孔和堆积胶之间的界面处积聚成一条清晰的条带。这些条带数字化图像的强度与加载到凝胶上的SWNTs量成正比，检测限为5 ng SWNTs。为了测试该方法，将培养的正常大鼠肾（NRK）细胞暴露于含有不同浓度BSA-SWNTs的培养基中，在不同时间和温度下摄取SWNTs。细胞裂解物样品的SDS-PAGE分析表明，当暴露于含有98μg/mL SWNTs的培养基中时，BSA-SWNTs以每天30 fg/细胞的速率通过液相内吞作用进入NRK细胞。
