Combination of on-chip field amplification and bovine serum albumin sweeping for ultrasensitive detection of green fluorescent protein.
Combination of on-chip field amplification and bovine serum albumin sweeping for ultrasensitive detection of green fluorescent protein.
复制标题
DOI：
10.1021/ac9007607
复制
发表时间：
2009-07-01
影响因子：
7.4
通讯作者：
Liu, Shaorong
中科院分区：
文献类型：
--
作者：
Pan, Qiong;Zhao, Meiping;Liu, Shaorong
报道了一种结合场放大进样(FasI)和牛血清白蛋白(BSA)扫描的高效芯片上预富集方法，用于在一个简单的跨通道微芯片设备上超灵敏地检测绿色荧光蛋白(GFP)。通过平衡流体动力流和电渗流(EOF)形成停滞的样品/运行缓冲液边界，GFP分子可以连续地注入样品加载通道并堆积。我们还证明，与常用的胶束相比，牛血清白蛋白是一种非常有效的蛋白质浓度的假固定相。FASI和牛血清白蛋白扫描相结合，GFP的浓缩系数为3570，检出限为8.4 pm。利用这种方法，我们分离出了GFP和GFP-胰岛素样生长因子-I(GFP-IGF-I)融合蛋白。整个分析(GFP浓缩、基质消除和电泳法分离)可以在5分钟内完成。此外，我们还成功地将该方法应用于检测大肠杆菌细胞中GFP的表达和单个大肠杆菌细胞中的GFP含量。
We report a highly effective on-chip preconcentration method by combining field-amplified sample injection (FASI) and bovine serum albumin (BSA) sweeping for ultrasensitive detection of green fluorescent protein (GFP) on a simple cross-channel microchip device. With the formation of a stagnant sample/running buffer boundary by balancing the hydrodynamic flow and the electroosmotic flow (EOF), GFP molecules can be continuously injected into the sample loading channel and stacked. We have also demonstrated that BSA is a very effective pseudo-stationary phase for sweeping concentration of proteins in comparison to the commonly used micelles. The combination of FASI and BSA sweeping yields a concentration factor of 3570 and a limit of detection of 8.4 pM for GFP. Using this method, we have separated GFP and GFP-insulin-like growth factor-I (GFP-IGF-I) fusion protein. The entire assay (GFP concentration, matrix elimination, and electrophoretic separation) can be completed within <5 min. Furthermore, we have successfully applied this method for the detection of GFP expression of E. coli cells and the GFP content in single E. coli cells.
登录
查看更多内容
影响因子：
4.1
作者：
Feng, Yong-Lai;Lian, Hongzhen;Zhu, Jiping
通讯作者：
Zhu, Jiping
影响因子：
2.9
作者：
Jung, B;Bharadwaj, R;Santiago, JG
通讯作者：
Santiago, JG
影响因子：
4.1
作者：
Markuszewski, MJ;Britz-McKibbin, P;Nishioka, T
通讯作者：
Nishioka, T
影响因子：
7.4
作者：
Palmer, J;Burgi, DS;Landers, JP
通讯作者：
Landers, JP
影响因子：
7.4
作者：
Shackman, Jonathan G.;Ross, David
通讯作者：
Ross, David