Enzymatic Hydrogelation-Induced Fluorescence Turn-Off for Sensing Alkaline Phosphatase in Vitro and in Living Cells
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- 作者：Ling Dong ; Qingqing Miao ; Zijuan Hai ; Yue Yuan ; Gaolin Liang
- 刊名：Analytical Chemistry
- 出版年：2015
- 出版时间：July 7, 2015
- 年：2015
- 卷：87
- 期：13
- 页码：6475-6478
- 全文大小：266K
- ISSN：1520-6882
文摘
Alkaline phosphatase (ALP)-catalyzed hydrogelation has been extensively explored and found wide applications. Spectroscopic and electrochemical approaches are commonly employed for the detection of ALP activity. Herein, by rational design of a fluorescence probe Fmoc-K(FITC)FFYp (P1) (where FITC is fluorescein), we incorporated sol鈥揼el transition with fluorescence 鈥渢urn-off鈥?and developed a new method for quantitative sensing ALP activity in vitro and in living cells. Under the catalysis of ALP, P1 was converted to hydrogelator Fmoc-K(FITC)FFY (1) which self-assembles into nanofibers to form Gel I. Accompanying this sol鈥揼el transition, the fluorescence emission of P1 was turned off. Our assay was employed to detect ALP activity over the range of 0鈥?.8 U/mL with a limit of detection (LOD) of 0.06 U/mL. ALP-inhibitor-treated cell imaging indicated that P1 could be applied for sensing ALP activity in living cells. Our method provides a new option for real time and quantitative sensing ALP activity in vitro and even in living cells.