Real-time Ratiometric Fluorescent Assay for Alkaline Phosphatase Activity with Stimulus Responsive Infinite Coordination Polymer Nanoparticles

Transcription

1 Supporting information Real-time Ratiometric luorescent Assay for Alkaline Phosphatase Activity with Stimulus Responsive Infinite Coordination Polymer Nanoparticles Jingjing Deng, Ping Yu, Yuexiang Wang, Lanqun Mao * Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, the Chinese Academy of Sciences (CAS), Beijing 119, China. * Corresponding Author. ax: , lqmao@iccas.ac.cn. S 1

2 S1. luorescent Spectra of the Supernatant Solution and the Suspension of Nanoparticles after Several Times of Washing (E x = 315 nm) Supernatant Solution Suspension igure S1. luorescent spectra of the dispersion (1 ml) containing courmarin@tb-gmp nanoparticles (.784 mg) after several times of washing. The curves show the fluorescent spectra of the supernatant of the dispersion (1 ml, black curve) and the courmarin@tb-gmp nanoparticles re-dispersed in water (1 ml, red curve). E x = 315 nm. The supernatant solution of courmarin@tb-gmp nanoparticles showed almost no fluorescent signal, indicating the free courmarin or courmarin absorbed on the surface of Tb-GMP network was clearly removed. S 2

3 S2. luorescent Spectra of Nanoparticles Dispersed in the Buffer Solution with Excitation at Different Wavelengths nm 3 nm 35 nm nm nm 6 32 nm nm 6 33 nm igure S2. luorescent spectra of the coumarin@tb-gmp ICP nanoparticles (.784 mg) dispersed in 1 ml Tris-HCl (5 mm, ph 8.) when excited at different wavelengths. S 3

4 S3. luorescent Spectra of Nanoparticles Dispersed in the Buffer with Excitation at 315 nm. 25 Coumarin 25 RhB AY(cationic dye) 25 2 Indigo blue (anionic dye) igure S3. luorescent spectra of the coumarin@tb-gmp ICP nanoparticles (.784 mg), RhB@Tb-GMP nanoparticles (.784 mg), AY@Tb-GMP nanoparticles (.784 mg) and Indigo@Tb-GMP nanoparticles (.784 mg) dispersed in 1 ml Tris-HCl (5 mm, ph 8.) buffer when excited at 315 nm. Herein, in order to establish the ratiometric ALP assay, the encapsulated guest molecules must be carefully screened. When being excited at 315 nm, Tb-GMP ICP exhibited very broad fluorescence emission from 498 nm to 628 nm, the guest molecules must meet the following requirements to avoid cross-interference. (1) They could be exited at 315 nm. (2) The emission spectra of the guest molecules do not overlap with that of the Tb-GMP. (3) The guest molecules should have a high fluorescent quantum yield. To choose S 4

5 the guest molecules, different kinds of fluorescent dyes such as rhodamine B (RhB), a cationic dye acridine yellow (AY) and an anionic dye indigo blue were incorporated into Tb-GMP network, respectively. After several times of washing, the fluorescent spectra of the buffer dispersion consisting of dye@tb-gmp nanoparticles were excited at 315 nm. As shown in igrue S3, only the fluorescent dye courmarin showed a clear fluorescent emission at 45 nm that did not overlap with the emission spectra of Tb-GMP. As a consequence, 7-amino-4-methyl coumarin (coumarin) was employed as guest molecules to form coumarin@tb-gmp nanoparticles for ratiometric ALP detection in this study. S 5

6 S4. luorescent Spectra and Photographs of the Buffer Dispersion Containing Nanoparticles with Different ALP Activity U/mL 1. U/mL 3.75 U/mL U/mL.2 U/mL.1 U/mL U/mL U/mL igure S4. luorescent spectra of the dispersion prepared by adding ALP (2 μl, 1.5 U, 1. U,.75 U,.5 U,.2 U,.1 U,.5 U,.25 U) into the dispersion of 1 ml Tris-HCl (5 mm, ph 8.) buffer containing coumarin@tb-gmp ICP nanoparticles (.784 mg). luorescent spectra were consecutively recorded every 1 min after the addition of ALP at 37 C. Inset: photographs of the dispersion at 2 min after the addition of different activities of ALP and excited by a 315 UV lamp. S 6

7 S5. luorescent Spectra of Buffer Dispersion Containing Nanoparticles with 2 mm Na 3 VO 4 for 2 min igure S5. luorescent spectra of the dispersion prepared by adding Na 3 VO 4 (2 µl, 2 mm) -treated ALP (2 µl,.2 U) into 1 ml Tris-HCl (5 mm, ph 8.) buffer containing coumarin@tb-gmp nanoparticles (.784 mg). luorescent spectra were consecutively recorded every 1 min after the addition of Na 3 VO 4 at 37 C for 2 min. S 7