| Vicinal thiol-containing proteins (VDPs) play a principal role in maintaining an appropriate redox state of the protein and respond to various diseases. Therefore, it is of considerable significance to detect and separate VDPs in biological systems. Based on the specific fluorescent probe for VDPs and magnetic nanoparticles, we synthesized multifunctional nanoparticles for VDPs separation. In addition, we designed and synthesized fluorescent probes for carbonic anhydrases (CA Ⅰ, CA Ⅱ and CA ⅩⅢ).1. Based on magnetic nanoparticles and the novel fluorescent probe reported recently by our research group for selective detection of VDPs, we designed and synthesized fluorescent magnetic material VM, which contains modified functional molecules covalently grafted to the surface of the core-shell magnetic nanoparticles Fe3O4@SiO2, for simultaneous detection and separation of VDPs. With particle size of50～60nm, VM has good solubility, dispersion, and typicall superparamagnetism. VM exhibits good selectivity towards VDPs, not only labeling them by fluorescence, but also responsing rapidly and sensitively for protein enrichment, separation and extraction.2. According to the property of carbonic anhydrases (CA Ⅰ, CA Ⅱ and CA ⅩⅢ) for acetate and diethyl phosphate hydrolysis, we designed and synthesized BODIPY-and naphthyl-fluorescent probes (E2, E3, Z2, and Z3) based on the mechanism of intramolecular charge transfer (ICT), for CA Ⅰ, CA Ⅱ and CA ⅩⅢ detection. The results show that, E2and E3could be hydrolyzed respectively by CA Ⅰ and CA Ⅱ, with the probe’s wavelength and fluorescence emission intensity changing before and after the reaction, so they could be used as selective ratiometric fluorescent probes to distinguish CA Ⅰ and CA Ⅱ by time resolution. And in CA ⅩⅢ system, the fluorescence intensity of E2and E3are significantly enhanced (2.0and3.5times). They exhibits fast response and high sensitivity for CA ⅩⅢ. |
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