The Preparation Of New Targeting Hydrophobin HFBI-RGD And Self-assembly Of BODIPY Fluorescent Probe

Tumor has been one of the most terrible killers threatening human life.How to diagnose and therapy tumor is the aim of researchers.NIR fluorescent probe can be widely applied due to the range of emission wavelengths from 650 nm to 900 nm,which decreases the interference from spontaneous fluorescence in vivo.BODIPY is widely used as singal part because of its high molar absorption coefficients,high quantum yields,insensitivity to the environment condition,strong photostability,large Stokes shif and is easy to modify and so on.However,there are some problems which low water solubility and poor membrane permeability,restrict the application of BODIPY in vivo.Based on the above problems,the BODIPY was self-assembled with new targeting hydrophobin HFBI-RGD to get fluorescent probe-BODIPY-HFBI-RGD to improve the water solubility and biocompatibility of the original BODIPY dye,at the same time,the BODIPY dye had better targeting to tumor.(1)Firstly,two methods of Escherichia coli and Pichia pastoris were used to purify a new targeting hydrophobin HFBI-RGD by genetic engineering.(2)Then,a novel structure of BODIPY dye was first synthesized and characterized by ~1HNMR and MS,the optical properties were measured by UV-absorption and fluorescence emission intensity.(3)Finally,BODIPY dye self-assembed with HFBI-RGD to get a NIR fluorescent probe BODIPY-HFBI-RGD,which optical and biological properties were measured by fluorescence emission intensity,UV-absorption,FT-IR spectra,SEM,TEM,MTT,flow cytometry,fluorescent imaging in vivo and intravital imaging.Experiment results demonstrated that BODIPY-HFBI-RGD had the excellent property of NIR fluorescence(720 nm)compared to original BODIPY dye,at the same time,it had the good water solubility and biocompatibility,especially for U87MG tumor cells with high integrin expression.