Establishment And Evaluation Of EGFR Single Domain Antibody-iRGD Dual-functionalized Nanoparticles For Tumor Targeting And Anti-tumor Efficiency

Purpose:Prepare EGFR single domain antibody-iRGD dual-functionalized nanoparticles.The anticancer activity and tumor targeting of the new drug delivery in vivo and in vitro were evaluated.Methods:The EGFR single domain antibody-iRGD recombinant protein was conjugated to the paclitaxel-silk fibroin nanoparticles(PTX-SF-NPs)with the carbodiimide-mediated coupling procedure.The basic characteristics were investigated with dynamic light scattering,transmission electron microscopy and Fourier transformed infrared spectroscopy.The conjugation efficiency was also quantitatively determined.The anticancer activity and tumor targeting of the new nanoparticles(A-PTX-SF-NPs)in vivo and in vitro were evaluated using high EGFR expressing cell line.Results:The diameter of A-PTX-SF-NPs was approximately 186nm,which was greater than PTX-SF-NPs.The FTIR spectrums of PTX-SF-NPs and A-PTX-SF-NPs were similar,meanwhile A-PTX-SF-NPs contained more carboxy group.The conjugation efficiency was 76.27%±12.29%approximately.In the cellular uptake assay,Coumarin-6 loaded NPs were used to coincubate with Hela cells,a high EGFR expressing cell line.The result demonstrated that the new nanoparticles were more targetable to cells with high EGFR expression.In vitro cytotoxicity study,A-PTX-SF-NPs exhibited significantly increased cytotoxicity compared with PTX-SF-NPs in Hela cells.A-PTX-SF-NPs group showed slower tumor growth smaller tumor volumes than PTX-SF-NPs in vivo antitumor efficacy assay.The real time NIR fluorescence imaging study demonstrated that A-PTX-SF-NPs accumulated more in tumor in tumor tissure than PTX-SF-NPs.Conclusions:The anticancer activity and tumor targeting of A-PTX-SF-NPs were superior to PTX-SF-NPs and may become a potential future clinical cancer treatment regimen.