Preparation of gelatin nanoparticles with EGFR selection ability via biotinylated-EGF conjugation for lung cancer targeting

Lung cancer is the most malignant cancer today, and specific drug delivery has been developed for superior outcome. In this study, gelatin nanoparticles (GPs) were firstly employed as native carriers. Second, NeutrAvidin ^FITC was then grafted on the particle surface (GP-Av); finally much more amount of biotinylated EGF were able to be conjugated with NeutrAvidin ^FITC forming ligand- binding nanoparticles (GP-Av-bEGF) to enhance the targeting efficiency. These nanoparticles were applied as EGFR-seeking agents to detect lung cancer cells. Results of particle characterization show that the modification process had no influence on size (230 nm). Round and smooth nanoparticles were observed by AFM. The surface property of nanoparticles was characterized by surface plasmon resonance (SPR) and flowcytometry analysis as well as by examining the interaction of the modified EGF on particle surface with the ability to recognize EGFR. The binding ability of GPs with or without EGF modification is different. SPR assay showed that EGF-conjugated particles (GP-Av-bEGF) have stronger and faster bonding signal than the unmodified one (GP-Av). Free EGF competition results from SPR and A549 cell (lung adenocarcinoma cells) culture also confirmed the EGF receptormediated endocytosis mechanism for nanoparticles with EGF-modified binding. The in vitro targeting ability was confirmed by the uptake rate of different cells via flow cytometry assay. GP-Av-bEGF resulted in higher entrance efficiency on A549 than on normal lung cells (HFL1) and U2-OS (osteosarcoma cells) due to A549 possessing more amounts of EGFR. The targeting ability of GP-Av-bEGF nanoparticles with specific EGFR tracing ability was proved, which holds promise for further anticancer drug applications.