The Preparation Of Anti-FGFR β CD/Starch Composite Microspheres And Research On Targeting

Epidermal growth factor receptor (EGFR) in a variety normal epithelial cells and cancer cells is excessively increased in the gene copy number, or expression and activation. This is closely related to the tumor occurrence and development. For a new molecular target for the anti-tumor therapy, at present, both at home and abroad, the polysaccharide cyclodextrin nanoparticles used as drug carrier and bind to the tumor-related antibody as targeted drug delivery system has hardly been reported.In this thesis, β-cyclodextrin (β-CD) and starch was used as materials which were biodegradable, biocompatible, non-toxic, non-immunogenicity to prepare β CD/starch nanospheres with inverse microemulsion, and activated with epichlorohydrin to introduce epoxy, cross-linked with the anti-EGFR antibody, and to formed anti-EGFR immune microspheres. The influences of the actived process with epoxy density (EPD) were investigated and the conditions for activated process were optimized. Fluorescence nanoparticles (FNP) were prepared using FITC-labeled anti-EGFR antibody and cross-linked polysaccharide. FNP were injected into experimental mice, and direct immunofluorescence experiments were conducted to study its immunofluorescence and active targeting. The immunofluorescence experiments of breast, thyroid cancer were examined with anti-EGFR immune microspheres targeting cancer EGFR.In response to the initial activation, when the amount of epichlorohydrin, NaOH concentration, reaction time and temperature were all increased, EPD value was also increased until reached to a certain value, and then trended downward. Combined with single factor and orthogonal experiment, key factor was the reaction temperature to impact composite nanoparticles activation. The influence order was temperature>the amount of epichlorohydrin>reaction time>NaOH concentration. With0.1g microspheres reaction system when epichlorohydrin was1.5mL, NaOH concentration2.0mol/L, reaction temperature50℃, reaction time6h time, EPD value reached to2.5μmol·g-1that was the highest level of activation, and β CD/starch composite nanoparticles can be cross-linked by anti-EGFR antibody. A bright yellow-green fluorescence was observated with fluorescent microscope. In37℃, the amount of anti-EGFR antibody cross-linked β CD/starch composite nanoparticles to unitage SM was2.7mg/g under the optimum cross-linking condition. For fluorescence nanoparticles were injected into mice, fluorescent reaction to liver targeting of EGFR showed targeted characteristics for the immune nanoparticles. The immunofluorescence on breast cancer and thyroid cancer cells was found with the immune nanoparticles. High degree of expression of EGFR in the cytoplasm of cancer tissues was agreed with cancer HE staining of the biopsy. The immune response of early and late developmental cancer sections showed a different intensity of fluorescence.The anti-EGFR antibody and glycoprotein of nanoparticles were covalently cross-linked, and maintain the original antigen binding specific activity. For the clinical application of targeting therapy of cancer and cancer detection using the immune nanoparticles, more studies should be needed.