Preparation Of Oxaliplatin Liposome Conjugated With Sevacizumab And Its Targeting To HCC Cells In Vitro

Objective: The purpose of this study is to prepare oxaliplatin liposomes modified by sevacizumab,an anti-VEGF monoclonal antibody,in order to increase the targeting of oxaliplatin to hepatocellular carcinoma cells and achieve high-efficiency and low-toxicity.The cell proliferation inhibition and targeting of the immunoliposome to the HCC cell line were evaluated by MTT in vitro and fluorescence-labeled liposome.Methods: The encapsulation rate of oxaliplatin immunoliposomes was determined by high performance liquid chromatography,and a suitable method for preparing liposomes is selected according to the encapsulation rate.The morphology,particle size,encapsulation rate,antibody coupling rate,antibody activity,in vitro release rate and other properties of the immunoliposomes prepared by this method were examined.Taking human hepatocellular carcinoma cells line Bel7402 and HepG2 as experimental subjects,the toxicity and uptake capacity of cells to oxaliplatin immunoliposomes were observed using MTT assay and fluorescence-labeled liposome,and the in vitro targeting of immunoliposomes was initially examined.Resaults: Oxaliplatin immunoliposomes were successfully prepared,with an average entrapment efficiency of 15.91%,an average particle size of 182.9 nm,and an average potential of-29.2 mV.The liposome was observed under electron microscope and the shape is spherical.The antibody coupling rate was 43.76% and the activity remained at 84.38%.After 96 hours,the cumulative release rate was 82.34%.The results of MTT cytotoxicity test showed that the carrier material has no obvious toxicity to the cells,and the cytotoxicity of oxaliplatin immunoliposome to hepatocellular carcinoma cells Bel7402 and HepG2 was stronger than that of ordinary oxaliplatin liposome and free oxaliplatin.Confocal microscopy observations showed that the cellular uptake of oxaliplatin immunolipids was higher than that of oxaliplatin liposomes.Preliminary determination of oxaliplatin immunoliposome has a certain targeting.Conclusions: In this study,oxaliplatin immunoliposomes were successfully prepared,and the immunoliposomes had a uniform particle size distribution and a low encapsulation efficiency,but the antibody activity was maintained at a high level.The oxaliplatin immunoliposome has the ability to actively target the HCC cell line Bel7402 and HepG2,and can inhibit the growth of the HCC cell line,and it is worth further study.