Preparation Of Multifunctional PLGA-ENO1mAb-H1 Nanoparticles And Its Experimental Study On Anti-cervical Cancer

Objective The experiment was performed by constructing ENO1-H1 monoclonal antibody(ENO1 m Ab-H1)loaded multifunctional PLGA nanoparticles(PLGA NPs)to achieve the intracellular delivery,and then investigated the therapeutic effect of ENO1m Ab-H1 nanoparticles on cervical cancer in vitro.Methods 1.The ENO1 monoclonal antibody was prepared by induced ascites in vivo.It was preliminarily purified through bitterness-ammonium sulfate precipitation from the ascites and further purified by protein A affinity chromatography.2.The FA-SS-PLGA copolymers was prepared by amidation reaction.3.Blank PLGA NPs,PLGA-ENO1m Ab-H1NPS and FA-SS-PLGA-ENO1m Ab-H1 NPs were prepared by double emulsion solvent volatilization method and characterized by corresponding instruments.4.To observe the uptake of drug-loaded nanoparticles in Hela cells using a fluorescence microscope.5.To investigate the inhibitory effect of ENO1 m Ab-H1 nanoparticles on glycolysis in Hela cells through the glycolysis kit.6.To evaluate the inhibitory effect of ENO1m Ab-H1 nanoparticles in Hela cells of cervical cancer by MTT test,scratch test and plate cloning formation experiment.Result 1.SDS-PAGE electrophoresis results showed ENO1m Ab-H1 had higher purity,and there were bands at the molecular weights of 50k Da and 25k Da corresponding to the positions of heavy chains and light chains of Ig G.2.FA-SS-PLGA copolymers were prepared successfully,and verified by FT-IR and ~1H-NMR.3.ENO1m Ab-H1 loaded multifunctional PLGA nanoparticles with a particle size less than 200nm were prepared successfully.4.The results of cell uptake experiments showed that the uptake of FA-PLGA NPs in Hela cells was higher than PLGA NPs.5.The inhibition of FA-SS-PLGA-ENO1m Ab-H1 nanoparticles on lactic acid and pyruvate in Hela cells was stronger than 3-Br PA(P<0.05).6.The cell proliferation experiment found that FA-SS-PLGA-ENO1m Ab-H1 targeting nanoparticles had a higher inhibitory effect on Hela cell proliferation than PLGA-ENO1m Ab-H1nanoparticles,and was weaker than 3-Br PA(P<0.05).In the scratch experiment,the migration inhibition effect of free ENO1m Ab-H1 showed the strongest in Hela cells after24h co-incubation,and followed by FA-SS-PLGA-ENO1m Ab-H1 NPs,PLGA-ENO1m Ab-H1 NPs and 3-Br PA(P<0.05).In the clone formation experiment,both FA-SS-PLGA-ENO1m Ab-H1 NPs and PLGA-ENO1m Ab-H1 NPs inhibited the formation of Hela cell clone balls compared with the negative control group(P<0.05).Conclusion 1.multifunctional PLGA-ENO1 m Ab-H1 NPs can inhibit the glycolysis in Hela cells of cervical cancer significantly.2.multifunctional PLGA-ENO1 m Ab-H1 NPs can inhibit the proliferation,migration and clone formation in Hela cells of cervical cancer.