携Gelsolin单抗靶向超声造影剂的制备及体外靶向实验

摘要

目的 制备一种以肿瘤淋巴转移相关特异性膜抗原蛋白——凝溶胶蛋白(Gelsolin)为靶点的纳米级高分子造影剂,并观察其体外寻靶及显影能力.方法 通过改良的双乳化法制备高分子PLGA-COOH造影剂,以碳二亚胺法将造影剂与Gelsolin单抗连接,制备出GSN-PLGA靶向超声造影剂.检测该造影剂的一般性质,评估Gelsolin单抗与造影剂表面连接情况,评价其体外寻靶能力并进行体外显影实验.结果 GSN-PLGA靶向超声造影剂平均粒径为(575.67±4.71)nm,表面电位(一11.46±1.19)mV,造影剂表面GSN单抗结合率达96.93％.体外摄取实验显示细胞表面高表达Gelsolin抗体的小鼠腹水型肝癌高淋巴转移细胞株(Hca-F细胞)可较多摄取GSN-PLGA靶向造影剂.体外显像实验显示靶向超声造影剂体外显影效果较好.结论 GSN-PLGA靶向超声造影剂可与高表达Gelsolin的Hca-F细胞特异性结合,且体外显影效果较好.%Objective To prepare a kind of Gelsolin-targeted ultrasound contrast agent (GSN-PLGA) and to explore its targeting and imaging effection in vitro.Methods The high molecular PLGA-COOH ultrasound contrast agents were prepared by a modified double emulsion technique and then conjugated with Gelsolin monoclonal antibody by carbodiimide technique.The physical property of contrast agent was determined.And the connectivity condition of ultrasound contrast agent with Gelsolin monoclonal antibody was estimated.The targeting ability and the effect of enhancing ultrasound imaging in vitro were explored.Results The average diameter of GSN-PLGA was (575.67 ± 4.71) nm.The potential was (-11.46±1.19) mV.And the binding rate of Gelsolin monoclonal antibody was 96.93％.In vitro experimentshowed more GSN-PLGA could be intaked by Hca-F cells and the ultrasound imaging cloud be enhanced greatly.Conclusion The GSN-PLGA nanoparticle can bind to Hca-F cells specifically and can enhance the ultrasound imaging greatly.