Dual-targeted microbubbles carrying both anti-CD34 monoclonal antibody and anti-ICAM-1 monoclonal antibody:preparation and investigation of its targeting performance in vitro

Abstract

Objective To prepare dual-targeted microbubbles carrying both anti-CD34 monoclonal antibody and anti-ICAM-1 monoclonal antibody,to identify its basic characteristics,and to investigate its targeting performance in vitro.Methods ICAM-1-targeted microbubbles (MgIcAM-1),CD34-targeted microbubbles(MBcD34) and dual-targeted (with both monoclonal antibodies) microbubbles (MBD) were prepared by biotin-avidin bridging chemistry method.The targeted microbubbles were observed under light microscrope and characterized by a Mastersizer 3000,DFY software,laser confocal microscopy,and flow cytometry.The targeting specificity and attachment capability of dual-targeted microbubbles to endothelial progenitor cells (EPCs) and damaged human umbilical vein endothelial cells (HUVECs) were assessed in vitro.Results The morphous,diameter,surface potential,concentration,and the antibody-binding rates of microbubbles were not statistically significant in MBIcAM-1,MBCD34,and MBD (P ＞0.05).Echo intensity in MBD significantly increased compared with MBICAM1,MBCD34,and MBiotin (P ＜ 0.01).The targeting experiment in vitro showed that the attachment rate of MBD to EPCs and damaged HUVECs significantly increased compared with MBiotin (P ＜0.01),while no remarkable difference in attachment rate was found between MBD and MBcD34 in EPCs,and between MBD and MBIcAM-1 in damaged HUVECs (P ＞0.05).Conclusions Dual-targeted microbubbles carrying both anti-CD34 monoclonal antibody and anti-ICAM-1 monoclonal antibody are prepared successfully.The study in vitro has proved that the dual-targeted microbubbles can specifically bind to both EPCs and damaged HUVECs. Key words: Ultrasonography; Microbubbles; Endothelial cells ; Intercellular adhesion molecule-1