靶向超声微泡MBICAM-1的制备及靶向实验研究

Abstract

目的 制备载抗ICAM-1(细胞间黏附分子-1)的靶向超声微泡MBICAM1,鉴定制备微泡的基本性质,研究其在体内及体外的靶向能力.方法 通过“生物素-亲和素”桥接法将抗ICAM-1结合到生物素化脂质微泡,制成MBICAM1;检测制备微泡的浓度、粒径,观察微泡的形态;检测抗ICAM-1与微泡的结合率;ICAM-1质粒转染Hela细胞,观察转染后的Hela细胞与MBICAM-1结合,验证MBICAM-1体外靶向功能;制备家兔左侧跟腱炎症模型,用制备的微泡对家兔双侧跟腱行造影检查,验证MBICAM-1体内靶向能力.结果 三种微泡平均粒径1.00～2.4 μm,浓度2.4×108～1010/ml;微泡的形态完整、规则,分布较匀;微泡和抗ICAM-1单抗的平均结合率为(86.5±5.3)％.MBICAM-1环绕在转染后的Hela细胞周围.MBICAM-1对家兔左侧跟腱造影呈高增强.结论 采用“生物素-亲和素”桥接法可制备MBICAM-1;MBICAM-1在体外、体内均具有靶向功能.

Objective To identify the basic properties of the prepared targeted ultrasound mierobubble MBICAM-1,and evaluated the targeting ability of MBICAM-1 in vitro and in vivo.Methods Prepared MBbiotm by biotin-avidin bridging chemistry method,with which binding the rabbit anti-ICAM-1 monoclonal antibody to the biotinylated lipid microbubbles surface.Detected the size and concentration of the prepared microbubbles and observed the morphology of them.Detecting the binding rate of rabbit anti-ICAM-1 monoclonal antibody and microbubbles.Transfected ICAM-1 plasmid into the Hela cells,observed the binding condition between the successfully transfected Hela cells and MBICAM-1 to ev alate the targeting ability of MBICAM-1 in vitro.Prepared rabbits left Achilles tendinitis model,used the microbubbles for CEUS,comparing the ultrasonography of bilateral Achilles tendon to evalate the targeting ability of MBICAM-1 in vivo.Results The average size of three kinds microbubbles was 1.00-2.4 μm,while the concentration was 2.4 ×108 10/ml.The three kinds of microbubbles were morphology completeness,rules and evenly distributed.The average binding rate of anti-ICAM-1 monoclonal antibody with microbubble was (86.5±5.3)％.MBICAM-1 binded with the surface of transfected Hela cells under microscope.MBICAM-1 showed high enhancement on the left Achilles tendon.Conclusions MBIcAM-1 was successfully prepared by biotin-avidin bridging chemistry method and have a good targeting ability both in vitro and in vivo.