静电吸附法制备抗αvβ_3靶向脂质微泡及体外寻靶实验

Preparation and in vitro evaluation of lipid-coated microbubbles targeted to αvβ_3 integrin

目的探讨静电吸附方法制备携带抗αvβ3单克隆抗体的靶向微泡的可能性,并初步评价该靶向微泡在体外的寻靶能力。方法将适量的负电荷脂质微泡造影剂与抗αvβ3单克隆抗体混合,利用静电吸附方法,制备亲黑色素瘤的靶向超声造影剂,并通过观察与黑色素瘤细胞的结合情况,验证该靶向微泡的靶向性。结果静电吸附方法能够使抗体与微泡结合。携带抗αvβ3单克隆抗体的靶向微泡粘附在黑色素瘤细胞表面的靶向微泡数目明显多于普通对照微泡。结论静电吸附法能够制备脂质靶向超声微泡,体外亲肿瘤细胞的寻靶试验验证了携带抗αvβ3单克隆抗体的靶向超声微泡具有一定的寻靶能力。

Objective To prepare microbubble ultrasound contrast agent specifically targeted to αvβ3 integrin using static electric binding.Methods Conjugation of αvβ3 integrin monoclonal antibody to negatively charged lipid microbubbles was tested by simply mixing, pH value adjusting,and incubation.Then the suspension was washed twice with phosphate- buffered saline （PBS）.The targeting ability of the microbubbles conjugated with αvβ3 integrin antibody was verified by adding the αvβ3 integrin antibody conjugated microbubbles to cultivated k1735 M2 melanin tumor cells. Results The αvβ3 targeted microbubbles were found to actively adhere to the melanin tumor cells. Much more targeted microbubbles were found around the tumor cells compared with the control group. Conclusions The conjugation of αvβ3 integrin monoclonal antibody to negatively charged hpid - wated microbubbles is possible by static electrical binding. The targeted microbubbles are apt to adhere to the melanoma cells in vitro.