van Zanten Jacoba, Doornbos-Van der Meer Berber, Audouy Sandrine, Kok Robbert Jan, de Leij Lou
Department of PLG/Medical Biology, Academic Hospital Groningen, PO Box 30.001, 9700 RB Groningen, The Netherlands.
Cancer Gene Ther. 2004 Feb;11(2):156-64. doi: 10.1038/sj.cgt.7700668.
In this study, we developed a nonviral, cationic, targeted DNA-carrier system by coupling SAINT/DOPE lipids to monoclonal antibodies. The two monoclonal antibodies used were both tumor specific, that is, MOC31 recognizes the epithelial glycoprotein EGP-2 present in carcinomas and Herceptin recognizes the HER-2/neu protein in breast and ovarian cancers. Coupling was performed under nonreducing conditions by covalent attachment. The coupling procedure appeared to be reproducible and the binding capacity of the antibody was not affected by linking them to the cationic lipid. Binding and transfection efficiency was assayed with target cells and nontarget cells. SAINT/DOPE lipoplexes as such appeared to be an effective transfection reagent for various cell lines. After coupling SAINT/DOPE to the monoclonal antibodies or F(ab)2 fragments, it was shown that the targeted MoAb-SAINT/DOPE lipoplexes preferably bound to target cells, compared to binding to the nontarget cells, especially for the Herceptin-SAINT/DOPE lipoplexes. More importantly, transfection of the target cells could also be improved with these targeted lipoplexes. In conclusion, we have shown that by using monoclonal antibody-coupled SAINT/DOPE lipoplexes cells targeted gene delivery can be achieved, and also a higher number of transfected target cells was seen.
在本研究中，我们通过将SAINT/DOPE脂质与单克隆抗体偶联，开发了一种非病毒、阳离子、靶向性DNA载体系统。所使用的两种单克隆抗体均具有肿瘤特异性，即MOC31识别癌组织中存在的上皮糖蛋白EGP-2，而赫赛汀识别乳腺癌和卵巢癌中的HER-2/neu蛋白。偶联在非还原条件下通过共价连接进行。偶联过程似乎具有可重复性，并且抗体与阳离子脂质连接后其结合能力不受影响。使用靶细胞和非靶细胞检测结合和转染效率。SAINT/DOPE脂质体本身似乎是一种对各种细胞系有效的转染试剂。将SAINT/DOPE与单克隆抗体或F(ab)2片段偶联后，结果表明，与非靶细胞相比，靶向性单克隆抗体-SAINT/DOPE脂质体更倾向于与靶细胞结合，尤其是对于赫赛汀-SAINT/DOPE脂质体。更重要的是，这些靶向性脂质体也能提高靶细胞的转染效率。总之，我们已经表明，通过使用单克隆抗体偶联的SAINT/DOPE脂质体可以实现细胞靶向基因递送，并且可以观察到更多数量的转染靶细胞。
