Rothdiener Miriam, Beuttler Julia, Messerschmidt Sylvia K E, Kontermann Roland E
Institute of Cell Biology and Immunology, University of Stuttgart, Stuttgart, Germany.
Methods Mol Biol. 2010;624:295-308. doi: 10.1007/978-1-60761-609-2_20.
Immunoliposomes generated by coupling of antibodies to the liposomal surface allow for an active tissue targeting, e.g., through binding to tumor cell-specific receptors. Instead of whole antibodies, single-chain Fv fragments (scFv), which represent the smallest part of an antibody containing the entire antigen-binding site, find increasing usage as targeting moiety. Here we provide protocols for the preparation of type II scFv immunoliposomes by the conventional coupling method as well as the post-insertion method. Furthermore protocols to analyze binding of these immunoliposomes to antigen-expressing cells as well as internalization through receptor-mediated endocytosis are included.
通过将抗体偶联到脂质体表面而产生的免疫脂质体能够实现主动组织靶向,例如通过与肿瘤细胞特异性受体结合。作为靶向部分,单链Fv片段(scFv)(代表抗体中包含整个抗原结合位点的最小部分)的使用越来越多,而不是使用完整抗体。在这里,我们提供了通过传统偶联方法以及后插入方法制备II型scFv免疫脂质体的方案。此外,还包括分析这些免疫脂质体与抗原表达细胞的结合以及通过受体介导的内吞作用进行内化的方案。