Targeting of Epidermal Growth Factor Receptor (EGFR)-Expressing Tumor Cells with Sterically Stabilized Affibody Liposomes (SAL)
Targeting of Epidermal Growth Factor Receptor (EGFR)-Expressing Tumor Cells with Sterically Stabilized Affibody Liposomes (SAL)
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DOI：
10.1021/bc900061v
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发表时间：
2009-06-01
影响因子：
4.7
通讯作者：
Kontermann, Roland E.
中科院分区：
文献类型：
--
作者：
Beuttler, Julia;Rothdiener, Miriam;Kontermann, Roland E.
Affibody 分子是源自蛋白 A 的 B 结构域的小而稳定的抗原结合分子。我们应用二价、高亲和力表皮生长因子受体 (EGFR) 特异性 Affibody 分子来生成靶向聚乙二醇化脂质体。这些空间稳定的亲和体脂质体 (SAL) 是通过将半胱氨酸修饰的亲和体分子与马来酰亚胺-PEG(2000)-DSPE 化学偶联并随后插入聚乙二醇化脂质体中而产生的。这些 SAL 对表达 EGFR 的肿瘤细胞系表现出强烈的选择性结合。结合取决于插入的亲和体分子-脂质缀合物的量，并且可以被可溶性 EGF 阻断。在 37 摄氏度下在人血浆中孵育 6 天后，仍保留约 30% 的结合活性。SAL 与细胞的结合导致脂质体的有效内化。使用负载米托蒽醌的脂质体，我们观察到与非靶向脂质体相比，SAL 对 EGFR 表达细胞的细胞毒性增强。总之，我们表明 SAL 可以很容易地从亲和体分子制备，因此可能适合开发用于靶向递送药物的载体系统。
Affibody molecules are small and stable antigen-binding molecules derived from the B domain of protein A. We applied a bivalent, high-affinity epidermal growth factor receptor (EGFR)-specific affibody molecule for the generation of targeted PEGylated liposomes. These sterically stabilized affibody liposomes (SAL) were produced by chemical coupling of the cysteine-modified affibody molecule to maleimide-PEG(2000)-DSPE and subsequent insertion into PEGylated liposomes. These SAL showed strong and selective binding to EGFR-expressing tumor cell lines. Binding was dependent on the amount of inserted affibody molecule-lipid conjugates and could be blocked by soluble EGF. Approximately 30% of binding activity was still retained after 6 days of incubation in human plasma at 37 degrees C. Binding of SAL to cells led to efficient internalization of the liposomes. Using mitoxantrone-loaded liposomes, we observed for SAL, compared to untargeted liposomes, an enhanced cytotoxicity toward EGFR-expressing cells. In summary, we show that SAL can be easily prepared from affibody molecules and thus may be suitable for the development of carrier systems for targeted delivery of drugs.