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egfr纳米抗体功能化牛奶外泌体用于药物靶向递送
首发时间：2022-01-25
摘要：肿瘤病灶部位的药物靶向递送是改善化疗药物临床疗效的一种重要策略。本文以牛奶来源的外泌体（milk exosomes，mexo）为药物载体，借助甘氨酸修饰的膜插入脂性分子nh2-gg-peg2000-dspe和sortase a酶介导的偶联反应，通过对mexo进行表皮生长因子受体（epidermal growth factor receptor, egfr）纳米抗体7d12的功能化修饰，构建了靶向肿瘤egfr的外泌体药物递送系统mexo-7d12。western blot、电镜形态、动态光衍射等表征结果显示mexo-7d12构建成功；细胞免疫荧光、流式细胞术等分析表明，mexo-7d12对抗肿瘤药物阿霉素（adriamycin，dox）的递送具有egfr靶向特性；细胞毒性评估结果显示，装载dox的mexo-7d12对高efgr表达肿瘤细胞具有较高的细胞毒性。本研究结果表明, 纳米抗体7d12功能化的牛奶外泌体载药体系mexo-7d12具有对肿瘤药物dox的靶向递送能力，其能够特异性靶向高egfr表达的肿瘤细胞并对其具有一定的选择性杀伤作用。
关键词： 靶向递送 牛奶外泌体 纳米靶向载药体系 表皮生长因子受体 7d12 阿霉素
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egfr nanobody-functionalized bovine milk exosomes for target-specific drug delivery
abstract：target-specific delivery of chemotherapy drugs to tumor site is an important path to improve the overall clinical outcomes. in this work, we reported an egfr-targeted drug delivery system using milk exosomes (mexo) functionalized with egfr nanobody 7d12. the mexo was first equipped with a glycine-modified membrane inserting molecule nh2-gg-peg2000-dspe through its hydrophobic tail, which was further fused with the nanobody 7d12 through a srta-mediated enzymatic ligation. the successful construction of 7d12-mexo was confirmed by western blotting analysis, electronic microscope, and dynamic light scattering (dls). immunocytochemistry and flow cytometry analysis of 7d12-mexo loaded with the antineoplastic drug adriamycin (dox) demonstrated that it can target and deliver dox to cells in response to the expression of egfr. finally, the cytotoxicity test showed that 7d12-mexo loaded with dox was more effective to kill tumor cells with high egfr expression. taken together, these results demonstrated that 7d12-fucnationalized mexo as a target-specific delivery system was able to deliver the cargo drug of dox to egfr expressing cell, and selectively kill the tumor cells.
keywords： target-specific delivery milk exosomes target target-specific nanoparticle egfr 7d12 dox
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