Anti-CD20 single chain variable antibody fragment-apolipoprotein A-I chimera containing nanodisks promote targeted bioactive agent delivery to CD20-positive lymphomas
Anti-CD20 single chain variable antibody fragment-apolipoprotein A-I chimera containing nanodisks promote targeted bioactive agent delivery to CD20-positive lymphomas
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DOI：
10.1139/bcb-2015-0009
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发表时间：
2015-08-01
期刊：
影响因子：
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通讯作者：
Ryan, Robert O.
中科院分区：
文献类型：
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作者：
Crosby, Natasha M.;Ghosh, Mistuni;Ryan, Robert O.
构建了包含α-CD20单链可变片段（scFv）抗体、间隔肽和人载脂蛋白（apo）A-I的融合蛋白并在大肠杆菌中表达。该嵌合体保留了 apoA-I 固有的脂质相互作用特性以及 scFv 的抗原识别特性。 scFv 中心点 apoA-I 被配制为纳米级重构高密度脂蛋白颗粒（称为纳米盘；ND）并与培养细胞一起孵育。 α-CD20 scFv中心点apoA-I ND与CD20阳性非霍奇金淋巴瘤(NHL)细胞(Ramos和Granta)结合，但不与CD20阴性T淋巴细胞(即Jurkat)结合。与 NHL 细胞的结合被利妥昔单抗（一种针对 CD20 的单克隆抗体）预孵育部分抑制。与用固有荧光疏水性多酚姜黄素配制的 α-CD20 scFv 中心点 apoA-I ND 孵育后，对 Granta 细胞进行共聚焦荧光显微镜分析，结果显示 α-CD20 scFv 中心点 apoA-I 定位于细胞表面，而姜黄素卸载并进入细胞。与对照温育相比，在与含有姜黄素的α-CD20 scFv中心点apoA-I ND温育后，培养的NHL细胞的活力降低。因此，姜黄素 ND 与 α-CD20 scFv 中心点 apoA-I 作为支架成分的配方赋予细胞靶向性和增强的生物活性剂递送，提供了一种最小化与化疗剂相关的毒性的策略。
A fusion protein comprising an alpha-CD20 single chain variable fragment (scFv) antibody, a spacer peptide, and human apolipoprotein (apo) A-I was constructed and expressed in Escherichia coli. The lipid interaction properties intrinsic to apoA-I as well as the antigen recognition properties of the scFv were retained by the chimera. scFv center dot apoA-I was formulated into nanoscale reconstituted high-density lipoprotein particles (termed nanodisks; ND) and incubated with cultured cells. alpha-CD20 scFv center dot apoA-I ND bound to CD20-positive non-Hodgkins lymphoma (NHL) cells (Ramos and Granta) but not to CD20-negative T lymphocytes (i.e., Jurkat). Binding to NHL cells was partially inhibited by pre-incubation with rituximab, a monoclonal antibody directed against CD20. Confocal fluorescence microscopy analysis of Granta cells following incubation with alpha-CD20 scFv center dot apoA-I ND formulated with the intrinsically fluorescent hydrophobic polyphenol, curcumin, revealed alpha-CD20 scFv center dot apoA-I localizes to the cell surface, while curcumin off-loads and gains entry to the cell. Compared to control incubations, viability of cultured NHL cells was decreased upon incubation with alpha-CD20 scFv center dot apoA-I ND harboring curcumin. Thus, formulation of curcumin ND with alpha-CD20 scFv center dot apoA-I as the scaffold component confers cell targeting and enhanced bioactive agent delivery, providing a strategy to minimize toxicity associated with chemotherapeutic agents.