Improved Doxorubicin Encapsulation and Pharmacokinetics of Ferritin-Fusion Protein Nanocarriers Bearing Proline, Serine, and Alanine Elements
Improved Doxorubicin Encapsulation and Pharmacokinetics of Ferritin-Fusion Protein Nanocarriers Bearing Proline, Serine, and Alanine Elements
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DOI：
10.1021/acs.biomac.5b01446
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发表时间：
2016-02-01
影响因子：
6.2
通讯作者：
Ceci, Pierpaolo
中科院分区：
文献类型：
--
作者：
Falvo, Elisabetta;Tremante, Elisa;Ceci, Pierpaolo
一种新型的基于人铁蛋白的纳米载体由 24 个修饰单体组成，能够自动组装成修饰的蛋白笼，并用作抗肿瘤有效负载的选择性载体。每个修饰单体均源自两个不同模块的基因融合，即人铁蛋白 (HFt) 的重链和富含脯氨酸 (13)、丝氨酸 (5) 和丙氨酸 (A) 残基的稳定/保护性 PAS 多肽序列。比较了两种分别含有长度为 40 和 75 个残基的 PAS 聚合物的基因融合蛋白构建体。它们是在大肠杆菌中以高产率生产和纯化的重组蛋白。两种制剂在体外以及小鼠血浆中均具有高度可溶性和稳定性。尺寸排阻色谱、动态光散射和透射电子显微镜结果表明，PASylated 铁蛋白完全组装且高度单分散。此外，两种 HFt-PAS 蛋白的产量均明显优于野生型 HFt。重要的是，PAS 序列显着延长了 HFt 在小鼠血液中的半衰期。最后，我们的负载阿霉素的纳米笼保留了药物的药理活性。总而言之，这些结果表明，两种开发的 HFt-PAS 融合蛋白都是未来在癌症治疗中应用的有前景的纳米载体。
A novel human ferritin-based nanocarrier, composed of 24 modified monomers able to auto-assemble into a modified protein cage, was produced and used as selective carrier of anti-tumor payloads. Each modified monomer derives from the genetic fusion of two distinct modules, namely the heavy chain of human ferritin (HFt) and a stabilizing/protective PAS polypeptide sequence rich in proline (13), serine (5), and alanine (A) residues. Two genetically fused protein constructs containing PAS polymers with 40- and 75-residue lengths, respectively, were compared. They were produced and purified as recombinant proteins in Escherichia coli at high yields. Both preparations were highly soluble and stable in vitro as well as in mouse plasma. Size exclusion chromatography, dynamic light scattering, and transmission electron microscopy results indicated that PASylated ferritins are fully assembled and highly monodispersed. In addition, yields significantly better for both HFt-PAS proteins than for wild-type HFt. Importantly, PAS sequences considerably prolonged the half-life of HFt in the mouse bloodstream. Finally, our doxorubicin-loaded nanocages preserved the pharmacological activity of the drug. Taken together, these results indicate that both of the developed HFt-PAS fusion proteins are promising nanocarriers for future applications in cancer therapy.