Complement Activation and Cell Uptake Responses Toward Polymer-Functionalized Protein Nanocapsules
Complement Activation and Cell Uptake Responses Toward Polymer-Functionalized Protein Nanocapsules
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DOI：
10.1021/bm300083e
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发表时间：
2012-04-01
影响因子：
6.2
通讯作者：
Wang, Szu-Wen
中科院分区：
文献类型：
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作者：
Molino, Nicholas M.;Bilotkach, Kateryna;Wang, Szu-Wen
自组装蛋白质纳米胶囊可以被工程化用于各种生物纳米技术应用。使用来自丙酮酸脱氢酶的E2亚基的十二面体支架，我们引入非天然表面半胱氨酸用于定点功能化。修饰的纳米颗粒的结构、组装和热稳定性与野生型支架（E2-WT）相当，并且在聚乙二醇（PEG）与这些半胱氨酸缀合后，纳米颗粒在高达79.7 +/- 1.8摄氏度下保持完整和稳定。颗粒的PEG化降低了人单核细胞衍生的巨噬细胞和MDA-MB-231乳腺癌细胞的摄取，随着PEG链长度的增加，摄取降低。体外C4消耗和C5 a产生测定产生97.6 +/- 10.8%的血清C4剩余和40.1 +/- 6.0 ng/mL E2-WT的C5 a，表明非PEG化E2纳米颗粒的补体激活较弱。PEG与这些颗粒的偶联适度增加了补体应答，得到79.7 +/- 6.0% C4剩余和87.6 +/- 10.1 ng/mL C5 a。我们的研究结果表明，PEG化的E2蛋白纳米胶囊可以调节细胞摄取和诱导低水平的补体激活，可能通过经典/凝集素途径。
Self-assembling protein nanocapsules can be engineered for various bionanotechnology applications. Using the dodecahedral scaffold of the E2 subunit from pyruvate dehydrogenase, we introduced non-native surface cysteines for site-directed functionalization. The modified nanoparticle's structural, assembly, and thermostability properties comparable to the wild-type scaffold (E2-WT), and after conjugation of poly(ethylene glycol) (PEG) to these cysteines, the nanoparticle remained intact and stable up to 79.7 +/- 1.8 degrees C. PEGylation of particles reduced uptake by human monocyte-derived macrophages and MDA-MB-231 breast cancer cells, with decreased uptake as PEG chain length is increased. In vitro C4-depletion and C5a-production assays yielded 97.6 +/- 10.8% serum C4 remaining and 40.1 +/- 6.0 ng/mL C5a for E2-WT, demonstrating that complement activation is weak for non-PEGylated E2 nanoparticles. Conjugation of PEG to these particles moderately increased complement response to give 79.7 +/- 6.0% C4 remaining and 87.6 +/- 10.1 ng/mL C5a. Our results demonstrate that PEGylation of the E2 protein nanocapsules can modulate cellular uptake and induce low levels of complement activation, likely via the classical/lectin pathways.