Polyvalent Hybrid Virus-Like Nanoparticles with Displayed Heparin Antagonist Peptides
Polyvalent Hybrid Virus-Like Nanoparticles with Displayed Heparin Antagonist Peptides
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DOI：
10.1021/acs.molpharmaceut.8b00135
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发表时间：
2018-08-01
影响因子：
4.9
通讯作者：
Udit, Andrew K.
中科院分区：
文献类型：
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作者：
Choi, Justin M.;Bourassa, Valerie;Udit, Andrew K.
随着新材料的开发以及环境和安全问题得到更充分的解决，纳米材料的潜在应用不断增长。特别是，病毒样颗粒（VLP）在医学和生物学中具有无数的应用，利用可靠、对称的自组装机制和利用可通过突变或生物共轭进行适当修饰的表面功能的能力。在此，我们描述了用作有效肝素拮抗剂的混合VLP的设计和应用，为有毒肝素解毒剂鱼精蛋白提供了替代品。利用双质粒系统生成包含野生型外壳蛋白和带有 C 端或 N 端阳离子肽延伸（4-28 个氨基酸）的第二外壳蛋白的 VLP。修饰的外壳蛋白的掺入范围为 8% 至 31%，而活化部分凝血活酶时间 (APTT) 测定显示了一系列肝素拮抗剂活性。值得注意的是，当根据不同掺入率导致的肽递送量进行检查时，VLP 似乎很大程度上遵循相似的趋势，递送的肽量与肝素拮抗剂活性更密切相关。具有最高掺入率和最佳抗肝素活性的颗粒显示出C端肽ARK(2)A(2)KA，其对应于与糖胺聚糖结合的Cardin-Weintraub共有序列。使用带有级分收集的肝素亲和色谱对该颗粒进行分析表明，在较高盐浓度下洗脱的颗粒具有较大比例的肽掺入。初步的双偏振干涉实验进一步支持了该颗粒与肝素之间的强烈相互作用。
The potential applications for nanomaterials continue to grow as new materials are developed and environmental and safety concerns are more adequately addressed. In particular, virus-like particles (VLPs) have myriad applications in medicine and biology, exploiting both the reliable, symmetric self-assembly mechanism and the ability to take advantage of surface functionalities that may be appropriately modified through mutation or bioconjugation. Herein we describe the design and application of hybrid VLPs for use as potent heparin antagonists, providing an alternative to the toxic heparin antidote protamine. A two-plasmid system was utilized to generate VLPs that contain both the wild-type coat protein and a second coat protein with either a C-or N-terminal cationic peptide extension (4-28 amino acids). Incorporation of the modified coat proteins varied from 8 to 31%, while activated partial thromboplastin time (APTT) assays revealed a range of the heparin antagonist activity. Notably, when examined on the basis of the quantity of peptide delivered due to the varied incorporation rates, it appeared that the VLPs largely followed a similar trend, with the quantity of peptide delivered more closely correlating with heparin antagonist activity. The particle with the highest incorporation rate and best antiheparin activity displayed the C-terminal peptide ARK(2)A(2)KA, which corresponds to the Cardin-Weintraub consensus sequence for binding to glycosaminoglycans. Analysis of this particle using heparin affinity chromatography with fraction collection revealed that particles eluting at higher salt concentration had a greater proportion of peptide incorporation. Preliminary dual polarization interferometry experiments further support a strong interaction between this particle and heparin.