A potential nanobiotechnology platform based on infectious bursal disease subviral particles.
A potential nanobiotechnology platform based on infectious bursal disease subviral particles.
复制标题
基于传染性法氏囊病亚病毒颗粒的潜在纳米生物技术平台。
DOI：
10.1039/c2ra00857b
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发表时间：
2012
期刊：
影响因子：
3.9
通讯作者：
Schillberg,Stefan
中科院分区：
文献类型：
--
作者：
Taghavian,Omid;Mandal,ManojK;Steinmetz,NicoleF;Rasche,Stefan;Spiegel,Holger;Fischer,Rainer;Schillberg,Stefan
我们描述了一种新的纳米生物技术平台，该平台基于来自传染性法氏囊病病毒（IBD-SVP）的亚病毒颗粒。当在酵母毕赤酵母中表达为异源蛋白时，主要病毒外壳蛋白VP 2组装成球形的23 nm SVP。我们从IL重组酵母培养物中回收了高达38 mg纯度> 95%的IBD-SVP。纯化的颗粒能够耐受高达20%浓度的有机溶剂（乙醇或二甲亚砜），它们耐受高达65 °C的温度，并且在宽pH范围（2.5-9.0）内保持稳定。我们实现了与赖氨酸残基的胺基以及与天冬氨酸和谷氨酸残基的羧基的生物缀合，从而允许用生物素官能化IBD-SVP。使用Alexa Fluor 488 N-羟基琥珀酰亚胺（NHS）酯（一种胺选择性荧光染料）测量表面胺基的可及性，结果显示约60个染料分子附着在每个颗粒的表面。因此，IBD-SVP可以作为一种强大的和通用的纳米支架来展示不同的功能配体。
We describe a novel nanobiotechnology platform based on subviral particles derived from Infectious bursal disease virus (IBD-SVPs). The major virus coat protein VP2 assembles into spherical, 23 nm SVPs when expressed as a heterologous protein in the yeast Pichia pastoris. We recovered up to 38 mg of IBD-SVPs at > 95% purity from 1 L of recombinant yeast culture. The purified particles were able to tolerate organic solvents up to 20% concentration (ethanol or dimethylsulfoxide), they resisted temperatures up to 65 °C and remained stable over a wide pH range (2.5–9.0). We achieved bioconjugation to the amine groups of lysine residues and to the carboxyl groups of aspartic and glutamic acid residues, allowing the functionalization of IBD-SVPs with biotin. The accessibility of surface amine groups was measured using Alexa Fluor 488 N-hydroxysuccinimide (NHS) ester, an amine-selective fluorescent dye, revealing that approximately 60 dye molecules were attached to the surface of each particle. IBD-SVPs can therefore be exploited as a robust and versatile nanoscaffold to display diverse functional ligands.