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Long term storage of virus templated fluorescent materials for sensing applications

机译:病毒模板荧光材料的长期存储,可用于传感应用

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摘要

Wild type, mutant, and chemically modified Cowpea mosaic viruses ( CPMV) were studied for long term preservation in the presence and absence of cryoprotectants. Viral complexes were reconstituted and tested via fluorescence spectroscopy and a UV/vis-based RNase assay for structural integrity. When viruses lyophilized in the absence of cryoprotectant were rehydrated and RNase treated, UV absorption increased, indicating that the capsids were damaged. The addition of trehalose during lyophilization protected capsid integrity for at least 7 weeks. Measurements of the fluorescence peak maximum of CPMV lyophilized with trehalose and reconstituted also indicate that the virus remained intact. Microarray binding assays indicated that CPMV particles chemically modified for use as a fluorescent tracer were intact and retained binding specificity after lyophilization in the presence of trehalose. Thus, we demonstrate that functionalized CPMV nanostructures can be stored for the long term, enabling their use in practical sensing applications.
机译:研究了野生型,突变型和化学修饰的Cow豆花叶病毒(CPMV)在存在和不存在冷冻保护剂的情况下的长期保存。重组病毒复合物,并通过荧光光谱和基于UV / vis的RNase测定结构完整性。当在没有冷冻保护剂的情况下冻干的病毒重新水化并用RNase处理时,紫外线吸收增加,表明衣壳被破坏。冻干过程中加入海藻糖至少可保护衣壳7周。海藻糖冻干并重构的CPMV的荧光峰值最大值的测量也表明该病毒保持完整。微阵列结合试验表明,在海藻糖存在下冻干后,经过化学修饰以用作荧光示踪剂的CPMV颗粒完整无损,并保留了结合特异性。因此,我们证明功能化的CPMV纳米结构可以长期存储,使其可以在实际的传感应用中使用。

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