BIONEM Laboratory, Department of Experimental and Clinical Medicine, University of Catanzaro "Magna Graecia", Germaneto (CZ), Italy.
Int J Nanomedicine. 2010 Nov 19;5:1005-15. doi: 10.2147/IJN.S12865.
Human serum has the potential to become the most informative source of novel biomarkers, but its study is very difficult due to the incredible complexity of its molecular composition. We describe a novel tool based on biodegradable nanoporous nanoparticles (NPNPs) that allows the harvesting of low-molecular-weight fractions of crude human serum or other biofluids. NPNPs with a diameter of 200 nm and pore size of a few nm were obtained by ultrasonication of nanoporous silicon. When incubated with a solution, the NPNPs harvest only the molecules small enough to be absorbed into the nanopores. Then they can be recovered by centrifugation and dissolved in water, making the harvested molecules available for further analyses.
Fluorescence microscopy, gel electrophoresis, and mass spectrometry were used to show the enrichment of low-molecular-weight fraction of serum under physiological conditions, with a cut-off of 13 kDa and an enrichment factor >50.
From these findings, we conclude that ability to tune pore size, combined with the availability of hundreds of biomolecule cross-linkers, opens up new perspectives on complex biofluid analysis, discovery of biomarkers, and in situ drug delivery.
人血清有可能成为新型生物标志物最具信息量的来源,但由于其分子组成的难以置信的复杂性,对其进行研究非常困难。我们描述了一种基于可生物降解的纳米多孔纳米粒子(NPNP)的新型工具,该工具允许从粗制人血清或其他生物流体中采集低分子量分数。通过超声处理纳米多孔硅获得直径为 200nm 且孔径为几纳米的 NPNP。当与溶液孵育时,NPNP 仅采集小到足以被吸收到纳米孔中的分子。然后可以通过离心回收并溶解在水中,从而可以进一步分析所采集的分子。
荧光显微镜、凝胶电泳和质谱法用于在生理条件下显示血清中低分子量部分的富集,截止值为 13kDa,富集因子>50。
从这些发现中,我们得出结论,能够调整孔径,再加上数百种生物分子交联剂的可用性,为复杂生物流体分析、生物标志物的发现和原位药物输送开辟了新的前景。