Department of Nanomedicine and BioMedical Engineering, The University of Texas Health Science Center at Houston, Texas 77030, USA.
ACS Nano. 2010 Jan 26;4(1):439-51. doi: 10.1021/nn901322d.
We present a fast, efficient, and reliable system based on mesoporous silica chips to specifically fractionate and enrich the low molecular weight proteome. Mesoporous silica thin films with tunable features at the nanoscale were fabricated using the triblock copolymer template pathway. Using different templates and concentrations in the precursor solution, various pore size distributions, pore structures, and connectivity were obtained and applied for selective recovery of low mass proteins. In combination with mass spectrometry and statistic analysis, we demonstrated the correlation between the nanophase characteristics of the mesoporous silica thin films and the specificity and efficacy of low mass proteome harvesting. In addition, to overcome the limitations of the prefunctionalization method in polymer selection, plasma ashing was used for the first time for the treatment of the mesoporous silica surface prior to chemical modification. Surface charge modifications by different functional groups resulted in a selective capture of the low molecular weight proteins from serum sample. In conclusion, our study demonstrates that the ability to tune the physicochemical properties of mesoporous silica surfaces, for a selective enrichment of the low molecular weight proteome from complex biological fluids, has the potential to promote proteomic biomarker discovery.
我们提出了一种快速、高效、可靠的基于介孔硅芯片的系统,用于特异性地分离和富集低分子量蛋白质组。采用三嵌段共聚物模板途径制备了具有纳米级可调特征的介孔硅薄膜。通过在前驱体溶液中使用不同的模板和浓度,获得了各种孔径分布、孔结构和连通性,并将其应用于低质量蛋白质的选择性回收。结合质谱和统计学分析,我们证明了介孔硅薄膜的纳米相特征与低质量蛋白质组收获的特异性和效果之间的相关性。此外,为了克服聚合物选择中预功能化方法的局限性,我们首次使用等离子体灰化法在化学修饰前对介孔硅表面进行处理。通过不同官能团进行表面电荷修饰,可从血清样品中选择性捕获低分子量蛋白质。总之,我们的研究表明,能够调整介孔硅表面的物理化学性质,从而从复杂的生物流体中选择性富集低分子量蛋白质组,有可能促进蛋白质组生物标志物的发现。