Fractionation of Serum Components Using Nanoporous Substrates

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• 作者：David Geho ; Mark Ming-Cheng Cheng ; Keith Killian ; Mark Lowenthal ; Sally Ross ; Kristina Frogale ; Jasper Nijdam ; Nicholas Lahar ; Don Johann ; Paul Herrmann ; Gordon Whiteley ; Mauro Ferrari ; Emanuel Petric
• 刊名：Bioconjugate Chemistry
• 出版年：2006
• 出版时间：May 2006
• 年：2006
• 卷：17
• 期：3
• 页码：654 - 661
• 全文大小：324K
• 年卷期：v.17,no.3(May 2006)
• ISSN：1520-4812

文摘

Numerous previously uncharacterized molecules resident within the low molecular weight circulatory proteomemay provide a picture of the ongoing pathophysiology of an organism. Recently, proteomic signatures composedof low molecular weight molecules have been identified using mass spectrometry combined with bioinformaticalgorithms. Attempts to sequence and identify the molecules that underpin the fingerprints are currently underway.The finding that many of these low molecular weight molecules may exist bound to circulating carrier proteinsaffords a new opportunity for fractionation and separation techniques prior to mass spectrometry-based analysis.In this study we demonstrate a method whereby nanoporous substrates may be used for the facile and reproduciblefractionation and selective binding of the serum-based biomarker material, including subcellular proteins foundwithin the serum. Aminopropyl-coated nanoporous silicon, when exposed to serum, can deplete serum of proteinsand yield a serum with a distinct, altered MS profile. Additionally, aminopropyl-coated, nanoporous controlled-pore glass beads are able to bind a subset of serum proteins and release them with stringent elution. The elutedproteins have distinct MS profiles, gel electrophoresis profiles, and differential peptide sequence identities, whichvary based on the size of the nanopores. These material surfaces could be employed in strategies for the harvestingand preservation of labile and carrier-protein-bound molecules in the blood.