Fractionation of serum components using nanoporous substrates

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 Petricoin, Lance Liotta

Research output: Contribution to journalArticle

34 Scopus citations

Abstract

Numerous previously uncharacterized molecules resident within the low molecular weight circulatory proteome may provide a picture of the ongoing pathophysiology of an organism. Recently, proteomic signatures composed of low molecular weight molecules have been identified using mass spectrometry combined with bioinformatic algorithms. 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 proteins affords 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 reproducible fractionation and selective binding of the serum-based biomarker material, including subcellular proteins found within the serum. Aminopropyl-coated nanoporous silicon, when exposed to serum, can deplete serum of proteins and 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 eluted proteins have distinct MS profiles, gel electrophoresis profiles, and differential peptide sequence identities, which vary based on the size of the nanopores. These material surfaces could be employed in strategies for the harvesting and preservation of labile and carrier-protein-bound molecules in the blood.

Original languageEnglish (US)
Pages (from-to)654-661
Number of pages8
JournalBioconjugate chemistry
Volume17
Issue number3
DOIs
StatePublished - May 1 2006

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Pharmacology
  • Pharmaceutical Science
  • Organic Chemistry

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