TY - JOUR
T1 - Fractionation of serum components using nanoporous substrates
AU - Geho, David
AU - Cheng, Mark Ming Cheng
AU - Killian, Keith
AU - Lowenthal, Mark
AU - Ross, Sally
AU - Frogale, Kristina
AU - Nijdam, Jasper
AU - Lahar, Nicholas
AU - Johann, Don
AU - Herrmann, Paul
AU - Whiteley, Gordon
AU - Ferrari, Mauro
AU - Petricoin, Emanuel
AU - Liotta, Lance
PY - 2006
Y1 - 2006
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=33646909377&partnerID=8YFLogxK
U2 - 10.1021/bc0503364
DO - 10.1021/bc0503364
M3 - Article
C2 - 16704202
AN - SCOPUS:33646909377
SN - 1043-1802
VL - 17
SP - 654
EP - 661
JO - Bioconjugate Chemistry
JF - Bioconjugate Chemistry
IS - 3
ER -