TY - JOUR
T1 - Efficacy of glycoprotein enrichment by microscale lectin affinity chromatography
AU - Madera, Milan
AU - Mann, Benjamin
AU - Mechref, Yehia
AU - Novotny, Milos V.
PY - 2008/8
Y1 - 2008/8
N2 - Reproducible and efficient affinity enrichment is increasingly viewed as an essential step in many investigations leading to the discovery of new biomarkers. In this work, we have evaluated the repeatability of lectin enrichment of glycoproteins from human blood serum through both qualitative and quantitative proteomic approaches. In a comprehensive evaluation of lectin binding, we have performed 30 separate microscale lectin affinity chromatography experiments, followed by a conventional sample purification, and LC-MS/MS analysis of the enriched glycoproteins. Two lectin affinity matrixes, both with Con A lectin, immobilized to the same solid support but differing in the amount of immobilized lectin, were investigated to characterize their binding properties. Both qualitative and quantitative data indicate acceptable repeatability and binding efficiency for the lectin materials received from two different commercial sources.
AB - Reproducible and efficient affinity enrichment is increasingly viewed as an essential step in many investigations leading to the discovery of new biomarkers. In this work, we have evaluated the repeatability of lectin enrichment of glycoproteins from human blood serum through both qualitative and quantitative proteomic approaches. In a comprehensive evaluation of lectin binding, we have performed 30 separate microscale lectin affinity chromatography experiments, followed by a conventional sample purification, and LC-MS/MS analysis of the enriched glycoproteins. Two lectin affinity matrixes, both with Con A lectin, immobilized to the same solid support but differing in the amount of immobilized lectin, were investigated to characterize their binding properties. Both qualitative and quantitative data indicate acceptable repeatability and binding efficiency for the lectin materials received from two different commercial sources.
KW - Glycoproteins
KW - LC-MS/MS
KW - Lectin affinity chromatography
KW - Quantitative glycoproteomics
UR - http://www.scopus.com/inward/record.url?scp=51049110528&partnerID=8YFLogxK
U2 - 10.1002/jssc.200800094
DO - 10.1002/jssc.200800094
M3 - Article
C2 - 18623281
AN - SCOPUS:51049110528
VL - 31
SP - 2722
EP - 2732
JO - Journal of Separation Science
JF - Journal of Separation Science
SN - 1615-9306
IS - 14
ER -