Tandem lectin affinity chromatography monolithic columns with surface immobilised concanavalin A, wheat germ agglutinin and Ricinus communis agglutinin-I for capturing sub-glycoproteomics from breast cancer and disease-free human sera.

Abstract

In this study, a liquid-phase separation platform consisting of tandem lectin affinity chromatography was introduced for the selective capturing of sub-glycoproteomics that are affected in cancers, e.g. breast cancer. The platform is comprised of three monolithic columns with surface immobilised lectins including concanavalin A (Con A), wheat germ agglutinin (WGA) and Ricinus communis agglutinin-I (RCA-I). While WGA and Con A have specificities directed towards the core portion of N-glycans on the glycoprotein surface, RCA-I specifically interacts with the non-reducing terminal moieties of the outer chain structures of N-glycans. The effects of the order in which the three lectin columns were arranged in the tandem columns format were evaluated. The most suitable order proved to be WGA → Con A → RCA-I (denoted as WCR) as far as the number of captured proteins was concerned. The WCR tandem columns allowed the capture of 113 and 112 proteins from disease-free and breast cancer sera, respectively, corresponding to 75 and 65 non-redundant proteins, respectively. Using mass spectral count ratios and Q-Q plots yielded a panel of 23 non-redundant differentially expressed proteins (i.e. a panel of 23 candidate markers), which should in principle be more representative of a pathophysiological state than a single marker candidate.

• Authors: Subhashini Selvaraju, Ziad El Rassi
• Journal: Journal of separation science (J Sep Sci) Vol. 35 Issue 14 Pg. 1785-95 (Jul 2012) ISSN: 1615-9314 [Electronic] Germany
• PMID: 22807361 (Publication Type: Evaluation Study, Journal Article, Research Support, N.I.H., Extramural)
• Copyright: © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Chemical References

• Glycoproteins
• Lectins
• Plant Lectins
• Ricinus communis agglutinin-1
• Wheat Germ Agglutinins
• Concanavalin A

Topics

• Breast Neoplasms (blood, chemistry, diagnosis)
• Chromatography, Affinity (instrumentation, methods)
• Concanavalin A (chemistry)
• Female
• Glycoproteins (blood, isolation & purification)
• Humans
• Kinetics
• Lectins (chemistry)
• Plant Lectins (chemistry)
• Wheat Germ Agglutinins (chemistry)