Tandem lectin affinity chromatography monolithic columns with surface immobilised concanavalin A, wheat germ agglutinin andRicinus communisagglutinin-I for capturing sub-glycoproteomics from breast cancer and disease-free human sera
| Autor: | Subhashini Selvaraju, Ziad El Rassi |
|---|---|
| Rok vydání: | 2012 |
| Předmět: |
Wheat Germ Agglutinins
Breast Neoplasms Filtration and Separation Chromatography Affinity Analytical Chemistry Lectins Concanavalin A Humans Glycoproteins chemistry.chemical_classification Lectin affinity chromatography Tandem biology Lectin Molecular biology Wheat germ agglutinin Glycoproteomics Kinetics Biochemistry chemistry Ricinus communis agglutinin biology.protein Female Plant Lectins Glycoprotein |
| Zdroj: | Journal of Separation Science. 35:1785-1795 |
| ISSN: | 1615-9306 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text