Iminodiacetic acid-modified magnetic poly(2-hydroxyethyl methacrylate)-based microspheres for phosphopeptide enrichment

0350470 - ÚMCH 2011 RIV NL eng J - Journal Article
Novotná, L. - Emmerová, T. - Horák, Daniel - Kučerová, Z. - Tichá, M.
Iminodiacetic acid-modified magnetic poly(2-hydroxyethyl methacrylate)-based microspheres for phosphopeptide enrichment.
Journal of Chromatography A. Roč. 1217, č. 51 (2010), s. 8032-8040. ISSN 0021-9673. E-ISSN 1873-3778
R&D Projects: GA AV ČR(CZ) KAN401220801; GA ČR GA203/09/0857; GA ČR GAP503/10/0664
Institutional research plan: CEZ:AV0Z40500505
Keywords : IMAC phosphopeptide separation * IDA-modified magnetic microspheres * Porcine pepsin A
Subject RIV: EE - Microbiology, Virology
Impact factor: 4.194, year: 2010

Magnetic non-porous hydrophilic poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) microspheres prepared by the dispersion polymerization and modified with iminodiacetic acid (IDA) were employed for the IMAC separation of phosphopeptides. Fe3+ and Ga3+ ions immobilized on IDA-modified magnetic microspheres were used for the enrichment of phosphopeptides from the proteolytic digests of two model proteins differing in their physico-chemical properties and phosphate group content: porcine pepsin A and bovine α-casein. The optimum conditions for phosphopeptide adsorption and desorption in both cases were investigated and compared. The phosphopeptides separated from the proteolytic digests were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The ability of the prepared Fe3+- and Ga3+-IDA-modified magnetic microspheres to capture phosphopeptides from complex mixtures was shown on an example of bovine milk proteolytic digest.
Permanent Link: http://hdl.handle.net/11104/0190470