Iminodiacetic acid-modified magnetic poly(2-hydroxyethyl methacrylate)-based microspheres for phosphopeptide enrichment.
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. Fe(3+) and Ga(3+) 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 Fe(3+)- and Ga(3+)-IDA-modified magnetic microspheres to capture phosphopeptides from complex mixtures was shown on an example of bovine milk proteolytic digest.
Novotna
Lenka
L
Institute of Pathological Physiology and CEH, First Faculty of Medicine, Charles University in Prague, U Nemocnice 5, 128 53 Praha 2, Czech Republic. lenka.novotna@LF1.cuni.cz
Emmerova
Tereza
T
Horak
Daniel
D
Kucerova
Zdenka
Z
Ticha
Marie
M
D000327:Adsorption; D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D007098:Imino Acids / Q000737:chemistry; D008280:Magnetics; D008863:Microspheres; D008892:Milk / Q000737:chemistry; D010748:Phosphopeptides / Q000032:analysis / Q000302:isolation & purification*; D011102:Polyhydroxyethyl Methacrylate / Q000737:chemistry*; D058105:Polymerization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D007098:Imino Acids; D010748:Phosphopeptides; D011102:Polyhydroxyethyl Methacrylate; C008109:iminodiacetic acid