Phosphate Anion Imprinted Cryogel Cartridges for Selective Preconcentration of Phosphorylated Amino Acids from Protein Lysate: An Alternative Sorbent for Proteome Analyses

Abstract

AbstractPhosphorylation are vital in many stages such as cellular communication in metabolism, protein regulation, cell division and apoptosis. Also, the rate of phosphorylation with many diseases, including cancer, is linear. Analysis of the phosphorylated proteins is classified as phosphoproteomic and samples of hydrolyzed protein are analyzed by mass spectrometry. Although there are generally 20 amino acids in biological systems; only three (tyrosine, threonine and serine) are phosphorylated based on the hydroxyl groups they have. Thus, the preconcentration of these amino acids, which are found to be relatively low in number and amount in phosphoproteomic analyses, is of great importance for diagnosing and monitoring of the treatment. Herein, it is aimed to develop phosphate‐imprinted cryogel cartridges for the preconcentration of phosphoproteomic lysates. Cryogel cartridges were synthesized in semi‐frozen medium and characterized by scanning electron microscopy (SEM). Phosphate adsorption conditions were optimized for cryogel cartridges and optimum adsorption conditions were determined as pH of 9.80, concentration of 125 ppm and temperature of 25 °C. The selectivity of cryogel cartridges to phosphate ion was determined using chloride and carbonate ions. According to the results, cryogel cartridges are selectively recognizing both phosphate ion and phosphorylated amino acids and are suitable for cheap, efficient and fast pre‐initiation studies with reusability.