title: Separation of peptides with an aqueous mobile phase by temperature-responsive chromatographic column.
abstract: Peptide separation technology is significant and is still an analytical challenge in proteomic studies. We report a simple preparation method for poly(N-isopropylacrylamide) grafted silica through the copolymerization of N-isopropylacrylamide with acetyl moieties immobilized on the silica surfaces. Differential scanning calorimetry results indicated that the prepared silica exhibited a sharp phase transition at 35.03°C. Silica grafted with poly(N-isopropylacrylamide) was evaluated as a temperature-responsive chromatography medium for the separation of peptides using 0.2 M NaCl solution as a mobile phase. Results indicated that at 10°C, the peptides were not resolved, but baseline separation with prolonged retention time at 50°C was attained. Particularly, a mixture of four peptides was efficiently separated within 8 min. The theoretical plate number of every peptide was more than 2500, and the resolutions were more than 3.40. The increased selectivity of the temperature-responsive column resulted from the temperature-modulated hydrophobic interaction with peptides. The retention times of these peptides were related to their hydrophobicities. This protocol provided a reliable set of chromatographic tool usable across all research and development applications that required isolation and analysis of peptides. It may represent a step forward in the complex analysis of hydrophobic and other proteins.
authors: School of Life Science, Beijing Institute of Technology, Beijing, P R China.
authors: Zongjian
authors: Z
authors: Liu
authors: Yanli
authors: Y
authors: Liang
authors: Fangfang
authors: F
authors: Geng
authors: Chen
authors: C
authors: Ge
authors: Kaleem
authors: K
authors: Ullah
authors: Fang
authors: F
authors: Lv
authors: Rongji
authors: R
authors: Dai
authors: Yukui
authors: Y
authors: Zhang
authors: Yulin
authors: Y
authors: Deng
mesh_terms: D000595:Amino Acid Sequence; D002152:Calorimetry, Differential Scanning; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D006358:Hot Temperature; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D010455:Peptides