title: Mixed-mode chromatography integrated with high-performance liquid chromatography for protein analysis and separation: Using bovine serum albumin and lysozyme as the model target.
abstract: A type of mixed-mode chromatography was integrated with high-performance liquid chromatography for protein analysis and separation. The chromatographic behavior was tested using bovine serum albumin and lysozyme as model proteins. For the mixed-mode column, the silica beads were activated with γ-(2,3-epoxypropoxy)-propytrimethoxysilane and coupled with 4-mercaptopyridine as the functional ligand. The effects of pH, salt, and the organic solvent conditions of the mobile phase on the retention behavior were studied, which provided valuable clues for separation strategy. When eluted with a suitable pH gradient, salt concentration gradient, and acetonitrile content gradient, the separation behavior of bovine serum albumin and lysozyme could be controlled by altering the conditions of the mobile phase. The results indicated this type of chromatography might be a useful method for protein analysis and separation.
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China.|National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi, China.
authors.forename: Hai-Feng
authors.initials: HF
authors.lastname: Xia
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China.
authors.forename: Bin-Bin
authors.initials: BB
authors.lastname: Don
authors.affiliation: National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi, China.
authors.forename: Meng-Jie
authors.initials: MJ
authors.lastname: Zheng
mesh_terms: D000097:Acetonitriles / Q000737:chemistry; D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D009113:Muramidase / Q000032:analysis* / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000032:analysis*; D012822:Silicon Dioxide / Q000737:chemistry; D012965:Sodium Chloride / Q000737:chemistry
chemical_list: D000097:Acetonitriles; D012710:Serum Albumin, Bovine; D012965:Sodium Chloride; D012822:Silicon Dioxide; D009113:Muramidase; C032159:acetonitrile
keywords: Mixed-mode chromatography; Protein separation; Resolution; Retention