title: Evaluation of steric exclusion chromatography on cryogel column for the separation of serum proteins.
abstract: Steric exclusion chromatography (SXC) is a new mode of protein chromatography, in which large proteins are retained on hydrophilic stationary phase surface due to the steric exclusion of polyethylene glycol (PEG) in the mobile phase, and thereafter the retained proteins can be eluted by reducing PEG concentration. In this work, SXC was evaluated on a polyacrylamide cryogel monolith. Microscopic observation of γ-globulin precipitates on the gel surface in SXC was reported for the first time. Due to the compact packing of protein precipitates on the stationary phase surface, the dynamic retention capacity of the cryogel monolith for γ-globulin reached 20 mg/mL bed volume, much higher than those of cryogel beds in adsorption-based chromatography. The effect of molecular weight and concentration of PEG, solution pH and salt concentration on protein retention capacity was in agreement with the earlier work on SXC. Because the cryogel monoliths with interconnected macropores (10-100 μm) allow much easy flow-through of viscous PEG buffer, the SXC can be operated at low back pressure. Hence, the cryogel monoliths are more suitable for SXC than other monoliths of narrow pores reported previously. In the separation of bovine serum proteins, albumin was recovered in the breakthrough fraction with high purity, and globulin was over eight times concentrated in the elution pool. This work has, thus, demonstrated the rapid serum protein separation and concentration by SXC on the cryogel monolith columns.
authors.affiliation: Department of Biochemical Engineering and Key Laboratory of Systems Bioengineering of the Ministry of Education, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China.
authors.forename: Chuan
authors.initials: C
authors.lastname: Wang
authors.affiliation: Department of Biochemical Engineering and Key Laboratory of Systems Bioengineering of the Ministry of Education, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China; Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin 300072, China.
authors.forename: Shu
authors.initials: S
authors.lastname: Bai
authors.affiliation: Department of Biochemical Engineering and Key Laboratory of Systems Bioengineering of the Ministry of Education, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China.
authors.forename: Shi-Peng
authors.initials: SP
authors.lastname: Tao
authors.affiliation: Department of Biochemical Engineering and Key Laboratory of Systems Bioengineering of the Ministry of Education, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China; Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin 300072, China. Electronic address: ysun@tju.edu.cn.
authors.forename: Yan
authors.initials: Y
authors.lastname: Sun
mesh_terms: D000180:Acrylic Resins / Q000737:chemistry; D000818:Animals; D001798:Blood Proteins / Q000302:isolation & purification*; D002417:Cattle; D002850:Chromatography, Gel / Q000295:instrumentation / Q000379:methods*; D059985:Cryogels / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D008970:Molecular Weight; D011092:Polyethylene Glycols / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000302:isolation & purification; D005719:gamma-Globulins / Q000737:chemistry / Q000302:isolation & purification; D000099166:Polyacrylamides
chemical_list: D000180:Acrylic Resins; D001798:Blood Proteins; D059985:Cryogels; D011092:Polyethylene Glycols; D012710:Serum Albumin, Bovine; D005719:gamma-Globulins; D000099166:Polyacrylamides
keywords: Cryogel monolith; Polyethylene glycol; Protein chromatography; Serum protein separation; Steric exclusion chromatography