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Development of Rigid Bidisperse Porous Microspheres for High‐Speed Protein Chromatography
Author(s) -
Wu Lei,
Bai Shu,
Sun Yan
Publication year - 2008
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp0201509
Subject(s) - chromatography , glycidyl methacrylate , porosimetry , packed bed , adsorption , chemical engineering , chemistry , porosity , polymerization , scanning electron microscope , monolithic hplc column , matrix (chemical analysis) , high performance liquid chromatography , materials science , porous medium , polymer , organic chemistry , composite material , engineering
Development of a high‐performance stationary phase is an essential demand for high‐speed separation of proteins by liquid chromatography. Based on a novel porogenic mode, that is, using superfine granules of calcium carbonate as solid porogen and a mixture of cyclohexanol and dodecanol as liquid porogen, a rigid spherical biporous poly(glycidyl methacrylate‐ co ‐ethylene dimethacrylate) matrix has been prepared by radical suspension‐polymerization. The epoxide groups of the matrix were modified with diethylamine to afford the ionizable weak base 1‐ N , N ‐diethylamino‐2‐hydeoxypropy functionalities that are required for ion exchange chromatography. Results from scanning electron microscopy and mercury intrusion porosimetry measurements revealed that the matrix contained two families of pores, that is, micropores (10–90 nm) and macropores (180–4000 nm). Furthermore, the biporous medium possesses specific surface area as high as 91.3 m 2 /g. Because of the presence of the macropores that provided convective flow channels for the mobile phase, the dynamic adsorption capacity was found to be as high as 54.6 mg/g wet bead at 300 cm/h, approximately 63.2% of its static capacity. In addition, the column efficiency and dynamic binding capacity decreased only slightly with mobile‐phase flow rate in the range of 300–3000 cm/h. These properties made the packed bed with the bidisperse porous matrix suitable for high‐speed protein chromatography.