Preparation of an ion-exchangeable polymer bead wrapped with bilayer membrane structures for high performance liquid chromatography |
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Authors: | Haratake Mamoru Hidaka Shuko Ono Masahiro Nakayama Morio |
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Institution: | Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki 852-8521, Japan |
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Abstract: | We synthesized a chromatographic packing material that has a non-covalently attached dihexadecyl phosphate (DHP) bilayer membrane structure on a CA08S, a nonporous-type cationic polymer bead with a diameter ranging from 11 to 14 μm. Confocal fluorescence microscopic and differential scanning calorimetric analyses of the DHP-CA08S complex revealed that the DHP bilayer membrane structures were formed on the surface of the CA08S polymer beads. When the functionality of the DHP-CA08S complex was evaluated in the ion-exchange HPLC of proteins, the retention behavior of the proteins on the DHP-CA08S complex column totally mirrored the anionic property of the DHP bilayer membrane surface, not the cationic property of the CA08S bead. Methylene blue (MB) was eluted from the DHP-CA08S complex column in the isocratic elution mode, but not at all from a CK08S column, a styrene-divinylbenzene based cation-exchange polymer. When the column temperature was elevated from 50 to 60 °C, the peak shape of MB on the DHP-CA08S complex column became fairly sharp without a change in its peak area, which mirrored the characteristic phase transition of the DHP bilayer membrane formed on the DHP-CA08S complex. |
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Keywords: | Cyc bovine heart cytochrome c DHP dihexadecyl phosphate DSC differential scanning calorimetry FITC-Dex fluoresceinisothiocyanate dextran HSA human serum albumin Lyz chicken egg white lysozyme MB methylene blue ODR octadecylrhodamine |
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