Silica containing primary hydroxyl groups for high-performance affinity chromatography |
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| Authors: | K Ernst-Cabrera M Wilchek |
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| Affiliation: | 1. INESC TEC, Sound and Music Computing Group, Porto, Portugal;2. School of Music Studies, Aristotle University of Thessaloniki, Greece;3. ATIC Research Group, ETSI Telecomunicación, University of Málaga, Spain;1. Department of Chemistry, Science and Research Branch, Islamic Azad University, Tehran, Iran;2. Department of Chemistry, Shahreza Branch, Islamic Azad University, Shahreza, Iran;1. Department of Analytical and Environmental Chemistry and Szentágothai Research Center, Ifjúság útja 6, H-7624 Pécs, Hungary;2. MTA–PTE Molecular Interactions in Separation Science Research Group, Ifjúság útja 6, H-7624 Pécs, Hungary;3. Waters Corporation, Milford, MA 01757, USA;4. Institute of Bioanalysis, Medical School, University of Pécs, Szigeti út 12, H-7624 Pécs, Hungary |
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| Abstract: | Primary hydroxyl groups were incorporated into silica by a four-step reaction procedure which includes modification of the silica surface with gamma-glycidoxypropyltrimethoxysilane, leading to an epoxide silica; hydrolysis with acid to yield a diol silica; oxidation of the diol silica with periodate to yield a silica resin with aldehyde functions; and reduction with sodium borohydride to obtain the primary hydroxyl-containing silica. The hydroxyl groups were activated with chloroformates or carbodiimidazole. Proteins were coupled under mild conditions in high yield to these activated silica resins. Columns containing these newly developed silica derivatives were used for the fast and efficient purification of antibodies on antigen-containing silica, as well as for the purification of trypsin on a trypsin inhibitor column (or vice-versa). The effect of pressure on association and dissociation of the affinity complex is discussed. |
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