Insulin adsorption on coated silica based supports grafted with N-acetylglucosamine by liquid affinity chromatography

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
H Lakhiari, D Muller

Abstract

Silica beads are coated with dextran carrying a calculated amount of positively charged diethylassminoethyl groups (DEAE) in order to neutralize negative charged silanol groups at the silica surface and in this way to minimize non specific interactions between silica surface and proteins in solution. Dextran-coated silica supports are potentially excellent stationary phases for high-performance liquid chromatography of proteins. These supports combine the advantages of polysaccharide phases with the excellent mechanical characteristics of silica. These supports (silica-dextran-DEAE = SID) are easily functionalized by grafting N-acetylglucosamine (GlcNAc) using conventional coupling methods. The performances of the support bearing GlcNAc are studied by high-performance liquid affinity chromatography (HPLAC) of insulin, the hypoglycemic peptide hormone of the human organism. The study shows that these supports exhibit a reversible and specific affinity towards insulin and allow separations with high purification yields. Moreover, the influence of different physico-chemical parameters (pH, NaCl and insulin concentration) on insulin retention on the support was analysed. This allowed us to optimize the conditions of adsorption and ...Continue Reading

References

Apr 1, 1977·Diabetes·D F Steiner
Jun 29, 1988·Journal of Chromatography·A SantarelliJ Jozefonvicz
Aug 4, 1989·Journal of Chromatography·F L ZhouJ Jozefonvicz
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Sep 8, 1995·Journal of Chromatography. a·H LakhiariJ Jozefonvicz
Feb 3, 1995·Journal of Chromatography. B, Biomedical Applications·H LakhiariJ Jozefonvicz
Apr 17, 1998·Journal of Chromatography. B, Biomedical Sciences and Applications·H LakhiariD Muller
Apr 18, 1998·Biochimica Et Biophysica Acta·H LakhiariJ Jozefonvicz

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Citations

Apr 26, 2005·Journal of Colloid and Interface Science·S H MollmannU Elofsson
Oct 11, 2011·European Journal of Pharmaceutics and Biopharmaceutics : Official Journal of Arbeitsgemeinschaft Für Pharmazeutische Verfahrenstechnik E.V·F DepreterK Amighi
Jul 9, 2009·Biotechnology Progress·Shaoling ZhangAkihiko Tanioka
Jul 15, 2009·Biotechnology Progress·Shaoling ZhangHidetoshi Matsumoto
Jan 29, 2011·Colloids and Surfaces. B, Biointerfaces·Müfrettin Murat SarıAdil Denizli

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