Abstract
The great interest of ion exchange and affinity chromatography for protein purification has prompted us to design new particles able to meet the technical requirements for industrial scale fractionation. Porous silica beads “Spherosil ® ” were impregnated with DEAE Dextran. Like other aminated macromolecules with a positive electric charge, it has a very strong affinity for the silica surface. In its presence, the strong and irreversible adsorption properties of silica completely disappear and DEAE Dextran gives the support the physicochemical properties of anion exchangers. After cross-linking with bisepoxy reagents, the DEAE Dextran coating is completely stable even in acid or alkaline solutions. Furthermore this new chromatographic support has outstanding mechanical properties which makes large columns very easy to prepare. The bed height is completely incompressible and allows very high flow-rates from 100 to 400 ml/cm2/h under reasonable pressures. For all these reasons, Spherosil-DEAE Dextran is well adapted to the large scale fractionation of proteins by ĭon-exchange (Tayot et al., 1978-a) and immunoaffinity chromatography (Tardy et al., 1978). We now describe the technique used for the preparation and attachment of the ganglioside GP41 to this spherosil derivative and its application to cholera toxin purification by affinity chromatography. A part of this work is described in a preliminary report (Tayot et al., 1978-b).
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
Similar content being viewed by others
References
CUATRECASAS P., PARIKH I. and HOLLENBERG M.D. (1973): Affinity chromatography and structural analysis of vibrio cholerae enterotoxin–Ganglioside agarose and the biological effects of ganglioside containing soluble polymers. Biochemistry 12, 4253–4264.
FREDMAN P. (1979): Quantitative isolation of gangliosides on a new form of glass bead ion-exchanger. This symposium.
HOLMGREN J., MÄNSSON J.E. and SVENNERHOLM L. (1974): Tissue receptor for cholera exotoxin: structural requirements of GMI ganglioside in toxin binding and inactivation. Medical Biology 52, 229–233.
HOLMGREN J., SVENNERHOLM A.M., LÖNNROTH I., FALL-PERSSON M., MARKMAN B. and LUNDBECK H. (1977): Development of improved cholera vaccine based on subunit toxoïd. Nature 269, 602–604.
PARIKH I., MARCH S. and CUATRECASAS P. (1974): Topics in the methodology of substitution reactions with agarose. In “Methods in Enzymology”JAKOBY-WILCHEK Eds, Vol. XXXIV, Academic Press (N.Y. and London), pp. 77–102.
SVENNERHOLM L. (1957): Quantitative estimation of sialic acids. II. A colorimetric resorcinol-hydrochloric acid method. Biochim. Biophys. Acta 24, 604–611.
TARDY M., TAYOT J.L., ROUMIANTZEFF M. and PLAN R. (1978): Immunoaffinity chromatography on derivatives of porous silica beads. Industrial extraction of antitetanus antibodies from placental blood and plasma. In “Chromatography of Synthetic and Biological Polymers”, EPTON R. Ed, Vol. II, Ellis Horwood, Chichester U.K., pp. 298–313.
TAYOT J.L. (1979): Specific agglutination between gangliosidescoated red cells and cholera toxin. This symposium (not printed).
TAYOT J.L., TARDY M., GATTEL P., PLAN R. and ROUMIANTZEFF M. (1978-a) Industrial Ton exchange chromatography of proteins on DEAE-Dextran derivatives of porous silica beads. In “Chromatography of Synthetic and Biological Polymers”, EPTON R. Ed, Vol. II, Ellis Horwood, Chichester U.K., pp. 95–110.
TAYOT J.L., TARDY M. and MYNARD M.C. (1978-b): Biospecific chromatography on new derivatives of porous silica beads. Coupling of ganglioside GM1 or anticholeragen antibody for purification of cholera toxin. In “Affinity Chromatography”, HOFFMANN-OSTENHOF 0. et al. Eds, Pergamon Press (Oxford and N.Y.), pp. 265–269.
WILSON M.B. and NAKANE P.K. (1976): The covalent coupling of proteins to periodate oxydized sephadex: a new approach to immunoadsorbent preparation. J. Immunol. Methods 12, 171–181.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1980 Plenum Press, New York
About this chapter
Cite this chapter
Tayot, JL., Tardy, M. (1980). Isolation of Cholera Toxin by Affinity Chromatography on Porous Silica Beads with Covalently Coupled Ganglioside GM1 . In: Svennerholm, L., Mandel, P., Dreyfus, H., Urban, PF. (eds) Structure and Function of Gangliosides. Advances in Experimental Medicine and Biology, vol 125. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-7844-0_41
Download citation
DOI: https://doi.org/10.1007/978-1-4684-7844-0_41
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4684-7846-4
Online ISBN: 978-1-4684-7844-0
eBook Packages: Springer Book Archive