By Sameh Magdeldin
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Many types of carriers with diverse properties were evolved for immobilization processes; however, the suitability of their application in individual cases needs a careful consideration and very often also laborious method adaptation. Preparing a suitable carrier may be seen as another problem and may incur additional costs for the whole immobilization procedure (Aehle, 2007; Brenda & Batista-Viera, 2006; Sheldon, 2007). 32 a. Affinity Chromatography Covalent binding For covalent binding of the protein to the carrier amino acid residues not involved in the reaction mechanism may be used.
2006; Sheldon, 2007). c. Ionic binding Ion-ion interactions, providing a stronger binding of immobilized molecules than physical adsorption, are utilized during immobilization process. This method also doesn´t change enzyme conformation in a substantial extent. , 2005). d. e. , 2004). 2. Principles of above mentioned methods are for illustration presented in the figure 1. Fig. 1. Immobilization methods exploiting binding to a carrier. 2 Enzyme molecules cross-linking In these methods bi- or multifunctional compounds are used for cross-linking of desired enzyme molecules (figure 2).
J Biomol Tech 13: 205-218 Hober S, Nord K, Linhult M (2007) Protein A chromatography for antibody purification. J Chromatogr B Analyt Technol Biomed Life Sci 848: 40-47 Huang W, Wang J, Bhattacharyya D, Bachas LG (1997) Improving the activity of immobilized subtilisin by site-specific attachment to surfaces. Anal Chem 69: 4601-4607 Jackson AJ, Xuan H, Hage DS (2010) Entrapment of proteins in glycogen-capped and hydrazide-activated supports. Anal Biochem 404: 106-108 Janson, J-C. (1984) Large-scale affinity purification — state of the art and future prospects.
Affinity chromatography by Sameh Magdeldin