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By Sameh Magdeldin

ISBN-10: 9535103253

ISBN-13: 9789535103257

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Involving of catalytic amino acids residues in the binding process with the subsequent loss of enzymatic activity or restricted accessibility of the active site for substrate molecule. These limitations were overcome in the method of site-specific enzyme immobilization exploiting the attachment of the enzyme due to defined binding sites. As a nice example of different immobilization outcomes a comparative study measured for β-galactosidase by Vishwanath and co-workers may serve. A conjugate of βgalactosidase with a polypeptide tail and unmodified β-galactosidase were immobilized.

Hirabayashi J, Hashidate T, Kasai K (2002) Glyco-catch method: A lectin affinity technique for glycoproteomics. 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.

Belongs to a group of immobilized metal affinity adsorption techniques. The attachment of polyhistidine tag to different protein partners was used in many cases for purification processes and also the interest of its possible employment in immobilizations is under intensive investigation. g. imidazole or histidine) or by usage of metal ions chelator like EDTA. This method, although used in many immobilization experiments, can in some cases suffer from different complications. For example leaching of the metal ions from the support can cause product contamination.

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Affinity chromatography by Sameh Magdeldin

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