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KMID : 1094720120170030584
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Biotechnology and Bioprocess Engineering
2012 Volume.17 No. 3 p.584 ~ p.590
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A one step separation of immunoglobulin g from bovine serum by pseudobioaffinity chromatography on histidine grafted to epoxy activated sepharose
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Elkak Assem
Yehya Tania
Salloub Ibrahim
Berry Fadwa
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Abstract
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The selective adsorption of proteins and macromolecules on activated matrix grafted with histidine has been shown to be dependent on certain separation parameters like, pH and buffer type. In the present study, the significant potential of the histidine ligand to separate bovine IgG from other bovine serum proteins has been demonstrated. The successful separation was carried out by pseudobioaffinty chromatography on histidine grafted-epoxy activated sepharose. The method was applied to sterile bovine serum. Bovine IgG was completely separated in the form of a single peak with 25 mM MES buffer containing 0.2 M NaCl at pH 5. The purity of the separated bovine IgG was determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions. Ouchterlony radial immunodiffusion (ODD) assay showed that bovine IgG was the main component present in the elution fraction.
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KEYWORD
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epoxy activated sepharose, histidine, pseudobioaffinity chromatography, bovine IgG, MES buffer
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