New PDF release: Affinity chromatography

By Sameh Magdeldin

ISBN-10: 9535103253

ISBN-13: 9789535103257

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6. g. exoglucanase and endoglucanase from Cellulomonas fimi or cellulase from Trichoderma harzianum, are able to specifically bind cellulose thanks to their cellulose-binding domain. These domains may be exploited as affinity tags for fusion proteins immobilization on cellulose supports, which are inert and exhibit only low non-specific affinity and are readily available. g. , Zymomonas mobilis extracellular invertase or Bacillus stearothermophilus L1 lipase. Alternative method using chemical coupling by glutaraldehyde was utilized for preparation of a conjugate containing glucose oxidase and cellulose-binding domain.

Nature 214: 1302-1304 Balbas P (2001) Understanding the art of producing protein and nonprotein molecules in Escherichia coli. Mol Biotechnol 19: 251-267 Bautz EK, Hall BD (1962) The isolation of T4-specific RNA on a DNA-cellulose column. Proc Natl Acad Sci U S A 48: 400-408 Affinity Chromatography: Principles and Applications 25 Beckett GJ, Hayes JD (1993) Glutathione S-transferases: biomedical applications. Adv Clin Chem 30: 281-380 Bottari P, Aebersold R, Turecek F, Gelb MH (2004) Design and synthesis of visible isotopecoded affinity tags for the absolute quantification of specific proteins in complex mixtures.

Both avidin and streptavidin have four subunits that can each bind one biotin molecule. To purify biotinylated biomolecules, streptavidin is immobilized onto a support material and used to extract the biotinylated molecules out of solution. Both avidin and streptavidin may be immobilized using aminereactive coupling chemistries. In addition, avidin can also be immobilized via its carbohydrate residues. Due to the strong interaction between biotin and (strept)avidin, harsh elution conditions are required to disrupt the binding.

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Affinity chromatography by Sameh Magdeldin

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