By Sameh Magdeldin
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One way is to insert a Affinity Chromatography: Principles and Applications 21 biotinylation sequence into a recombinant protein. Biotin protein ligase can then be used to add biotin in a post-translational modification step (Cronan & Reed, 2000). Biotin, also known as vitamin H or vitamin B7, is a relatively small cofactor present in cells. In affinity chromatography it is often used an affinity tag due to its very strong interactions with avidin and streptavidin. One advantage of using biotin as an affinity tag is that it has a minimal effect on the activity of a large biomolecule due to its small size (244 Da).
Nat Protoc 1: 637-640 Young JL, Webb BA (1978) Two methods for the separation of human alpha-fetoprotein and albumin. (A) Affinity chromatography using Blue Sepharose CL-6B and (B) ampholyte displacement chromatography. Anal Biochem 88: 619-623 Zachariou M (2008) Affinity chromatography: methods and protocols. Preface. Methods Mol Biol 421: vii-viii 2 Affinity Interactions as a Tool for Protein Immobilization Eva Benešová and Blanka Králová The Institute of Chemical Technology, Prague Czech Republic 1.
J Proteome Res 9: 4982-4991 Tyutyulkova S, Paul S (1995) Purification of antibody light chains by metal affinity and protein L chromatography. Methods Mol Biol 51: 395-401 Udomsinprasert R, Pongjaroenkit S, Wongsantichon J, Oakley AJ, Prapanthadara LA, Wilce MC, Ketterman AJ (2005) Identification, characterization and structure of a new Delta class glutathione transferase isoenzyme. Biochem J 388: 763-771 Wang G, De J, Schoeniger JS, Roe DC, Carbonell RG (2004) A hexamer peptide ligand that binds selectively to staphylococcal enterotoxin B: isolation from a solid phase combinatorial library.
Affinity chromatography by Sameh Magdeldin