Synthesis and Evaluation of Dye-Ligand Affinity Adsorbents for Protein Purification

Autor: Georgios Premetis, Evangelia G. Chronopoulou, Nikolaos E. Labrou, Nikolaos Georgakis, Christina Varotsou, Elisavet Ioannou
Rok vydání: 2020
Předmět:
Zdroj: Methods in Molecular Biology ISBN: 9781071607749
DOI: 10.1007/978-1-0716-0775-6_15
Popis: Dye-ligand affinity chromatography is a widely used technique in protein purification. The utility of the reactive dyes as affinity ligands results from their unique chemistry, which confers wide specificity toward a large number of proteins. They are commercially available, inexpensive, stable and can easily be immobilized. Significant factors that contribute to the successful operation of a dye-ligand chromatography include matrix type, dye-ligand density, adsorption along with elution conditions and flow rate. The present chapter provides protocols for the synthesis of dye-ligand affinity adsorbents as well as protocols for screening, selection, and optimization of a given dye-ligand purification step. The purification of the glutathione transferases from Phaseolus vulgaris on Cibacron Blue 3GA-Sepharose affinity adsorbent is given as an example.
Databáze: OpenAIRE