Protein purification by dye-ligand chromatography

Application of dye-ligand chromatography for purification of coagulation factor VIII

“…Комерційні реактивні барвники містять різні домішки, які можуть вплинути на їхні біохімічні властивості й, відповідно, можливість використання барвників [2]. Ре-активні барвники очищають за допомогою низки хроматографічних процедур, таких як тонкошарова хроматографія, високоефективна рідинна хроматографія та ко-лонкова на силікагелі або сефадексі [8].…”

High‐Performance Liquid Chromatography of Biological Macromolecules

“…372 In general, protein elution from dye ligands can take place by salt gradients, sometimes with altered pH, but enzymes are often eluted more efficiently and at higher purity if a biospecific eluent component such as a substrate 373 or substrate analog, 374 an inhibitor, 371,375 or a product or product analog 376 is added to the eluent either continuously at a low concentration or in pulses. Cofactors such as nucleotides are the most common biospecific displacers, and sometimes an oligonucleotide may also be needed to develop a biospecific solution phase interaction.…”

A potential generic downstream process using Cibracon Blue resin at very high loading capacity produces a highly purified monoclonal antibody preparation from cell culture harvest

“…Triazine dyes, first developed in the 1950s for the textile industry, have been widely used for large scale enzyme production but have found limited application in biopharmaceutical production [1]. This lack of usage in the pharmaceutical industry is principally due to a concern of toxicity associated with dye leakage resulting in dye presence in drug [2,3].…”

“…and operated for multiple cycles before replacing the adsorbent [4]. Cibracon Blue (CB), a synthetic polycyclic dye, binds to a wide variety of proteins [1,5]. The interaction is either through a strong affinity association between protein and CB due to its mimicry of nucleotide cofactors, as in case of oxidoreductases, transferases, kinases and dehydrogenases [6], an interaction of dye at the enzyme active site resulting in competitive inhibition of substrate binding [7,8,9], or an interaction distal from the active site through multiple interaction involving hydrogen bonding, Van der Waals forces, and hydrophobic, and ionic bonding [10].…”