Dye-affinity techniques for bioprocessing: Recent developments

Abstract

Textile or triazine dyes play an important role as affinity ligands in protein purification. Each step of the protein purification protocol can be divided into three stages, partitioning between two phases, separation of these phases and recovery of the target protein from the enriched phase. Now developments in dye-affinity techniques are discussed emphasizing the innovations in all three stages of the protein purification process. Dye-affinity chromatography has become a routine step in protein purification. New dyes have been developed and used successfully in both traditional chromatographic mode and new modes like affinity precipitation, polymer aqueous two-phase partitioning or expanded bed chromatography. The specificity of dye techniques has been increased by both purposeful designing of new dyes and decreasing non-specific protein-dye interactions with polymer shielding. One can envisage further development and ramification of dye-affinity techniques in protein purification.

Discriminatory protein binding by a library of 96 new affinity resins: A novel dye-affinity chromatography tool-kit

“…It is often therefore considered as a pseudo-affinity ligand. Few structural variants of trizine-based ligands as well as different spacer lengths are described to generate novel affinity resins [16][17][18]. However there have been no systematic efforts to generate structural variants of CB3GA, and to establish a structure-activity relationship.…”

“…Several directed and empirical attempts were made in the past to generate novel triazine-based dye-affinity resins [16][17][18]. The issues of selectivity were addressed by preparing synthetic ligands, synthesizing tailor made ligands based on molecular modeling and using other combinatorial approaches.…”

High‐Performance Liquid Chromatography of Biological Macromolecules

“…The same authors also reviewed new developments in dye affinity for bioprocessing, dealing specifically with new dyes and conditions for the protein -ligand attachment and detachment steps. 387 An interesting extension of this work is their use of water-soluble polymers for protein displacement in dye -ligand chromatography. 388 It was demonstrated that porcine muscle lactate dehydrogenase (LDH) could be efficiently displaced from a blue agarose gel by poly(N-vinyl pyrrolidinone) and poly(N-vinyl caprolactam).…”

Purification of lysozyme by multistage affinity filtration

“…broth feeding (selective adsorption), contaminant washing and desired protein elution. Affinity filtration shows great potential for application in large-scale and continuous processes [8,9]. Another important advantage of affinity filtration over affinity chromatography is its ability to process a crude broth without a prior clarification step.…”

“…Cibacron Blue 3GA (CB) was chosen as an affinity ligand and was immobilized to TSK gel to produce an affinity adsorbent. This inexpensive, stable and group-specific dye ligand is well suited for the large-scale purification of proteins [9]. The feasibility of a multistage affinity filtration process was investigated by the purification of lysozyme from solutions of lysozyme-BSA mixture and chicken egg white.…”