Affinity chromatography with pseudobiospecific ligands on high-performance supports for purification of proteins of biotechnological interest

Abstract

High-performance affinity matrices were obtained by attaching pseudobiospecific ligands to hollow-fibre membranes. The neutral protease contained in FlavourzymeTM was purified to homogeneity with Yellow 4R-HE affinity hollow-fibre membranes. Immobilisation of Red HE-3B allowed purification of a milk-clotting enzyme obtained by solid-state culture of Mucor bacilliformis. Copper immobilisation through iminodiacetic acid allowed fractionation of Biocon Bioconcentrated PlusTM to separate the pectinesterase-containing fraction. The productivity of the developed processes - 1900, 94 and 750 U/ml.min, respectively - was 10- to 15-fold higher than that achieved with the same ligands immobilised on agarose-based soft gels, mainly due to the shortening of the purification processes

Development of New Strategy for Non-Antibiotic Therapy: Bovine Lactoferrin Has a Potent Antimicrobial and Immunomodulator Effects

“…The limitation of this technology for large scale applications lies with its high cost and its relatively low yield [30]. However, affinity membranes with immobilized triazinic dyes have not achieved yet good acceptance in the biotechnological industry, mainly because of their low capacity for proteins in comparison with the same ligands immobilized on soft gels [31,32] and the dye leaching in the elution and regeneration steps [33]. Although, under equilibrium conditions, membranes show an acceptable chromatographic performance for Lf purification from bovine colostrum, better than the obtained with d-Sepharose, as a model of soft gels [34], the main problems affecting industrial utilization of adsorptive dye membranes, such as low capacity, dye leaching and pressure drop along the fiber axis need to be overcome.…”

Improved hollow-fibre membranes for dye-affinity chromatography

“…From these results, it is evident that the hydrophilicity of the grafted polymer improves elution of the adsorbed protein to a significant extent. This aspect is interesting in membrane chromatography where elution of adsorbed proteins is not always as efficient as desired [6].…”

“…However, dye affinity hollow-fibre membranes show a protein adsorption capacity lower than that of soft-gel matrices [6]. Increasing maximum capacity is a key factor for improving the performance of this methodology.…”

Solution-blown core–shell hydrogel nanofibers for bovine serum albumin affinity adsorption

“…It is usually carried out in packed-bed chromatography columns lled with so gels. 1 However, classical bead-based column chromatography exhibits drawbacks such as slow intraparticle diffusion, large pressure drop through the column, low product throughput and high cost, etc. 10 With the development of membrane technology, ligand molecules are introduced into the surfaces of membranes to specically capture ligate, while letting other molecules pass through.…”