BIA Separations (Slovenia)

About: BIA Separations (Slovenia) is a company organization based out in Ljubljana, Slovenia. It is known for research contribution in the topics: Monolithic HPLC column & Monolith. The organization has 84 authors who have published 152 publications receiving 4883 citations.

Monolithic bioreactors for macromolecules.

Abstract: Enzymes immobilized on solid-phase matrices have found various applications in biotechnology, molecular biology and molecular diagnostics and can serve as industrial catalysts and as specific reagents for analytical procedures. A wide range of supports have been utilized for immobilization among which particle-based supports are the most commonly implemented. Type of support used for immobilization is one of the key considerations in practical application due to different immobilization efficiency, ligand utilization and the mass transfer regime. The mass transfer between the mobile and the particulate stationary phase is often a bottleneck for the entire process due to slow pore diffusion of large molecules. In contrast, monoliths due to their structure enable almost flow-independent properties. Consequently, the overall behavior of the immobilized ligand reflects its intrinsic reaction kinetics. Therefore, such an immobilized system is expected to allow higher throughput because of higher enzyme efficiency, especially pronounced for macromolecular substrates having low mobility. In this work, different methods for immobilization of enzymes on Convective Interaction Media monolithic supports are presented. In particular, enzymes acting on macromolecular substrates, such as trypsin, deoxyribonuclease and ribonuclease, are described in detail. Immobilized efficiency is evaluated for different immobilization procedures in terms of biologic activity and long-term stability. Finally, their performance on real samples is demonstrated.

Scale-Up of Plasmid DNA Downstream Process Based on Chromatographic Monoliths.

Abstract: Purification of high-quality plasmid DNA in large quantities is a crucial step in its production for therapeutic use and is usually conducted by different chromatographic techniques. Large-scale preparations require the optimization of yield and homogeneity, while maximizing removal of contaminants and preserving molecular integrity. The advantages of Convective Interaction Media® (CIM®) monolith stationary phases, including low backpressure, fast separation of macromolecules, and flow-rate-independent resolution qualified them to be used effectively in separation of plasmid DNA on laboratory as well as on large scale. A development and scale-up of plasmid DNA downstream process based on chromatographic monoliths is described and discussed below. Special emphasis is put on the introduction of process analytical technology principles and tools for optimization and control of a downstream process.