Monolithic chromatography supports

ITQB/IBET-Seminar

Title: Monolithic chromatography supports – technology platform for the analysis and production of new generation of drug candidates like plasmid DNA and viral vector particles.

Speaker: Aleš Štrancar-MD PhD

Affiliation: BIA Separations, Teslova 30, SI-1000 Ljubljana

Host: Cristina Peixoto (ACTU)

Abstract:

Developments in gene therapy and threat of pandemics highlight the fact that therapeutics based on very large biomolecules and nanoparticles are undergoing a rapid expansion. Production of plasmid DNA and virus particles on industrial scale is becoming an imperative, in first instance, the purification and in-process control.

Short monolithic columns represent a novel generation of chromatographic supports. In contrast to conventional particle supports, where the void volume between individual porous particles is unavoidable, these supports consist of a single highly interconnected monolith with large and small open flow-through channels (large channels are of several µm in size). Due to the monolith structure, the target molecules are transported to the active sites on the stationary phase by convection resulting in very short separation times. In addition most of the active sites are easily accessible resulting in a very high binding capacity for large molecules (e.g. 10 mg pDNA/ml support) and nanoparticles. Beside the speed and capacity also the yield of active biomolecule can largely be improved.

One of the successful application of these columns on industrial scale is cGMP production of pDNA for gene therapy purposes. The 15 fold increase of productivity (in comparison to porous particles based supports) of the purification part of the production process have been reported1. Several other examples of virus purification will be presented as well.

Rapid analysis of adenoviruses to support the development of high titre manufacturing platform and PAT will be presented as an example of successful introduction for in-process control2.

1J. Urthaler et al; Application of monoliths for plasmid DNA purification: Development and transfer to production. J. Chromatogr. A, 2005, 1065, 93.

2P. Ball et all; Rapid high-performance liquid chromatographic analysis of adenovirus type 5 particles with a prototype anion-exchange analytical monolith column. J. Chromatogr. A, 2009, 1216  2725.