Relation of structure to performance characteristics of monolithic and perfusive stationary phases

Trilisky E. I. , Koku H. , Czymmek K. J. , Lenhoff A. M.

JOURNAL OF CHROMATOGRAPHY A, vol.1216, no.36, pp.6365-6376, 2009 (Journal Indexed in SCI) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 1216 Issue: 36
  • Publication Date: 2009
  • Doi Number: 10.1016/j.chroma.2009.07.005
  • Page Numbers: pp.6365-6376
  • Keywords: Perfusion, Monolith, Protein, Virus, SEM, TEM, Binding capacity, Permeability, Porosity, Pore size distribution, Breakthrough curve, PORE-SIZE DISTRIBUTIONS, ION-EXCHANGE CHROMATOGRAPHY, UPTAKE RATES, PURIFICATION, ADSORPTION, AFFINITY, BIOMOLECULES, ADSORBENTS, VECTORS, SEPARATION


Commercially available polymer-based monolithic and perfusive stationary phases were evaluated for their applicability in chromatography of biologics. Information on bed geometry, including that from electron microscopy (EM), was used to interpret and predict accessible volumes, binding capacities, and pressure drops. For preparative purification of biologics up to at least 7 nm in diameter, monoliths and perfusive resins are inferior to conventional stationary phases due to their low binding capacities (20-30 g/L for BSA). For larger biologics, up to several hundred nanometers in diameter, calculations from EM images predict a potential increase in binding capacity to nearly 100 g/L. The accessible volume for adenovirtis calculated from the EM images matched the experimental value. While the pores of perfusive resins are essentially inaccessible to adenovirus under binding conditions, under non-adsorbing conditions the accessible intrabead porosity is almost as large as the interbead porosity. Modeling of breakthrough curves showed that the experimentally observed slow approach to full saturation can be explained by the distribution of pore sizes. (C) 2009 Elsevier B.V. All rights reserved.