In the development of biopharmaceutical protein therapeuticdrugs, the extent of aggregation must be minimized in order to reduce thepotential for aggregate-induced immunogenicity. Size exclusion chromatographyprovides a rapid and simple means to assess the extent of aggregation.Traditional SEC separations typically require long analysistimes, primarily because the slow diffusion of large molecular weight analyteswithin the porous media requires operation at low linear velocities. By usingsmaller chromatographic particles, speed and resolution can be improved owingto faster mass transfer of analyte between particle and bulk fluid.  We have recently expanded the scope ofthe ethylene-bridged hybrid (BEH) technology by re-engineering ourparticle-making process to create porous particles with almost a two-foldincrease in pore volume compared with the original BEH particle.  This newly designed chromatographicmedia has been shown to maintain the mechanical strength requirements necessaryof a 1.7 micron particle for UPLC applications. This presentation will discuss the benefits in speed andresolution that can be obtained with UPLC-SEC for biomolecule separations.  The effect of particle size, pore volumeand pore size distribution on chromatographic resolution will be evaluated, aswell as the effect of ionic strength on SEC performance.  In addition, the effect of instrumentdispersion on size-exclusion separations will be discussed.  Examples will be given of size-basedseparations, including separations of monoclonal antibody monomers fromaggregates.

Separations of biomolecules by UPLC-SEC

240th National Meeting (2010)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.