The multivariate use of vibrational spectroscopy for chemical characterisation of chromatography media

Abstract

Chromatography media used in purification of biomolecules are extensively tested with various methods to ensure high quality products. The spherical and porous polymer particles are derivatised to give different chromatographic properties. The variations in, for instance, ligand content and chromatographic function are generally tested with a number of conventional analytical methods (mainly wet-chemical and functional methods) and these are time consuming. However, by using vibrational spectroscopy, chemical information can be efficiently obtained from the spectral data. In combination with traditional analytical data, multivariate models can be constructed. These models can then be used as a tool for determination (prediction) of, for example, ligand content. In the present work, prediction of ionic capacity and content of allyl groups are exemplified by Raman spectroscopy of two different types of agarose-based media (Sepharose™ Fast Flow Sepharose™). The conclusion is that vibrational spectroscopy is a simple, fast and highly informative tool for both qualitative and quantitative characterisation of adsorbents used in chromatography of biomolecules.