Methods for characterization of biochromatography media

Abstract

Chromatographic methods represent the most powerful techniques for purification of biopharmaceutical compounds. Quite often, the question arises which chromatographic medium should be chosen for a particular purification task or which technique should be applied to obtain the required information for a process, respectively. The present review aims to guide through these questions by presenting experimental and modeling techniques that allow a detailed characterization and comparison of chromatography media as well provide a guideline of techniques for process development. The first section provides basic information on chromatographic theory, types of chromatographic media, and different types of techniques. The second section governs description of experimental techniques including some advises for laboratory practice. The third section presents and discusses selected references from literature. Within this article, the main focus is on traditional laboratory techniques but also automated high-throughput screening methods will briefly be discussed.

Characterization of lysozyme adsorption in cellulosic chromatographic materials using small-angle neutron scattering

“…Consequently, protein loading of these particles can be thought of as a partitioning into the three-dimensional cellulosic structure, instead of as adsorption at an extended twodimensional surface. This structural similarity to protein sorption in polymer-derivatized materials supports the high static binding capacity and rapid uptake of protein observed in HyperCel TM [5,14,15].…”

“…Chromatography is the dominant large-scale separation technique for proteins, such that improvements in efficiency and the development of non-chromatographic alternatives are a subject of current research [1][2][3][4]. Virtually all protein purification processes are developed around at least one chromatographic step [5], with an average of roughly three chromatographic steps per process. About 40% of these steps are ion-exchange chromatography [6].…”

Capture and intermediate purification of recombinant antibodies with combined precipitation methods

“…Currently, the state-of-the-art process for the biopharmaceutical production of monoclonal antibodies (mAbs) [2] consists of seven to nine discontinuous processing steps, including column-based polishing and mAb capturing [3][4][5][6]. In recent years, the development of downstream processing has mainly focused on improving the column-based purification steps [7][8][9]. Due to the cost of producing high affinity chromatography materials, most development work has focused on improving the binding capacity and stability of protein A resins [10,11].…”

A microscale method of protein extraction from bacteria: Interaction of Escherichia coli with cationic microparticles

“…DNA and endotoxins, both highly negatively charged, showed strong binding, and could be separated from the protein in a step desorption process with defined salt concentrations. As known from conventional ion exchange chromatography, proteins may have significantly different binding capacities, despite similar sizes and theoretical isoelectric points (Hahn, 2012). This phenomenon was observed for GFP and SOD.…”