Posters

CIM (Convective Interaction Media) represent a new generation of chromatographic supports. In contrast to conventional particle supports, where the void volume between individual porous particles is unavoidable, CIM supports consist of a single monolith with open channels. In this way, molecules to be separated are transported into the pores by convection, resulting in short separation times.

CIMsupports proved to be very efficient for extremely fast separations of proteins in ion exchange, hydrophobic interaction and affinity chromatography mode. Recently, the successful separation of DNA as well as some smaller molecules like e.g. peptides and oligonucleotides were also performed.

All the above mentioned separations were carried out on an analytical scale with the use of 0.34 mL CIM discs. The scale-up of monolithic units was limited mainly due to the problems associated to the mechanical stability, poor sample distribution and higher backpressures. The change from the axial to radial flow enables the design of the so-called 8 and 80mLCIM tubes. They were basically designed for very fast purification of macromolecules.

In this work we present some basic characteristics of these newly developed units in terms of separation and binding capacity. In addition, some practical examples will be given and discussed as well.

Isolation and purification of proteins, peptides and polynucleotides as well as fractionation of biological mixtures are of great importance both for the solution of theoretical problems in chemistry and biology and the
realization of practical plans connected, in particular, with the production of medicines on the basis of large biomolecules. An important problem in the production of biological substances for medicine is to work out the step of their isolation and fine purification, e.g. creation of high performance separation methods, particularly, the chromatographic techniques. Here, fast and efficient affinity separations based on dynamical interaction of biocomplements play very important role.

High Performance Membrane (Monolith) Chromatography (HPMC) allows to solve all problems of High Liquid Chromatography (HPLC) demonstrating a number of number of distinct advantages. A small thickness of separation layer and opened structure of throughput channels where the separation takes place cause minimum difussion resistance for normal mass transport of the substances as well as low working back pressure and thus, the possibility of use of high elution flow rates.

In order to enable the detection of low abundance proteins from human plasma, it is necessary to remove high abundance proteins. Among them, human serum albumin and immonoglobulin G represent more than 75 % of all abundance proteins. There are many strategies developed for an efficient removal of these two main proteins, the majority of them rely on highly selective, yet expensive affinity techniques. In this work an affinity monolithic column was used for the depletion of IgG. For the removal of HSA we tested an alternative - complementary approach, where an ion-exchange mode was used as one of the depletion steps. the results were compared to the ones obtained by by using the prseudoaffinity columns.