(745f) Synthesis, Functionalization and Application of Novel Porous Microclusters for Protein Separation

Separation of various biomolecules from fermentation broth can be very efficiently done by high performance liquid chromatography (HPLC).

The separation speed and efficiency strongly depends on the active surface and functionalization of the stationary phases used. Although spherical particles packed into a chromatographic column is the simplest solution, they are not always the optimal one. On one hand, chromatographic columns packed with small particles provide sufficient surface for good separation, however due to small size and therefore high pressure drop, their application is restricted to very low flow rates. On the other hand, large particles could ensure low pressure drop and therefore rather high flow rates, up to tens of ml per minute, however their active surface is limited.

Objective of this work is to synthesize highly porous particles to maintain low pressure drop, however with a sufficient number of active sites available for separation. The presented approach is based on an aggregation and breakage process to form porous microclusters combined with “reactive gelation” to increase mechanical rigidity of the prepared microclusters. Atom transfer radical polymerization is used to growth functional brushes on the internal surface of formed microclusters.

The effects of parameters such as size and crosslinking degree of primary particles, as well as shear rate applied during microcluster preparation are investigated. Various analytical techniques, namely light scattering, N₂ sorption, mercury porosimetry, micro- and nano-tomography, size-exclusion chromatography are used to characterize the size and internal structure of the obtained microclusters, which are subsequently applied for protein separation, with their performance evaluated against other commercial stationary phases.