Membrane based separations are emerging as a superior alternative to packed bed chromatographic processes by virtue of high surface area, reduced diffusion distance, low operating pressure, and higher cost effectiveness. Their application can be particularly interesting for large scale isolation of therapeutic proteins if the membranes are made more selective for the protein of interest. In this paper, an affinity membrane was designed and prepared for isolating plasminogen activators (PAs) from mono- and multi- component systems. PAs are therapeutic agents for thrombovascular disorders. Regenerated cellulose (RC) membranes were chemically modified with para-aminobenzamidine (pABA) via spacer arms of different lengths. The membrane containing pABA linked through 14-atoms spacer arm was found to bind up to 5.5-times higher amounts of PA as compared to membranes linked to pABA through shorter (5-, and 7- atoms) spacer arms, from pure PA solution as well as the conditioned cell culture media. A 40-fold purification was achieved in a single step separation of PA from HEK-293 conditioned media using these affinity membranes, irrespective of the length of the spacer arm. An extensive regeneration procedure allowed the preservation of approximately 90% of the PA binding capacity of the membranes even after five cycles of use.

Para-aminobenzamidine linked affinity membranes for plasminogen activator purification

243rd National Meeting (2012)

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