Preparation of Membrane Receptor Proteins Utilising Large Scale Micro-carrier Culture and a 'Pearl Freezing Factory' to Finalise the Product

摘要

Bioreactor culture of adherent cells offers clear advantages for scale-up of cells for membrane preparations. However, harvesting cells from micro-carriers is time-consuming. In addition, enzymatic treatment may lead to loss of ligand binding and functional activity. To address this, we investigated scalable procedures to streamline cell harvesting and membrane preparation avoiding the use of enzymatic treatment. The concept is based on wash of cell-loaded micro-carriers in the fermentor, harvest andfreezing of the cells directly without enzymatic treatment. Starting from the thawed carrier-cell material we compared enzymatically treated with non treated membrane preparations with respect to yield of functionally active receptors.