The growing market of biopharmaceuticals and the constant developments in upstream fermentation have generated a strong demand for new downstream purification methods. Magnetic separation in combination with functional magnetic particles has been known for many years as a promising candidate for a direct capturing tool in protein purification but the lack of suitable GMP-compliant purification equipment has prevented the launch of this technology in large scale bioprocessing. To tackle this bottle-neck, the principle of a "rotor-stator" high-gradient magnetic separator is fully redesigned to meet the rigorous requirements of modern cGMP biotechnology purification processes. In order to fulfill regulatory requirements, the separation chamber is reengineered to allow effective cleaning and sterilization in place while maintaining excellent separation capacities and efficiencies. Two kinds of commercially available magnetic particles are used to validate key performance data and determine system related parameters in order to calculate process performance figures for process optimization of the new magnetic separation device. With separation capacities of over 400 g of magnetic particles per liter of separation chamber volume and separation efficiencies as well as recovery rates over 99%, the system is able to process more than 200 l crude feedstock per day and capture more than 1.6 kg target compounds.
Magnetic separation processes are known as integrated bioanalytical protein purification method since decades and are well described. However, use of magnetic separation processes in a regulated industrial production environment has been prevented by the lack of suitable process equipment and prejudice against the productivity of the process and its qualification for cleaning‐in‐place operation. With the aim of overcoming this prejudice, a comprehensive process development approach is presented, based on a GMP‐compliant magnetic separator, including an optimization of the batch adsorption process, implementation into a technical‐scale, and the development and validation of cleaning routines for the device. By the implementation of a two‐step counter‐current binding process, it was possible to raise the yields of the magnetic separation process even for very low concentrated targets in a vast surplus of competing proteins, like the hormone equine chorionic gonadotropin in serum, from 74% to over 95%. For the validation of the cleaning process, a direct surface swabbing method combined with a total organic carbon analysis was established for the determination of two model contaminants. The cleanability of the process equipment was proven for both model contaminants by reliably meeting the 10 ppm criteria.
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