In-situ product removal (ISPR) of functional proteins from prokaryotic cultivations by high specific magnetic separation

摘要

The objective of this work is to demonstrate the use of functionalized magnetic particles as a basis technology for high specific in-situ separation of proteins. His-tagged phytase and subtilisin, extracellular produced by Escherichia coli and Bacillus licheniformis respectively, were chosen as model products. Two different kinds of magnetizable filter chambers were used to perform solid-liquid separation of magnetic particles and the crude fermentation broth. The filter systems differed with regard to lattice geometry, total chamber volume and the nature of the magnetic field generation (electromagnet and permanent magnet, respectively). Analysis of particle capacity (with respect to the target products) and stability showed that the used magnetic particles are well suited regarding both product capacity and purity. Furthermore the reusability of the particles could be verified by multiple loading and elution cycles. Kmagnetic separation;; bio separation;; HGMF;; functionalized magnetic particles