RESUMO
In downstream processing of protein therapeutics, ion exchange (IEX) chromatography is a powerful tool for removing byproducts whose isoelectric point (pI) is appreciably different from that of the product. Although in theory for a given case cation exchange (CEX) and anion exchange (AEX) chromatography should be equally effective for separation, in reality they may show different effectiveness. In the current work, with a case study, we demonstrated that AEX is more effective than CEX chromatography at removing the associated byproducts. In addition, we screened AEX resins and loading conditions to achieve best separation. Finally, we demonstrated that effective separation was achieved with the selected resin/condition, and chromatography performance was comparable between runs conducted at low and high load densities, suggesting that the developed process was relatively robust. The procedure described in this work can be used as a general approach for selecting resin and loading condition that allow for effective and robust removal of byproduct that binds weaker than the product to the selected type of column.
Assuntos
Resinas de Troca Aniônica , Cromatografia por Troca Iônica/métodos , Resinas de Troca Aniônica/química , Ânions , Cátions/químicaRESUMO
The introduction of the epothilone polyketide synthase (PKS) into Myxococcus xanthus has enabled the heterologous production of epothilone D (1) on a large scale. To isolate this valuable product from the fermentation medium, an economical, scalable, and high-yielding purification process was developed. With the crystallization of 1 from a binary solvent system that consisted of ethanol and water, the product was recovered as white crystals with a final purity of > or =97% (w/w). This is the first reported crystallization of 1.