The process uses differential precipitation in combination with microfiltration to remove soluble and insoluble impurities from the protein or peptide. The two MF steps facilitates purification of proteins and peptides by reducing both soluble and insoluble impurities that may be associated with the protein or peptide while providing for the exchange the protein or peptide into a suitable buffer for use or for downstream purification. The two MF steps may comprise a single upstream purification step in a process for producing a purified or substantially purified protein or peptide product that is performed before further downstream purification of the protein or peptide. The two MF steps comprise a single downstream purification step for purification of a protein or peptide, which may be performed subsequent to obtaining the protein or peptide from cell culture or in vitro synthesis to produce a purified or substantially purified protein or peptide product. The present invention provides a process for producing proteins and peptides comprising two tandem microfiltration (MF) or diafiltration steps. 2002), efficient scale-up remains a significant challenge. Although models such as pore blockage, pore constriction, and cake filtration have been developed to aid in mitigation of fouling (Ho and Zydney 2000 Palacio et al. Fouling is typically quantified as decreased flux at constant TMP or increased TMP at constant flux. The most common operational challenge for MF processes is filter fouling due to deposition of insoluble material on the membrane surface or inside membrane pores, which increases flow resistance (Marichal-Gallardo and Alvarez 2012 van Reis and Zydney 2007). However, performance upon scale-up is difficult to predict, heavily dependent on the nature of the feedstock, and often must be optimized empirically (Roush and Lu 2008 Saxena et al. Most commonly, MF unit operations are scaled on constant load factor (volume or mass per filter area), membrane channel geometry, path length, crossflow velocity, and transmembrane pressure (TMP) (Marichal-Gallardo and Alvarez 2012). Successful scale-up is a key challenge in implementation of microfiltration processes.
MF processes have been classically employed in harvest and primary recovery, whereas UF processes have been largely used as polishing steps for buffer exchange or concentration (Marichal-Gallardo and Alvarez van Reis and Zydney 2001). TFF is typically categorized as microfiltration (MF pore sizing in microns) or ultrafiltration (UF pore sizing in nominal molecular weight limit). Tangential flow filtration (TFF) is a robust and versatile separation technique used in biopharmaceutical manufacturing. The present invention relates to the use of two tandem microfiltration (MF) steps in a process for making recombinant insulin. BACKGROUND OF THE INVENTION (1) Field of the Invention
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