Cation exchange chromatography performed in overloaded mode is effective in removing viruses during the manufacturing of monoclonal antibodies

Abstract

Viral safety is a critical concern with regard to monoclonal antibody (mAb) products produced in mammalian cells such as Chinese hamster ovary cells. Manufacturers are required to ensure the safety of such products by validating the clearance of viruses in downstream purification steps. Cation exchange (CEX) chromatography is widely used in bind/elute mode as a polishing step in mAb purification. However, bind/elute modes require a large volume of expensive resin. To reduce the production cost, the use of CEX chromatography in overloaded mode has recently been investigated. The viral clearance ability in overloaded mode was evaluated using murine leukemia virus (MLV). Even under high‐load conditions such as 2,000 g mAb/L resin, MLV was removed from mAb solutions. This viral clearance ability was not significantly affected by resin type or mAb type. The overloaded mode can also remove other types of viruses such as pseudorabies virus and reovirus Type 3 from mAb solutions. Based on these results, this cost‐effective overloaded mode is comparable to the bind‐elute mode in terms of viral removal.

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

“…Viral safety is a critical concern for therapeutic proteins such as mAb produced using mammalian cells such as CHO cells. It was also reported that CEX carried out in overloaded mode was able to remove viruses during the manufacture of mAbs (Masuda et al, 2019). Hydrophobic interaction chromatography (HIC) is based on the relative hydrophobicity of the protein molecules.…”

1Progress, applications, challenges and prospects of protein purification technology

“… Radhakrishnan et al (2022) isolated a leucine-rich lumen binding protein of 24 kDa from Solanum trilobatum leaves by ammonium sulfate precipitation and ion exchange chromatography, which was found to have antibacterial activity and edible properties, and can be used for the clinical treatment of S. aureus and V. cholerae infections to alleviate the bacterial drug resistance. Ion exchange chromatography can also remove various impurities such as target protein variants, host cell residual proteins, DNA, culture medium components, endotoxin, and viruses ( Saraswat et al, 2013 ; Tripathi, 2016 ; Kimia et al, 2019 ; Masuda et al, 2019 ). In recent years, some improved ion exchange chromatography methods have been proposed.…”

Combinatorial peptides: A library that continuously probes low‐abundance proteins

“…This mechanism is verified on all types of adsorbents used in chromatography. Examples have been published on hydrophobic resins [28] as well as on anion- [29] and cation-exchangers [30]. It has been demonstrated as being very effective for the enrichment of certain low-abundance proteins in lectin affinity chromatography, [31] and in the detection of protein impurity traces [32] as well as in the removal of host cell proteins from purified antibodies [33].…”