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Evaluation of a New Cation Exchanger for Viral Clearance in Processing of Therapeutic Proteins

IP.com Disclosure Number: IPCOM000237476D
Publication Date: 2014-Jun-18
Document File: 3 page(s) / 273K

Publishing Venue

The IP.com Prior Art Database

Abstract

The present study concerns an evaluation of a new cation exchanger for viral clearance of therapeutic proteins such as antibodies or antibody-related proteins (fragments, fusion proteins, conjugates etc.). A bacteriophage has been used as a model virus. The new cation exchanger is based on highly crosslinked agarose beads of 50 micrometer average diameter (d50v), onto which a sulfonate- and pyrrolidone-functional copolymer has been tethered by grafting, producing a strong cation exchanger with an ionic capacity of 37-63 micromol H+/ml. Similar constructions, including methods of manufacturing, are described in (10).

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Evaluation of a new cation exchanger for viral clearance in processing of therapeutic proteins

Background

Therapeutic proteins, such as monoclonal antibodies (mAbs) are often expressed in mammalian cells, e.g. CHO cells, and are recovered and further purified in processes including a series of chromatography steps. The first step is typically a capture step (a Protein A column in the case of mAbs), which is then followed by one or two steps involving ion exchange chromatography, hydrophobic interaction chromatography and/or multimodal chromatography (1). It is common practice to include a strong cation exchange step in the process, typically with the purpose of removing antibody aggregates, host cell proteins, leached Protein A, viruses, undesirable antibody variants etc. (2-6). The cation exchange step is often performed in bind-elute mode, but may also be performed in flow-through or weak partitioning mode (3). Clearance of adventitious viruses occurs both in the chromatography steps and in a virus filtration step normally included in the process (9). For a high overall clearance, the viral clearance in each individual step should be as high as possible.

Cation exchangers are known to provide good viral clearance, e.g. in antibody processing (7,8).

Outline

The present study concerns an evaluation of a new cation exchanger for viral clearance of therapeutic proteins such as antibodies or antibody-related proteins (fragments, fusion proteins, conjugates etc.). A bacteriophage has been used as a model virus. The new cation exchanger is based on highly crosslinked agarose beads of 50 micrometer average diameter (d50v), onto which a sulfonate- and pyrrolidone-functional copolymer has been tethered by grafting, producing a strong cation exchanger with an ionic capacity of 37-63 micromol H+/ml. Similar constructions, including methods of manufacturing, are described in (10).

The performance of the new cation exchanger is illustrated by the experimental results below. The data are retrieved at an early stage and the experiments can be further optimized.

Experiments


1. Bind-elute removal of host cell proteins, fragments and aggregates

A monoclonal antibody Protein A eluate was buffer exchanged into 50 mM Na-acetate pH 5.0 + 50 mM NaCl (A-buffer), spiked with bacteriophage stock solution and applied on a column packed with the new cation exchanger. After loading of 80 g mAb per L resin and washing with A-buffer, the column was eluted with a 20 column volum...