Antibody Expression and Production
Engineered antibodies currently represent over 30% of biopharmaceuticals in clinical trials and their total worldwide sales continue to increase significantly. The importance of antibody applications is reflected in their increasing clinical and industria
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Recovery and Purification of Antibody XueJun Han, Arthur Hewig, and Ganesh Vedantham
Abstract Monoclonal antibody drugs have become a large portion of protein therapeutics and many antibody molecules are being evaluated at various stages of clinical trials in the biopharmaceutical industry. This review article summarizes the state of the art antibody purification techniques. The main focus is chromatographic techniques that include protein A, ion exchange and HIC. For each technique, the mechanisms of antibody binding, factors affecting binding capacity, resin selection, as well as typical process parameters and chromatograms are discussed in detail. Cell culture clarification, filtration, alternative antibody purification techniques, and viral clearance strategies are discussed briefly. The goal of this article is to provide a broad coverage of antibody purification technology. Readers are referred to extensive articles for further in-depth reading.
14.1 Introduction Monoclonal antibodies (mAbs) have unique properties that have made them the most prevalent therapeutics in the biopharmaceutical industry, presently accounting for a large proportion of recombinant protein drug candidates in clinical development (Walsh, 2004). Two of these unique properties are their specificity for disease targets and wide range of targets. In the last 2 decades, many mAbs have received marketing approval by the regulatory agencies for a variety of indications such as non-Hodgkin’s lymphoma, rheumatoid arthritis, and colorectal cancer (Shukla and Kandula, 2009). Antibody therapies may involve frequent high doses. To meet the demand for some indications, several hundred kilograms of antibody product per year may be required. Recent advances in cell line selection, growth and production media, and feeding strategies have led to antibody expression levels as high as 5 g L–1 in a 12-day fed-batch process (Jagschies et al., 2006), with some reporting 10 g L–1 through longer cell culture duration (Luan et al., 2006). The combination of high titer and large bioreactors will result in >100 kg batch sizes. Consequently, purification costs are now greater than cell culture costs and process bottlenecks have moved to downstream (Gottschalk, 2008). The current focus for antibody X. Han, A. Hewig, and G. Vedantham (B) Amgen Inc., Seattle, WA 98119, USA e-mail: [email protected] M. Al-Rubeai (ed.), Antibody Expression and Production, Cell Engineering 7, C Springer Science+Business Media B.V. 2011 DOI 10.1007/978-94-007-1257-7_14,
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purification process development is to streamline process development activities, reduce manufacturing cost, and increase throughput in manufacturing facilities. The primary considerations for downstream process development are product purity and process yield. The process needs to remove process related contaminants and product related impurities. Process related contaminants include cells, cell debris, host cell protein (HCP), DNA, endotoxin, leached Protein A, as well as chemical reagent
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