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Optimization of a platform process operating space for a monoclonal antibody susceptible to reversible and irreversible aggregation using a solution stability screening approach

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Platform manufacturing processes are widely adopted to simplify and standardize the development and manufacturing of monoclonal antibodies (mAbs). However, there are mAbs that do not conform to a platform design due to instability or other protein properties leading to a negative impact on product quality or process performance (non-platform mAb).

Journal of Chromatography A, 1597 (2019) 100–108 Contents lists available at ScienceDirect Journal of Chromatography A journal homepage: www.elsevier.com/locate/chroma Optimization of a platform process operating space for a monoclonal antibody susceptible to reversible and irreversible aggregation using a solution stability screening approach Adrian Man a , Haibin Luo a , Sophia V Levitskaya b , Nathaniel Macapagal a , Kelcy J Newell a,∗ a b Purification Process Sciences, AstraZeneca, One MedImmune Way, Gaithersburg, MD, 20878, USA Analytical Sciences, AstraZeneca, One MedImmune Way, Gaithersburg, MD, 20878, USA a r t i c l e i n f o Article history: Received 19 September 2018 Received in revised form 12 March 2019 Accepted 13 March 2019 Available online 20 March 2019 Keywords: Monoclonal antibody purification Automation High throughput process development Aggregation Scale-down Reversible self-association a b s t r a c t Platform manufacturing processes are widely adopted to simplify and standardize the development and manufacturing of monoclonal antibodies (mAbs) However, there are mAbs that not conform to a platform design due to instability or other protein properties leading to a negative impact on product quality or process performance (non-platform mAb) Non-platform mAbs typically require prolonged development times and significant deviations from the platform process to address these issues due to the need to sequentially optimize individual process steps In this study, we describe an IgG2 mAb (mAb A) that is susceptible to aggregation and reversible self-association (RSA) under platform conditions In lieu of a sequential optimization approach, we evaluated the solution stability of mAb A across the platform operating space (solution stability screen) This screening design was used to identify interacting parameters that affected the non-platform mAb stability A subsequent response surface design was found to predict an acceptable operating space that minimized aggregate formation and RSA across the entire process This information guided the selection of optimal parameters best suited to avoid destabilizing conditions for each process step Substantial time savings was achieved by focusing development around these factors including protein concentration, buffer pH, salt concentration, and excipient type In addition, this work enabled the optimization of a cation exchange chromatography step that removed aggregate without yield losses due to the presence of reversible aggregation The final optimized process derived from this study resulted in an increase in ˜ yield of 30% over the original process while maintaining the same level of aggregate clearance to match product quality Solution stability screening is readily adapted to high throughput technologies to minimize material requirements and accelerate analytical data availability Implementation of high throughput approaches will further expedite process development and enable enhanced selection of candidate drugs by including process development objectives © 2019 The Authors Published by Elsevier B.V This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/) Introduction Monoclonal antibodies (mAbs) have emerged as a rapidly growing class of therapeutic since the mid-1990s According to the FDA database there are more than 70 modern antibody-based therapeutic agents approved in the US and more than 500 additional ∗ Corresponding author E-mail address: newellkj@medimmune.com (K.J Newell) products are currently in clinical development [1] Many biopharmaceutical companies have adopted platform mAb purification processes to simplify process development and manufacture of mAbs [2] Fig illustrates a common platform process with commonly used steps including; (1) affinity purification capture, (2) low pH virus inactivation, (3) anion exchange chromatography for process related impurity and virus removal, (4) cation exchange chromatography for process and product related impurity removal (5) virus filtration, and (6) formulation that utilizes (a) ultrafiltration and (b) diafiltration to generate drug substance https://doi.org/10.1016/j.chroma.2019.03.021 0021-9673/© 2019 The Authors Published by Elsevier B.V This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4 0/) A Man et al / J Chromatogr A 1597 (2019) 100–108 101 Table Summary of non-ideal observations for a mAb in the platform process Step Protein A Chromatography Low pH Inactivation Anion Exchange Chromatography Cation Exchange Chromatography Virus Filtration 6A UF1 6B UF2 a b c Condition pH Conductivity (mS/cm) Protein conc (mg/mL) 3.6-4.5 0.5-1.5 10-25 3.4-3.6

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