High throughput determination of cleaning solutions to prevent the fouling of an anion exchange resin

• Published in: Biotechnology and bioengineering Vol. 113; no. 6; pp. 1251 - 1259
• Main Authors: Elich, Thomas, Iskra, Timothy, Daniels, William, Morrison, Christopher J.
• Format: Journal Article
• Language: English
• Published: United States Blackwell Publishing Ltd 01.06.2016; Wiley Subscription Services, Inc
• Subjects: anion exchange chromatography; Anion Exchange Resins - analysis; Anion Exchange Resins - chemistry; Antibodies, Monoclonal - chemistry; Automation; Bioengineering; Biotechnology; Chromatography; Chromatography, Ion Exchange - instrumentation; Chromatography, Ion Exchange - methods; Cleaning; Cleaning agents; Cleaning compounds; Detergents - analysis; Detergents - chemistry; Equipment Contamination - prevention & control; Equipment Design; Equipment Failure Analysis; Fouling; Heat treatment; high throughput process development; High-Throughput Screening Assays - instrumentation; High-Throughput Screening Assays - methods; miniaturized column chromatography; monoclonal antibody; Polymers; purification; resin fouling; Resins; high throughput process development; anion exchange chromatography; monoclonal antibody; miniaturized column chromatography; purification; resin fouling
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/bit.25881

ABSTRACT Effective cleaning of chromatography resin is required to prevent fouling and maximize the number of processing cycles which can be achieved. Optimization of resin cleaning procedures, however, can lead to prohibitive material, labor, and time requirements, even when using milliliter scale chromatography columns. In this work, high throughput (HT) techniques were used to evaluate cleaning agents for a monoclonal antibody (mAb) polishing step utilizing Fractogel® EMD TMAE HiCap (M) anion exchange (AEX) resin. For this particular mAb feed stream, the AEX resin could not be fully restored with traditional NaCl and NaOH cleaning solutions, resulting in a loss of impurity capacity with resin cycling. Miniaturized microliter scale chromatography columns and an automated liquid handling system (LHS) were employed to evaluate various experimental cleaning conditions. Cleaning agents were monitored for their ability to maintain resin impurity capacity over multiple processing cycles by analyzing the flowthrough material for turbidity and high molecular weight (HMW) content. HT experiments indicated that a 167 mM acetic acid strip solution followed by a 0.5 M NaOH, 2 M NaCl sanitization provided approximately 90% cleaning improvement over solutions containing solely NaCl and/or NaOH. Results from the microliter scale HT experiments were confirmed in subsequent evaluations at the milliliter scale. These results identify cleaning agents which may restore resin performance for applications involving fouling species in ion exchange systems. In addition, this work demonstrates the use of miniaturized columns operated with an automated LHS for HT evaluation of chromatographic cleaning procedures, effectively decreasing material requirements while simultaneously increasing throughput. Biotechnol. Bioeng. 2016;113: 1251–1259. © 2015 Wiley Periodicals, Inc. Microliter scale chromatography columns and an automated liquid handling system were employed in a high throughout manner to evaluate cleaning solutions for a monoclonal antibody anion exchange polishing step which could not be fully restored with traditional NaCl and NaOH cleaning agents. By using this strategy, the authors determined an effective cleaning procedure with minimal material requirements. The optimal cleaning procedure was verified at milliliter scale, confirming that microliter scale chromatography columns provide representative performance in cleaning applications.