Improving Downstream Process Related Manufacturability Based on Protein Engineering—A Feasibility Study

• Published in: Engineering in life sciences Vol. 24; no. 9; pp. e202400019 - n/a
• Main Authors: Capito, Florian, Wong, Ting Hin, Faust, Christine, Brand, Kilian, Dittrich, Werner, Sommerfeld, Mark, Tiwari, Garima, Langer, Thomas
• Format: Journal Article
• Language: English
• Published: Germany John Wiley & Sons, Inc 01-09-2024; John Wiley and Sons Inc; Wiley-VCH
• Subjects: anion exchange chromatography; Anion exchanging; Antibodies; Biological activity; Charge distribution; Charge exchange; Chromatography; Cloning; codon usage; Design; Design for manufacturability; Diabetes; Engineering; Fc receptors; Fc‐fusion protein; Feasibility studies; Fusion protein; Impurities; Insulin; Isoelectric points; Manufacturability; Optimization; Peptides; Plasma; Production costs; Protein engineering; Protein expression; Protein purification; protein surface charge; Proteins; Surface charge; Viruses; anion exchange chromatography; Fc‐fusion protein; protein surface charge; codon usage; manufacturability
• ISSN: 1618-0240; 1618-2863
• DOI: 10.1002/elsc.202400019

ABSTRACT
While bioactivity and a favorable safety profile for biotherapeutics is of utmost importance, manufacturability is also worth of consideration to ease the manufacturing process. Manufacturability in the scientific literature is mostly related to stability of formulated drug substances, with limited focus on downstream process‐related manufacturability, that is, how easily can a protein be purified. Process‐related impurities or biological impurities like viruses and host cell proteins (HCP) are present in the harvest which have mostly acid isoelectric points and need to be removed to ensure patient safety. Therefore, during molecule design, the surface charge of the target molecule should preferably differ sufficiently from the surface charge of the impurities to enable an efficient purification strategy. In this feasibility study, we evaluated the possibility of improving manufacturability by adapting the surface charge of the target protein. We generated several variants of a GLP1‐receptor‐agonist‐Fc‐domain‐FGF21‐fusion protein and demonstrated proof of concept exemplarily for an anion exchange chromatography step which then can be operated at high pH values with maximal product recovery allowing removal of HCP and viruses. Altering the surface charge distribution of biotherapeutic proteins can thus be useful allowing for an efficient manufacturing process for removing HCP and viruses, thereby reducing manufacturing costs.