Application of platform process development approaches to the manufacturing of Mabcalin™ bispecifics

• Published in: Journal of biotechnology 2023-11, Vol.377, p.13-22
• Main Authors: Wachter, Stefanie, Angevin, Thibaut, Bubna, Niket, Tan, Adelene, Cichy, Adam, Brown, David, Wolfe, Leslie S., Sappington, Ryan, Lilla, Edward, Berry, Luke, Grismer, Dane, Orth, Christian, Blanusa, Milan, Mostafa, Sigma, Kaufmann, Hitto, Felderer, Karin
• Format: Article
• Language: English
• Subjects: anion exchange; Anticalin® protein; Biologics manufacturing; bioreactors; Bispecifics; cation exchange; class; drugs; Fusion proteins; hydrophobic interaction chromatography; Mabcalin™ bispecifics; monoclonal antibodies; Process development; therapeutics; ultrafiltration
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/j.jbiotec.2023.10.003

Bispecific biotherapeutics offer potent and highly specific treatment options in oncology and immuno-oncology. However, many bispecific formats are prone to high levels of aggregation and instability, leading to prolonged development timelines, inefficient manufacturing, and high costs. The novel class of Mabcalin™ molecules consist of Anticalin® proteins fused to an IgG and are currently being evaluated in pre-clinical and clinical studies. Here, we describe a robust high-yield manufacturing platform for these therapeutic fusion proteins providing data up to commercially relevant scales. A platform upstream process was established for one of the Mabcalin bispecifics and then applied to other clinically relevant drug candidates with different IgG target specificities. Process performance was compared in 3 L bioreactors and production was scaled-up to up to 1000 L for confirmation. The Mabcalin proteins’ structural and biophysical similarities enabled a downstream platform approach consisting of initial protein A capture, viral inactivation, mixed-mode anion exchange polishing, second polishing by cation exchange or hydrophobic interaction chromatography, viral filtration, buffer exchange and concentration by ultrafiltration/diafiltration. All three processes met their target specifications and achieved comparable clearance of impurities and product yields across scales. The described platform approach provides a fast and economic path to process confirmation and is well comparable to classical monoclonal antibody approaches in terms of costs and time to clinic. •Mabcalin proteins are novel bispecifics consisting of IgG-Anticalin protein fusions.•Scalable upstream and downstream manufacturing processes for bispecific molecules.•Platform for faster development timelines of bispecifics with reduced costs.•Establishing a high-titer bispecific platform yielding consistent product quality.