Landscape of Manufacturing Process of ATMP Cell Therapy Products for Unmet Clinical Needs

Abstract

Immune cell therapies have been studied in numerous clinical trials using Advanced Therapy Medicinal Products (ATMP) against a number of diseases having no or inadequate alternative therapies available, for example, various cancer types, cerebral stroke, cardiac infarction, severe autoimmune disorders, or chronic infections. Despite the enormous number of positive observation in ex vivo or animal studies, convincing results in clinical studies remain scanty. The chapter presents a survey and reveals that the manufacturing of immune cells especially for clinical trials is until today primarily performed using archaic, scarcely controlled, and incomparable processes and methods. A deeper characterization of ex vivo expanded immune cells is urgently needed not only on the level of a few receptors and ligands on the cell surface but also with respect to the ever-contained subtypes in an expanded immune cell population, the pattern of secreted effector molecules, and their amounts over time and influences from in vivo components on them.

Mass Production of Highly Active NK Cells for Cancer Immunotherapy in a GMP Conform Perfusion Bioreactor

“…CD3 + T cell depleted PBMCs were isolated and ex vivo expanded in a proprietary cultivation process developed by Zellwerk GmbH (Diederichs et al, 2009; Reichardt et al, 2013; Pörtner et al, 2017). Briefly, anonymized blood products for research purposes (whole blood) from five donors were obtained, mixed with RosetteSep™ Human CD3 Depletion Cocktail and incubated for 30 min.…”

“…The optimization of appropriate expansion processes depends on a variety of factors including the starting material, additives like cytokines as well as the culture system itself. Meanwhile, various different and extremely heterogeneous expansion processes have been described (Childs and Berg, 2013; Granzin et al, 2015; Pörtner et al, 2017). Techniques for expansion of immune cells include simple culture flasks, multi-layered flasks (such as Millicell, Millipore, or BD Multi-flask etc.…”

“…Techniques for expansion of immune cells include simple culture flasks, multi-layered flasks (such as Millicell, Millipore, or BD Multi-flask etc. ), microcarrier techniques as well as special culture systems (e.g., G-Rex, WAVE-type bioreactors, Zellwerk's Z®RP Cell Breeder, Miltenyi's CliniMACS Prodigy®, the TERUMO Quantum system) (Pörtner et al, 2017, 2019). All the equipment materials are single use products which is of advantage with respect to handling and sterility.…”

“…Within the studies discussed here, an innovative perfusion process for expansion of human NK cells was developed and evaluated. The process is based on the Z®RP platform of Zellwerk and the belonging meander type bioreactors, which provide sophisticated features for mass production of different immune cells and allow cell culture and isolation within a functionally closed environment (Figure 1; Diederichs et al, 2009; Lavrentieva et al, 2013; Reichardt et al, 2013; Neumann et al, 2014; Pörtner et al, 2017). Bioreactors of the Z®RP system can be operated in the GMP Breeder.…”

“…The ratio of medium circulation and fresh medium flow is automatically regulated over time by a chosen algorithm guaranteeing a consistent homogenous supply with nutrients and gasses as well as a precise regulation of pO 2 , pH and temperature in the medium. In preliminary studies, this system has been successfully used for expansion of NK cells, T cells and tumor infiltrating lymphocytes (TILs) up to more than 10 10 cells in a single closed cultivation run with expansion factors of 5,000–50,000 (Pörtner et al, 2017). Further optimization was dedicated to coating of seeding areas with specific matrices/antibodies to promote suppression or expansion of several immune cell species including T cell subsets and CD56 + NK cells (Pörtner et al, 2019).…”