Challenges and advances in scale-up of label-free downstream processing for allogeneic cell therapies

Purifying stem cell‐derived red blood cells: a high‐throughput label‐free downstream processing strategy based on microfluidic spiral inertial separation and membrane filtration

“…Membrane filtration alone is less effective in processing the mRBC than the combined process consisting of processing in spiral microchannel followed by filtration. Particle separation by means of filtration is a widely applied technique within field of bioprocessing (Masri et al, 2017). Membrane filtration uses an average pore size where particles larger than the pore size cannot pass through.…”

“…Currently, only a limited number of fluorophore‐conjugated antibody reagents are suitable for clinical processing (McIntyre, Flyg, & Fong, 2010) and the adverse effects of introducing these probes into patients are unknown, but it is generally recognized that they could potentially trigger immune and toxic responses (Willoughby et al, 2016). Various alternatives to FACS and MACS for cellular therapies, such as mRBC production, have been proposed and were recently reviewed (Masri, Hoeve, Sousa, & Willoughby, 2017). Recent work on deterministic lateral displacement (Campos‐Gonzalez et al, 2018) and inertial vortexes (Pritchard et al, 2019) have demonstrated application to CAR‐T cell processing and inertial focussing has been used to isolate, enrich, and purify stem cells (Hur, Brinckerhoff, Walthers, Dunn, & Di Carlo, 2012; Lee et al, 2018; Song et al, 2017), to obtain desired subpopulation (Lee et al, 2011, 2014; Poon et al, 2015), to isolate single cells from clusters (Nathamgari et al, 2015), for nonviable cell removal (Kwon, Yao, Hamel, & Han, 2018) as well as microcarrier scaffold removal (Moloudi et al, 2018).…”

Automation in cell and gene therapy manufacturing: from past to future

“…When manufacturing such highly complex products, it is highly important to acknowledge that any change, regardless of how minor in the culture environment as determined by the manufacturing process may result in the alteration of product quality which is a key determinant of its safety and efficacy (Morrow et al 2017). In order to comply with these requirements, it is suggested that the manufacturing process should be simple enough to allow reproducibility and of a short duration to minimise costs associated with resources and labour (Masri et al 2017).…”

Investigation of particle movement in irregularly shaped channels in inertial fluidics for scale up applications in bioprocessing

“…As a result, the designed spiral could potentially be stacked up to deliver on a more relevant, industrial scale (~1-5 L/min) in practice. This proposed inertial-based filtration method offers beneficial attributes such as a closed system, scalability, and continuous mode (vs. batch processing) that are appreciated for large-scale cell manufacturing in downstream processing [174,185].…”

“…Furthermore, as for patients suffering from impaired immune system, non-sterile particulates can also cause adverse effects[165,173]. Unfortunately, in contrast to protein-based products, it is not possible to carry out downstream filtration and purification step using membrane technology, as cell sizes fall within the range of that of the particulates[166,172,174]. Patient safety, therefore, is a major challenge which needs to be addressed to pave the way for successful development, and ultimately commercialization of cell therapy products[175].In this chapter, the inertial microfluidic technology is reviewed while concentrating on particle focusing dynamics.…”