Automation of cellular therapy product manufacturing: results of a split validation comparing CD34 selection of peripheral blood stem cell apheresis product with a semi-manual vs. an automatic procedure

Abstract

BackgroundAutomation of cell therapy manufacturing promises higher productivity of cell factories, more economical use of highly-trained (and costly) manufacturing staff, facilitation of processes requiring manufacturing steps at inconvenient hours, improved consistency of processing steps and other benefits. One of the most broadly disseminated engineered cell therapy products is immunomagnetically selected CD34+ hematopoietic “stem” cells (HSCs).MethodsAs the clinical GMP-compliant automat CliniMACS Prodigy is being programmed to perform ever more complex sequential manufacturing steps, we developed a CD34+ selection module for comparison with the standard semi-automatic CD34 “normal scale” selection process on CliniMACS Plus, applicable for 600 × 106 target cells out of 60 × 109 total cells. Three split-validation processings with healthy donor G-CSF-mobilized apheresis products were performed; feasibility, time consumption and product quality were assessed.ResultsAll processes proceeded uneventfully. Prodigy runs took about 1 h longer than CliniMACS Plus runs, albeit with markedly less hands-on operator time and therefore also suitable for less experienced operators. Recovery of target cells was the same for both technologies. Although impurities, specifically T- and B-cells, were 5 ± 1.6-fold and 4 ± 0.4-fold higher in the Prodigy products (p = ns and p = 0.013 for T and B cell depletion, respectively), T cell contents per kg of a virtual recipient receiving 4 × 106 CD34+ cells/kg was below 10 × 103/kg even in the worst Prodigy product and thus more than fivefold below the specification of CD34+ selected mismatched-donor stem cell products. The products’ theoretical clinical usability is thus confirmed.ConclusionsThis split validation exercise of a relatively short and simple process exemplifies the potential of automatic cell manufacturing. Automation will further gain in attractiveness when applied to more complex processes, requiring frequent interventions or handling at unfavourable working hours, such as re-targeting of T-cells.

Genetic Engineering and Manufacturing of Hematopoietic Stem Cells

“…It has been reported that a medium recovery of 71% and medium purity of 97% of CD34 + cells, with a median of 0.04% residual CD3 + cells could be achieved from either BM or MPB by using the CliniMACS device 23 . Similar results were obtained with the automated CliniMACS Prodigy, although CD34 + cell recovery and depletion of CD3 + cells may be lower than with the semi-automated CliniMACS Plus instrument 24, 25…”

Automated Enrichment, Transduction, and Expansion of Clinical-Scale CD62L+ T Cells for Manufacturing of Gene Therapy Medicinal Products

“…reporting equivalent results for one split run. 10 Bunos et al . 12 demonstrated the enrichment of CMV-reactive T cells from donor apheresis products in five clinical scale runs reaching 64–93% purity of CD3 + T cells.…”

“…To date, clinical-scale expansion and transduction of engineered T cells or other effector cells for Phase I/II trials is a complex multistep hands-on process and a standardized automated manufacturing process observing good manufacturing practice (GMP) conditions is still a challenge. Currently, the use of a closed, automated system—the CliniMACS ® Prodigy (Miltenyi Biotec, Bergisch Gladbach, Germany)—has been successfully demonstrated as a short-term process for the purification of CD34 + progenitor cells 10 , 11 or cytomegalovirus (CMV)-specific T cells, 12 as well as for long-term NK cell expansion. 13 …”

Transplantation of gene-modified haematopoietic stem cells: Application and clinical considerations

“…These devices are certified for cell therapy applications and globally accepted by regulatory authorities. [14,15] However, the system is costly and has low throughput, which will have its challenges if we are to foresee roll-out of ex vivo gene-modified cell therapies for diseases such as HIV and sickle cell disease where the patient burden is in the millions and is often in lower-to-middle income countries. There is thus an opportunity to address these limitations with more efficient process developments, lower cost and higher throughput equipment solutions.…”