title: 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: BACKGROUND
  Automation 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).
  
  
  METHODS
  As 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 × 10(6) target cells out of 60 × 10(9) total cells. Three split-validation processings with healthy donor G-CSF-mobilized apheresis products were performed; feasibility, time consumption and product quality were assessed.
  
  
  RESULTS
  All 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 × 10(6) CD34+ cells/kg was below 10 × 10(3)/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.
  
  
  CONCLUSIONS
  This 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.
authors.affiliation: Department of Cellular Therapeutics (GMP), German Red Cross Blood Service Baden-Württemberg-Hesse, Institute Frankfurt, Frankfurt, Germany.
authors.forename: Christiane
authors.initials: C
authors.lastname: Hümmer
authors.affiliation: Department of Cellular Therapeutics (GMP), German Red Cross Blood Service Baden-Württemberg-Hesse, Institute Frankfurt, Frankfurt, Germany.
authors.forename: Carolin
authors.initials: C
authors.lastname: Poppe
authors.affiliation: Department of Cellular Therapeutics (GMP), German Red Cross Blood Service Baden-Württemberg-Hesse, Institute Frankfurt, Frankfurt, Germany.
authors.forename: Milica
authors.initials: M
authors.lastname: Bunos
authors.affiliation: Department of Cellular Therapeutics (GMP), German Red Cross Blood Service Baden-Württemberg-Hesse, Institute Frankfurt, Frankfurt, Germany.
authors.forename: Belinda
authors.initials: B
authors.lastname: Stock
authors.affiliation: Department of Cellular Therapeutics (GMP), German Red Cross Blood Service Baden-Württemberg-Hesse, Institute Frankfurt, Frankfurt, Germany.
authors.forename: Eva
authors.initials: E
authors.lastname: Wingenfeld
authors.affiliation: Miltenyi Biotec GmbH, Bergisch-Gladbach, Germany.
authors.forename: Volker
authors.initials: V
authors.lastname: Huppert
authors.affiliation: Miltenyi Biotec GmbH, Bergisch-Gladbach, Germany.
authors.forename: Juliane
authors.initials: J
authors.lastname: Stuth
authors.affiliation: Miltenyi Biotec GmbH, Bergisch-Gladbach, Germany.
authors.forename: Kristina
authors.initials: K
authors.lastname: Reck
authors.affiliation: Miltenyi Biotec GmbH, Bergisch-Gladbach, Germany.
authors.forename: Mike
authors.initials: M
authors.lastname: Essl
authors.affiliation: Department of Cellular Therapeutics (GMP), German Red Cross Blood Service Baden-Württemberg-Hesse, Institute Frankfurt, Frankfurt, Germany.|Institute for Transfusion Medicine and Immunohematology, Goethe University Medical Center, Frankfurt, Germany.
authors.forename: Erhard
authors.initials: E
authors.lastname: Seifried
authors.affiliation: Department of Cellular Therapeutics (GMP), German Red Cross Blood Service Baden-Württemberg-Hesse, Institute Frankfurt, Frankfurt, Germany. hbonig@uw.edu.|Institute for Transfusion Medicine and Immunohematology, Goethe University Medical Center, Frankfurt, Germany. hbonig@uw.edu.|Department of Medicine, Division of Hematology, University of Washington, Seattle, WA, USA. hbonig@uw.edu.
authors.forename: Halvard
authors.initials: H
authors.lastname: Bonig
mesh_terms: D018952:Antigens, CD34 / Q000378:metabolism*; D001331:Automation; D001781:Blood Component Removal / Q000379:methods*; D064987:Cell- and Tissue-Based Therapy; D005434:Flow Cytometry; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D015203:Reproducibility of Results
chemical_list: D018952:Antigens, CD34
keywords: Allogeneic; Automation; CD34; Cell therapy; Clean room; CliniMACS; Good manufacturing practice; Haplo-identical; Immunomagnetic; Prodigy; Stem cell transplantation