Fig. 1

Overview of the study approach. A CliniMACS Prodigy TS 320 setup for the automated LP-TCRαβ-19-45RA Depletion procedure. A brief description of the components and preparation steps of the tubing set is added in Additional file 1: Figure S1. Starting material (mobilized apheresis products, mob LP) is automatically split for CD45RA and for TCRαβ/CD19 depletion which are performed sequentially and which results in two separate cell products. Target Cell Bag 1 (TCB1) contains a cell product depleted of naïve T cells and other CD45RA+ leukocyte subsets. Target Cell Bag 2 (TCB2) includes TCRγδ+ cells, HSPCs, NK cells, among others. Copyright © 2023 Miltenyi Biotec B.V. & Co. KG. All rights reserved. B Diagram showing the different use cases of the depletion software. C Workflow diagram of the manufacturing procedure with timeline and quality controls. White, dashed and gray shading mark hands-on, automatic-supervised and autonomous preparation steps, respectively. D Graphical description and schedule of experimental setup. All processes consisted of sequential CD45RA and TCRαβ/CD19 depletion. Ten depletion procedures from six mob LP (red) were performed. Four thereof were split runs to compare immediate processing vs. processing after 48 h storage at 2–6 °C to analyze the effect of apheresis product aging (mob LP, patterned red) on manufacturing performance and product quality (TCB1, patterned green and TCB2, patterned blue, respectively). Additionally, the effect of aging on the depleted products (TCB1, light green and TCB2, light blue, respectively) was assessed after storage at 2–6 °C for 48 h. Normal scale (5/10 runs NS) and large scale (5/10 runs LS) depletion reagents were used to test both, for depletion performance and final product quality. Final products from four split runs were cryopreserved to test how freeze–thaw procedure affects the final product quality at the end of the of their shelf life (TCB1, light green with snowflake and TCB2, light blue with snowflake, respectively, as well as TCB1, patterned green with snowflake and TCB2, patterned blue with snowflake, resepctively)