Reproducibility of GMP-compliant production of therapeutic stressed peripheral blood mononuclear cell-derived secretomes a novel class of biological medicinal products

摘要

Experimental setup. PBMCs were enriched by Ficoll-assisted centrifugation, exposed to 60 Gy γ-irradiation, and cultured in GMP medium for 24 h. Secretome was filtered, virus-inactivated, and lyophilized (GMP PBMC ). To produce placebo control, GMP medium alone without cells was treated equally. Major biomolecular components present in PBMC (EVs, proteins, and lipids) were analyzed. A set of physico-chemical-biological tests reflecting key effects mediated by PBMC was selected to determine the reproducibility of 3 PBMC secretome batches produced under GMP. In parallel, tests were performed with one placebo batch. Selected proteins were quantified and functional assays were performed to compare variability of individual donor secretomes, small, and large secretome batches. Stability of 2 PBMC batches was assessed after storage at various ambient temperatures for up to 6 months. EVs, extracellular vesicles; GMP, good manufacturing practice; P, phosphorylation; PBMC , secretome of γ-irradiated, peripheral blood mononuclear cells