Validation of a method evaluating T cell metabolic potential in compliance with ICH Q2 (R1)

Abstract Background Metabolic cell features are able to give reliable information on functional state. Thus, metabolic potential assessment of T cells in malignancy setting represents a promising area, especially adoptive therapy procedures. Easy set up and convenient Seahorse technology have recently been proposed by Agilent Technologies it could be used monitor potential. However, this method demonstrates an inter-assay variability lacks practices standardization. Results We aimed overcome these shortcomings thanks lymphoblastic derived JURKAT line seeding each experiment standardize the process. adapted XF Cell MitoStress Kit protocol, consisting evaluation basal, stressed maximal glycolysis oxidative phosphorylation related parameters, through sequential addition oligomycin carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP) glucose containing medium. Data were acquired analyzed XFe96 analyzer. Indeed, we validated light ICH Q2 (R1) guidelines. confirm specificity accuracy method. also demonstrated precision, linearity range our experimental conditions. Conclusion The validation incorporation as control material contributes improve technology’s robustness. These results lay groundwork for implementation optimize based cellular products production process monitoring.