Tips and Tricks for Validation of Quality Control Analytical Methods in Good Manufacturing Practice Mesenchymal Stromal Cell Production

Figure 3

Adventitious viruses validation results. Specificity of the method was assessed spiking MSC () with a high and low viral loads (HL and LL) of adenovirus (AdV), enterovirus (HEV), cytomegalorovirus (CMV), and Epstein-Barr virus (EBV) (a–c). (a) Results obtained from the qualitative analysis of fifteen respiratory viruses. The samples spiked with HEV were treated (_1) or not (_2) with protease K. Positive controls (CTRLs +) were represented by virus alone (in set A AdV and in set C HEV). A PCR internal control (PCRC) and a whole process control (WPC) were visible in set A and set C. (b–c) Real-time PCR analysis expressed as quantity (gEq/reaction) of CMV and EBV in all the spiked MSC and in the positive controls (CMV and EBV). (d, e) To assess detection limit and accuracy, CBMSC () were spiked with a lowest viral load of CMV and EBV. Ct of different quantities of DNA plasmid (standard) was plotted with Ct of positive controls and spiked MSC.

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