TY  - JOUR
A1  - Bittorf, Patrick
A1  - Bergmann, Thorsten
A1  - Merlin, Simone
A1  - Olgasi, Chistina
A1  - Pullig, Oliver
A1  - Sanzenbacher, Ralf
A1  - Zierau, Martin
A1  - Walles, Heike
A1  - Follenzi, Antonia
A1  - Braspenning, Joris
T1  - Regulatory-Compliant Validation of a Highly Sensitive qPCR for Biodistribution Assessment of Hemophilia A Patient Cells
T2  - Molecular Therapy - Methods & Clinical Development
N2  - The investigation of the biodistribution profile of a cell-based medicinal product is a pivotal prerequisite to allow a factual benefit-risk assessment within the non-clinical to clinical translation in product development. Here, a qPCR-based method to determine the amount of human DNA in mouse DNA was validated according to the guidelines of the European Medicines Agency and the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use. Furthermore, a preclinical worst-case scenario study was performed in which this method was applied to investigate the biodistribution of 2 x 10\(^6\) intravenously administered, genetically modified, blood outgrowth endothelial cells from hemophilia A patients after 24 h and 7 days. The validation of the qPCR method demonstrated high accuracy, precision, and linearity for the concentration interval of 1:1 x 10\(^3\) to 1:1 x 10\(^6\) human to mouse DNA. The application of this method in the biodistribution study resulted in the detection of human genomes in four out of the eight investigated organs after 24 h. After 7 days, no human DNA was detected in the eight organs analyzed. This biodistribution study provides mandatory data on the toxicokinetic safety profile of an actual candidate cell-based medicinal product. The extensive evaluation of the required validation parameters confirms the applicability of the qPCR method for non-clinical biodistribution studies.
KW  - outgrowth endothelial cells
KW  - real time PCR
KW  - in vivo
KW  - gene therapy
KW  - factor-VIII
KW  - murine
KW  - quantification
KW  - establishment
KW  - phenotype
KW  - xenotransplantation
Y1  - 2020
UR  - https://opus.bibliothek.uni-wuerzburg.de/frontdoor/index/index/docId/23028
UR  - https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-230284
VL  - 18
ER  -