Replication Data for Exploring the fate of antibody-encoding pDNA after intramuscular electroporation in mice

This dataset contains raw data contributing to the publication 'Exploring the fate of antibody-encoding pDNA after intramuscular electroporation in mice'. This publication is about DNA-based antibody therapy, which seeks to administer the encoding nucleotide sequence, rather than the antibody protein. To further improve the in vivo monoclonal antibody (mAb) expression, a better understanding of what happens after the administration of the encoding plasmid DNA (pDNA) is required. This study reports the quantitative evaluation and localization of the administered pDNA over time and its association with corresponding mRNA levels and systemic protein concentrations. pDNA encoding the murine anti-HER2 4D5 mAb was administered to BALB/c mice via intramuscular injection followed by electroporation. Muscle biopsies and blood samples were taken at different time points (up to 3 months). In muscle, pDNA levels decreased 90% between 24 hours and one week post treatment. In contrast, mRNA levels remained stable over time. 4D5 antibody plasma concentrations reached peak levels at week two followed by a slow decrease (50% after 12 weeks). Evaluation of pDNA localization revealed that extranuclear pDNA was cleared fast whereas the nuclear fraction remained relatively stable. This is in line with the observed mRNA and protein levels over time, and indicates that only a minor fraction of the administered pDNA is ultimately responsible for the observed systemic mAb levels. Therefore, efforts to increase the protein levels upon pDNA-based gene therapy should focus on strategies to increase both cellular entry and migration of the pDNA into the nucleus. The currently applied methodology can be used to guide the design and evaluation of novel plasmid-based vectors or alternative delivery methods, to achieve a robust and prolonged protein expression.

Publication year:2023

Accessibility:open

Publisher:KU Leuven RDR

License:CC-BY-4.0

Format:czi, xlsx

Keywords: Antibody gene transfer, DNA localization, DNA scope, DNA/RNA quantification, Intramuscular electroporation, Plasmid DNA