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Exploring the Fate of Antibody-Encoding pDNA after Intramuscular Electroporation in Mice
Marie-Lynn Cuypers1, Nick Geukens2, Kevin Hollevoet2
1Laboratory for Therapeutic and Diagnostic Antibodies, Department of Pharmaceutical and Pharmacological Sciences, KU Leuven-University of Leuven, O&N II Herestraat 49 Box 820, 3000 Leuven, Belgium.
Pharmaceutics
|April 28, 2023
Summary
DNA-based antibody therapy uses plasmid DNA (pDNA) for monoclonal antibody (mAb) expression. Durable expression depends on nuclear pDNA uptake, guiding future gene therapy strategies for enhanced protein levels.
Area of Science:
- Biotechnology
- Molecular Biology
- Immunotherapy
Background:
- DNA-based antibody therapy administers nucleotide sequences instead of proteins for in vivo monoclonal antibody (mAb) expression.
- Improving in vivo mAb expression requires understanding plasmid DNA (pDNA) fate post-administration.
Purpose of the Study:
- Quantitatively evaluate and localize administered pDNA over time.
- Correlate pDNA levels with mRNA and systemic protein concentrations.
- Identify factors influencing durable mAb expression.
Main Methods:
- Administered pDNA encoding anti-HER2 4D5 mAb to mice via intramuscular injection and electroporation.
- Collected muscle biopsies and blood samples at various time points up to 3 months.
- Quantified pDNA and mRNA levels, and measured plasma antibody concentrations.
Main Results:
- pDNA levels decreased significantly within one week, while mRNA remained stable.
- Peak antibody plasma concentrations were observed at week two, followed by a gradual decline.
- Extranuclear pDNA was rapidly cleared, whereas nuclear pDNA fraction remained stable, correlating with sustained mRNA and protein levels.
Conclusions:
- Durable protein expression in DNA-based antibody therapy is dependent on nuclear uptake of pDNA.
- Strategies to enhance gene therapy should focus on improving cellular entry and nuclear migration of pDNA.
- The methodology can guide the development of novel vectors and delivery methods for sustained protein expression.
