Supercoiled plasmid quality assessment by analytical arginine-affinity chromatography

Area of Science:

• Biotechnology
• Analytical Chemistry
• Molecular Biology

Background:

• Supercoiled plasmids (sc pDNA) are crucial for gene-based therapies like gene therapy and DNA vaccination.
• Efficiently assessing sc pDNA purity and yield is vital for therapeutic distribution.

Purpose of the Study:

• To develop a method for quantifying supercoiled plasmid DNA (sc pDNA) isoforms.
• To establish a quality control procedure for sc pDNA biopharmaceuticals.

Main Methods:

• An arginine affinity methodology utilizing UV detection was developed.
• The method leverages amino acid-nucleic acid interactions for selective quantification.
• Analytical experiments were conducted within a 12-minute timeframe.

Main Results:

• The method accurately quantifies sc pDNA in a concentration range of 2–150 μg/mL.
• It demonstrates accuracy, precision, reproducibility, and linearity.
• The technique distinguishes between different plasmid isoforms, including sc pDNA.

Conclusions:

• This arginine affinity method offers a selective and accurate way to quantify biologically active sc pDNA.
• It enables precise quality analysis and ensures the purity of plasmid products for therapeutic applications.
• The method provides a significant advancement over existing techniques for pDNA quantification.