Breaking barriers in the sensitive and accurate mass determination of large DNA plasmids by mass photometry.

ABSTRACT: DNA plasmids (pDNAs) are essential for gene cloning and protein expression, whereby engineered plasmids serve as vectors to insert foreign DNA into host cells, enabling mass production of proteins and vaccines. Due to the rapidly increasing use and application of a wide variety of pDNA (e.g., CRISPR-based gene editing, RNA therapeutics, and DNA vaccines), analytical methods to characterize their key attributes are vital. Here we explore mass photometry (MP) to analyze pDNAs and find that it completely fails using standard procedures as developed for MP on proteins, with masses underestimated by 30%-40%. While DNA can be measured by using coated glass slides, the large double-stranded DNA (dsDNA) particles diffract light beyond the diffraction limit, rendering most landing events unusable.

SUBMITTER: Ebberink EHTM 

PROVIDER: S-EPMC12328894 | biostudies-literature | 2025 Sep

REPOSITORIES: biostudies-literature