Plasmid DNA (pDNA) molecules are employed in several applications including transient protein production, transgene delivery for gene therapy, and DNA vaccines. Therapeutic plasmid DNAs have already shown promise in diseases including cancer. We have generated novel antibiotic-derived microbeads and monoliths that exploit the natural binding affinity of antibiotics towards nucleic acids. Aminoglycoside molecules were crosslinked with PEG and emulsion polymerization methods were employed for the formation microbeads using, approximately 10 μm in diameter. Surface area, amine content, and surface morphology of these anion-exchange microbeads were determined, and the binding capacity of these materials was determined using a model plasmid that encodes for luciferase protein. Maximum adsorption capacity of these parental microbeads was found to be ~1.2 mg of pDNA/mL of resin. Quaternization of amines in the parental microbeads resulted in a 7-fold enhancement in pDNA binding capacity. Several desorption buffers were screened to facilitate efficient desorption of plasmid DNA from the microbeads, and the role of hydrophobic modifiers on pDNA desorption was also investigated. In addition to pDNA binding, the microbeads were investigated for their ability to simultaneously lyse mammalian cells and bind DNA for whole-cell extraction applications. Recent studies involve development of cylindrical aminoglycoside-derived monoliths for pDNA binding and elution. Our results indicate the developments of novel anion-exchange materials for use in DNA biotechnology.

Aminoglycoside antibiotic derived anion-exchange microbeads and monoliths for plasmid DNA binding and in-situ DNA capture

249th National Meeting (2015)

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