title: Aminoglycoside antibiotic-derived anion-exchange microbeads for plasmid DNA binding and in situ DNA capture.
abstract: Plasmid DNA (pDNA) therapeutics are being investigated for gene therapy and DNA vaccines against diseases including cancer, cystic fibrosis and AIDS. In addition, several applications in modern biotechnology require pDNA for transient protein production. Here, we describe the synthesis, characterization, and evaluation of microbeads ("Amikabeads") derived from the aminoglycoside antibiotic amikacin for pDNA binding and in situ DNA capture from mammalian cells. The parental aminoglycoside-derived microbeads (Amikabeads-P) acted as anion-exchange materials, and demonstrated high capacities for binding pDNA. Binding of pDNA was significantly enhanced following quaternization of the amines on the microbeads (Amikabeads-Q). Amikabeads were further employed for the disruption and extraction of DNA from mammalian cells, indicating their utility for in situ DNA capture. Our results indicate that Amikabeads are a novel material, with multiple reactive groups for further conjugation, and can have several applications in plasmid DNA biotechnology.
authors.affiliation: Harrington Biomedical Engineering, School of Biological and Health Systems Engineering ‡Chemical Engineering, School for Engineering of Matter, Transport, and Energy, Arizona State University , Tempe, Arizona 85287, United States.
authors.forename: Taraka Sai Pavan
authors.initials: TS
authors.lastname: Grandhi
authors.forename: Amrita
authors.initials: A
authors.lastname: Mallik
authors.forename: Kevin N.
authors.initials: KN
authors.lastname: Lin
authors.forename: Bhavani
authors.initials: B
authors.lastname: Miryala
authors.forename: Thrimoorthy
authors.initials: T
authors.lastname: Potta
authors.forename: Yifan
authors.initials: Y
authors.lastname: Tian
authors.forename: Kaushal
authors.initials: K
authors.lastname: Rege
mesh_terms: D000617:Aminoglycosides / Q000737:chemistry*; D000837:Anion Exchange Resins / Q000737:chemistry*; D000900:Anti-Bacterial Agents / Q000737:chemistry*; D045744:Cell Line, Tumor; D002852:Chromatography, Ion Exchange / Q000295:instrumentation*; D003584:Cytological Techniques; D004247:DNA / Q000302:isolation & purification*; D006801:Humans; D008863:Microspheres; D010316:Particle Size; D010957:Plasmids / Q000302:isolation & purification
chemical_list: D000617:Aminoglycosides; D000837:Anion Exchange Resins; D000900:Anti-Bacterial Agents; D004247:DNA
keywords: DNA biotechnology; bioseparations; chromatography; microgel; plasmid DNA binding; whole cell genomic extraction