title: Two methods for quantifying DNA extracted from poly(lactide-co-glycolide) microspheres.
abstract: DNA can be formulated with synthetic polymers such as poly(lactide-co-glycolide) (PLG) to generate microparticles. Researchers have used either UV spectroscopy or fluorometry with PicoGreen((R)) dye to quantify PLG-encapsulated DNA. While the sensitivity of DNA detection and quantification by PicoGreen is higher ( approximately 12 pg/ml) compared to UV ( approximately 0.5 microg/ml), each method as an analytical tool has limitations. The premise of this work addresses the usefulness and limitations of each method to determine encapsulation efficiencies in PLG microspheres post-process, and to quantify release of DNA from microspheres during in vitro release experiments. In addition, assay conditions for accurate and reproducible extraction of DNA from PLG microspheres using a biphasic (aqueous/organic) solvent system are described. It was also determined that residual poly(vinyl alcohol) and DNA isoforms (linear, nicked, supercoiled) affected PicoGreen/DNA fluorescence values.
authors: ZYCOS Inc., 44 Hartwell, Lexington, MA 02421, USA.
authors: S P
authors: SP
authors: Barman
authors: L
authors: L
authors: Lunsford
authors: P
authors: P
authors: Chambers
authors: M L
authors: ML
authors: Hedley
mesh_terms: D004247:DNA / Q000008:administration & dosage / Q000032:analysis*; D019344:Lactic Acid / Q000008:administration & dosage*; D008863:Microspheres; D010957:Plasmids; D011100:Polyglycolic Acid / Q000008:administration & dosage*; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011108:Polymers / Q000008:administration & dosage*; D011142:Polyvinyl Alcohol / Q000494:pharmacology
chemical_list: D011108:Polymers; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D011142:Polyvinyl Alcohol; D004247:DNA