Biological potency of microsphere encapsulated plasmid DNA.
We evaluated the utility of three in vitro methods to monitor the biological potency of PLGA encapsulated DNA. For each assay we also determined whether the biological activity was influenced by the structural profile of DNA isomers. Collectively, the results indicate that all three methods can be used to evaluate the biological activity of DNA extracted from PLGA microspheres, but they are differentially sensitive to the structural changes of plasmid DNA that can occur during microencapsulation and microsphere storage. More specifically, mammalian cell transfection followed by an enzyme assay affords an accurate determination of DNA potency over time and is less influenced by DNA isoform than bacterial transformation. Cell-free transcription/translation systems can also be utilized, and the results of this assay are influenced by DNA isoform. Finally, bacterial transformation was found to be more sensitive to DNA isoform than the other assay methods.
Hao
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ZYCOS Inc., 44 Hartwell Ave, Lexington, MA 02421, USA.
McKeever
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Hedley
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ML
D000818:Animals; D019556:COS Cells; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics; D004339:Drug Compounding; D004356:Drug Storage; D005057:Eukaryotic Cells; D005456:Fluorescent Dyes; D008863:Microspheres; D009930:Organic Chemicals; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics; D014176:Protein Biosynthesis / Q000235:genetics; D013056:Spectrophotometry, Ultraviolet; D013696:Temperature; D014158:Transcription, Genetic / Q000235:genetics; D014162:Transfection; D014169:Transformation, Bacterial / Q000235:genetics
D005456:Fluorescent Dyes; D009930:Organic Chemicals; C099476:PicoGreen; D004247:DNA