| Stability of poly(L-lysine)-complexed plasmid DNA during mechanical stress and DNase I treatment. | |
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MedLine Citation:
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PMID: 10578502 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The aim of this study was to investigate the formation and stability of complexes between plasmid DNA (pDNA) and poly(L-lysine) (PLL). Formation of pDNA/PLL complexes with various ratios was determined by a fluorescence spectrophotometric method using fluorescamine. The effects of sonication, vortexing, and exposure to DNase I on the stability of free pDNA and pDNA/PLL complexes are discussed. A linear correlation between PLL added and PLL bound was obtained with overall reaction efficiency of 84.2-92.6%. Sonication degraded both free and PLL-complexed pDNA within 15 sec of vortexing. However, vortexing did not alter the stability of free and complexed pDNA. Dramatic increase in the protection of pDNA in pDNA/PLL complexes was observed in the DNase I digestion experiment; 68.1-89.0% of total pDNA in the pDNA/PLL complexes was protected from DNase I digestion compared to only 19.2% of total pDNA that remained undegraded after DNase I treatment of free pDNA. An increase in the PLL/pDNA ratio led to an increase in the protection of supercoiled pDNA; 15.5-38.2% of supercoiled pDNA pin PLL/pDNA complexes was protected after DNase I treatment. The results show that complexation of pDNA with PLL can stabilize the supercoiled structure of pDNA for the development of biodegradable microspheres as a delivery system for pDNA. Stability of pDNA/PLL complex can be monitored by PicoGreen dye and fluorescence densitometric assay methods. |
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Authors:
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Y Capan; B H Woo; S Gebrekidan; S Ahmed; P P DeLuca |
Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Pharmaceutical development and technology Volume: 4 ISSN: 1083-7450 ISO Abbreviation: Pharm Dev Technol Publication Date: 1999 |
Date Detail:
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Created Date: 2000-01-19 Completed Date: 2000-01-19 Revised Date: 2006-05-01 |
Medline Journal Info:
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Nlm Unique ID: 9610932 Medline TA: Pharm Dev Technol Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 491-8 Citation Subset: IM |
Affiliation:
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University of Kentucky, College of Pharmacy, Faculty of Pharmaceutical Sciences, Lexington 40536, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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DNA
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chemistry* Densitometry Deoxyribonuclease I / chemistry* Drug Stability Electrophoresis, Agar Gel Fluorescent Dyes Organic Chemicals Plasmids / chemistry* Polylysine / chemistry* Solutions Spectrometry, Fluorescence Stress, Mechanical* Ultrasonics |
| Chemical | |
Reg. No./Substance:
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0/Fluorescent Dyes; 0/Organic Chemicals; 0/PicoGreen; 0/Solutions; 25104-18-1/Polylysine; 9007-49-2/DNA; EC 3.1.21.1/Deoxyribonuclease I |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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