Biophysical Characterization of PEI/DNA Complexes
The main goal of this study was to determine the effects of polyethylenimine (PEI) molecular weight and structure (750kDa, 25kDa, 2kDa branched, and 25kDa linear PEI) and the nitrogen/phosphate (N/P) molar ratio on the physical properties and transfection efficiencies of PEI/DNA complexes. Fourier t...
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| Veröffentlicht in: | Journal of pharmaceutical sciences 2003-08, Vol.92 (8), p.1710-1722 |
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| creator | Choosakoonkriang, Sirirat Lobo, Brian A. Koe, Gary S. Koe, Janet G. Middaugh, C.Russell |
| description | The main goal of this study was to determine the effects of polyethylenimine (PEI) molecular weight and structure (750kDa, 25kDa, 2kDa branched, and 25kDa linear PEI) and the nitrogen/phosphate (N/P) molar ratio on the physical properties and transfection efficiencies of PEI/DNA complexes. Fourier transform infrared spectroscopy revealed that DNA remained in the B conformation when complexed to all PEIs. Unique alterations in the circular dichroism spectra of DNA were observed in the presence of each PEI, whereas differential scanning calorimetry measurements showed that all PEIs examined destabilized supercoiled DNA at N/P |
| doi_str_mv | 10.1002/jps.10437 |
| format | Article |
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Fourier transform infrared spectroscopy revealed that DNA remained in the B conformation when complexed to all PEIs. Unique alterations in the circular dichroism spectra of DNA were observed in the presence of each PEI, whereas differential scanning calorimetry measurements showed that all PEIs examined destabilized supercoiled DNA at N/P<3/1, but not at higher ratios. Isothermal titration calorimetry revealed the existence of protonation changes at low ionic strength due to possible shifts in pKa of the ionizable groups of PEI during complex formation. Twenty-five kilodalton branched and 25kDa linear PEI complexes showed the highest transfection efficiencies at an N/P ratio of 6:1 in COS-7 and CHO-K1 cells, respectively. These investigations have detected alterations in the physical and colloidal properties of the complexes that were sensitive to polymer structure, molecular weight, and polymer/DNA ratio, but these properties did not directly correlate with their transfection efficiencies. To further probe any possible relationship between these parameters and activity, a more refined biophysical analysis of any subpopulations in these samples that may differ in transfection activity is suggested, although the existence of such species remains unknown. © 2003 Wiley-Liss, Inc. and the American Pharmacists Association</description><identifier>ISSN: 0022-3549</identifier><identifier>EISSN: 1520-6017</identifier><identifier>DOI: 10.1002/jps.10437</identifier><identifier>PMID: 12884257</identifier><identifier>CODEN: JPMSAE</identifier><language>eng</language><publisher>Hoboken: Elsevier Inc</publisher><subject>Animals ; Biological and medical sciences ; Biophysical Phenomena ; Biophysics ; Cercopithecus aethiops ; CHO Cells ; circular dichroism ; COS Cells ; Cricetinae ; differential scanning calorimetry ; DNA - analysis ; DNA - chemistry ; DNA - genetics ; DNA delivery ; FTIR ; gene delivery ; General pharmacology ; Genetic Therapy - methods ; isothermal titration calorimetry ; Medical sciences ; Pharmaceutical technology. Pharmaceutical industry ; Pharmacology. Drug treatments ; physical characterization ; plasmid DNA ; Polyethyleneimine - analysis ; Polyethyleneimine - chemistry ; polyethylenimine ; Spectroscopy, Fourier Transform Infrared - methods ; Transfection - methods</subject><ispartof>Journal of pharmaceutical sciences, 2003-08, Vol.92 (8), p.1710-1722</ispartof><rights>2003 Wiley-Liss, Inc.</rights><rights>Copyright © 2003 Wiley‐Liss, Inc.</rights><rights>2003 INIST-CNRS</rights><rights>Copyright 2003 Wiley-Liss, Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4687-5d8c1e8b5c8f32420a536a2522a33d1410b8586ede0dd7c43a11b89533af799d3</citedby><cites>FETCH-LOGICAL-c4687-5d8c1e8b5c8f32420a536a2522a33d1410b8586ede0dd7c43a11b89533af799d3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fjps.10437$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fjps.10437$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=15036969$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12884257$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Choosakoonkriang, Sirirat</creatorcontrib><creatorcontrib>Lobo, Brian A.</creatorcontrib><creatorcontrib>Koe, Gary S.</creatorcontrib><creatorcontrib>Koe, Janet G.</creatorcontrib><creatorcontrib>Middaugh, C.Russell</creatorcontrib><title>Biophysical Characterization of PEI/DNA Complexes</title><title>Journal of pharmaceutical sciences</title><addtitle>J. Pharm. Sci</addtitle><description>The main goal of this study was to determine the effects of polyethylenimine (PEI) molecular weight and structure (750kDa, 25kDa, 2kDa branched, and 25kDa linear PEI) and the nitrogen/phosphate (N/P) molar ratio on the physical properties and transfection efficiencies of PEI/DNA complexes. Fourier transform infrared spectroscopy revealed that DNA remained in the B conformation when complexed to all PEIs. Unique alterations in the circular dichroism spectra of DNA were observed in the presence of each PEI, whereas differential scanning calorimetry measurements showed that all PEIs examined destabilized supercoiled DNA at N/P<3/1, but not at higher ratios. Isothermal titration calorimetry revealed the existence of protonation changes at low ionic strength due to possible shifts in pKa of the ionizable groups of PEI during complex formation. Twenty-five kilodalton branched and 25kDa linear PEI complexes showed the highest transfection efficiencies at an N/P ratio of 6:1 in COS-7 and CHO-K1 cells, respectively. These investigations have detected alterations in the physical and colloidal properties of the complexes that were sensitive to polymer structure, molecular weight, and polymer/DNA ratio, but these properties did not directly correlate with their transfection efficiencies. To further probe any possible relationship between these parameters and activity, a more refined biophysical analysis of any subpopulations in these samples that may differ in transfection activity is suggested, although the existence of such species remains unknown. © 2003 Wiley-Liss, Inc. and the American Pharmacists Association</description><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Biophysical Phenomena</subject><subject>Biophysics</subject><subject>Cercopithecus aethiops</subject><subject>CHO Cells</subject><subject>circular dichroism</subject><subject>COS Cells</subject><subject>Cricetinae</subject><subject>differential scanning calorimetry</subject><subject>DNA - analysis</subject><subject>DNA - chemistry</subject><subject>DNA - genetics</subject><subject>DNA delivery</subject><subject>FTIR</subject><subject>gene delivery</subject><subject>General pharmacology</subject><subject>Genetic Therapy - methods</subject><subject>isothermal titration calorimetry</subject><subject>Medical sciences</subject><subject>Pharmaceutical technology. Pharmaceutical industry</subject><subject>Pharmacology. Drug treatments</subject><subject>physical characterization</subject><subject>plasmid DNA</subject><subject>Polyethyleneimine - analysis</subject><subject>Polyethyleneimine - chemistry</subject><subject>polyethylenimine</subject><subject>Spectroscopy, Fourier Transform Infrared - methods</subject><subject>Transfection - methods</subject><issn>0022-3549</issn><issn>1520-6017</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp10E1PwjAYwPHGaATRg1_AcPHgYdKXdeuOOF6EEFx8icema7tQBLa0qOCntzqUi57apL-nzb8AnCN4jSDEnXnl_CYk8QFoIophEEEUH4KmP8MBoWHSACfOzSGEEaT0GDQQZizENG4CdGPKarZ1RopFO50JK-RaW_Mh1qZctcuinfVHnd60207LZbXQG-1OwVEhFk6f7dYWeBr0H9PbYHI3HKXdSSDDiMUBVUwizXIqWUFwiKGgJBKYYiwIUShEMGeURVppqFQsQyIQyllCCRFFnCSKtMBVfa-0pXNWF7yyZinsliPIv7K5z-bf2d5e1LZ6zZda7eWu04PLHRDOlxZWrKRxe0chiZIo8a5Tu3ez0Nv_X-Tj7OHn6aCeMG6tN78Twr7wKCYx5c_TIR8P7mHWy1JOvSe11_7v3oy23EmjV1IrY7Vcc1WaPwI_AbkYjRY</recordid><startdate>200308</startdate><enddate>200308</enddate><creator>Choosakoonkriang, Sirirat</creator><creator>Lobo, Brian A.</creator><creator>Koe, Gary S.</creator><creator>Koe, Janet G.</creator><creator>Middaugh, C.Russell</creator><general>Elsevier Inc</general><general>Wiley Subscription Services, Inc., A Wiley Company</general><general>Wiley</general><general>American Pharmaceutical Association</general><scope>BSCLL</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope></search><sort><creationdate>200308</creationdate><title>Biophysical Characterization of PEI/DNA Complexes</title><author>Choosakoonkriang, Sirirat ; Lobo, Brian A. ; Koe, Gary S. ; Koe, Janet G. ; Middaugh, C.Russell</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4687-5d8c1e8b5c8f32420a536a2522a33d1410b8586ede0dd7c43a11b89533af799d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Biophysical Phenomena</topic><topic>Biophysics</topic><topic>Cercopithecus aethiops</topic><topic>CHO Cells</topic><topic>circular dichroism</topic><topic>COS Cells</topic><topic>Cricetinae</topic><topic>differential scanning calorimetry</topic><topic>DNA - analysis</topic><topic>DNA - chemistry</topic><topic>DNA - genetics</topic><topic>DNA delivery</topic><topic>FTIR</topic><topic>gene delivery</topic><topic>General pharmacology</topic><topic>Genetic Therapy - methods</topic><topic>isothermal titration calorimetry</topic><topic>Medical sciences</topic><topic>Pharmaceutical technology. Pharmaceutical industry</topic><topic>Pharmacology. Drug treatments</topic><topic>physical characterization</topic><topic>plasmid DNA</topic><topic>Polyethyleneimine - analysis</topic><topic>Polyethyleneimine - chemistry</topic><topic>polyethylenimine</topic><topic>Spectroscopy, Fourier Transform Infrared - methods</topic><topic>Transfection - methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Choosakoonkriang, Sirirat</creatorcontrib><creatorcontrib>Lobo, Brian A.</creatorcontrib><creatorcontrib>Koe, Gary S.</creatorcontrib><creatorcontrib>Koe, Janet G.</creatorcontrib><creatorcontrib>Middaugh, C.Russell</creatorcontrib><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><jtitle>Journal of pharmaceutical sciences</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Choosakoonkriang, Sirirat</au><au>Lobo, Brian A.</au><au>Koe, Gary S.</au><au>Koe, Janet G.</au><au>Middaugh, C.Russell</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Biophysical Characterization of PEI/DNA Complexes</atitle><jtitle>Journal of pharmaceutical sciences</jtitle><addtitle>J. Pharm. Sci</addtitle><date>2003-08</date><risdate>2003</risdate><volume>92</volume><issue>8</issue><spage>1710</spage><epage>1722</epage><pages>1710-1722</pages><issn>0022-3549</issn><eissn>1520-6017</eissn><coden>JPMSAE</coden><abstract>The main goal of this study was to determine the effects of polyethylenimine (PEI) molecular weight and structure (750kDa, 25kDa, 2kDa branched, and 25kDa linear PEI) and the nitrogen/phosphate (N/P) molar ratio on the physical properties and transfection efficiencies of PEI/DNA complexes. Fourier transform infrared spectroscopy revealed that DNA remained in the B conformation when complexed to all PEIs. Unique alterations in the circular dichroism spectra of DNA were observed in the presence of each PEI, whereas differential scanning calorimetry measurements showed that all PEIs examined destabilized supercoiled DNA at N/P<3/1, but not at higher ratios. Isothermal titration calorimetry revealed the existence of protonation changes at low ionic strength due to possible shifts in pKa of the ionizable groups of PEI during complex formation. Twenty-five kilodalton branched and 25kDa linear PEI complexes showed the highest transfection efficiencies at an N/P ratio of 6:1 in COS-7 and CHO-K1 cells, respectively. These investigations have detected alterations in the physical and colloidal properties of the complexes that were sensitive to polymer structure, molecular weight, and polymer/DNA ratio, but these properties did not directly correlate with their transfection efficiencies. To further probe any possible relationship between these parameters and activity, a more refined biophysical analysis of any subpopulations in these samples that may differ in transfection activity is suggested, although the existence of such species remains unknown. © 2003 Wiley-Liss, Inc. and the American Pharmacists Association</abstract><cop>Hoboken</cop><pub>Elsevier Inc</pub><pmid>12884257</pmid><doi>10.1002/jps.10437</doi><tpages>13</tpages></addata></record> |
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| subjects | Animals Biological and medical sciences Biophysical Phenomena Biophysics Cercopithecus aethiops CHO Cells circular dichroism COS Cells Cricetinae differential scanning calorimetry DNA - analysis DNA - chemistry DNA - genetics DNA delivery FTIR gene delivery General pharmacology Genetic Therapy - methods isothermal titration calorimetry Medical sciences Pharmaceutical technology. Pharmaceutical industry Pharmacology. Drug treatments physical characterization plasmid DNA Polyethyleneimine - analysis Polyethyleneimine - chemistry polyethylenimine Spectroscopy, Fourier Transform Infrared - methods Transfection - methods |
| title | Biophysical Characterization of PEI/DNA Complexes |
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