A Fluorescence-Based Method for Determining the Surface Coverage and Hybridization Efficiency of Thiol-Capped Oligonucleotides Bound to Gold Thin Films and Nanoparticles
Using a fluorescence-based method, we have determined the number of thiol-derivatized single-stranded oligonucleotides bound to gold nanoparticles and their extent of hybridization with complementary oligonucleotides in solution. Oligonucleotide surface coverages of hexanethiol 12-mer oligonucleotid...
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| Veröffentlicht in: | Analytical chemistry (Washington) 2000-11, Vol.72 (22), p.5535-5541 |
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| container_title | Analytical chemistry (Washington) |
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| creator | Demers, Linette M Mirkin, Chad A Mucic, Robert C Reynolds, Robert A Letsinger, Robert L Elghanian, Robert Viswanadham, Garimella |
| description | Using a fluorescence-based method, we have determined the number of thiol-derivatized single-stranded oligonucleotides bound to gold nanoparticles and their extent of hybridization with complementary oligonucleotides in solution. Oligonucleotide surface coverages of hexanethiol 12-mer oligonucleotides on gold nanoparticles (34 ± 1 pmol/cm2) were significantly higher than on planar gold thin films (18 ± 3 pmol/cm2), while the percentage of hybridizable strands on the gold nanoparticles (1.3 ± 0.3 pmol/cm2, 4%) was lower than for gold thin films (6 ± 2 pmol/cm2, 33%). A gradual increase in electrolyte concentration over the course of oligonucleotide deposition significantly increases surface coverage and consequently particle stability. In addition, oligonucleotide spacer sequences improve the hybridization efficiency of oligonucleotide-modified nanoparticles from ∼4 to 44%. The surface coverage of recognition strands can be tailored using coadsorbed diluent oligonucleotides. This provides a means of indirectly controlling the average number of hybridized strands per nanoparticle. The work presented here has important implications with regard to understanding interactions between modified oligonucleotides and metal nanoparticles, as well as optimizing the sensitivity of gold nanoparticle-based oligonucleotide detection methods. |
| doi_str_mv | 10.1021/ac0006627 |
| format | Article |
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Oligonucleotide surface coverages of hexanethiol 12-mer oligonucleotides on gold nanoparticles (34 ± 1 pmol/cm2) were significantly higher than on planar gold thin films (18 ± 3 pmol/cm2), while the percentage of hybridizable strands on the gold nanoparticles (1.3 ± 0.3 pmol/cm2, 4%) was lower than for gold thin films (6 ± 2 pmol/cm2, 33%). A gradual increase in electrolyte concentration over the course of oligonucleotide deposition significantly increases surface coverage and consequently particle stability. In addition, oligonucleotide spacer sequences improve the hybridization efficiency of oligonucleotide-modified nanoparticles from ∼4 to 44%. The surface coverage of recognition strands can be tailored using coadsorbed diluent oligonucleotides. This provides a means of indirectly controlling the average number of hybridized strands per nanoparticle. The work presented here has important implications with regard to understanding interactions between modified oligonucleotides and metal nanoparticles, as well as optimizing the sensitivity of gold nanoparticle-based oligonucleotide detection methods.</description><identifier>ISSN: 0003-2700</identifier><identifier>EISSN: 1520-6882</identifier><identifier>DOI: 10.1021/ac0006627</identifier><identifier>PMID: 11101228</identifier><identifier>CODEN: ANCHAM</identifier><language>eng</language><publisher>Washington, DC: American Chemical Society</publisher><subject>Atoms & subatomic particles ; Biological and medical sciences ; Diverse techniques ; Fundamental and applied biological sciences. 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Chem</addtitle><description>Using a fluorescence-based method, we have determined the number of thiol-derivatized single-stranded oligonucleotides bound to gold nanoparticles and their extent of hybridization with complementary oligonucleotides in solution. Oligonucleotide surface coverages of hexanethiol 12-mer oligonucleotides on gold nanoparticles (34 ± 1 pmol/cm2) were significantly higher than on planar gold thin films (18 ± 3 pmol/cm2), while the percentage of hybridizable strands on the gold nanoparticles (1.3 ± 0.3 pmol/cm2, 4%) was lower than for gold thin films (6 ± 2 pmol/cm2, 33%). A gradual increase in electrolyte concentration over the course of oligonucleotide deposition significantly increases surface coverage and consequently particle stability. In addition, oligonucleotide spacer sequences improve the hybridization efficiency of oligonucleotide-modified nanoparticles from ∼4 to 44%. The surface coverage of recognition strands can be tailored using coadsorbed diluent oligonucleotides. This provides a means of indirectly controlling the average number of hybridized strands per nanoparticle. The work presented here has important implications with regard to understanding interactions between modified oligonucleotides and metal nanoparticles, as well as optimizing the sensitivity of gold nanoparticle-based oligonucleotide detection methods.</description><subject>Atoms & subatomic particles</subject><subject>Biological and medical sciences</subject><subject>Diverse techniques</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gold</subject><subject>Gold - chemistry</subject><subject>Metals</subject><subject>Molecular and cellular biology</subject><subject>Nucleic Acid Hybridization</subject><subject>Oligonucleotides - chemistry</subject><subject>Particle Size</subject><subject>Spectrometry, Fluorescence - methods</subject><subject>Surface Properties</subject><issn>0003-2700</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0UFv0zAUB_AIgVgZHPgCyAKBxCFgO03sHLdu3ZA2hlhhR-vFeW69pXaxE0T5RnxLXFp1Ehw42fL7-S8_vyx7zug7Rjl7D5pSWlVcPMhGrOQ0r6TkD7NROi1yLig9yJ7EeEspY5RVj7MDttlwLkfZryMy7QYfMGp0GvNjiNiSS-wXviXGB3KCPYalddbNSb9Acj0EAxrJxH_HAHMk4Fpyvm6Cbe1P6K135NQYq22KWxNvyGxhfZdPYLVKwVednXs36A59b1uM5NgP6X7vyZnv2o11ZGq7ZfwT-xGcX0HobfLxafbIQBfx2W49zL5MT2eT8_zi6uzD5Ogih5KyPtdgTFtCjdCgrmtdN_VYiqZgVZlK1FAARCrSjzVNo-tWN5oWNa84N01dtLI4zN5sc1fBfxsw9mpp0-d0HTj0Q1SCjwspOP8vZEKMJS_qBF_-BW_9EFxqQnEmZFWUFU3o7Rbp4GMMaNQq2CWEtWJUbaas9lNO9sUucGiW2N7L3VgTeLUDEDV0JoDTNu6dlJTJTUy-VTb2-GNfhXCnKlGIUs0-Xaubr58vJzcnUzVO_vXWg473Lfz7vN8K9Mv2</recordid><startdate>20001115</startdate><enddate>20001115</enddate><creator>Demers, Linette M</creator><creator>Mirkin, Chad A</creator><creator>Mucic, Robert C</creator><creator>Reynolds, Robert A</creator><creator>Letsinger, Robert L</creator><creator>Elghanian, Robert</creator><creator>Viswanadham, Garimella</creator><general>American Chemical Society</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><scope>7QF</scope><scope>7QO</scope><scope>7QQ</scope><scope>7SC</scope><scope>7SE</scope><scope>7SP</scope><scope>7SR</scope><scope>7TA</scope><scope>7TB</scope><scope>7TM</scope><scope>7U5</scope><scope>7U7</scope><scope>7U9</scope><scope>8BQ</scope><scope>8FD</scope><scope>C1K</scope><scope>F28</scope><scope>FR3</scope><scope>H8D</scope><scope>H8G</scope><scope>H94</scope><scope>JG9</scope><scope>JQ2</scope><scope>KR7</scope><scope>L7M</scope><scope>L~C</scope><scope>L~D</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20001115</creationdate><title>A Fluorescence-Based Method for Determining the Surface Coverage and Hybridization Efficiency of Thiol-Capped Oligonucleotides Bound to Gold Thin Films and Nanoparticles</title><author>Demers, Linette M ; Mirkin, Chad A ; Mucic, Robert C ; Reynolds, Robert A ; Letsinger, Robert L ; Elghanian, Robert ; Viswanadham, Garimella</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a501t-caffd5a9eabec99c9b9487b3165caf0f0aaee07c00bbbc9dcbc0392622fb93d83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Atoms & subatomic particles</topic><topic>Biological and medical sciences</topic><topic>Diverse techniques</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gold</topic><topic>Gold - chemistry</topic><topic>Metals</topic><topic>Molecular and cellular biology</topic><topic>Nucleic Acid Hybridization</topic><topic>Oligonucleotides - chemistry</topic><topic>Particle Size</topic><topic>Spectrometry, Fluorescence - methods</topic><topic>Surface Properties</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Demers, Linette M</creatorcontrib><creatorcontrib>Mirkin, Chad A</creatorcontrib><creatorcontrib>Mucic, Robert C</creatorcontrib><creatorcontrib>Reynolds, Robert A</creatorcontrib><creatorcontrib>Letsinger, Robert L</creatorcontrib><creatorcontrib>Elghanian, Robert</creatorcontrib><creatorcontrib>Viswanadham, Garimella</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><collection>Aluminium Industry Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>Ceramic Abstracts</collection><collection>Computer and Information Systems Abstracts</collection><collection>Corrosion Abstracts</collection><collection>Electronics & Communications Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Materials Business File</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Engineering Research Database</collection><collection>Aerospace Database</collection><collection>Copper Technical Reference Library</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Materials Research Database</collection><collection>ProQuest Computer Science Collection</collection><collection>Civil Engineering Abstracts</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Computer and Information Systems Abstracts Academic</collection><collection>Computer and Information Systems Abstracts Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Analytical chemistry (Washington)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Demers, Linette M</au><au>Mirkin, Chad A</au><au>Mucic, Robert C</au><au>Reynolds, Robert A</au><au>Letsinger, Robert L</au><au>Elghanian, Robert</au><au>Viswanadham, Garimella</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A Fluorescence-Based Method for Determining the Surface Coverage and Hybridization Efficiency of Thiol-Capped Oligonucleotides Bound to Gold Thin Films and Nanoparticles</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>2000-11-15</date><risdate>2000</risdate><volume>72</volume><issue>22</issue><spage>5535</spage><epage>5541</epage><pages>5535-5541</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><coden>ANCHAM</coden><abstract>Using a fluorescence-based method, we have determined the number of thiol-derivatized single-stranded oligonucleotides bound to gold nanoparticles and their extent of hybridization with complementary oligonucleotides in solution. Oligonucleotide surface coverages of hexanethiol 12-mer oligonucleotides on gold nanoparticles (34 ± 1 pmol/cm2) were significantly higher than on planar gold thin films (18 ± 3 pmol/cm2), while the percentage of hybridizable strands on the gold nanoparticles (1.3 ± 0.3 pmol/cm2, 4%) was lower than for gold thin films (6 ± 2 pmol/cm2, 33%). A gradual increase in electrolyte concentration over the course of oligonucleotide deposition significantly increases surface coverage and consequently particle stability. In addition, oligonucleotide spacer sequences improve the hybridization efficiency of oligonucleotide-modified nanoparticles from ∼4 to 44%. The surface coverage of recognition strands can be tailored using coadsorbed diluent oligonucleotides. This provides a means of indirectly controlling the average number of hybridized strands per nanoparticle. The work presented here has important implications with regard to understanding interactions between modified oligonucleotides and metal nanoparticles, as well as optimizing the sensitivity of gold nanoparticle-based oligonucleotide detection methods.</abstract><cop>Washington, DC</cop><pub>American Chemical Society</pub><pmid>11101228</pmid><doi>10.1021/ac0006627</doi><tpages>7</tpages></addata></record> |
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| ispartof | Analytical chemistry (Washington), 2000-11, Vol.72 (22), p.5535-5541 |
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| subjects | Atoms & subatomic particles Biological and medical sciences Diverse techniques Fundamental and applied biological sciences. Psychology Gold Gold - chemistry Metals Molecular and cellular biology Nucleic Acid Hybridization Oligonucleotides - chemistry Particle Size Spectrometry, Fluorescence - methods Surface Properties |
| title | A Fluorescence-Based Method for Determining the Surface Coverage and Hybridization Efficiency of Thiol-Capped Oligonucleotides Bound to Gold Thin Films and Nanoparticles |
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