Selection of DNA aptamers against insulin and construction of an aptameric enzyme subunit for insulin sensing
We selected DNA aptamers against insulin and developed an aptameric enzyme subunit (AES) for insulin sensing. The insulin-binding aptamers were identified from a single-strand DNA library which was expected to form various kinds of G-quartet structures. In vitro selection was carried out by means of...
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| Veröffentlicht in: | Biosensors & bioelectronics 2009, Vol.24 (5), p.1116-1120 |
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| container_title | Biosensors & bioelectronics |
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| creator | Yoshida, Wataru Mochizuki, Eriko Takase, Madoka Hasegawa, Hijiri Morita, Yo Yamazaki, Hiroki Sode, Koji Ikebukuro, Kazunori |
| description | We selected DNA aptamers against insulin and developed an aptameric enzyme subunit (AES) for insulin sensing. The insulin-binding aptamers were identified from a single-strand DNA library which was expected to form various kinds of G-quartet structures.
In vitro selection was carried out by means of aptamer blotting, which visualizes the oligonucleotides binding to the target protein at each round. After the 6th round of selection, insulin-binding aptamers were identified. These identified insulin-binding aptamers had a higher binding ability than the insulin-linked polymorphic region (ILPR) oligonucleotide, which can be called a “natural” insulin-binding DNA aptamer. The circular-dichroism (CD) spectrum measurement of the identified insulin-binding DNA aptamers indicated that the aptamers would fold into a G-quartet structure. We also developed an AES by connecting the best identified insulin-binding aptamer with the thrombin-inhibiting aptamer. Using this AES, we were able to detect insulin by measuring the thrombin enzymatic activity without bound/free separation. |
| doi_str_mv | 10.1016/j.bios.2008.06.016 |
| format | Article |
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In vitro selection was carried out by means of aptamer blotting, which visualizes the oligonucleotides binding to the target protein at each round. After the 6th round of selection, insulin-binding aptamers were identified. These identified insulin-binding aptamers had a higher binding ability than the insulin-linked polymorphic region (ILPR) oligonucleotide, which can be called a “natural” insulin-binding DNA aptamer. The circular-dichroism (CD) spectrum measurement of the identified insulin-binding DNA aptamers indicated that the aptamers would fold into a G-quartet structure. We also developed an AES by connecting the best identified insulin-binding aptamer with the thrombin-inhibiting aptamer. Using this AES, we were able to detect insulin by measuring the thrombin enzymatic activity without bound/free separation.</description><subject>Aptameric enzyme subunit</subject><subject>Aptamers, Nucleotide - chemistry</subject><subject>Biological and medical sciences</subject><subject>Biosensing Techniques - instrumentation</subject><subject>Biosensing Techniques - methods</subject><subject>Biotechnology</subject><subject>Equipment Design</subject><subject>Equipment Failure Analysis</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>G-quartet library</subject><subject>Insulin - analysis</subject><subject>Insulin - chemistry</subject><subject>Insulin detection</subject><subject>Insulin-binding aptamer</subject><subject>Reproducibility of Results</subject><subject>Sensitivity and Specificity</subject><subject>Spectrometry, Fluorescence - instrumentation</subject><subject>Spectrometry, Fluorescence - methods</subject><issn>0956-5663</issn><issn>1873-4235</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc1u1DAURi0EosPAC7BA3sAu6bXj2InEpip_lSq6aPeWY99UHiX2YCdI5enxaIZhBxtb-nS-K_seQt4yqBkwebmrBx9zzQG6GmRdomdkwzrVVII37XOygb6VVStlc0Fe5bwDAMV6eEkuWCeFUIxtyHyPE9rFx0DjSD99v6Jmv5gZU6bm0fiQF1qOdfKBmuCojSVJ67lgwh_eW4rh19OMNK_DGvxCx5jO3Ywh-_D4mrwYzZTxzenekocvnx-uv1W3d19vrq9uKys6tlRGWei5k1bwEZxsOWsktqMDDkYNfWs4jIoJ4-QwNNIxybEXapC9tGZQotmSD8ex-xR_rJgXPftscZpMwLhmLWUHouua_4IcuJJdxwvIj6BNMeeEo94nP5v0pBnogwy90wcZ-iBDg9QlKqV3p-nrMKP7WzltvwDvT4DJ1kxjMsH6fObKtxulis8t-XjksOzsp8eks_UYLDqfijztov_XO34DRHqpMg</recordid><startdate>2009</startdate><enddate>2009</enddate><creator>Yoshida, Wataru</creator><creator>Mochizuki, Eriko</creator><creator>Takase, Madoka</creator><creator>Hasegawa, Hijiri</creator><creator>Morita, Yo</creator><creator>Yamazaki, Hiroki</creator><creator>Sode, Koji</creator><creator>Ikebukuro, Kazunori</creator><general>Elsevier B.V</general><general>Elsevier</general><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>7QO</scope><scope>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>2009</creationdate><title>Selection of DNA aptamers against insulin and construction of an aptameric enzyme subunit for insulin sensing</title><author>Yoshida, Wataru ; Mochizuki, Eriko ; Takase, Madoka ; Hasegawa, Hijiri ; Morita, Yo ; Yamazaki, Hiroki ; Sode, Koji ; Ikebukuro, Kazunori</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c481t-a7c092d6c42f0d652136e5fd020a7b95a20f714ad6bb36d162e947b696cab743</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>Aptameric enzyme subunit</topic><topic>Aptamers, Nucleotide - chemistry</topic><topic>Biological and medical sciences</topic><topic>Biosensing Techniques - instrumentation</topic><topic>Biosensing Techniques - methods</topic><topic>Biotechnology</topic><topic>Equipment Design</topic><topic>Equipment Failure Analysis</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>G-quartet library</topic><topic>Insulin - analysis</topic><topic>Insulin - chemistry</topic><topic>Insulin detection</topic><topic>Insulin-binding aptamer</topic><topic>Reproducibility of Results</topic><topic>Sensitivity and Specificity</topic><topic>Spectrometry, Fluorescence - instrumentation</topic><topic>Spectrometry, Fluorescence - methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Yoshida, Wataru</creatorcontrib><creatorcontrib>Mochizuki, Eriko</creatorcontrib><creatorcontrib>Takase, Madoka</creatorcontrib><creatorcontrib>Hasegawa, Hijiri</creatorcontrib><creatorcontrib>Morita, Yo</creatorcontrib><creatorcontrib>Yamazaki, Hiroki</creatorcontrib><creatorcontrib>Sode, Koji</creatorcontrib><creatorcontrib>Ikebukuro, Kazunori</creatorcontrib><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>Biotechnology Research Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Biosensors & bioelectronics</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Yoshida, Wataru</au><au>Mochizuki, Eriko</au><au>Takase, Madoka</au><au>Hasegawa, Hijiri</au><au>Morita, Yo</au><au>Yamazaki, Hiroki</au><au>Sode, Koji</au><au>Ikebukuro, Kazunori</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Selection of DNA aptamers against insulin and construction of an aptameric enzyme subunit for insulin sensing</atitle><jtitle>Biosensors & bioelectronics</jtitle><addtitle>Biosens Bioelectron</addtitle><date>2009</date><risdate>2009</risdate><volume>24</volume><issue>5</issue><spage>1116</spage><epage>1120</epage><pages>1116-1120</pages><issn>0956-5663</issn><eissn>1873-4235</eissn><abstract>We selected DNA aptamers against insulin and developed an aptameric enzyme subunit (AES) for insulin sensing. The insulin-binding aptamers were identified from a single-strand DNA library which was expected to form various kinds of G-quartet structures.
In vitro selection was carried out by means of aptamer blotting, which visualizes the oligonucleotides binding to the target protein at each round. After the 6th round of selection, insulin-binding aptamers were identified. These identified insulin-binding aptamers had a higher binding ability than the insulin-linked polymorphic region (ILPR) oligonucleotide, which can be called a “natural” insulin-binding DNA aptamer. The circular-dichroism (CD) spectrum measurement of the identified insulin-binding DNA aptamers indicated that the aptamers would fold into a G-quartet structure. We also developed an AES by connecting the best identified insulin-binding aptamer with the thrombin-inhibiting aptamer. Using this AES, we were able to detect insulin by measuring the thrombin enzymatic activity without bound/free separation.</abstract><cop>Kidlington</cop><pub>Elsevier B.V</pub><pmid>18644711</pmid><doi>10.1016/j.bios.2008.06.016</doi><tpages>5</tpages></addata></record> |
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| subjects | Aptameric enzyme subunit Aptamers, Nucleotide - chemistry Biological and medical sciences Biosensing Techniques - instrumentation Biosensing Techniques - methods Biotechnology Equipment Design Equipment Failure Analysis Fundamental and applied biological sciences. Psychology G-quartet library Insulin - analysis Insulin - chemistry Insulin detection Insulin-binding aptamer Reproducibility of Results Sensitivity and Specificity Spectrometry, Fluorescence - instrumentation Spectrometry, Fluorescence - methods |
| title | Selection of DNA aptamers against insulin and construction of an aptameric enzyme subunit for insulin sensing |
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