One round of SELEX for the generation of DNA aptamers directed against KLK6
Identifieur interne : 000057 ( Main/Exploration ); précédent : 000056; suivant : 000058One round of SELEX for the generation of DNA aptamers directed against KLK6
Auteurs : Steve Arnold [Royaume-Uni] ; Georgios Pampalakis [Grèce] ; Kalliopi Kantiotou [Grèce] ; Dilson Silva [Brésil] ; Celia Cortez [Brésil] ; Sotiris Missailidis [Royaume-Uni] ; Georgia Sotiropoulou [Grèce]Source :
- Biological Chemistry [ 1431-6730 ] ; 2012-05-01.
Abstract
Kallikrein-related peptidase 6 (KLK6) is an active serine protease that has been implicated in common pathologies, including neurodegenerative disorders such as Parkinson and Alzheimer disease and certain types of cancer. Antibodies, either polyclonal or monoclonal, that exhibit specificity for distinct members of the extended kallikrein family, including KLK6, were developed. With the exception of KLK3/PSA, the identification and generation of aptamers, as potential new tools with improved characteristics demanded for therapeutic and diagnostic applications, has not been explored for KLKs. Here, we report for the first time the identification of novel DNA aptamers against KLK6 that were isolated using a modified systemic evolution of ligands by exponential enrichment technique. The identified aptamers were characterized using fluorescence spectroscopy, competition ELISA, and quartz crystal microbalance, and two aptamers (008 and 022) were found to exhibit high affinity (Kd in the low nanomolar range) for KLK6. Aptamers were tested for their ability to bind to serum albumin, to demonstrate their specificity for their target, and the possible involvement of such proteins in the transport of aptamers into the bloodstream. The developed aptamers are expected to assist the development of novel diagnostic, biosensing, and therapeutic strategies.
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DOI: 10.1515/hsz-2011-0253
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Janeiro</region></placeName></affiliation></author><author><name sortKey="Missailidis, Sotiris" sort="Missailidis, Sotiris" uniqKey="Missailidis S" first="Sotiris" last="Missailidis">Sotiris Missailidis</name><affiliation wicri:level="1"><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Department of Chemistry, The Open University, Milton, Keynes MK7 6AA</wicri:regionArea><wicri:noRegion>Keynes MK7 6AA</wicri:noRegion></affiliation></author><author><name sortKey="Sotiropoulou, Georgia" sort="Sotiropoulou, Georgia" uniqKey="Sotiropoulou G" first="Georgia" last="Sotiropoulou">Georgia Sotiropoulou</name><affiliation wicri:level="1"><country xml:lang="fr">Grèce</country><wicri:regionArea>Department of Pharmacy, University of Patras, Rion-Patras 26500</wicri:regionArea><wicri:noRegion>Rion-Patras 26500</wicri:noRegion></affiliation></author></analytic><monogr></monogr><series><title level="j">Biological Chemistry</title><idno type="ISSN">1431-6730</idno><idno 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Parkinson and Alzheimer disease and certain types of cancer. Antibodies, either polyclonal or monoclonal, that exhibit specificity for distinct members of the extended kallikrein family, including KLK6, were developed. With the exception of KLK3/PSA, the identification and generation of aptamers, as potential new tools with improved characteristics demanded for therapeutic and diagnostic applications, has not been explored for KLKs. Here, we report for the first time the identification of novel DNA aptamers against KLK6 that were isolated using a modified systemic evolution of ligands by exponential enrichment technique. The identified aptamers were characterized using fluorescence spectroscopy, competition ELISA, and quartz crystal microbalance, and two aptamers (008 and 022) were found to exhibit high affinity (Kd in the low nanomolar range) for KLK6. Aptamers were tested for their ability to bind to serum albumin, to demonstrate their specificity for their target, and the possible involvement of such proteins in the transport of aptamers into the bloodstream. The developed aptamers are expected to assist the development of novel diagnostic, biosensing, and therapeutic strategies.</div></front></TEI><affiliations><list><country><li>Brésil</li><li>Grèce</li><li>Royaume-Uni</li></country><region><li>État de Rio de Janeiro</li></region><settlement><li>Rio de Janeiro</li></settlement></list><tree><country name="Royaume-Uni"><noRegion><name sortKey="Arnold, Steve" sort="Arnold, Steve" uniqKey="Arnold S" first="Steve" last="Arnold">Steve Arnold</name></noRegion><name sortKey="Missailidis, Sotiris" sort="Missailidis, Sotiris" uniqKey="Missailidis S" first="Sotiris" last="Missailidis">Sotiris Missailidis</name></country><country name="Grèce"><noRegion><name sortKey="Pampalakis, Georgios" sort="Pampalakis, Georgios" uniqKey="Pampalakis G" first="Georgios" last="Pampalakis">Georgios Pampalakis</name></noRegion><name sortKey="Kantiotou, Kalliopi" sort="Kantiotou, Kalliopi" uniqKey="Kantiotou K" first="Kalliopi" last="Kantiotou">Kalliopi Kantiotou</name><name sortKey="Sotiropoulou, Georgia" sort="Sotiropoulou, Georgia" uniqKey="Sotiropoulou G" first="Georgia" last="Sotiropoulou">Georgia Sotiropoulou</name></country><country name="Brésil"><region name="État de Rio de Janeiro"><name sortKey="Silva, Dilson" sort="Silva, Dilson" uniqKey="Silva D" first="Dilson" last="Silva">Dilson Silva</name></region><name sortKey="Cortez, Celia" sort="Cortez, Celia" uniqKey="Cortez C" first="Celia" last="Cortez">Celia Cortez</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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