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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Le, Thao T. Scott, Steven Cass, Anthony E. G. |
| Description | Country affiliation: United kingdom Author Affiliation: Le TT ( Department of Chemistry, Imperial College London, SW7 2AZ, UK.) |
| Abstract | This paper describes the measurement of the binding affinities of two bifunctional RNA aptamers to their respective ligands. The aptamers comprise either a theophylline or malachite green binding sequence fused to a streptavidin binding sequence. These bifunctional aptamers are shown to bind simultaneously to both the small ligand and to streptavidin whether in free solution or on gold surfaces. Binding isotherms for both interactions were measured by different physiochemical techniques: surface plasmon resonance, fluorescence spectroscopy and dynamic light scattering. Both qualitatively and quantitatively there is little difference in binding affinities between the bifunctional aptamers and their monofunctional components. The respective K(d) values for streptavidin binding in the monofunctional aptamer and in the theophylline bifunctional aptamer were 12 nM and 65 nM, respectively whilst the K(d) values for theophylline binding in the monofunctional aptamer and the streptavidin bifunctional aptamer were 300 nM and 120 nM. These results are consistent with treating each aptamer sequence as a module that can be combined with others without significant loss of function. This allows for the use of streptavidin based immobilization strategies without either the cost of biotinylated dNTPs or the variable yields associated with the chemical biotinylation of RNA. |
| File Format | HTM / HTML |
| ISSN | 00032670 |
| Volume Number | 761 |
| e-ISSN | 18734324 |
| Journal | Analytica Chimica Acta |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2013-01-25 |
| Publisher Place | Netherlands |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Analytical Discipline Chemistry Aptamers, Nucleotide Chemistry Metabolism Rosaniline Dyes Streptavidin Theophylline Base Sequence Binding Sites Ligands Light Models, Molecular Molecular Sequence Data Nucleic Acid Conformation Scattering, Radiation Spectrometry, Fluorescence Surface Plasmon Resonance Journal Article Research Support, Non-u.s. Gov't |
| Content Type | Text |
| Resource Type | Article |
| Subject | Spectroscopy Environmental Chemistry Analytical Chemistry Biochemistry |
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