Simple and rapid colorimetric detection of cofactors of aptazymes using noncrosslinking gold nanoparticle aggregation

Bioorganic and Medicinal Chemistry Letters

Volumn 18, Issue 24, 2008, Pages 6517-6520

  Ogawa, Atsushi ^a   Maeda, Mizuo ^a  

^a INST OF PHYS AND CHEMICAL RESEARCH   (Japan)

Author keywords

Aptazyme; Biosensor; Gold nanoparticle

Indexed keywords

APTAZYME; ENZYME; GOLD; NANOPARTICLE; RIBOZYME; THEOPHYLLINE; UNCLASSIFIED DRUG;

ARTICLE; BIOSENSOR; COLORIMETRY; CONCENTRATION RESPONSE; DNA BINDING; MOLECULAR INTERACTION; RNA BINDING;

BIOSENSING TECHNIQUES; CAFFEINE; CHEMISTRY, PHARMACEUTICAL; COLORIMETRY; CROSS-LINKING REAGENTS; DNA; DOSE-RESPONSE RELATIONSHIP, DRUG; DRUG DESIGN; GOLD; METAL NANOPARTICLES; MOLECULAR CONFORMATION; NANOTECHNOLOGY; NUCLEIC ACID HYBRIDIZATION; SENSITIVITY AND SPECIFICITY; THEOPHYLLINE;

EID: 56249146373     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2008.10.051     Document Type: Article

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24. We optimized this cleaved RNA-releasing temperature. Although no change was detected at 40 °C, a color change from red to purple was detected at 50 °C. The color changed from red to colorless at 55 °C. This indicates that Tm between the cleaved RNA and the parent aptazyme is 50-55 °C. On the other hand, because aggregation occurred at 55 °C, Tm between the cleaved RNA and the probe-DNA on AuNPs is estimated well above 55 °C.
25. To verify that the AuNPs precipitate certainly by hybridization between the probe-DNA and the cleaved RNA, we performed the aggregation experiments using noncomplementary probe-DNA (on AuNPs)/cleaved RNA (from the aptazyme) pairs. As a result, using these noncomplementary pairs, aggregation did not occur even in the presence of cofactors and the cleaved RNA. Because the complementary but non-cleaved aptazyme or the cofactor did not also induce aggregation, it is clear that only the complementary cleaved RNA can induce aggregation by hybridizing to the probe-DNA on AuNPs.
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