Selection of DNA aptamers against insulin and construction of an aptameric enzyme subunit for insulin sensing

Yoshida, Wataru; Mochizuki, Eriko; Takase, Madoka; Hasegawa, Hijiri; Morita, Yo; Yamazaki, Hiroki; Sode, Koji; Ikebukuro, Kazunori

doi:10.1016/j.bios.2008.06.016

Cited by 122 publications

(88 citation statements)

References 24 publications

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“…͔; cancer biomarkers; [22][23][24][25] HIVassociated proteins; [26][27][28][29] food-borne pathogens; 30 and other biologically important biomolecules such as insulin, 31,32 immunoglobulin E ͑IgE͒, 33 and thrombin. 34 They exhibit protein binding affinities that are comparable to those of corresponding antibodies.…”

mentioning

confidence: 99%

Direct detection of aptamer-thrombin binding via surface-enhanced Raman spectroscopy

et al. 2010

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Abstract. In this study, we exploit the sensitivity offered by surfaceenhanced Raman scattering ͑SERS͒ for the direct detection of thrombin using the thrombin-binding aptamer ͑TBA͒ as molecular receptor. The technique utilizes immobilized silver nanoparticles that are functionalized with thiolated thrombin-specific binding aptamer, a 15-mer ͑5Ј-GGTTGGTGTGGTTGG-3Ј͒ quadruplex forming oligonucleotide. In addition to the Raman vibrational bands corresponding to the aptamer and blocking agent, new peaks ͑mainly at 1140, 1540, and 1635 cm −1 ͒ that are characteristic of the protein are observed upon binding of thrombin. These spectral changes are not observed when the aptamer-nanoparticle assembly is exposed to a nonbinding protein such as bovine serum albumin ͑BSA͒. This methodology could be further used for the development of label-free biosensors for direct detection of proteins and other molecules of interest for which aptamers are available.

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confidence: 99%

Direct detection of aptamer-thrombin binding via surface-enhanced Raman spectroscopy

et al. 2010

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“…We have developed two types of AES for the detection of adenosine (14) and insulin. (15) We found the lower limit of adenosine detection was 25 μM. Given that the enzymatic activity of thrombin is not very high, it is possible to construct more sensitive AES by utilizing other enzymes.…”

Section: Proof Of Concept Of Using Various Proteinaceous Enzymes For mentioning

confidence: 85%

“…For example, a targetbinding aptamer is inactivated by the hybridization of a partially complementary strand attached at an enzyme-inhibiting aptamer. (14,15) Thus, in the absence of target molecules, the enzyme-inhibiting aptamer of AES folds into the structure, binds to the enzyme, and inhibits its enzymatic activity. In the presence of target molecules, the target-binding aptamer folds into the original structure, which leads to the disruption of the structure of the enzyme-inhibiting aptamer and a decrease in the inhibitory activity.…”

Section: Proof Of Concept Of Using Various Proteinaceous Enzymes For mentioning

confidence: 99%

Sensitive and Homogeneous Detection Systemwith Aptamer-based Biosensor

2016

Sensors and Materials

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Rapid and highly sensitive methods for detecting target molecules are required to develop biomarker-based diagnoses that help medication or therapy. Thus, the development of homogeneous sensing methods with high sensitivity can provide the basis for detecting a biomarker in routine diagnostics and on site. Oligonucleotide aptamers, ligands that consist of nucleic acids, have been selected for numerous targets such as proteins and small compounds, and are applicable to biosensing platforms. In particular, attractive features of aptamers enable us to construct valuable sensing methods for clinical uses without the need to use nanomaterials. Recent discoveries of novel signal transducing elements derived from an enzymatic and a fluorescent oligonucleotide also allow for the development of homogeneous aptasensors. In this review, we describe recent progress in the development of aptamer-based biosensors with high sensitivity.

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“…If the target molecule activates enzymatic activity, we can quantify the target molecule via an enzyme activity measurement. We named this sensing system the Aptameric Enzyme Subunit (AES) Yoshida et al, 2009;Yoshida et al, 2006aYoshida et al, , b, 2008). An AES consists of two aptamers: an enzyme-inhibiting aptamer and a target moleculebinding aptamer.…”

Section: Enzyme Activity Regulation By the Target Moleculementioning

confidence: 99%