Fluorescence polarization immunoassays and related methods for simple, high-throughput screening of small molecules

Smith, David S.; Eremin, Sergei A.

doi:10.1007/s00216-008-1897-z

Cited by 228 publications

(155 citation statements)

References 104 publications

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“…Since the discovery of aptamers in 1990s, aptamer-based assays for small molecules have drawn increasing attentions in environmental sensing, food safety, and clinical analysis due to the unique features of aptamers, such as easy generation, facile labeling, good thermal stability, small size, and target-binding induced structure change (Citartan et al, 2012;Feng et al, 2014;Juskowiak, 2011;Kim and Gu 2014;Li et al, 2015;Liu et al, 2009;McKeague and Derosa, 2012). Taking advantage of fluorescence anisotropy (FA) or fluorescence polarization (FP) in sensitivity, reproducibility, and simplicity (Lea and Simeonov, 2011;Gradinaru et al, 2010;Le et al, 2002;Smith and Eremin, 2008), the aptamer-based FA/FP sensors are attractive Liu et al, 2009;Ruta et al, 2009;Zhang et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

Aptamer fluorescence anisotropy sensors for adenosine triphosphate by comprehensive screening tetramethylrhodamine labeled nucleotides

Zhao

Wang

2015

Biosensors and Bioelectronics

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a b s t r a c tWe previously reported a fluorescence anisotropy (FA) approach for small molecules using tetramethylrhodamine (TMR) labeled aptamer. It relies on target-binding induced change of intramolecular interaction between TMR and guanine (G) base. TMR-labeling sites are crucial for this approach. Only terminal ends and thymine (T) bases could be tested for TMR labeling in our previous work, possibly causing limitation in analysis of different targets with this FA strategy. Here, taking the analysis of adenosine triphosphate (ATP) as an example, we demonstrated a success of conjugating TMR on other bases of aptamer adenine (A) or cytosine (C) bases and an achievement of full mapping various labeling sites of aptamers. We successfully constructed aptamer fluorescence anisotropy (FA) sensors for adenosine triphosphate (ATP). We conjugated single TMR on adenine (A), cytosine (C), or thymine (T) bases or terminals of a 25-mer aptamer against ATP and tested FA responses of 14 TMR-labeled aptamer to ATP. The aptamers having TMR labeled on the 16th base C or 23rd base A were screened out and exhibited significant FA-decreasing or FA-increasing responses upon ATP, respectively. These two favorable TMRlabeled aptamers enabled direct FA sensing ATP with a detection limit of 1 mM and the analysis of ATP in diluted serum. The comprehensive screening various TMR labeling sites of aptamers facilitates the successful construction of FA sensors using TMR-labeled aptamers. It will expand application of TMR-G interaction based aptamer FA strategy to a variety of targets.

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Section: Introductionmentioning

confidence: 99%

Aptamer fluorescence anisotropy sensors for adenosine triphosphate by comprehensive screening tetramethylrhodamine labeled nucleotides

Zhao

Wang

2015

Biosensors and Bioelectronics

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“…Fluorescence anisotropy (FA)/fluorescence polarization is a powerful technique, and is not only useful for rapid and quantitative measurements of diverse molecular interactions but also for development of rapid assays [13][14][15][16][17][18][19]. FA analysis is a real-time solutionbased equilibrium method without the need for separation, showing unique advantage of no disturbance in the reaction equilibrium.…”

Section: Introductionmentioning

confidence: 99%

Screening interaction between ochratoxin A and aptamers by fluorescence anisotropy approach

et al. 2013

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By taking advantage of the intrinsic fluorescence of ochratoxin A (OTA), we present a fluorescence anisotropy approach for rapid analysis of the interactions between OTA and aptamers. The specific binding of OTA with a 36-mer aptamer can induce increased fluorescence anisotropy (FA) of OTA as the result of the freedom restriction of OTA and the increase of molecular volume, and the maximum FA change is about 0.160. This FA approach enables an easy way to investigate the effects of buffer compositions like metal ions on the affinity binding. FA analysis shows the interaction between OTA and aptamer is greatly enhanced by the simultaneous presence of Ca 2+ and Na + , while the binding affinity of aptamer decreases more than 18-fold when only Ca 2+ exists, and the binding is completely lost when Ca 2+ is absent. Crucial region of the aptamer for binding can be mapped through FA analysis and aptamer mutation. The demonstrated FA approach maintains the advantages of FA in simplicity, rapidity, and robustness. This investigation will help the development of aptamerbased assays for OTA detection in optimizing the binding conditions, modification of aptamers, and rational design.

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“…[14] Biochemical assays can be heterogeneous or homogenous, heterogeneous assays requiring multiple steps, compared to simpler homogeneous assays that are simply 'mix and read' type systems. Detection techniques in HTS are predominantly fluorescence and luminescence based techniques, including standard fluorescence and luminescence assays as well as extensions of these such as fluorescence resonance energy transfer (FRET) [25], fluorescence polarization (FP) [26], homogeneous time resolved fluorescence (HTRF) [27] and fluorescence correlation spectroscopy (FCS) [28]. Assays are developed to detect specific binding processes or to probe particular pathways, networks or phenotypes.…”

Section: Current High Throughput Screening Techniques: Could Vibratiomentioning

confidence: 99%

Vibrational spectroscopy as a tool for studying drug-cell interaction: Could high throughput vibrational spectroscopic screening improve drug development?

Jamieson

Byrne²

2017

Vibrational Spectroscopy

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. (2017). Vibrational spectroscopy as a tool for studying drug-cell interaction: could high throughput vibrational spectroscopic screening improve drug development? Vibrational Spectroscopy, vol. 91, July, pp. 16-30. doi.org/10.1016/ j.vibspec.2016 Vibrational spectroscopy as a tool for studying drug-cell interaction: could high throughput vibrational spectroscopic screening improve drug development? AbstractVibrational spectroscopy is currently widely explored as a tool in biomedical applications. An area at the forefront of this field is the use of vibrational spectroscopy for disease diagnosis, ultimately aiming towards spectral pathology. However, while this field shows promising results, moving this technique into the clinic faces the challenges of widespread clinical trials and legislative approval. While spectral pathology has received a lot of attention, there are many other biomedical applications of vibrational spectroscopy, which could potentially be translated to applications with greater ease. A particularly promising application is the use of vibrational spectroscopic techniques to study the interaction of drugs with cells. Many studies have demonstrated the ability to detect biochemical changes in cells in response to drug application, using both infrared and Raman spectroscopy. This has shown potential for use in high throughput screening (HTS) applications, for screening of efficacy and mode of action of potential drug candidates, to speed up the drug discovery process. HTS is still a relatively new and growing area of research and, therefore, there is more potential for new techniques to move into and shape this field. Vibrational spectroscopic techniques come with many benefits over the techniques used currently in HTS, primarily based on fluorescence assays to detect specific binding interactions or phenotypes. They are label free, and an infrared or Raman spectrum provides a wealth of biochemical information, and therefore could reveal not only information about a specific interaction, but about how the overall biochemistry of a cell changes in response to application of a drug candidate. Therefore, drug mode of action could be elucidated. This review will investigate the potential for vibrational spectroscopy, particularly FTIR and Raman spectroscopy, to benefit the field of HTS and improve the drug development process. In addition to FTIR and Raman spectroscopy, surface enhanced Raman spectroscopy (SERS), coherent anti-Stokes Raman spectroscopy (CARS) and stimulated Raman spectroscopy (SRS), will be investigated as an alternative tool in the HTS process.

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Fluorescence polarization immunoassays and related methods for simple, high-throughput screening of small molecules

Cited by 228 publications

References 104 publications

Aptamer fluorescence anisotropy sensors for adenosine triphosphate by comprehensive screening tetramethylrhodamine labeled nucleotides

Aptamer fluorescence anisotropy sensors for adenosine triphosphate by comprehensive screening tetramethylrhodamine labeled nucleotides

Screening interaction between ochratoxin A and aptamers by fluorescence anisotropy approach

Vibrational spectroscopy as a tool for studying drug-cell interaction: Could high throughput vibrational spectroscopic screening improve drug development?

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