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Development of Functional Fluorescent Molecular Probes for the Detection of Biological Substances.

Suzuki Y, Yokoyama K - Biosensors (Basel) (2015)

Bottom Line: This review is confined to sensors that use fluorescence to transmit biochemical information.Fluorescence is, by far, the most frequently exploited phenomenon for chemical sensors and biosensors.To achieve selective (bio)molecular recognition based on these fluorescence phenomena, various fluorescent elements such as small organic molecules, enzymes, antibodies, and oligonucleotides have been designed and synthesized over the past decades.

View Article: PubMed Central - PubMed

Affiliation: Health Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1-1 Higashi, Tsukuba 305-8566, Japan. suzuki-yoshio@aist.go.jp.

ABSTRACT
This review is confined to sensors that use fluorescence to transmit biochemical information. Fluorescence is, by far, the most frequently exploited phenomenon for chemical sensors and biosensors. Parameters that define the application of such sensors include intensity, decay time, anisotropy, quenching efficiency, and luminescence energy transfer. To achieve selective (bio)molecular recognition based on these fluorescence phenomena, various fluorescent elements such as small organic molecules, enzymes, antibodies, and oligonucleotides have been designed and synthesized over the past decades. This review describes the immense variety of fluorescent probes that have been designed for the recognitions of ions, small and large molecules, and their biological applications in terms of intracellular fluorescent imaging techniques.

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Chemical structure of fluorescent probe for the detection of protein.
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biosensors-05-00337-f014: Chemical structure of fluorescent probe for the detection of protein.

Mentions: Suzuki et al. reported fluorescent molecular probes possessing cyanopyranyl moieties or zinc (II) complexes that were designed and synthesized to detect proteins via non-specific binding (compounds 4, 5, 6, and 7 in Figure 14) [83,84,85,86]. These fluorescent probes did not produce any fluorescent emission in the absence of protein. On the other hand, the fluorescence spectra of these probes showed a large Stokes shift and dramatic increase in intensity after the addition of BSA. The fluorescence intensities of the probes were plotted as a function of the protein concentration. A good linear relationship was observed for up to 1 mg/mL of protein and the detection limit was found to be 100 ng/mL at the given assay conditions. Similar results were observed for the measurements of not only BSA, but also other proteins (for example BGG). The responses of these probes were not affected by the presence of various non-protein substances (such as inorganic salts and chelating agents). Moreover, high performance staining for 1D and 2D SDS-PAGE was carried out using a novel protein-binding fluorophore. In order to achieve the high-throughput analysis of proteins for SDS-PAGE, the general protocols for in-gel protein staining (SDS-PAGE, fixation, staining, washing, and detection) were simplified in order to produce a straightforward and rapid protocol. As a result, protein staining could be performed in a minimum of only 30 min in this study. The protein-to-protein variation was low, and the detection limit was 0.4 ng/band of BSA (signal-to-noise ratio was 3.0), which was as sensitive as the short-protocol silver staining methods.


Development of Functional Fluorescent Molecular Probes for the Detection of Biological Substances.

Suzuki Y, Yokoyama K - Biosensors (Basel) (2015)

Chemical structure of fluorescent probe for the detection of protein.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493553&req=5

biosensors-05-00337-f014: Chemical structure of fluorescent probe for the detection of protein.
Mentions: Suzuki et al. reported fluorescent molecular probes possessing cyanopyranyl moieties or zinc (II) complexes that were designed and synthesized to detect proteins via non-specific binding (compounds 4, 5, 6, and 7 in Figure 14) [83,84,85,86]. These fluorescent probes did not produce any fluorescent emission in the absence of protein. On the other hand, the fluorescence spectra of these probes showed a large Stokes shift and dramatic increase in intensity after the addition of BSA. The fluorescence intensities of the probes were plotted as a function of the protein concentration. A good linear relationship was observed for up to 1 mg/mL of protein and the detection limit was found to be 100 ng/mL at the given assay conditions. Similar results were observed for the measurements of not only BSA, but also other proteins (for example BGG). The responses of these probes were not affected by the presence of various non-protein substances (such as inorganic salts and chelating agents). Moreover, high performance staining for 1D and 2D SDS-PAGE was carried out using a novel protein-binding fluorophore. In order to achieve the high-throughput analysis of proteins for SDS-PAGE, the general protocols for in-gel protein staining (SDS-PAGE, fixation, staining, washing, and detection) were simplified in order to produce a straightforward and rapid protocol. As a result, protein staining could be performed in a minimum of only 30 min in this study. The protein-to-protein variation was low, and the detection limit was 0.4 ng/band of BSA (signal-to-noise ratio was 3.0), which was as sensitive as the short-protocol silver staining methods.

Bottom Line: This review is confined to sensors that use fluorescence to transmit biochemical information.Fluorescence is, by far, the most frequently exploited phenomenon for chemical sensors and biosensors.To achieve selective (bio)molecular recognition based on these fluorescence phenomena, various fluorescent elements such as small organic molecules, enzymes, antibodies, and oligonucleotides have been designed and synthesized over the past decades.

View Article: PubMed Central - PubMed

Affiliation: Health Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1-1 Higashi, Tsukuba 305-8566, Japan. suzuki-yoshio@aist.go.jp.

ABSTRACT
This review is confined to sensors that use fluorescence to transmit biochemical information. Fluorescence is, by far, the most frequently exploited phenomenon for chemical sensors and biosensors. Parameters that define the application of such sensors include intensity, decay time, anisotropy, quenching efficiency, and luminescence energy transfer. To achieve selective (bio)molecular recognition based on these fluorescence phenomena, various fluorescent elements such as small organic molecules, enzymes, antibodies, and oligonucleotides have been designed and synthesized over the past decades. This review describes the immense variety of fluorescent probes that have been designed for the recognitions of ions, small and large molecules, and their biological applications in terms of intracellular fluorescent imaging techniques.

Show MeSH
Related in: MedlinePlus