Fluorescent DNA Nanotags Featuring Covalently Attached Intercalating Dyes: Synthesis, Antibody Conjugation, and Intracellular Imaging

Stadler, Andreas; Santos, Junriz O Delos; Stensrud, Elizabeth S; Dembska, Anna; Silva, Gloria L.; Liu, Shengpeng; Shank, Nathaniel; Kunttas-Tatli, Ezgi; Sobers, Courtney J; Gramlich, Philipp M. E.; Carell, Thomas; Peteanu, Linda A.; McCartney, Brooke M.; Armitage, Bruce A.

doi:10.1021/bc100485f

Cited by 27 publications

(18 citation statements)

References 52 publications

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“…Each Ab2 is usually allowed to conjugate with 3 5 RBITC dyes because higher conjugations can cause internal quenching between the dyes and influence the immunogenic activity of Ab2. 27 After the conjugation, the anti-PSA Ab2 showed a typical RBITC emission peak at around 580 nm, demonstrating that RBITC had been successfully labled onto Ab2 (Figure S8 in Supporting Information). Later, we compared the detection performance of the RBITC labeled Ab2 with our activatable Ab2-RBITC-AuNPs probes (30 nm AuNPs were applied as the core).…”

Section: Resultsmentioning

confidence: 95%

Gold Nanoparticle-Based Activatable Probe for Sensing Ultralow Levels of Prostate-Specific Antigen

et al. 2013

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It is still in high demand to develop extremely sensitive and accurate clinical tools for biomarkers of interest for early diagnosis and monitoring of diseases. In this report, we present a highly sensitive and compatible gold nanoparticle (AuNP)-based fluorescence activatable probe for sensing ultra-low levels of prostate-specific antigen (PSA) in patient serum samples. The limit of detection of the newly-developed probe for PSA was pushed down to 0.032 pg/mL, which is more than two orders of magnitude lower than that of the conventional fluorescence probe. The ultrahigh sensitivity of this probe was attributed to the high loading efficiency of the dyes on AuNP surfaces and high fluorescence quenching unquenching abilities of the dye-AuNP pairs. The efficiency and robustness of this probe was investigated in patient serum samples, demonstrating the great potential of this probe in real-world applications. Keywordsfluorescence activatable probe; gold nanoparticle; prostate specific antigen (PSA); Rhodamine B isothiocyanate (RBITC)We report a fluorescence-based activatable immunoassay for ultrasensitive detection of cancer biomarkers in a panel of serum samples. A biomarker is defined as the "molecular signature" of a physiological or disease process at certain stages and is therefore particularly useful for diagnosing disease, monitoring disease progression and evaluating therapeutic response. 1,2 Many diseases such as cancers are much more susceptible to treatment when diagnosed in the early stages, indicating that earlier diagnosis of the disease may enable improved therapeutic intervention. 3 However, the levels of biomarkers are often low in * To whom correspondence should be addressed. Xiaoyuan Chen (shawn.chen@nih.gov). Supporting InformationDetails of characterizing gold nanoparticles and various conjugates described here can be found in the Supporting Information. This material is available free of charge via the Internet at http://pubs.acs.org.The authors declare no competing financial interest. NIH Public AccessAuthor Manuscript ACS Nano. Author manuscript; available in PMC 2014 June 25.Published in final edited form as: ACS Nano. 2013 June 25; 7(6): 5568-5576. doi:10.1021/nn401837q. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author Manuscript biological samples from patients. 4,5 It is therefore extremely important to develop ultrasensitive biosensors for biomarkers of interest.The current gold standard for clinical biomarker detection is the enzyme-linked immunosorbent assay (ELISA). However, due to its moderate sensitivity, ELISA usually allows detection only after biomarker levels have already reached critical threshold concentrations, at which point the disease has already advanced remarkably. Thus, ultrasensitive assays are urgently required for detection of biomarkers in the clinic. Nanotechnology is already playing an increasingly important role in the design of ultrasensitive biosensors. 68 In the past few decades, many efforts have been made to create signal transducers ...

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Section: Resultsmentioning

confidence: 95%

Gold Nanoparticle-Based Activatable Probe for Sensing Ultralow Levels of Prostate-Specific Antigen

et al. 2013

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“…28 Recently, we have successfully applied both these strategies for incorporation of porphyrins into different positions in DNA 29. It should be noted that our synthetic procedure developed for the dye TO–N 3 is more convenient than syntheses of several other thiazole orange derivatives:22a, 23 hundreds of milligrams of material can be obtained without the need for tedious and expensive reverse‐phase chromatography on silica gel.…”

Section: Resultsmentioning

confidence: 99%

Artificial Photosynthesis at Dynamic Self‐Assembled Interfaces in Water

Hansen

Troppmann

König

2015

Chemistry A European J

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Artificial photosynthesis is one of the big scientific challenges of today. Self-assembled dynamic interfaces, such as vesicles or micelles, have been used as microreactors to mimic biological photosynthesis. These aggregates can help to overcome typical problems of homogeneous photocatalytic water splitting. Microheterogeneous environments organize catalyst-photosensitizer assemblies at the interface in close proximity and thus enhance intermolecular interactions. Thereby vesicles and micelles may promote photoinitiated charge separation and suppress back electron transfer. The dynamic self-assembled interfaces solubilize non-polar compounds and protect sensitive catalytic units and intermediates against degradation. In addition, vesicles provide compartmentation that was used to separate different redox environments needed for an overall water splitting system. This Minireview provides an overview of the applications of micellar and vesicular microheterogeneous systems for solar energy conversion by photosensitized water oxidation and hydrogen generation.

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“…Intercalators can be either attached covalently to the nucleic acid sequence or interact with the duplex non-covalently (Rye and Glazer 1995;Karlsen et al 2012). Non-nucleosidic analogues of nucleic acids such as IPN, UNA, and TINA contain a covalently attached intercalating dye within the oligonucleotide probe which is labeled during oligonucleotide synthesis or postsynthetically (Filichev and Pedersen 2005;El-Sayed et al 2012;Stadler et al 2011). The resulting probes were found to be useful in simultaneous targeting and detection of double-stranded DNA in various fashions.…”

Section: Intercalating Dyesmentioning

confidence: 99%

Fluorescent Nucleic Acid Analogues in Research and Clinical Diagnostics

Astakhova

2015

RNA Technologies

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Fluorescent DNA Nanotags Featuring Covalently Attached Intercalating Dyes: Synthesis, Antibody Conjugation, and Intracellular Imaging

Cited by 27 publications

References 52 publications

Gold Nanoparticle-Based Activatable Probe for Sensing Ultralow Levels of Prostate-Specific Antigen

Gold Nanoparticle-Based Activatable Probe for Sensing Ultralow Levels of Prostate-Specific Antigen

Artificial Photosynthesis at Dynamic Self‐Assembled Interfaces in Water

Fluorescent Nucleic Acid Analogues in Research and Clinical Diagnostics

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