scholarly journals Modular Fluorescent Nanoparticle DNA Probes for Detection of Peptides and Proteins

2021 ◽  
Author(s):  
Cassandra M. Stawicki ◽  
Torri E. Rinker ◽  
Markus Burns ◽  
Sonal S. Tonapi ◽  
Rachel P. Galimidi ◽  
...  
Keyword(s):  
Fluorescent Labeling ◽  
Dna Aptamer ◽  
Covalent Attachment ◽  
Polystyrene Nanoparticles ◽  
Laboratory Equipment ◽  
Nanoparticle Probes ◽  
Biological Studies ◽  
Fluorescent Nanoparticle ◽  
Aptamer Probe ◽  
Barriers To Use

Fluorescently labeled antibody and aptamer probes are used in biological studies to characterize binding interactions, measure concentrations of analytes, and sort cells. Fluorescent nanoparticle labels offer an excellent alternative to standard fluorescent labeling strategies due to their enhanced brightness, stability and multivalency; however, challenges in functionalization and characterization have impeded their use. This work introduces a straightforward approach for preparation of fluorescent nanoparticle probes using commercially available reagents and common laboratory equipment. Fluorescent polystyrene nanoparticles, Thermo Fisher FluoSpheres™, were used in proof-of-principle studies. Particle passivation was achieved by covalent attachment of amine-PEG-azide to carboxylated particles, neutralizing the surface charge from -47 to -17 mV. A conjugation-annealing handle and DNA aptamer probe was attached to the azide-PEG nanoparticle surface either through reaction of pre-annealed handle and probe or through a stepwise reaction of the nanoparticles with the handle followed by aptamer annealing. Nanoparticles functionalized with DNA aptamers targeting histidine tags and VEGF protein had high affinity (EC50s ranging from 2-7 nM) and specificity, and were more stable than conventional labels. This protocol for preparation of nanoparticle probes relies solely on commercially available reagents and common equipment, breaking down the barriers to use of nanoparticles in biological experiments.

scholarly journals Modular fluorescent nanoparticle DNA probes for detection of peptides and proteins

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Cassandra M. Stawicki ◽  
Torri E. Rinker ◽  
Markus Burns ◽  
Sonal S. Tonapi ◽  
Rachel P. Galimidi ◽  
...  
Keyword(s):  
Fluorescent Labeling ◽  
Dna Aptamer ◽  
Covalent Attachment ◽  
Laboratory Equipment ◽  
Nanoparticle Probes ◽  
Biological Studies ◽  
Thermo Fisher Scientific ◽  
Fluorescent Nanoparticle ◽  
Aptamer Probe ◽  
Barriers To Use

AbstractFluorescently labeled antibody and aptamer probes are used in biological studies to characterize binding interactions, measure concentrations of analytes, and sort cells. Fluorescent nanoparticle labels offer an excellent alternative to standard fluorescent labeling strategies due to their enhanced brightness, stability and multivalency; however, challenges in functionalization and characterization have impeded their use. This work introduces a straightforward approach for preparation of fluorescent nanoparticle probes using commercially available reagents and common laboratory equipment. Fluorescent polystyrene nanoparticles, Thermo Fisher Scientific FluoSpheres, were used in these proof-of-principle studies. Particle passivation was achieved by covalent attachment of amine-PEG-azide to carboxylated particles, neutralizing the surface charge from − 43 to − 15 mV. A conjugation-annealing handle and DNA aptamer probe were attached to the azide-PEG nanoparticle surface either through reaction of pre-annealed handle and probe or through a stepwise reaction of the nanoparticles with the handle followed by aptamer annealing. Nanoparticles functionalized with DNA aptamers targeting histidine tags and VEGF protein had high affinity (EC50s ranging from 3 to 12 nM) and specificity, and were more stable than conventional labels. This protocol for preparation of nanoparticle probes relies solely on commercially available reagents and common equipment, breaking down the barriers to use nanoparticles in biological experiments.

scholarly journals Enzyme-free amplified detection of cellular microRNA by light-harvesting fluorescent nanoparticle probes

2021 ◽  
Vol 179 ◽  
pp. 113084 ◽  
Author(s):  
Sylvie Egloff ◽  
Nina Melnychuk ◽  
Andreas Reisch ◽  
Sophie Martin ◽  
Andrey S. Klymchenko
Keyword(s):  
Light Harvesting ◽  
Nanoparticle Probes ◽  
Fluorescent Nanoparticle

scholarly journals Using DNA Aptamer Probe for Immunostaining of Cancer Frozen Tissues

2015 ◽  
Vol 87 (3) ◽  
pp. 1919-1924 ◽  
Author(s):  
Ying Pu ◽  
Zhenxu Liu ◽  
Yi Lu ◽  
Peng Yuan ◽  
Jun Liu ◽  
...  
Keyword(s):  
Dna Aptamer ◽  
Aptamer Probe

scholarly journals Fluorescent Nanoparticle Probes for Cancer Imaging

2005 ◽  
Vol 4 (6) ◽  
pp. 593-602 ◽  
Author(s):  
Swadeshmukul Santra ◽  
Debamitra Dutta ◽  
Glenn A. Walter ◽  
Brij M. Moudgil
Keyword(s):  
Early Stage ◽  
Cancer Imaging ◽  
Cellular Level ◽  
Design Synthesis ◽  
Nanoparticle Probes ◽  
Non Invasive ◽  
Fluorescent Nanoparticle ◽  
Nanoparticle Design ◽  
Inexpensive Technique

Optical imaging technique has strong potential for sensitive cancer diagnosis, particularly at the early stage of cancer development. This is a sensitive, non-invasive, non-ionizing (clinically safe) and relatively inexpensive technique. Cancer imaging with optical technique however greatly relies upon the use of sensitive and stable optical probes. Unlike the traditional organic fluorescent probes, fluorescent nanoparticle probes such as dye-doped nanoparticles and quantum dots (Qdots) are bright and photostable. Fluorescent nanoparticle probes are shown to be very effective for sensitive cancer imaging with greater success in the cellular level. However, cancer imaging in an in vivo setup has been recently realized. There are several challenges in developing fluorescent nanoparticle probes for in vivo cancer imaging applications. In this review, we will discuss various aspects of nanoparticle design, synthesis, surface functionalization for bioconjugation and cancer cell targeting. A brief overview of in vivo cancer imaging with Qdots will also be presented.

scholarly journals Analytical Ancestry: “Firsts” in Fluorescent Labeling of Nucleosides, Nucleotides, and Nucleic Acids

2009 ◽  
Vol 55 (4) ◽  
pp. 670-683 ◽  
Author(s):  
Larry J Kricka ◽  
Paolo Fortina
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Fluorescent Dyes ◽  
Fluorescent Labeling ◽  
Covalent Attachment ◽  
Detection Methods ◽  
Fluorescent Label ◽  
Nucleic Acid Detection ◽  
Fluorescent Labels ◽  
Fluorescent Properties

Abstract Background: The inherent fluorescent properties of nucleosides, nucleotides, and nucleic acids are limited, and thus the need has arisen for fluorescent labeling of these molecules for a variety of analytical applications. Content: This review traces the analytical ancestry of fluorescent labeling of nucleosides, nucleotides, and nucleic acids, with an emphasis on the first to publish or patent. The scope of labeling includes (a) direct labeling by covalent labeling of nucleic acids with a fluorescent label or noncovalent binding or intercalation of a fluorescent dye to nucleic acids and (b) indirect labeling via covalent attachment of a secondary label to a nucleic acid, and then binding this to a fluorescently labeled ligand binder. An alternative indirect strategy involves binding of a nucleic acid to a nucleic acid binder molecule (e.g., antibody, antibiotic, histone, antibody, nuclease) that is labeled with a fluorophore. Fluorescent labels for nucleic acids include organic fluorescent dyes, metal chelates, carbon nanotubes, quantum dots, gold particles, and fluorescent minerals. Summary: Fluorescently labeled nucleosides, nucleotides, and nucleic acids are important types of reagents for biological assay methods and underpin current methods of chromosome analysis, gel staining, DNA sequencing and quantitative PCR. Although these methods use predominantly organic fluorophores, new types of particulate fluorophores in the form of nanoparticles, nanorods, and nanotubes may provide the basis of a new generation of fluorescent labels and nucleic acid detection methods.

A ZnS-Nanoparticle-Label-Based Electrochemical Codeine Sensor

2017 ◽  
Vol 872 ◽  
pp. 173-177 ◽  
Author(s):  
Wei Xiong ◽  
Su Fang Wu ◽  
Fu Sheng Liao ◽  
Nian Hong ◽  
Hao Fan ◽  
...  
Keyword(s):  
Electrochemical Sensor ◽  
Modified Electrode ◽  
Sedative Effect ◽  
Dna Aptamer ◽  
Dna Aptamers ◽  
Moderate Pain ◽  
Current Signal ◽  
Electrochemical Signal ◽  
Aptamer Probe

Codeine (3-methylmorphine) is an opiate that is widely used to treat mild or moderate pain and cough suppression. It is the second predominant alkaloid in opium with a mild sedative effect. In the present study, we describe an electrochemical sensor for codeine detection by using the DNA aptamers against codeine. In the sensing protocol, a dually-labeled DNA aptamer probe was designed to be labeled at one end with HS, and at its another end with dabcyl as an electrochemical tag to produce electrochemical signal via recognization occurrence. One special electrochemical marker was prepared by modifying ZnS nanoparticle with-cyclodextrins (ab. ZnS-CDs), which employed as electrochemical signal provider and would conjunct with the codeine probe modified electrode through the host–guest recognition of CDs to dabcyl. With codeine adding, aptamer folding allows the ZnS-CDs into soultion that caused an increase of current signal. This sensor has the ability to detect 37pM codeine. Our study demonstrates that the biosensor has good specificity and stability. It can be used to detect codeine.

scholarly journals Self-Assembled MoS2/ssDNA Nanostructures for the Capacitive Aptasensing of Acetamiprid Insecticide

2021 ◽  
Vol 11 (4) ◽  
pp. 1382 ◽  
Author(s):  
Maroua Hamami ◽  
Noureddine Raouafi ◽  
Hafsa Korri-Youssoufi
Keyword(s):  
Lipoic Acid ◽  
Dna Sequences ◽  
Electrochemical Impedance ◽  
Dna Aptamer ◽  
Covalent Attachment ◽  
Food Contaminants ◽  
Real Sample Analysis ◽  
Self Assembled ◽  
The Stability ◽  
Self Assembled Layer

The aim of this work is to detect acetamiprid using electrochemical capacitance spectroscopy, which is widely used as a pesticide in agriculture and is harmful to humans. We have designed aptasensing platform based on the adsorption of a DNA aptamer on lipoic acid-modified MoS2 nano-sheets. The biosensor takes advantage of the high affinity of single-stranded DNA sequences to MoS2 nano-sheets. The stability of DNA on MoS2 nano-sheets is assured by covalent attachment to lipoic acid that forms self-assembled layer on MoS2 surface. The biosensor exhibits excellent capacitance performances owing to its large effective surface area making it interesting material for capacitive transduction system. The impedance-derived capacitance varies with the increasing concentrations of acetamiprid that can be attributed to the aptamer desorption from the MoS2 nanosheets facilitating ion diffusion into MoS2 interlayers. The developed device showed high analytical performances for acetamiprid detection on electrochemical impedance spectroscopy EIS- derived capacitance variation and high selectivity toward the target in presence of other pesticides. Real sample analysis of food stuff such as tomatoes is demonstrated which open the way to their use for monitoring of food contaminants by tailoring the aptamer.

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