Assembling DNA through Affinity Binding to Achieve Ultrasensitive Protein Detection

Zhang, Hongquan; Li, Feng; Dever, Brittany; Wang, Chuan; Li, Xing‐Fang; Le, X. Chris

doi:10.1002/anie.201210022

Cited by 160 publications

(133 citation statements)

References 58 publications

Supporting

Mentioning

129

Contrasting

Order By: Relevance

“…179 This also allows ultrahigh sensitivity. [180][181][182] It is important to couple these methods in a practically feasible format for biomedical diagnosis. Addressing these challenges will fuel the growth of the aptamer field in the future.…”

Section: Summary and Future Directionsmentioning

confidence: 99%

Aptamer-based biosensors for biomedical diagnostics

Zhou

Huang

Ding

et al. 2014

Analyst

475

290

View full text Add to dashboard Cite

Aptamers are single-stranded nucleic acids that selectively bind to target molecules. Most aptamers are obtained through a combinatorial biology technique called SELEX. Since aptamers can be isolated to bind to almost any molecule of choice, can be readily modified at arbitrary positions and they possess predictable secondary structures, this platform technology shows great promise in biosensor development. Over the past two decades, more than one thousand papers have been published on aptamer-based biosensors. Given this progress, the application of aptamer technology in biomedical diagnosis is still in a quite preliminary stage. Most previous work involves only a few model aptamers to demonstrate the sensing concept with limited biomedical impact. This Critical Review aims to summarize progresses that might enable practical applications of aptamer for biological samples. First, general sensing strategies based on the unique properties of aptamers are summarized. Each strategy can be coupled to various signaling methods. Among these, a few detection methods including fluorescence lifetime, flow cytometry, upconverting nanoparticles, nanoflare technology, magnetic resonance imaging, electronic aptamer-based sensors, and lateral flow devices have been discussed in more detail since they are more likely to work in a complex sample matrix. The current limitations of this field include the lack of high quality aptamers for clinically important targets. In addition, the aptamer technology has to be extensively tested in a clinical sample matrix to establish reliability and accuracy. Future directions are also speculated to overcome these challenges.3

show abstract

Section: Summary and Future Directionsmentioning

confidence: 99%

Aptamer-based biosensors for biomedical diagnostics

Zhou

Huang

Ding

et al. 2014

Analyst

475

290

View full text Add to dashboard Cite

show abstract

“…One successful option is to translate protein amount into a nucleic acid reporter. 6–8 This technique lends itself to multiplexability and high sensitivity, since nucleic acid output sequences can encode target identities and are amplifiable.…”

mentioning

confidence: 99%

Protein Quantification Using Controlled DNA Melting Transitions in Bivalent Probe Assemblies

Kim

Bezerra

et al. 2015

Anal. Chem.

View full text Add to dashboard Cite

Homogenous protein assays, despite the potential for mix-and-read workflows, have eluded widespread acceptance due to interferences in biological matrices and limited multiplexability. Here, we employ standard qPCR instrumentation for thermofluorimetric analysis of bivalent probe (TFAB) assemblies, allowing protein levels to be quantitatively translated into multiplexable DNA melting transitions within 30 min. As protein-bound bivalent probes are thermodynamically more stable than unbound probes, differential thermal analysis can remove background analytically, without physical separation. Using either antibody-oligonucleotides or aptamers as probes, TFAB is validated for protein quantification in buffer, human serum, and human plasma and for assaying hormone secretions from endocrine cells. The direct optical method exhibits superior scalability, allowing detection of only 1 amol of protein in microfluidic channels of 100 pL volume. Overall, we demonstrate TFAB as a robust and generalizable homogeneous protein assay with superior performance in biological matrices.

show abstract

“…In this proximity hybridization process, the local concentrations of P1 and P2 were increased, thus raising the melting temperature for the intramolecular hybridization (P1 and P2), favoring the strand displacement reaction between P2 and Output. 15 As a consequence, the Output was released from P1, triggering the strand displacement cycle. And, the subsequent amplification process was triggered meanwhile.…”

Section: Principle Of the Biosensor For Pdgf-bb Detectionmentioning

confidence: 99%

“…10,11 Different from the toehold-mediated strand displacement, 12,13 which has been extensively employed to design DNA assembly, a proximity hybridization-induced DNA assembly homogeneous assay has been proposed. 14,15 The method was based on the target-binding process to promote strand displacement and reinforce the hybridization of two short DNA sequences, which cannot form stable duplex at normal circumstances. Through converting the target binding event into detectable DNA assembly, proximity hybridization strategy can be combined with various signal amplification strategies, such as enzyme-free cyclic assembly of hairpins, 16 ligation-mediated strand recycling, 17 and binding-induced self-assembly of DNAzyme 18 to build ultrasensitive biosensors.…”

Section: Introductionmentioning

confidence: 99%

Proximity hybridization-mediated isothermal exponential amplification for ultrasensitive electrochemical protein detection

Yu¹,

Su²,

Zhu³

et al. 2017

IJN

View full text Add to dashboard Cite

In this study, we fabricated a novel electrochemical biosensing platform on the basis of target-triggered proximity hybridization-mediated isothermal exponential amplification reaction (EXPAR) for ultrasensitive protein analysis. Through rational design, the aptamers for protein recognition were integrated within two DNA probes. Via proximity hybridization principle, the affinity protein-binding event was converted into DNA assembly process. The recognition of protein by aptamers can trigger the strand displacement through the increase of the local concentrations of the involved probes. As a consequence, the output DNA was displaced, which can hybridize with the duplex probes immobilized on the electrode surface subsequently, leading to the initiation of the EXPAR as well as the cleavage of duplex probes. Each cleavage will release the gold nanoparticles (AuNPs) binding sequence. With the modification of G-quadruplex sequence, electrochemical signals were yielded by the AuNPs through oxidizing 3,3′,5,5′-tetramethylbenzidine in the presence of H 2 O 2 . The study we proposed exhibited high sensitivity toward platelet-derived growth factor BB (PDGF-BB) with the detection limit of 52 fM. And, this method also showed great selectivity among the PDGF isoforms and performed well in spiked human serum samples.

show abstract

Assembling DNA through Affinity Binding to Achieve Ultrasensitive Protein Detection

Cited by 160 publications

References 58 publications

Aptamer-based biosensors for biomedical diagnostics

Aptamer-based biosensors for biomedical diagnostics

Protein Quantification Using Controlled DNA Melting Transitions in Bivalent Probe Assemblies

Proximity hybridization-mediated isothermal exponential amplification for ultrasensitive electrochemical protein detection

Contact Info

Product

Resources

About