A quantum-dot based protein module for in vivo monitoring of protease activity through fluorescence resonance energy transfer

Biswas, Payal; Cella, Lakshmi N.; Kang, Seung-Gul; Mulchandani, Ashok; Yates, Marylynn V.; Chen, Wilfred

doi:10.1039/c1cc10648a

Cited by 43 publications

(39 citation statements)

References 29 publications

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“…One of such examples is the present pandemic of human immunodeficiency virus (HIV/AIDS), which originated in west-central Africa during the early twentieth century, and has caused nearly 30 million deaths since the first case of AIDS was reported on June 5, 1981 (De Cock et al, 2011). At present, the majority of the developed methods for HIV detection are based on the detection of the presence of antibodies that the patient’s body makes against HIV (Bhimji et al, 2013; Laird et al, 2013), direct molecular recognition of HIV and its components such as specific nucleic acid sequences or antigens (Zhang et al, 2013; Palmer et al, 2003; Yan et al, 2011), or measurement of the activity of HIV-1 protease (HIV-1 PR) (Esseghaier et al, 2013; Biswas et al, 2011; Davis et al, 2009), many of which are often laborious and time-consuming, and/or require the use of labels or sophisticated instruments.…”

Section: Introductionmentioning

confidence: 99%

Real-time label-free measurement of HIV-1 protease activity by nanopore analysis

Wang

Yang

Zhou

et al. 2014

Biosensors and Bioelectronics

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A label-free method for the measurement of the activity of HIV-1 protease is developed by real-time monitoring of the cleavage of a peptide substrate by HIV-1 protease in a nanopore. The method is rapid and sensitive: picomolar concentrations of HIV-1 protease could be detected in ~10 minutes. Simulated clinical samples are analyzed, and the activity of HIV-1 protease could be accurately detected. Our developed nanopore sensor design strategy should find useful application in the development of stochastic sensors for other proteases of medical, pharmaceutical, and biological importance.

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Section: Introductionmentioning

confidence: 99%

Real-time label-free measurement of HIV-1 protease activity by nanopore analysis

Wang

Yang

Zhou

et al. 2014

Biosensors and Bioelectronics

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“…In optical biosensors, due to the wide range of compounds that can be employed in their construction, to result in selective recognition systems that converts the optical signal by several techniques, significant development have been achieved in the past twenty years. In particular, biosensors based on fluorescent responses greatly benefit from supramolecular compounds used as recognition systems and transducers, since they are able to significantly enhance charge transfer processes and this property can be exploited to label the analyte and, hence, enable detection via fluorescence imaging or enabling FRET, which can be efficiently detected using special biosensors [7][8][9].…”

Section: Supramolecular Compounds For Optical Biosensorsmentioning

confidence: 99%

Supramolecular Materials for Optical and Electrochemical Biosensors

Martins¹,

Ribeiro²,

FlavioColmati³

et al. 2015

Biosensors - Micro and Nanoscale Applications

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It is incontestable that the interactions and bonds that keep molecules united to generate unique supramolecular compounds, with individual properties, morphologies and behaviour, are of special dynamics and singular forces. Therefore, it is necessary to discuss and consider the types of interactions that may occur in a determined system, their dynamics and number, which directly act on the energetic balance that strengthen the union between participants and give rise to a supramolecule.In this chapter, a number of such supramolecular systems that find application as any component of a biosensor are presented and discussed, considering intermolecular interaction forces that confer them shape, function and unique properties. To better understand their structural dynamics and the mechanisms through which they can be used in biosensing, a brief explanation on the interaction thermodynamics, types of intermolecular interactions that compete against each other and the energetic equilibrium that originate and stabilize supramolecular systems is given. To explain how this balance of forces can be extensively exploited to develop methods to produce supramolecular compounds, an overview on supramolecular strategies is presented and their contribution is explored in each example presented in this text, to evidence the importance of planning and developing methodologies of preparation, based on

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“…In the presence of HIV-1 Pr, the probe is cleaved and the QD and Alexa dye are separated resulting in the disruption of FRET. The protein module (H-MA-ET) containing the MA/CA cleavage sequence was purified and labeled with Alexa 568 as reported previously (Biswas et al, 2011), and subsequently conjugated to DHLA-capped CdSe-ZnS QDs (QD 545 ) because of the favorable spectral overlap with Alexa 568. Successful conjugation was first confirmed by agarose gel analysis and a clear shift in the QD mobility was detected (Fig.…”

Section: Probe Design and Fret Characterizationsmentioning

confidence: 99%

Quantitative assessment of in vivo HIV protease activity using genetically engineered QD‐based FRET probes

Cella

Biswas

Yates

et al. 2014

Biotech & Bioengineering

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HIV protease plays a central role in its life cycle leading to release of functional viral particles. It has been successfully used as a therapeutic target to block HIV infection. Several protease inhibitors (PIs) are currently being employed as a part of anti-HIV therapy. However, the constant genetic drift in the virus leads to accumulation of mutations in both cleavage site and the protease, resulting in resistance and failure of therapy. We reported the use of a quantum dot (QD)-based protein probe for the in vivo monitoring of HIV-1 protease activity based on fluorescence resonance energy transfer. In the current study, we demonstrate the utility of this approach by quantifying the in vivo cleavage rates of three known protease and cleavage site mutations in the presence or absence of different PIs. The changes in IC50 values for the different PIs were similar to that observed in patients, validating our assay as a rapid platform for PI screening.

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A quantum-dot based protein module for in vivo monitoring of protease activity through fluorescence resonance energy transfer

Cited by 43 publications

References 29 publications

Real-time label-free measurement of HIV-1 protease activity by nanopore analysis

Real-time label-free measurement of HIV-1 protease activity by nanopore analysis

Supramolecular Materials for Optical and Electrochemical Biosensors

Quantitative assessment of in vivo HIV protease activity using genetically engineered QD‐based FRET probes

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