Impedimetric biosensor for assay of caspase-3 activity and evaluation of cell apoptosis using self-assembled biotin-phenylalanine network as signal enhancer

Xia, Ning; Huang, Yaliang; Cui, Zhiyue; Liu, Shudi; Deng, Dehua; Liu, Lin; Wang, Jianxiu

doi:10.1016/j.snb.2020.128436

Cited by 28 publications

(14 citation statements)

References 40 publications

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“…The increase in inhibitor concentration induced the enhancement in the SPR signal, indicating that DEVD-FMK is an effective Cas-3 inhibitor. The half-maximal inhibitory concentration (IC 50 ) was estimated to be 98 nM, which is in agreement with that measured by other methods [30,36]. Thus, the method has bright prospects for the screening of protease inhibitors.…”

Section: Detection Of Cas-3 and Its Inhibitorsupporting

confidence: 84%

“…Then, the resultant solution was incubated with the peptide substrate. For real sample assays, HeLa cells were cultured and the cell lysates were extracted with our reported procedures [30,31]. Then, the peptide substrate was incubated with the diluted cell lysates to react for 2 h. Finally, the reaction mixture was delivered to the flow cell, followed by injection of SA-IgG to the channel after the baseline stable was attained.…”

Section: Inhibitor Detection and Cell Lysate Analysismentioning

confidence: 99%

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Surface Plasmon Resonance for Protease Detection by Integration of Homogeneous Reaction

Xia

Liu

2021

Biosensors

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The heterogeneous assays of proteases usually require the immobilization of peptide substrates on the solid surface for enzymatic hydrolysis reactions. However, immobilization of peptides on the solid surface may cause a steric hindrance to prevent the interaction between the substrate and the active center of protease, thus limiting the enzymatic cleavage of the peptide. In this work, we reported a heterogeneous surface plasmon resonance (SPR) method for protease detection by integration of homogeneous reaction. The sensitivity was enhanced by the signal amplification of streptavidin (SA)-conjugated immunoglobulin G (SA-IgG). Caspase-3 (Cas-3) was determined as the model. A peptide labeled with two biotin tags at the N- and C-terminals (bio-GDEVDGK-bio) was used as the substrate. In the absence of Cas-3, the substrate peptide was captured by neutravidin (NA)-covered SPR chip to facilitate the attachment of SA-IgG by the avidin-biotin interaction. However, once the peptide substrate was digested by Cas-3 in the aqueous phase, the products of bio-GDEVD and GK-bio would compete with the substrate to bond NA on the chip surface, thus limiting the attachment of SA-IgG. The method integrated the advantages of both heterogeneous and homogeneous assays and has been used to determine Cas-3 inhibitor and evaluate cell apoptosis with satisfactory results.

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Section: Detection Of Cas-3 and Its Inhibitorsupporting

confidence: 84%

Section: Inhibitor Detection and Cell Lysate Analysismentioning

confidence: 99%

Surface Plasmon Resonance for Protease Detection by Integration of Homogeneous Reaction

Xia

Liu

2021

Biosensors

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“…Finally, the peptide-covered electrode was rinsed with 50% ethanol and used for the detection of Mpro. In this work, the CPPPP segment is an effective linker and spacer for peptide immobilization to attain high cleavage efficiency [ 32 , 33 ].…”

Section: Methodsmentioning

confidence: 99%

Colorimetric and Electrochemical Methods for the Detection of SARS-CoV-2 Main Protease by Peptide-Triggered Assembly of Gold Nanoparticles

Feng

Liu

et al. 2022

Molecules

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Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) main protease (Mpro) has been regarded as one of the ideal targets for the development of antiviral drugs. The currently used methods for the probing of Mpro activity and the screening of its inhibitors require the use of a double-labeled peptide substrate. In this work, we suggested that the label-free peptide substrate could induce the aggregation of AuNPs through the electrostatic interactions, and the cleavage of the peptide by the Mpro inhibited the aggregation of AuNPs. This fact allowed for the visual analysis of Mpro activity by observing the color change of the AuNPs suspension. Furthermore, the co-assembly of AuNPs and peptide was achieved on the peptide-covered electrode surface. Cleavage of the peptide substrate by the Mpro limited the formation of AuNPs/peptide assembles, thus allowing for the development of a simple and sensitive electrochemical method for Mpro detection in serum samples. The change of the electrochemical signal was easily monitored by electrochemical impedance spectroscopy (EIS). The detection limits of the colorimetric and electrochemical methods are 10 and 0.1 pM, respectively. This work should be valuable for the development of effective antiviral drugs and the design of novel optical and electrical biosensors.

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“…This method had been successfully used to detect the activity of caspase-3 in HeLa cells treated with different anticancer agents. (Xia et al, 2020). Recently, Yildirim and coworkers developed a simple and sensitive method to monitor specific protease activity in biological solutions.…”

Section: Detectionmentioning

confidence: 99%

Supramolecular Nanomedicines of In-Situ Self-Assembling Peptides

Zhang

Gao

2022

Front. Chem.

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Nanomedicines provide distinct clinical advantages over traditional monomolecular therapeutic and diagnostic agents. Supramolecular nanomedicines made from in-situ self-assembling peptides have emerged as a promising strategy in designing and fabricating nanomedicines. In-situ self-assambly (SA) allows the combination of nanomedicines approach with prodrug approach, which exhibited both advantages of these strategies while addressed the problems of both and thus receiving more and more research attention. In this review, we summarized recently designed supramolecular nanomedicines of in-situ SA peptides in the manner of applications and design principles, and the interaction between the materials and biological environments was also discussed.

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Impedimetric biosensor for assay of caspase-3 activity and evaluation of cell apoptosis using self-assembled biotin-phenylalanine network as signal enhancer

Cited by 28 publications

References 40 publications

Surface Plasmon Resonance for Protease Detection by Integration of Homogeneous Reaction

Surface Plasmon Resonance for Protease Detection by Integration of Homogeneous Reaction

Colorimetric and Electrochemical Methods for the Detection of SARS-CoV-2 Main Protease by Peptide-Triggered Assembly of Gold Nanoparticles

Supramolecular Nanomedicines of In-Situ Self-Assembling Peptides

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