scholarly journals Quantitative and Label-Free Technique for Measuring Protease Activity and Inhibition using a Microfluidic Cantilever Array

Nano Letters ◽  
2008 ◽  
Vol 8 (9) ◽  
pp. 2968-2974 ◽  
Author(s):  
Digvijay A. Raorane ◽  
Mark D. Lim ◽  
Fanqing Frank Chen ◽  
Charles S. Craik ◽  
Arun Majumdar
Keyword(s):  
Protease Activity ◽  
Label Free ◽  
Cantilever Array

A Label-Free Gold Nanocluster Fluorescent Probe for Protease Activity Monitoring

2014 ◽  
Vol 14 (6) ◽  
pp. 4029-4035 ◽  
Author(s):  
Jiajing Zhang ◽  
Zhen Zhang ◽  
Xin Nie ◽  
Zhenjiang Zhang ◽  
Xiaochun Wu ◽  
...  
Keyword(s):  
Fluorescent Probe ◽  
Protease Activity ◽  
Activity Monitoring ◽  
Label Free ◽  
Gold Nanocluster ◽  
Free Gold

Label-free detection of single-stranded DNA binding protein based on a cantilever array

Talanta ◽  
2013 ◽  
Vol 109 ◽  
pp. 173-176 ◽  
Author(s):  
Hui Hou ◽  
Xiaojing Bai ◽  
Chunyan Xing ◽  
Baoping Lu ◽  
Jinhui Hao ◽  
...  
Keyword(s):  
Dna Binding ◽  
Binding Protein ◽  
Dna Binding Protein ◽  
Label Free ◽  
Single Stranded Dna ◽  
Label Free Detection ◽  
Cantilever Array

scholarly journals Multiple label-free biodetection and quantitative DNA-binding assays on a nanomechanical cantilever array

2002 ◽  
Vol 99 (15) ◽  
pp. 9783-9788 ◽  
Author(s):  
R. McKendry ◽  
J. Zhang ◽  
Y. Arntz ◽  
T. Strunz ◽  
M. Hegner ◽  
...  
Keyword(s):  
Dna Binding ◽  
Label Free ◽  
Binding Assays ◽  
Cantilever Array

Label-free detection of kanamycin using aptamer-based cantilever array sensor

2014 ◽  
Vol 56 ◽  
pp. 112-116 ◽  
Author(s):  
Xiaojing Bai ◽  
Hui Hou ◽  
Bailin Zhang ◽  
Jilin Tang
Keyword(s):  
Label Free ◽  
Array Sensor ◽  
Label Free Detection ◽  
Cantilever Array

scholarly journals Protease Substrate-Independent Universal Assay for Monitoring Digestion of Native Unmodified Proteins

2021 ◽  
Vol 22 (12) ◽  
pp. 6362
Author(s):  
Emmiliisa Vuorinen ◽  
Salla Valtonen ◽  
Nazia Hassan ◽  
Randa Mahran ◽  
Huda Habib ◽  
...  
Keyword(s):  
Protease Activity ◽  
Microtiter Plate ◽  
Probe Method ◽  
Label Free ◽  
Target Class ◽  
Signaling Mechanism ◽  
Peptide Bonds ◽  
Time Resolved ◽  
Drug Candidates ◽  
Catalytic Function

Proteases are a group of enzymes with a catalytic function to hydrolyze peptide bonds of proteins. Proteases regulate the activity, signaling mechanism, fate, and localization of many proteins, and their dysregulation is associated with various pathological conditions. Proteases have been identified as biomarkers and potential therapeutic targets for multiple diseases, such as acquired immunodeficiency syndrome, cardiovascular diseases, osteoporosis, type 2 diabetes, and cancer, where they are essential to disease progression. Thus, protease inhibitors and inhibitor-like molecules are interesting drug candidates. To study proteases and their substrates and inhibitors, simple, rapid, and sensitive protease activity assays are needed. Existing fluorescence-based assays enable protease monitoring in a high-throughput compatible microtiter plate format, but the methods often rely on either molecular labeling or synthetic protease targets that only mimic the hydrolysis site of the true target proteins. Here, we present a homogenous, label-free, and time-resolved luminescence utilizing the protein-probe method to assay proteases with native and denatured substrates at nanomolar sensitivity. The developed protein-probe method is not restricted to any single protein or protein target class, enabling digestion and substrate fragmentation studies with the natural unmodified substrate proteins. The versatility of the assay for studying protease targets was shown by monitoring the digestion of a substrate panel with different proteases. These results indicate that the protein-probe method not only monitors the protease activity and inhibition, but also studies the substrate specificity of individual proteases.

scholarly journals Detection of Sub-Nanomolar Concentration of Trypsin by Thickness-Shear Mode Acoustic Biosensor and Spectrophotometry

Biosensors ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 117
Author(s):  
Ivan Piovarci ◽  
Sopio Melikishvili ◽  
Marek Tatarko ◽  
Tibor Hianik ◽  
Michael Thompson
Keyword(s):  
Gold Nanoparticles ◽  
Protease Activity ◽  
Limit Of Detection ◽  
Colorimetric Assay ◽  
Enzymatic Reaction ◽  
Disease Diagnosis ◽  
Shear Mode ◽  
Label Free ◽  
Thickness Shear Mode ◽  
Thickness Shear

The determination of protease activity is very important for disease diagnosis, drug development, and quality and safety assurance for dairy products. Therefore, the development of low-cost and sensitive methods for assessing protease activity is crucial. We report two approaches for monitoring protease activity: in a volume and at surface, via colorimetric and acoustic wave-based biosensors operated in the thickness-shear mode (TSM), respectively. The TSM sensor was based on a β-casein substrate immobilized on a piezoelectric quartz crystal transducer. After an enzymatic reaction with trypsin, it cleaved the surface-bound β-casein, which increased the resonant frequency of the crystal. The limit of detection (LOD) was 0.48 ± 0.08 nM. A label-free colorimetric assay for trypsin detection has also been performed using β-casein and 6-mercaptohexanol (MCH) functionalized gold nanoparticles (AuNPs/MCH-β-casein). Due to the trypsin cleavage of β-casein, the gold nanoparticles lost shelter, and MCH increased the attractive force between the modified AuNPs. Consequently, AuNPs aggregated, and the red shift of the absorption spectra was observed. Spectrophotometric assay enabled an LOD of 0.42 ± 0.03 nM. The Michaelis–Menten constant, KM, for reverse enzyme reaction has also been estimated by both methods. This value for the colorimetric assay (0.56 ± 0.10 nM) is lower in comparison with those for the TSM sensor (0.92 ± 0.44 nM). This is likely due to the better access of the trypsin to the β-casein substrate at the surface of AuNPs in comparison with those at the TSM transducer.

Peptide-templated gold nanoclusters as a novel label-free biosensor for the detection of protease activity

RSC Advances ◽  
2014 ◽  
Vol 4 (27) ◽  
pp. 13753-13756 ◽  
Author(s):  
Yi Gu ◽  
Qian Wen ◽  
Yongqing Kuang ◽  
Lijuan Tang ◽  
Jianhui Jiang
Keyword(s):  
Protease Activity ◽  
Gold Nanoclusters ◽  
Label Free ◽  
Activity Assay ◽  
Fluorescent Biosensor

A novel label-free fluorescent biosensor platform has been developed for protease activity assay using peptide-templated gold nanoclusters (AuNCs).

Label-free protein assay based on a nanomechanical cantilever array

2002 ◽  
Vol 14 (1) ◽  
pp. 86-90 ◽  
Author(s):  
Y Arntz ◽  
J D Seelig ◽  
H P Lang ◽  
J Zhang ◽  
P Hunziker ◽  
...  
Keyword(s):  
Label Free ◽  
Free Protein ◽  
Protein Assay ◽  
Cantilever Array

Plasma-polymerized antifouling biochips for label-free measurement of protease activity in cell culture media

2019 ◽  
Vol 281 ◽  
pp. 527-534 ◽  
Author(s):  
Jisoo Park ◽  
Gae Baik Kim ◽  
Andreas Lippitz ◽  
Young Mi Kim ◽  
Donggeun Jung ◽  
...  
Keyword(s):  
Cell Culture ◽  
Protease Activity ◽  
Culture Media ◽  
Label Free ◽  
Cell Culture Media

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

2014 ◽  
Vol 62 ◽  
pp. 158-162 ◽  
Author(s):  
Liang Wang ◽  
Yujing Han ◽  
Shuo Zhou ◽  
Xiyun Guan
Keyword(s):  
Real Time ◽  
Protease Activity ◽  
Label Free ◽  
Hiv 1
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