enzyme detection
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Author(s):  
Michael G. Christiansen ◽  
Matej ◽  
Vizovišek ◽  
Simone Schuerle

Enzymes are appealing diagnostic targets because of their centrality in human health and disease. Continuous efforts spanning several decades have yielded methods for magnetically detecting the interactions of enzymes with exogenous molecular substrates. Nevertheless, measuring enzymatic activity in vivo remains challenging due to background noise, insufficient selectivity, and overlapping enzymatic functions. Magnetic micro- and nanoagents are poised to help overcome these issues by offering possible advantages such as site-selective sampling, modular architectures, new forms of magnetic detection, and favorable biocompatibility. Here, we review relevant control and detection strategies and consider examples of magnetic enzyme detection demonstrated with micro- or nanorobotic systems. Most cases have focused on proteolytic enzymes, leaving ample opportunity to expand to other classes of enzymes. Enzyme-responsive magnetic micro- and nanoagents hold promise for lowering barriers of translation and enabling preemptive, point-of-care medical applications. Expected final online publication date for the Annual Review of Control, Robotics, and Autonomous Systems, Volume 5 is May 2022. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.


2021 ◽  
Vol 13 (11) ◽  
pp. 12928-12940
Author(s):  
Dipankar Das ◽  
Qasim F. M. Alhusaini ◽  
Kawaljit Kaur ◽  
Mohammad Raoufi ◽  
Holger Schönherr

Biosensors ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 25
Author(s):  
Kawaljit Kaur ◽  
Winny Chelangat ◽  
Sergey I. Druzhinin ◽  
Nancy Wangechi Karuri ◽  
Mareike Müller ◽  
...  

There is a growing demand for rapid and sensitive detection approaches for pathogenic bacteria that can be applied by non-specialists in non-laboratory field settings. Here, the detection of the typical E. coli enzyme β-glucuronidase using a chitosan-based sensing hydrogel-coated paper sensor and the detailed analysis of the reaction kinetics, as detected by a smartphone camera, is reported. The chromogenic reporter unit affords an intense blue color in a two-step reaction, which was analyzed using a modified Michaelis–Menten approach. This generalizable approach can be used to determine the limit of detection and comprises an invaluable tool to characterize the performance of lab-in-a-phone type approaches. For the particular system analyzed, the ratio of reaction rate and equilibrium constants of the enzyme–substrate complex are 0.3 and 0.9 pM−1h−1 for β-glucuronidase in phosphate buffered saline and lysogeny broth, respectively. The minimal degree of substrate conversion for detection of the indigo pigment formed during the reaction is 0.15, while the minimal time required for detection in this particular system is ~2 h at an enzyme concentration of 100 nM. Therefore, this approach is applicable for quantitative lab-in-a-phone based point of care detection systems that are based on enzymatic substrate conversion via bacterial enzymes.


2021 ◽  
Author(s):  
Jiangui Mao ◽  
Xi Yang ◽  
Yiling Liu ◽  
Yuan Gong ◽  
Yun-Jiang Rao

2020 ◽  
Vol 17 ◽  
Author(s):  
Seyyed Mojtaba Mousavi ◽  
Seyyed Alireza Hashemi ◽  
Sargol Mazraedoost ◽  
Ahmad Gholami ◽  
Khadije Yousefi ◽  
...  

Background: There is a vital need to advance a cheap, quick, and robust strategy to detect biological proteins since these biomolecules are regularly utilized as biomarkers responsible for diagnosing many diseases such as malignancies. Graphene quantum dots (GQDs) have attracted extensive consideration of researchers in various fields because of their particular optical properties and extraordinary execution in photovoltaic gadgets, photocatalysis, and biological imaging. These nanomaterials adequately improve the sensor performance for their reproducibility, selectivity just as effectively. Graphene quantum dots (GQDs) comprise discrete highlights, starting as attractive fluorophores and superb electro-impulses inferable from their photographic soundness, water-solvency, biocompatibility, non-poisonous quality, and making them an ideal contender for a wide range of new biomedical applications. Methods: All online published studies and online content related to the detection of biological proteins and immunological assays using graphene quantum dots from January 2000 to 2020 are reviewed. This review begins with a rundown of the new methodologies and various structures of graphene quantum dots. In the next step, the detailed description is followed on their applications in the fields of protein and immune detection for drug probing applications. Results: After providing a brief review for chemical and biological synthesis of GQD, in this review, we categorized the detection method of biological proteins using GQD into four main categories including, GQD fluorescence sensors for serum enzyme detection, ultra-trace detection of biological using GQDs, GQD-based immunological assays (immunosensors) especially for cancer and cardiac biomarkers, protein probing for theragnostic purposes. Conclusion: Through this review, we discuss the current trends and future application of graphene quantum dots and their role in proteins and other biomolecules detection, diagnosis in diseases, and their importance in theragnostic applications.


2020 ◽  
Vol 38 (18) ◽  
pp. 5205-5211
Author(s):  
Jiangui Mao ◽  
Xi Yang ◽  
Yiling Liu ◽  
Yanqiong Wang ◽  
Gang-Ding Peng ◽  
...  
Keyword(s):  

2020 ◽  
Vol 25 (9) ◽  
pp. 1026-1037
Author(s):  
Jan Tykvart ◽  
Václav Navrátil ◽  
Michael Kugler ◽  
Pavel Šácha ◽  
Jiří Schimer ◽  
...  

The DNA-linked inhibitor antibody assay (DIANA) has been recently validated for ultrasensitive enzyme detection and for quantitative evaluation of enzyme inhibitor potency. Here we present its adaptation for high-throughput screening of human carbonic anhydrase IX (CAIX), a promising drug and diagnostic target. We tested DIANA’s performance by screening a unique compound collection of 2816 compounds consisting of lead-like small molecules synthesized at the Institute of Organic Chemistry and Biochemistry (IOCB) Prague (“IOCB library”). Additionally, to test the robustness of the assay and its potential for upscaling, we screened a pooled version of the IOCB library. The results from the pooled screening were in agreement with the initial nonpooled screen with no lost hits and no false positives, which shows DIANA’s potential to screen more than 100,000 compounds per day. All DIANA screens showed a high signal-to-noise ratio with a Z′ factor of >0.89. The DIANA screen identified 13 compounds with Ki values equal to or better than 10 µM. All retested hits were active also in an orthogonal enzymatic assay showing zero false positives. However, further biophysical validation of identified hits revealed that the inhibition activity of several hits was caused by a single highly potent CAIX inhibitor, being present as a minor impurity. This finding eventually led us to the identification of three novel CAIX inhibitors from the screen. We confirmed the validity of these compounds by elucidating their mode of binding into the CAIX active site by x-ray crystallography.


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