scholarly journals FRET-Based Detection of Enzymatic Reaction of Botulinum on Microfluidic Device

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Young Min Bae ◽  
Seung Oh Jin ◽  
Insoo Kim ◽  
Ki Young Shin
Keyword(s):  
Microfluidic Device ◽  
Botulinum Toxin A ◽  
Enzymatic Reaction ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Reaction Chamber ◽  
Main Channel ◽  
Side Channel ◽  
Peptide Substrate ◽  
Electric Voltage

A microfluidic device was implemented to detect the enzymatic reaction of botulinum toxin A (BTA) using Förster resonance energy transfer (FRET). The microfluidic device comprised a main channel having two loading zones, a reaction chamber and a side channel perpendicular to the main channel. The reaction chamber defined by weir in the main channel was packed with microbeads. The movement of the peptide substrate and the BTA in the microfluidic device was controlled by electrophoresis, and the enzymatic reaction of the BTA was detected through the changes of the fluorescence intensity in the reaction chamber. As a result, it was observed that the enzymatic reaction was affected by the electric voltage applied for the movement of the BTA and the peptide and improved by packing the microbeads in the reaction chamber. The microfluidic device provides the tool to investigate the proteolysis of the substrate by the BTA.

scholarly journals Peptide substrate screening for the diagnosis of SARS-CoV-2 using fluorescence resonance energy transfer (FRET) assay

2021 ◽  
Vol 188 (4) ◽  
Author(s):  
Hani A. Alhadrami ◽  
Ahmed M. Hassan ◽  
Raja Chinnappan ◽  
Hind Al-Hadrami ◽  
Wesam H. Abdulaal ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Fluorescence Resonance Energy Transfer ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Peptide Substrate ◽  
Fluorescence Resonance

scholarly journals Practical Application of Novel Test Methods to Evaluate the Potency of Botulinum Toxin: A Comparison Analysis among Widely Used Products in Korea

Toxins ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 833
Author(s):  
Ji-Yeon Hong ◽  
Jong-Hee Kim ◽  
Jung-Eun Jin ◽  
Sun-Hye Shin ◽  
Kui-Young Park
Keyword(s):  
Botulinum Toxin A ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Test Methods ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protein Levels ◽  
Onabotulinumtoxin A ◽  
Botulinum Neurotoxins ◽  
Reliable Methods

The safe and effective dosing of botulinum neurotoxins (BoNTs) requires accurate and reliable methods to measure their potency. Several novel methods have been introduced over the past decade; however, only few studies have compared the potency of BoNT products with that of the LD50 and other alternative assays. Therefore, the objective of this study was to comparatively evaluate widely used BoNT products using various test methods. Four types of BoNTs (prabotulinumtoxin A, onabotulinumtoxin A, neubotulinumtoxin A, and letibotulinumtoxin A) were used in this study. The estimated potency was assessed using the LD50 assay, and the total BoNT type A protein levels were measured using the enzyme-linked immunosorbent assay (ELISA). The in vitro efficacy of the BoNTs was determined using fluorescence resonance energy transfer (FRET) and surface plasmon resonance (SPR) assays. The results showed differences in the total amount of BoNT protein and the cleavage activity of SNAP-25 within all types of BoNTs. The SPR study seemed to be useful for evaluating the potency by specifically measuring intact 19S neurotoxin, and these results provide new insights for assessing different BoNT products.

Thermal Analysis of a Modular, Microfluidic Device for the Rapid Detection of Stroke

2012 ◽  
Author(s):  
Brooks B. Lowrey ◽  
Daniel S. Park ◽  
Michael C. Murphy
Keyword(s):  
Microfluidic Device ◽  
Rapid Detection ◽  
Solid Phase ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Cell Lysis ◽  
Single Pair ◽  
Cell Capture ◽  
Stroke Event ◽  
Thermal Isolation

A modular, microfluidic device was designed to aid in the rapid detection and treatment of stroke. The device modules process whole blood for the detection of genes expressed in response to a stroke event using a series of assay steps including: cell capture using antibodies, thermal lysis, solid-phase reversible immobilization (SPRI) of nucleic acids, reverse transcription (RT), ligase detection reaction (LDR), and single pair fluorescence resonance energy transfer (spFRET) readout. Cell lysis, RT, and LDR require temperatures of 90°C, 37°C, 65°C and 95°C respectively, therefore strict thermal isolation constraints between thermal zones in the device modules were necessary. Thermal isolation was accomplished using 2 mm and 1 mm air gaps between the fluidic modules and heating elements in the polymer device. Finite element mathematical models (ANSYS v. 12.1, Houston, PA) were used to characterize the thermal zones in two 2D simulations. Section 1 simulation results showed ±1.5°C in cell lysis, ±1.6°C in RT, and ±1.6°C in the denaturation section of LDR. Section 2 simulation showed ±2.6°C in denaturation and ±0.4°C in the annealing/extension zone of LDR.

Creation of a recombinant peptide substrate for fluorescence resonance energy transfer-based protease assays

2006 ◽  
Vol 358 (2) ◽  
pp. 298-300 ◽  
Author(s):  
Lin Zhang ◽  
Heather L. Lawson ◽  
Vallathucherry C. Harish ◽  
Jason D. Huff ◽  
Mary Ann Knovich ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Fluorescence Resonance Energy Transfer ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Peptide Substrate ◽  
Recombinant Peptide ◽  
Fluorescence Resonance

scholarly journals High-Resolution Temperature−Concentration Diagram of α-Synuclein Conformation Obtained from a Single Förster Resonance Energy Transfer Image in a Microfluidic Device

2009 ◽  
Vol 81 (16) ◽  
pp. 6929-6935 ◽  
Author(s):  
Virginia Vandelinder ◽  
Allan Chris M. Ferreon ◽  
Yann Gambin ◽  
Ashok A. Deniz ◽  
Alex Groisman
Keyword(s):  
Energy Transfer ◽  
High Resolution ◽  
Microfluidic Device ◽  
Resonance Energy Transfer ◽  
Resonance Energy
Sign in / Sign up

Export Citation Format

Share Document