Synthesis of new hydrophilic rhodamine based enzymatic substrates compatible with droplet-based microfluidic assays

2017 ◽  
Vol 53 (39) ◽  
pp. 5437-5440 ◽  
Author(s):  
Johan Fenneteau ◽  
Dany Chauvin ◽  
Andrew D. Griffiths ◽  
Clément Nizak ◽  
Janine Cossy
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Enzymatic Assay ◽  
Microfluidic Assays

The modification of rhodamine core with a hydrophilic tag yields new fluorogenic enzymatic substrates compatible with high-throughput screening using droplet-based enzymatic assay.

scholarly journals Determining Appropriate Substrate Conversion for Enzymatic Assays in High-Throughput Screening

2003 ◽  
Vol 8 (6) ◽  
pp. 694-700 ◽  
Author(s):  
Ge Wu ◽  
Yue Yuan ◽  
C. Nicholas Hodge
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Single Point ◽  
Enzymatic Assay ◽  
Order Kinetic ◽  
Assay Sensitivity ◽  
High Substrate ◽  
Substrate Conversion ◽  
Order Kinetic Model ◽  
Point Data

It is generally accepted that the conversion of substrate should be kept at less than 10% of the total substrate used when studying enzyme kinetics. However, 10% or less substrate conversion often will not produce sufficient signal changes required for robust high-throughput screening (HTS). To increase the signal-to-background ratio, HTS is often performed at higher than 10% substrate conversion. Because the consequences of high substrate conversion are poorly understood, the screening results are sometimes questioned by enzymologists. The quality of an assay is judged by the ability to detect an inhibitor under HTS conditions, which depends on the robustness of the primary detection signal (Z factor) and the sensitivity to an inhibitor. The assay sensitivity to an inhibitor is reflected in the observed IC50 value or percent inhibition at a fixed compound concentration when single-point data are collected. The major concern for an enzymatic assay under high substrate conversion is that the sensitivity of the screen may be compromised. Here we derive the relationship between the IC 50 value for a given inhibitor and the percentage of substrate conversion using a first-order kinetic model under conditions that obey Henri-Michaelis-Menten kinetics. The derived theory was further verified experimentally with a cAMP-dependent protein kinase. This model provides guidance for assay developers to choose an appropriate substrate conversion in designing an enzymatic assay, balancing the needs for robust signal and sensitivity to inhibitors.

scholarly journals An Enzymatic Assay for High-Throughput Screening of Cytidine-Producing Microbial Strains

PLoS ONE ◽  
2015 ◽  
Vol 10 (3) ◽  
pp. e0121612 ◽  
Author(s):  
Huina Dong ◽  
Yongfei Liu ◽  
Xin Zu ◽  
Ning Li ◽  
Feiran Li ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Enzymatic Assay ◽  
Microbial Strains

scholarly journals A rapid enzymatic assay for high‐throughput screening of adenosine‐producing strains

2015 ◽  
Vol 8 (2) ◽  
pp. 230-238 ◽  
Author(s):  
Huina Dong ◽  
Xin Zu ◽  
Ping Zheng ◽  
Dawei Zhang
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Enzymatic Assay

High-throughput screening program for botanical extracts

Planta Medica ◽  
2012 ◽  
Vol 78 (11) ◽  
Author(s):  
L Hingorani ◽  
NP Seeram ◽  
B Ebersole
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Screening Program ◽  
Botanical Extracts

High throughput screening of microbial biodiversity for the discovery of novel cosmeceutical agents

Planta Medica ◽  
2015 ◽  
Vol 81 (16) ◽  
Author(s):  
K Georgousaki ◽  
N DePedro ◽  
AM Chinchilla ◽  
N Aliagiannis ◽  
F Vicente ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Microbial Biodiversity

High throughput screening to investigate Brazilian Cerrado biome plant extract bank chemical diversity

Planta Medica ◽  
2016 ◽  
Vol 81 (S 01) ◽  
pp. S1-S381
Author(s):  
LS Espindola ◽  
RG Dusi ◽  
KR Gustafson ◽  
J McMahon ◽  
JA Beutler
Keyword(s):  
High Throughput ◽  
Plant Extract ◽  
High Throughput Screening ◽  
Chemical Diversity ◽  
Brazilian Cerrado ◽  
Cerrado Biome

High-throughput screening revealed a clinically relevant drug to induce sperm motility

2014 ◽  
Author(s):  
Clair Cochrane ◽  
Halil Ruso ◽  
Anthony Hope ◽  
Rosemary G Clarke ◽  
Christopher Barratt ◽  
...  
Keyword(s):  
Sperm Motility ◽  
High Throughput ◽  
High Throughput Screening
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