Optimization of a non-radioactive high-throughput assay for decarboxylase enzymes.
Assay Drug Dev Technol
; 8(2): 175-85, 2010 Apr.
Article
en En
| MEDLINE
| ID: mdl-20085486
ABSTRACT
Herein, we describe the optimization of a linked enzyme assay suitable for high-throughput screening of decarboxylases, a target family whose activity has historically been difficult to quantify. Our approach uses a commercially available bicarbonate detection reagent to measure decarboxylase activity. The assay is performed in a fully enclosed automated screening system under inert nitrogen atmosphere to minimize perturbation by exogenous CO2. Receiver operating characteristic (ROC) analysis following a pilot screen of a small library of approximately 3,600 unique molecules for inhibitors of Trypanosoma brucei ornithine decarboxylase quantitatively demonstrates that the assay has excellent discriminatory power (area under the curve = 0.90 with 95% confidence interval between 0.82 and 0.97).
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Carboxiliasas
Tipo de estudio:
Prognostic_studies
Idioma:
En
Revista:
Assay Drug Dev Technol
Año:
2010
Tipo del documento:
Article
País de afiliación:
Estados Unidos