Development of a Platform to Enable Fully Automated Cross-Titration Experiments.
SLAS Technol
; 22(2): 195-205, 2017 04.
Article
em En
| MEDLINE
| ID: mdl-27864339
ABSTRACT
In the triage of hits from a high-throughput screening campaign or during the optimization of a lead compound, it is relatively routine to test compounds at multiple concentrations to determine potency and maximal effect. Additional follow-up experiments, such as agonist shift, can be quite valuable in ascertaining compound mechanism of action (MOA). However, these experiments require cross-titration of a test compound with the activating ligand of the receptor requiring 100-200 data points, severely limiting the number tested in MOA assays in a screening triage. We describe a process to enhance the throughput of such cross-titration experiments through the integration of Hewlett Packard's D300 digital dispenser onto one of our robotics platforms to enable on-the-fly cross-titration of compounds in a 1536-well plate format. The process handles all the compound management and data tracking, as well as the biological assay. The process relies heavily on in-house-built software and hardware, and uses our proprietary control software for the platform. Using this system, we were able to automate the cross-titration of compounds for both positive and negative allosteric modulators of two different G protein-coupled receptors (GPCRs) using two distinct assay detection formats, IP1 and Ca2+ detection, on nearly 100 compounds for each target.
Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Titulometria
/
Avaliação Pré-Clínica de Medicamentos
/
Automação Laboratorial
Idioma:
En
Revista:
SLAS Technol
Ano de publicação:
2017
Tipo de documento:
Article
País de afiliação:
Estados Unidos