Development of a Platform to Enable Fully Automated Cross-Titration Experiments
| Autor: | Bharti Gajera, Albert Rauch, Paul Zuck, Sylvie Jezequel-Sur, Michael F.A. Finley, Jason Cassaday, Jeffrey D. Hermes, Victor N. Uebele, Brian Squadroni |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
Automation
Laboratory 0301 basic medicine Engineering Dose-Response Relationship Drug business.industry High-throughput screening Drug Evaluation Preclinical Titrimetry Computational biology 01 natural sciences High-Throughput Screening Assays Receptors G-Protein-Coupled 0104 chemical sciences Computer Science Applications 010404 medicinal & biomolecular chemistry 03 medical and health sciences Medical Laboratory Technology 030104 developmental biology Fully automated Humans Titration business Cells Cultured Simulation |
| Zdroj: | SLAS Technology. 22:195-205 |
| ISSN: | 2472-6303 |
| Popis: | 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 Ca |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|