Automation of a Phospho-STAT5 Staining Procedure for Flow Cytometry for Application in Drug Discovery

Abstract

Drug discovery often requires the screening of compound libraries on tissue cultured cells. Some major targets in drug discovery belong to signal transduction pathways, and PerFix EXPOSE* allows easy flow cytometry phospho assays. We thus investigated the possibility to further simplify and automate this assay, to allow the direct screening of drugs targeting signaling pathways. We show here the sensitivity of this fully automated assay on human growth hormone (hGH)-driven JAK/STAT5-activated IM-9 cells, and we discuss the throughput of this system, which is compatible with medium-throughput drug screening. Because the kit works directly on whole blood samples, ex-vivo assays are also possible with this approach, which could allow for the screening of drugs under more physiological conditions.

A Semi-automated Approach to Preparing Antibody Cocktails for Immunophenotypic Analysis of Human Peripheral Blood

“…Although it takes time to set up and entails an upfront investment to purchase the instrument, in the long run such factors will be compensated for by the increased productivity and reproducibility of assays. In fact, a few groups previously reported the successful integration of the automated liquid handler into their workflow of immunophenotyping or similar applications 21,22 . Automated solutions for flow cytometry analysis are also available from commercial sources (FACS SPA III, Automated Cocktail Preparation Workstation, and FlowStainer).…”

High-Throughput Flow Cytometry in Drug Discovery

“…Several automation systems with or without HTFC integrated have been presented previously. [6][7][8][9][10] In this special issue are three papers that provide details on the upstream sample preparation automation systems and HTFC sample acquisition systems used at the Genomics Institute of the Novartis Research Foundation (GNF), AstraZeneca and Novartis Institute for Biomedical Research. The system presented by Joslin et al 5 is a fully automated HTFC GNF system with sample preparation and HTFC sample acquisition integrated, enabling phenotypic drug discovery at GNF with a throughput of 50,000 wells/ day using 384-well or 1536-well plates.…”

Application of High-Throughput Flow Cytometry in Early Drug Discovery: An AstraZeneca Perspective

“…In recent years, an increasing amount of reports have described diverse applications of different HTFC platforms using 96-, 384-, and 1536-well formats at various phases of drug discovery in pharma and biotech companies as well as in academic institutes. 3,8,10–33 These reports demonstrate that HTFC technology has played, and will continue to play, an important role in early drug discovery. This article describes the deployment of HTFC platforms within AstraZeneca and MedImmune to support our early drug discovery process from target identification (TI), through hit identification (HI) and lead identification (LI), to lead optimization (LO).…”