For Research Use Only. Not for Use in Diagnostic Procedures.
Dr. Sheraz Gul, European Screening Port, Hamburg, Germany
The European ScreeningPort Team
The European ScreeningPort (ESP) is a public-private partnership which works for academic institutions to create and operate drug discovery programs that address novel therapeutic targets. The key competency of the ESP lies in early stage (target-to-lead) small molecule discovery, using a screening approach.
The aim of the facility is to accelerate the translation of promising results on the causes of diseases, generated in basic research, into new therapeutics. These processes have until recently only been available within the Pharmaceutical industry. The ESP has a proven track record of successful project delivery for major German academic institutions, and internationally they have active projects with academics in Canada, The Netherlands, New Zealand, Switzerland and the United Kingdom.
The ESP operates a state-of-the-art infrastructure for in-vitroand in-silico screening, which includes access to up to 400,000 “ready-to-screen” compounds. The compound collections have been designed to address a range of target classes including those with specific relevance to neurological disease (ion channels, GPCR’s, kinases, etc), and protein::protein interactions such as those associated with intracellular signalling pathways. Typical projects integrate medicinal chemistry insight with in-vitro and in-silico structural and disease biology knowledge, to identify molecules which have the best chance of being translated from the laboratory to the clinical setting.
Dr. Sheraz Gul
Dr. Sheraz Gul (pictured) is Vice President and Head of Biology at ESP, and manages the assay development and screening of academic targets. Here, he describes the instrumentation used within the facility.
“Our infrastructure is based around the fully integrated PerkinElmer robotic cell::explorer™automated high content screening system, which forms the centrepiece of the ESPs in-vitroscreening facility (Figure 1). The cell::explorer system handles 96 to 1536 well microplates with integrated nano-litre scale dispensing, industry standard liquid handling for larger volumes, robust plate handling and sensitive multi-mode optical based readouts. We provide mainstream optical assay readouts commonly found within the pharmaceutical industry, including high content imaging, AlphaScreen® assays and time-resolved FRET. Our aim is always to develop the most physiologically relevant assay readouts, for example using native protein substrates or primary cells rather than synthetic peptides or engineered cells which may be easy to handle but bear little relation to the underlying disease biology”.
Figure 1: ESP’s in-vitro screening platform is composed of four different workstations based around the fully integrated robotic cell::explorer system. The workstations allow for compound storage and dispensing (Compound Workstation), plate incubation (Hotel Workstation), high content imaging (Opera Workstation) and multi-mode reading of plates (Reader Workstation).
"Approximately 40% of the academic targets we are investigating involve hit finding to identify small molecules which selectively disrupt protein::protein or protein::DNA interactions. PerkinElmer’s AlphaScreen technology has proved to be a versatile platform to address these target classes, and we use a “toolbox” of donor and acceptor bead combinations to rapidly identify optimal assay formats. The introduction of the Enspire® Multimode Plate Reader by PerkinElmer promises to make this platform cost-effective for academic laboratories and allow them to develop assays which could more readily be scaled-up to larger platforms such as ours".
"To follow up on hits identified using AlphaScreen technology, we employ high content imaging to directly visualize cellular processes and interactions. To reduce complexity we need to focus on the molecule of interest, which we mostly do by direct or indirect staining using labelled antibodies. We have set up automated proximity ligation analysis (PLA) assays to monitor the localization of transcription factors and examine their impact on the cell cycle (Figure 2)".
Figure 2: Schematic representation of the high-content proximity ligation assay (PLA) workflow established at ESP in cooperation with the Center for Integrative Psychiatry in Kiel. Cells are plated in a 96 or 384 well high-content format and then analyzed for multiple biological modalities combined with a single cell PLA signal, localizing the protein of interest. The biological modality represented in the figure is the correlation of DNA content (estimated from the relative intensity of nuclear staining of individual cells) with cell cycle phases. This unique image and multidimensional data analysis extracts biological insights at unprecedented resolution.
"The in-situ PLA technology can localize and quantify individual protein interactions and modifications which we visualise using PerkinElmer’s confocal Opera® High Content Screening System. Information on the cell cycle state of individual cells can also be inferred using the Opera system, based on the relative intensity of nuclear DNA staining. Our in-situ PLA technology and subsequent data handling and processing are all performed with medium throughput in an industrial environment and thus can be automated for secondary screening purposes".
About Sheraz Gul
Prior to joining the European ScreeningPort, Sheraz worked for 7 years at GlaxoSmithKline and has a wide range of experience in drug discovery, ranging from target validation, development of assays for screening, compound profiling, lead optimisation and candidate characterization. He has a BSc, PhD and 5 years postdoctoral research experience from the University of London, and is the co-author of several high ranking publications, including the Enzyme Assays: Essential Data handbook.