Technologieplattform

Jens Peter von Kries

Screening Unit

[Translate to Deutsch:] Portrait

Die Screening-Einheit bietet eine Hochdurchsatz-Technologieplattform für das Screening von Substanzbibliotheken (ca. 60.000 cpds) und für genomweite RNA-Interferenz unter Verwendung automatisierter Mikroskope oder anderer Methoden. Das Labor verwaltet die ChemBioNet-Screening-Sammlung (20.000 cpds), die gemeinsam mit Partnerplattformen in Europa genutzt wird.


Automatisiertes Screening von Wirkstoffen

Substanzen finden und neue Targets identifizieren: Die Screening Unit ist eine frei zugängliche Technologieplattform für automatisierte systematische Testungen („Screenings“). Wir verwenden entweder Substanzbibliotheken (~70.000 chemische Substanzen) oder genomweite RNA-Interferenz (RNAi)-Bibliotheken (Mensch, Maus, Nematoden), die spezifische RNA-Moleküle zur Hemmung der Übersetzung jedes einzelnen Gens in sein entsprechendes Protein enthalten. Wir unterstützen die Forschenden des FMP und anderer, kooperierender Institutionen bei der Testentwicklung, Prozessautomatisierung, beim Screening und bei der automatischen Datenanalyse. Darüber hinaus identifizieren wir neue Screening-Techniken und implementieren diese für den Einsatz. Derzeit unterstützen wir Substanz-Screening-Projekte, bei der Entwicklung und Optimierung von Testverfahren zum Hochdurchsatz-Screening (Silke Radetzki), und bei der automatisierten Datendokumentation und Analyse (Martin Neuenschwander). Zudem haben wir in unserer Unit das genomweite RNAi- und CRISPR-CAS-Screening (Katina Lazarow) etabliert, um die Identifizierung neuer zellulärer Zielstrukturen („Targets“) für die pharmakologische Interferenz in Krankheiten zu ermöglichen. Seit 2019 etablieren wir 3D-Analysen mit konfokalen Mikroskopen, um in der Tradition von Rudolf Virchow (Zelluläre Pathologie) morphologische Muster in der Reaktion von Zellen auf toxische Substanzen zu identifizieren (Christopher Wolff).

 

Gruppenleiter

Open Access Plattform

Project Application

You may contact us via < screening@fmp-berlin.de >

For effective support and data documentation you need to fill in the requested information and send us the project application form

Assay Setup

You may visit the Screening Unit, where we can help you in assay transfer to the 384well format and test your assay set up together for HTS usage. You will then be able to update the project application form with the revised protocol. Afterwards the materials for your next visit and primary screen plus validation are calculated.

Primary Screen

We perform the primary screen together with one person from your team, the data are documented and analyzed automatically. You receive the raw data, statistic analysis, and statistics-based hit lists containing also structural data.

Review of hit lists

If desired, you may revise the hit lists. You send us a list of 352 compounds via e-mail.

Cherry picking

We provide you with a 384-well plate containing your requested 352 hits, (4 µl 5mM compound in DMSO).

Quality control of 352 hit compounds via LC-MS Analysis

A small aliquot (0.2 µl) of your hit picking plate is taken and diluted in ACN/H2O for the LC-MS analysis. We analyze the samples in ACN/H2O (1:1, 25 µM) with our Agilent TOF mass spectrometer and determine the purity via UV absorption at 254 nm.

IC50 validation

IC50 validation (using nine serial dilutions of the compounds in duplicate) is carried out by us, the data are analyzed automatically. You receive for a maximum of 352 "hit" compounds IC50 reports containing MS analysis data, frequent hitter information, data from biophysical and cytotox profiling. To our experience less then 10% of the compounds from the hit list of 352 can be expected to be true hits.

Review of IC50 data

Our chemists revise with you the list of validated compounds and suggest proposals, which compounds might be selected for further characterization and optimization of leads.

Please bear in mind that all steps need only to be performed once per project!

The Screening Unit can assist your project with know-how gained from about 170 previous screens. But the data documentation, analysis and follow up studies are in the responsibility of the PI of the project.

RNAi and CRISPR-based Screening

The facility offers scientists from the FMP and MDC access to high-throughput screening with arrayed miRNA, siRNA and sgRNA libraries on the basis of a scientific collaboration.

Broad range of assays
State of the art equipment supports a broad range of assays in high-throughput formats, ranging from basic reporter assays to high-content screening campaigns. Instrumentation covers automated fluorescence microscopy, flow cytometry, luminescence and real-time kinetic fluorescence reading (see Technologies).

Full support in high-throughput screening
We operate on the basis of a scientific collaboration and offer support through all stages of a screening campaign: assay development and adaption to high-throughput screening, the actual wet-lab screen, data acquisition and quality assessment, statistical analysis as well as hit candidate selection.

Data processing
We are constantly advancing our in-house analysis, database and reporting tools to keep the platform updated. The integration of freely available open-source tools (e.g. KNIME Analytics Platform) and the development of our own analysis workflows enables us to meet the specific requirements of a multitude of biological questions.

Tools for Assay Setup

Kontakt

Kontakt Sekretariat

Dajana Baudach

Sekretariat, Sun Gruppe,
Sekretariat Nazaré Gruppe


Forschungsbereich

Chemische Biologie