FCET

Frankfurt Competence center for Emerging Therapeutics

Biomedical basic research has a rich tradition in Frankfurt. With FCET, we are bringing together cutting-edge technology platforms to consolidate advanced biomedical research, facilitate interdisciplinary collaboration, and develop new therapies.

Located at the Riedberg and Niederrad campi, FCET is a hub for various specialized units focusing on medicinal chemistry, RNA, protein chemistry, biochemistry, biophysics, cellular and phenotypic analysis, proteomics, genomics, and computational biomedicine.

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FCET office

Cellular and Phenotypic Screening

Head scientist: Alexandra Stolz
Our phenotypic live-cell screens are performed with (modified, preferable adherent) cell lines and are image-based. Modifications include fluorescent proteins/reporters, stably expressed by the cell line. Treatment of choice may include arrayed media add-ons, arrayed knock-down of proteins (siRNA libraries), arrayed compound treatment (chemical libraries), as well as adding specific dyes required for assay specific analysis. Treatments may require the module Automated Liquid Handling that allows the handling of cells requiring biosafety level 1.
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Automated liquid handling
  • Multidrop Combi Reagent Dispenser (8-chanel-dispenser)
  • Echo 550 automated liquid handler
FACS analysis & cell sorting
  • Benchtop SH800SFP Cell Sorter
Phenotypic live-cell screening and High-Content Screening
  • IncuCyte S3
  • Yokogawa Quantitative Confocal Image Cytometer CQ1

 

3D Tissue Models

Head scientist: Maike Windbergs
We offer complex 3D human in vitro models of different organ systems, including the intestine, the lung, the skin as well as the blood-brain barrier. The models can be employed for a wide range of scientific research questions, such as permeation, biocompatibility or immunogenicity testing. Based on the adaptable composition, we can simulate physiological and pathophysiological conditions, emulating inflamed or infected tissues, or incorporating immune cells to study, e.g., wound healing. Besides using bio-relevant culture media and culture substrates simulating the in vivo extracellular matrix, we deploy microfluidic organ-on-a-chip tissue models for specific research questions. Down-stream analyses of single cells or the whole multicellular 3D tissue models, including different microscopy techniques (label-free Raman, confocal laser scanning, as well as electron microscopy) and molecular biological assays (e.g. ELISA, RTq-PCR) allow for further in-depth characterization of the in vitro models.
 
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Cell Culture
  • Primary human cells and human cell lines
  • Co-culture systems
  • Immunocompetent and wound healing models
  • Infection and Inflammation models
  • Microfluidic-based models
Instrumentation
  • Imaging and molecular analyses
  • Nanoindentation
  • Aerosol deposition
  • Impedance spectroscopy

 

Protein Production

Head scientists: Mohit Misra, Sebastian Mathea
We offer the possibility to express proteins in bacterial system (E. coli), eukaryotic expression system insect cells (Spodoptera frugiperda; SF9 cells) or mammalian cells (Expi293 cells).
Protein Production comprehends culture of cells expressing the protein of interest, protein extraction and purification, validation of protein purity and size, assessment of protein homogeneity, optional functional assays (activity, thermal stability) and crystallization.
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Expression systems
  • Bacterial: E. coli
  • Eukaryotic: insect cells (SF9 cells), mammalian cells (Expi293 cells)
Protein purification
Optional
  • Functional assays (activity, thermal stability)
  • Crystallisation

 

Genomics Screening

Head scientists: Manuel Kaulich and Koraljka Husnjak

Gene perturbation is a powerful tool for dissecting genotype-to-phenotype relationships. Genome-wide and tailored CRISPR-Cas9 gRNAs and libraries have been used to identify essential and pathway-specific genes. This platform produces project-tailored reagents and libraries using 3Cs technology.

The quantitative phenotypic profiling of genes/sequences allows for the identification of i) single and synergistic gene essentiality and ii) novel pathways and complex components.

Available libraries:

Name Species Genes/gRNAs
Kinome mouse and human 713 / 3140
FDA drug targets human 690 / 3036
Autophagy human 199 / 876
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CRISPR-Cas9 gRNA library production
  • Assembly of target gene list
  • Design of 2-6 gRNAs per gene
  • Cloning using 3Cs technology into lenti-, retro- or AVV-backbone
  • Delivery as DNA, viral particles or transduced cells
Available genetic screen formats:
  • Drop-out
  • Enrichment
  • Drug synergizing or sensitizing
  • Biochemical reporter/FACS enrichment/depletion

 

Quantitative Proteomics

Head scientist: Christian Münch

Proteomics is a powerful method for evaluating drug-protein interactions. It is based on evaluating the quantity of thousands of cellular proteins that are produced upon treatment with chemicals.

We can process cell pellets or lysates of cells treated with varying chemicals or concentrations of chemicals.

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Sample processing:
  • From protein extraction to peptide quantification
  • TMT-labelling and clean-ups
  • Fractionation
  • LC-MS3 analysis on mass spectrometer
  • Raw file processing for quality control and primary data output
Instrumentation:
  • ThermoFisher QExactive HF with EASY 1200
  • ThermoFisher Fusion Lumos with EASY 1200
  • ThermoFisher Ascend with Vanquish Neo

 

Computational Biomedicine

Head scientist: Ram Bhaskara
We deploy a multidisciplinary approach leveraging vast datasets (in-house databases), cutting-edge algorithms, and high-performance computing to accelerate drug discovery and harness the power of advanced computational techniques. Through the integration of high-throughput genomics, proteomics, and structural biology data we (i) identify novel drug targets, (ii) predict drug interactions, (iii) prioritize targets, and (iv) optimize drug candidates, enabling the exploration of innovative therapeutic modalities using data-driven approaches. By applying state-of-the-art integrative modelling approaches in combination with physics-based simulation methods, we then decode complex disease processes and identify molecular vulnerabilities providing insights into novel drugging strategies.
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Our approaches:
  • Integrative modeling
  • Biomolecular simulations
  • Data integration

 

DARPin / Sybody Selection

Head scientists: Volker Dötsch, Katharina Holzhüter

DARPins (Designed Ankyrin Repeat Proteins) are small (14-18 kDa), very stable proteins. Certain positions can be randomized to create by a combination of in vitro selection strategies (ribosome display and phage display) of high-affinity binders against folded domains/proteins. Due to the absence of disulfide bridges, DARPins can be used for intracellular applications, for example, as highly selective and affine binders or fused to E3 ligases as bio-PROTACS. In addition to DARPin libraries, we have Sybody (nanobody) libraries.

We can run selection experiments for DARPin and Sybodies to create high-affinity binders.

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Generation of DARPins/Sybodies:
  • Run ribosome display with DARPin libraries with different numbers of helical modules (1, 2 or 3) followed by two rounds of phage display
  • Run ribosome display with Sybody libraries ( with short, medium or long loops) followed by two rounds of phage display