Mechanistic Cell Biology In Vitro Proof-of-Concept Studies

Cell-Based Assays to Determine Biological Pathways and Genetic Drivers for Precision Oncology

Sound, in vitro mechanism-of-action data are needed to progress your compound towards the clinic. The right assay format is determined by your anti-cancer drug candidate and its target. We support you in determining the effects of your compound on selected biological pathways.

Downstream Signalling Responses in Cancer Cell Lines and Primary Material

The discovery process of novel compounds with improved properties can be facilitated using a variety of assays. A typical workflow consists of 1. Selection of cell lines; 2. Generation of cell lines; 3. Assays to determine drug activity; 4. Confirmation in primary material.

selection of cell lines

1

Selection of
cell lines through bioinformatics analysis

Selection of cell lines on the basis of expression of the target or tissue type.
Our bioinformatics experts extract information on mRNA levels for 200 genetically characterized cancer cell lines. This allows comparison of cell lines with high expression versus low expression. Other selection tools are mutation data, protein phosphorylation data, and sensitivity data to certain drugs.

Knock Down Cell Lines

2

Generation of
cell lines to test against control cell lines

Cell pools are generated for screening against control cell lines. For instance, via generation of resistance by dose escalation, gene transfection, knock down.
You can choose from 200 genetically characterized cancer cell lines. Confirmation is done by DNA sequencing, quantitative real-time PCR and western blot.

The Activity Of Your Drug Candidate

3

Assays to measure
the activity of
your drug candidate

The activity of your drug candidate will be determined in the most suitable assay.
This can be downstream phosphorylation of target proteins via Western blot followed by determination of dose response curves using fluorescence based assays to reveal the IC50 of the compound, apoptosis assays, accumulation of metabolites etc.

Primary Material

4

Confirm the
Mechanism of Action
in primary material

Primary material can be used in confirmatory studies, providing additional proof of the mechanism of action.

  • Expression analysis by quantitative real-time PCR, flow cytometry or immuno blot

  • Modulation of expression by siRNA

  • Generation of resistant cell lines

  • Confirmation of genetic drivers by DNA sequencing and qPCR

  • Apoptosis assays, readout caspase 3/7 or flow cytometry

  • Colony formation assays

  • Assays based on fluorescence, chemiluminescence, TR-FRET or Alpha technology

  • Choose from 200 genetically characterized cancer cell lines

  • Primary material for confirmatory studies

Differences in PD-L1 gene expression between primary samples, as shown by qPCR. PD-L1 expression in samples 04 and 05 is low compared to ‘reference’ sample 01 (> 10-fold difference). Expression values are calculated using ΔΔCq method, fold change is 2^(ΔCq PS 01 -ΔCq PS 0X)
Differences in PD-L1 gene expression between primary samples, as shown by qPCR. PD-L1 expression in samples 04 and 05 is low compared to ‘reference’ sample 01 (>10-fold difference).

Modulation of Biological Pathways and Target Proteins

Our cancer biologists and immunologists are in close contact with the sponsor to determine the most suitable assay format for the study and share scientific insights.

PD-L1 expression in Primary Sample, TP53 Mutant
Analysis of PD-L1 protein expression by flowcytometry.
A: Primary sample, TP53 mutant; B: Primary sample, TP53 wild type, NRAS mutant; C: Cancer cell line SKOV-3. The grey areas represent the isotype control.

In Vitro Studies Enhance Discovery of Precision Oncology Drugs

It is our mission to help you find a mechanistic hypothesis before entering the clinic. We have generated science-based technologies that help you to identify the mechanism of preclinical and clinical drug candidates. Our technologies provide in vitro proof of concept in the preparation of in vivo studies and clinical trials.

Deliverables You Can Count On

We deliver reproducible results that are adapted to your specific needs with a short delivery time. Direct contacts between the sponsor and our staff result in efficient scientific and technical support. We provide cost-free consultation on next steps in research if desired by the sponsor.

Quality. Flexibility. Short Turnaround Time.

back to Home