QMCF Technology for generation of cell based assays
QMCF Technology is a time saving tool for screening target proteins in cell based assays for signal pathway studies and high-content screening applications.
Stable maintenance, segregation and replication of QMCF expression plasmids in QMCF cells allow generation of stable expression system within 2-3 weeks after transfection. Generation of expression cell banks enables to obtain ready-to-monitor cell cultures.
QMCF Technology based assay system consists of:
- CHO and U2OS based modified adherent QMCF cell lines and
- compatible QMCF expression plasmids that are stably maintained by QMCF cell lines
- target visualization by
immunofluorescence and fluorescent protein based universal detection
systems
The whole process to develop QMCF assay can be described as follows:
1) fusion of target protein encoding sequence to detection marker (optional)
2) insertion of target protein encoding sequence into the QMCF expression plasmid. A number of different expression cassettes allow to modulate expression level of the target molecule
3)
transfection of the expression plasmid into QMCF adherent cell line by electroporation or
chemical transfection with high efficiency (Figure 1: 48 hours)
4) cultivation of the cell culture under selective conditions enables maximal plasmid maintenance and target gene expression (Figure 1: 14 days)
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Figure 1: Percentage of target gene expressing cells after transfection 48 hrs and 14 days in adherent QMCF/U2OS and QMCF/CHO cells |
5) generation of expression cell banks allows to quickly obtain ready-to-monitor cell cultures for further studies
6) optimization of target molecule based screening
The objectives of the process:
- selection of most applicable cell based assay target protein
- achievement of novel cell based assay for high throughput compound screening
