Developing Cell-Based Assays

Cascaded protein interactions form the foundation of all signaling pathways, many of which are involved in multiple human diseases. These interactions are sensitively and precisely regulated by various post-translation modifications such as phosphorylation, acetylation, ubiquitination, etc. Many action points of the protein modifications have been targeted during drug development. From a survey we have conducted on assays aimed at these targets, we found that most of the assays are based on the enzymatic reactions, e.g. phosphorylation-specific ELISA, and chemically modified FRET, which require pre-assembled reagent kits which are hard to apply to different targets and different cell models.

Fluorescent Protein based FRET might be the optimal choice to develop a versatile cell-based assay. Since signaling pathways rely on hierarchical protein-protein interactions, the most direct and precise way to study cell signaling pathways would be to detect the interactions between a target protein and its immediate downstream protein. Furthermore, different upstream signals can activate the same set of target proteins in different post-modification patterns, resulting in specific activation of downstream responding factors. These signal flows may be individually monitored by using FRET based assay redesigned and validated for each downstream pathway.

Allele’s scientists can develop cell-based assays with in-depth understanding of protein interactions within the context of human genome, such as the SH2, SH3 and PTB domains in tyrosine kinase signaling, the F-box, BTB-box, SOCS, WDR, and LRR domains in the ubiquitin proteasome system, etc. Additionally, Allele’s cell-based assays can be carried on world’s most powerful lentivirus packaging platform, suitable for virtually all different cell lines and primary cells.

New Product of the Week 061410 to 062010: Rat monoclonal antibody against GFP, strong signal for GFP labeling

Promotion of the Week 061410 to 062010: Mouse LIF-bFGF expressing feeder cells for stem cell culture (ABP-SC-BLIFM05), one vial free to go with any iPS lentivirus/retrovirus kit.

Tags: BTB-box, cell based assays, F-box, FRET, LRR domain, pathway, signaling pathway, SOCS, WDR domain