Barcoded Ba/F3 cells stably expressing onco-kinase
Ba/F3 cell is characterized by its dependence on mIL3. Once an onco-kinase is introduced into Ba/F3 cells, the onco-kinase can drive the cell growth and the cell become mIL-3 independent. The establishment of Ba/F3 cells stably expressing onco-kinase is quite straight forward, and the chemo-response data from these cells are very reliable, so Ba/F3 cells with onco-kinase are widely used in kinase inhibitor screening and characterization.

For small molecule inhibitors development, one challenge is to avoid or understand its off-target activities. At enzymatic level, kinome profiling is the routine approach to study off-target activities. However, at cellular level, there is no high-throughput way to screen the off-target activities.

In order to improve the screening throughput, Truwaybio developed the Barcoded Ba/F3 Kinase Panel (please refer to Nat Biotechnol . 2016 April ; 34(4): 419–423. doi:10.1038/nbt.3460.). The construction process of barcoded Ba/F3 kinase panel is depicted as the figure below. Each member in the panel carries a unique barcode sequence and a corresponding kinase, then the cells are mixed in a fixed ratio and used for screening. After treatment, the genomic DNA from the survived cells and the barcode DNA sequences are amplified by PCR. Then the PCR products are quantified via XMAP and the relative activities of the test article on each kinase are calculated and reported.

Using the panel, you can test the cellular kinase inhibition activities of the test article on 150 kinases and mutants in one well in a 6-well plate. It is much more high through put and cost effective than routine cell proliferation assay.





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