In Vitro Assays Selectively Modulating Imatinib Impact

ATP-mimicking ligand is in a fixed phase within a chromatographic column. Thus, the affinity of the test compound is measured relative to that of the fixed phase ligand through its ability to displace the latter and hence enable the kinase display bacteriophage to elute from the column. 

5 In Vitro Assays Selectively Modulating Imatinib Impact 

In vitro colorimetric assays were performed over a 1 pM to 100 (xM range in ligand concentration to assess the inhibition of phosphorylating activity by antibody recognition of phosphorylated peptide substrates [14]. The IC50 (50% inhibition concentration) for imatinib/Abl is 1 (xM, while the WBZ 4/Abl value is above 100 (xM (Fig. 8.7a). The active recombinant Abl kinase and its substrate (Abl-tide) were incubated in the presence of various WBZ 4 or imatinib concentrations and ATP (100 nM). Phosphorylation of Abl-tide peptide was detected by spectrophotometry following incubation with phospho-Abl-tide antibodies. 

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