Site Specific Biotinylated Protein Kinase is Easy-to-Use Solution in the SPR Technology for Kinase Inhibitor Evaluation
In characterizing kinase inhibitors for drug discovery research, biophysical assays are increasingly of value for the measurement of parameters such as the rates of association (ka) and dissociation (kd) and the dissociation constant (KD). The Surface Plasmon Resonance (SPR) is the gold standard method to determine KD values for protein and small molecule interaction. So far, the amine coupling, avidin-biotin interaction and antibody capturing has been utilized to immobilize the protein on the sensor chip. However, the immobilization using amine coupling and the preparation of the biotinylated protein have been one of the bottlenecks. The amine coupling requires acidic condition which leads to lose or lowers kinase activity or disrupts steric structure. The chemical biotinylation probing is also problematic, because it is difficult to control the sites and numbers of the biotinylation and additional purification is also required before use. In addition, antibody capturing lowers the sensitivity because of the larger molecular weight.
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