Establishment of high throughput screening model for epidermal growth factor receptor tyrosine kinase inhibitor

Objective:To establish a high throughput screening(HTS) model for epidermal growth factor(EGF) receptor tyrosine kinase(RTK) inhibitor.Methods: The bioactive EGFR-RTK was expressed by genetic engineering technology.The binding activity and bioactivity of the compounds were examined by surface plasmon resonance and ELISA.Results: The bioactive EGFR-RTK protein was successfully expressed in the prokaryotic expression system and was immobilized on the sensor chip.The equilibrium constants(Kd) between TKI(EI 188) and RTK was 5.00×10-7 mol·L-1,and the IC50 was 12.37 μmol·L-1,which was consistent with expected.With this model we screened 31 compounds and found that 6 compounds had binding activity and inhibitory activity.Conclusion: A novel HTS model of EGFR-TKI has been successfully established using surface plasmon resonance biosensor and ELISA,which lays a foundation for discovery of new TKIs.