为了建立体外蛋白酪氨酸磷酸酶SHP2抑制剂的高通量筛选模型，筛选潜在的SHP2抑制剂，通过应用大肠杆菌系统克隆表达GST-SHP2 融合蛋白，经过GST-Sepharose 亲和层析分离得到纯化的GST-SHP2 蛋白，建立384 孔板的高通量筛选模型，对48000个有明确结构的小分子化合物进行体外筛选，筛选出75个活性化合物对SHP2的抑制作用大于50%，确定3个化合物具有较高的抑制活性。该筛选模型灵敏、稳定，对SHP2抑制剂药物研发打下了基础。
The aims of this study is to set up a high throughput screening model for SHP2 inhibitors and screen the potential inhibitors. By the E. coil expression system GST-SHP2 fusion protein is cloned and over expressed. High-purity SHP2 protein is purified though GSTSepharose column. After establishing the high through screening system with the colorimetric assay of SHP2, a library of 48000 pure compounds is screened using the 384 micro- plate. Among them 75 compounds have inhibitory effects over 50%. Ultimately, three inhibitors are identified as SHP2 inhibitors with high activity. The high throughput screening is a highly sensitive, inexpensive, and operationally simple assay method in identifying SHP2 inhibitors.
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