利用多克隆抗体有效识别中草药中抗表皮生长因子抑制剂(英文)

The Novel Application of Polyclonal Antibodies in Recognizing Anti-EGFR Inhibitors Directly from Herb

用一种已知的抗表皮生长因子受体抑制剂(piceatannol)作为半抗原与载体牛血清白蛋白(BSA)连接后免疫制备相应的多克隆抗体(PcAb).利用该多克隆抗体来模拟酶制成亲和色谱柱,从一种藏药粗提物中将包括该半抗原在内的几种结构不同的抗表皮生长因子受体抑制剂识别出来.研究采用前沿亲和色谱-质谱联用技术对样品进行分析,可以直接从中药复杂体系中识别出有效成分并进行鉴定,实现中药有效成分的筛选与鉴定一体化技术.

A polyclonal antibody (PcAb) prepared with piceatannol, a known inhibitor against the epidermal growth factor receptor (EGFR), was adopted as a stationary phase in the chromatographic system. Using the antibody to mimic the enzyme, several anti-EGFR inhibitors were recognized directly from a crude extract of Tibetan herb. Frontal affinity chromatography (FAC) was used here for analyzing molecular interactions between the analytes and an immobilized ligand (in this case the PcAb conjugated to Sepharose CL-4B) by calculating the extent of retardation of the elution front. By combining FAC with mass spectrometry (MS) in the current study, a very efficient and straightforward procedure was developed for analyzing the binding properties of different inhibitors. The novel effective screening of anti-EGFR inhibitors using PcAb from natural resources affords us a new feasible approach for the discovery of lead compounds.