针对癌症关键蛋白翻译后修饰的靶向蛋白质组学CE-MS方法研究

表皮生长因子受体（EGFR）作为一种重要的糖基化跨膜蛋白，已被证实与多种恶性肿瘤的发生有着密切关系。根据EGFR变异的致癌机理，人们开发了多种能与EGFR特异性结合的靶向抗癌药物。但是，这些针对EGFR的不同基因突变体的药物在使用过程中对不同癌症类型及个体的药效差异较大，且均发生了耐药情况。虽然这些药效差异性和耐药性很可能来源于EGFR蛋白结构的变化以及翻译后修饰的异常，但尚无一种可行的分析方法来研究这类微区特定蛋白质结构和修饰上的变化对疾病发展和其药物响应的影响。本课题拟采用亲和富集的方式，通过与EGFR特异性结合的新设计抗体以及已有单抗药物抓取样本细胞中的EGFRs，研究无鞘液接口毛细管电泳质谱联用技术（CESI-MS）分离检测不同EGFR结构和修饰变体及其含量比例的方法，希望通过此技术分离分析各类癌症样本中的EGFR蛋白变异情况，为针对EGFR的靶向抗体药物及精确治疗提供指导。

The epidermal growth factor receptor (EGFR or ErbB1) is an essential transmembrane protein which play an important role in the pathogenesis and progression of different carcinoma types, like lung cancer and breast cancer. During last thirty years, small molecule drugs which can inhibit the tyrosine kinase of EGFR and monoclonal antibodies which block the activation of EGFR and ErbB2 have been developed. Although the mutations of EGFR gene in cancer are clear now, the changes on the protein itself are still unknown. These changes may be connected with the drug resistence of the tumor cells. In this work, capillary electrophoresis (CE) coupled with mass spectrometry (MS) and affinity enrichment will be applied to investigate the structure and post-translational modification of EGFR in different types of cancer, and monitor it changes during the anti-cancer treatment. This work will provide scientists a new perspective on the changes of EGFR in the progression of cancer and help people design new anti-cancer drugs.