细胞膜色谱法（CMC）已得到广泛应用和良好评价。但其细胞膜固定相无法只反映细胞膜外表面特性，会造成药物与细胞膜受体作用强度的测定偏差。针对该问题，本项目在CMC基础之上提出新的生物亲和色谱构想-固定化细胞色谱法。应用细胞生物学技术和色谱技术结合离体药理学实验方法，利用高表达FGFR4细胞结合微载体培养技术，以免疫细胞化学细胞固定技术制备固定化细胞固定相；建立固定化高表达FGFR4细胞色谱法，用于模拟药物与细胞的相互作用。以其为核心技术与LC-MS结合建立在线二维分析系统，提高活性筛选的定性与定量水平。并应用于秦巴山区特有药用植物“七药”中可特异性作用于高表达FGFR4细胞的活性成分筛选；结合分子生物学、离体药理学等技术对活性成分作用进行验证。项目的完成将丰富生物亲和色谱内容，拓展色谱技术在药物分析中的应用；为中药活性成分筛选分析提供新技术和方法。

Cell membrane chromatography (CMC) was found in 1996 and was been wide application and well evaluated. However, the applicant study the theory and develop new method on CMC since 1997, and found that CMC is difficult to distinguish the inner or outer surface of the cell membrane. CMC cannot reflect the real interaction between ligand and cell, and the further application of CMC is limited. On the basis of the theory of CMC, we put forward a new biological affinity chromatography conception – immobilized cell chromatography (CC), and the technique and method of cell biology, chromatography and in vitro pharmacology will be used. In this project, the FGFR4 over expressed cell line is cultured on micro carrier. Then, immobilized cell stationary phase will be prepared using modified immunocytochemistry cell fixation technique. Using the stationary phase, the cell chromatography will established. To improve the identification and qualification of cell chromatography for the screening efficiency, the cell chromatography will be hyphenated with LC/MS to established a two dimensional CC-LC/MS system. The system will be used to screen the active components from the “Qiyao” a kind of folk herb medicine in “Qin-Ba” mountain area. The screened compound will be test by the method of cell biology, and in vitro pharmacology and ensure their effective. The establish of CC method and the results of the project will provide a new methodological basis for material study of traditional Chinese medicine, and fundamental basis for higher efficiency screening method.