Department of Medicinal Chemistry, Center for Frontier Research in Medicinal Science, Kyoto Pharmaceutical University, Kyoto 607-8412, Japan.
Anal Biochem. 2012 Jan 15;420(2):185-7. doi: 10.1016/j.ab.2011.09.022. Epub 2011 Sep 29.
Toward future applications to the discovery of drugs against membrane receptors on pathological cells, an intact-cell-based surface plasmon resonance (SPR) methodology has been developed. The injection of a suspension of epidermal carcinoma A431 cells (5×10(7)cells/ml), as an analyte, generated clear SPR responses to epidermal growth factor (EGF) immobilized on the sensor chip. Because the responses were competitively reduced by the free ligand EGF, added to the analyte cell suspension, they certainly reflect the specific interaction of the immobilized EGF with the extracellular region of its receptor, which is highly expressed on the surface of the A431 cells.
为了将这项技术应用于针对病理细胞上的膜受体的药物研发,我们开发了一种基于完整细胞的表面等离子体共振(SPR)方法。将表皮癌细胞 A431 的悬浮液(5×10(7)个细胞/ml)作为分析物注入,会在传感器芯片上固定的表皮生长因子(EGF)上产生清晰的 SPR 响应。由于自由配体 EGF 的加入竞争性地减少了分析物细胞悬浮液中的响应,这肯定反映了固定化 EGF 与其受体的细胞外区域的特异性相互作用,而这种受体在 A431 细胞表面高度表达。