目的建立与优化非小细胞肺癌(non-small cell lung cancer,NSCLC)患者尿液全蛋白双向凝胶电泳技术(2-DE)。方法采用4种不同的方法制备尿液蛋白质样品,以非线性预制胶条进行第一向等电聚焦,12%的SDS-PAGE进行第二向电泳,银染显色。结果以乙腈沉淀蛋白后经过超滤制备的蛋白质样品,在300μg上样量时,可获得图像清晰、分辨率高、重复性好的双向电泳图谱。结论建立了较稳定的非小细胞肺癌患者尿蛋白质组双向凝胶电泳技术。 Objective To establish and optimize urine full-protein two-dimensional gel electrophoresis (2-DE) in patients with non-small cell lung cancer (NSCLC). Methods Urine protein samples were prepared by four different methods. The first-dimension isoelectric focusing was carried out by using non-linear preformed strips. The second-dimension electrophoresis was performed on 12% SDS-PAGE and silver staining was performed. Results The protein samples prepared by ultrafiltration after the protein was precipitated by acetonitrile showed clear, high resolution and good reproducibility of two-dimensional gel electrophoresis at 300 μg. Conclusions A more stable two-dimensional gel electrophoresis of urine proteome was established in patients with non-small cell lung cancer.