关键词: 蛋白质组学, 毛细管电泳, 肺癌, 蛋白质混合物, 聚丙烯酰胺凝胶电泳, 蓝绿温和胶电泳

Protein mixture (denatured and native) differences of extracted from lung cancer and adjacent normal tissue by established method of capillary electrophoresis (CE)-laser-induced fluorescence (LIF) detection of protein was detected using the fluorescein isothiocyanate (FITC) as a derivative agent. When detected by the 1×TBE electrophoresis buffer solution (pH 10.0 of denaturing gel electrophoresis and pH 8.3 of native gel electrophoresis containing 2 mg/L Coomassie brilliant blue), separation voltage of 15 kV, column temperature of 15 ℃, electric injection (10 kV×10 s), and excitation wavelength/emission wavelength of 488/520 nm, lung cancer and adjacent normal tissue protein mixture samples obtained a better separation and there was significant difference. Comparing with the current commonly used analysis methods, the SDS-denatured polyacrylamide gel electrophoresis (SDS-PAGE) as well as the Blue Native polyacrylamide gel electrophoresis (BN-PAGE), the BN-PAGE results showed that compared to normal tissue, the lung cancer tissue had more significant protein type differences. But the SDS-PAGE results showed that some differences of protein expression had significant difference between the lung cancer and adjacent normal tissue, and mainly concentrated in 20～116 kDa. CE-LIF detection results were roughly the same with PAGE, and CE-LIF is more sensitivey than PAGE and it can more accurately reflect protein differences of lung cancer and adjacent normal tissue. We concluded that the CE-LIF could be used to detect protein differences. And the time was shorter, effect was better, and analysis of active protein reflected its advantage of providing a strong driving force-voltage and a well temperature stability by strong motivation.

Key words: proteomics, capillary electrophoresis, lung cancer, protein mixture, polyacrylamide gel electrophoresis, blue native polyacrylamide gel electrophoresis