肺癌组织蛋白质混合物检测方法学研究  

Study on Method of Protein Complex Detection of Lung Cancer

作　　者：刘勇[1,2] 王荣[1,2] 高岚[2] 贾正平[1,2] 辛晓婷[2] 谢华[1] 马骏[1] 郭志强[1] 

机构地区：[1]兰州军区兰州总医院全军临床药理基地,兰州730050 [2]兰州大学生命科学学院,兰州730000

出　　处：《分析化学》2010年第9期1287-1292,共6页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(Nos20775089;30970312)资助项目

摘　　要：采用无胶筛分毛细管电泳-激光诱导荧光法(NGS-CE-LIF)检测蛋白质,考察了分离条件对分离的影响,并对提取的肺癌组织蛋白质混合物进行分析,与毛细管电泳紫外检测(CE-UV)及常规蛋白质检测手段聚丙烯酰胺凝胶电泳(PAGE)进行比较。异硫氰酸酯(FITC)衍生蛋白质,筛分介质为0.05%(w/V)聚环氧乙烷(PEO,Mr=300000),以TBE缓冲液(pH10.0)为电极缓冲液,分离电压为15kV,柱温15℃,氩离子激光器(λex=488nm,λem=520nm)检测。在此条件下可对6.5～200kDa范围的蛋白质进行分离,分离效果较好,15min内完成分离,理论塔板数(N)均值为9.12×104/m,检出限为0.28mg/L。结果表明,本方法具有筛分介质浓度调节简便,分离效果好,时间短等优点,具有一定的实际应用价值;对提取肺癌组织蛋白质混合物进行检测,检测效率和效果优于CE-UV及PAGE。An experimental method of protein detection by non-gel sieving capillary electrophoresis with laser induced fluorescence（NGS-CE-LIF） was established and improved.The method was applied to the analysis of protein complex of lung cancer and compared to capillary electrophoresis with UV detection（CE-UV） and polyacrylamide gel electrophoresis（PAGE）.The protein was derived by fluorescein isothiocyanate.The optimized conditions such as 0.05%（w/V） poly（ethylene oxide）（300000） as sieving medium,laboratory buffer TBE（pH 10.0） as electrophoresis buffer,separation voltage of 15 kV,temperature of 15 ℃ and argon ion laser（λex=488 nm,λem=520 nm） as detector were employed.Under the conditions,the proteins under the range of 6.5-200 kDa were completely separated in 15 min with good separation efficiency.The average theoretical plate number（N） is 9.12×104 /m,the detection limit is 2.8×10-4 g/L.The results showed that the improved method has the advantages of good separation efficiency,short separation time and the concentration of sieving medium was easy to adjust.The detection rate and efficiency of the method for the protein complex detection of lung cancer are better than those of CE-UV and PAGE.

关 键 词：肺癌 无胶筛分毛细管电泳 聚环氧乙烷 异硫氰酸酯 激光诱导荧光检测 蛋白质 

分 类 号：R734.2[医药卫生—肿瘤]