反相高效液相色谱法分析肝癌血清蛋白质组  

Establishment of reversed-phase high-performance liquid chromatography for serum proteomics

作　　者：罗婵[1] 黄丽华[1] 母昭德[1] 

机构地区：[1]重庆医科大学药学院药物分析教研室,重庆400016

出　　处：《重庆医科大学学报》2010年第6期894-896,共3页Journal of Chongqing Medical University

基　　金：重庆市自然科学基金项目资助(编号:CSTC;2008BB5398)

摘　　要：目的:建立肝癌血清蛋白质组RP-HPLC技术筛选肝癌血清差异蛋白质。方法:8例肝癌和20例正常组血清样品采用Agilent高丰度蛋白质分离系统洗脱白蛋白等高丰度蛋白质后,样品采用色谱柱ZORBAX 300SB-C18(250mm×4.6mm,5μm)分离,以A=0.1%三氟乙酸溶液,B=0.09%三氟乙酸乙腈为流动相梯度洗脱,在波长280nm处检测。结果:在肝癌血清和健康志愿者血清的RP-HPLC图谱中存在一组疏水性较强的差异蛋白质。结论:建立的血清蛋白质RP-HPLC技术可以发现肝癌发生发展过程中血清蛋白表达谱质的变化,从而为肝癌的早期诊断及治疗打下基础。Objective：To improve the early diagnosis of hepatocellular carcinoma（HCC）,a reversed-phase high-performance liquid chromatography（RP-HPLC）was employed to identify the differential proteins of serum related to HCC. Methods：The sera from 20 healthy volunteers and 8 HCC patients were collected. After the depletion of six serum proteins of the highest abundance with the Multiple Affinity Removal System （MARS） technologies from Agilent,the sera were subjected to RP-HPLC.The chromatographic conditions were： analytical column：ZORBAX 300SB-C18 （250 mm×4.6 mm,5 μm）;column temperature,25℃;the elution was performed with linear gradient elution of A：0.1% trifluoroacetic acid water solution and B：0.09% trifluoroacetic acid ACN solution at a flow rate of 0.75 ml/min. The detection wavelength was 280 nm. The time procedure was：1%B was held 3 min,then the gradient ran from 1 to 100% B in 100 min, then was held for another 10 min,at last,the gradient ran from 100 to 1%B in 5 min and was held for another 5 min. Results：There was a group of differential proteins of strong hydrophobicity in the serum between healthy persons and patients. Conclusion：This method can be useful in detecting protein expression alteration resulting from the carcinogenesis and development of HCC,and newly discovered biomarkers might be an aid in the early diagnosis and therapy of HCC.

关 键 词：血清蛋白质组学 蛋白质组 RP-HPLC 

分 类 号：R446.6[医药卫生—诊断学]