Establishment of reversed-phase high-performance liquid chromatography for serum proteomics
- VernacularTitle:反相高效液相色谱法分析肝癌血清蛋白质组
- Author:
Chan LUO
- Publication Type:Journal Article
- Keywords:
Serum proteomics;
Proteome;
RP-HPLC
- From:
Journal of Chongqing Medical University
2003;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
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 ?m);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.
