The gold standard of assessing liver fibrosis is liver biopsy, which is invasive and not without risk. Therefore, searching for noninvasive serologic biomarkers for liver fibrosis is an importantly clinical issue. Methods: A total of 16 healthy volunteers and 45 patients with chronic hepatitis C virus (HCV) were enrolled (F0: n = 16, F1: n = 7, F2: n = 17, F3: n = 8 and F4: n = 13, according to the METAVIR classification). Three serum samples of each fibrotic stage were analyzed by two-dimension difference gel electrophoresis (2D-DIGE). The differential proteins were identified by. | Ho et al. Journal of Biomedical Science 2010 17 58 http content 17 1 58 a NSC Tha cost of publication In Journal of Blomodlcal Science Is bome by tlM National Science Council Taiwan JOURNAL OF BIOMEDICAL SCIENCE RESEARCH Open Access Novel biomarkers predict liver fibrosis in hepatitis C patients alpha 2 macroglobulin vitamin D binding protein and apolipoprotein AI Ai-Sheng Ho41 Chun-Chia Cheng42 3 Shui-Cheng Lee3 Meng-Lun Liu1 Jing-Ying Lee1 Wen-Ming Wang4 5 6 and Chia- Chi Wang 7 Abstract Background The gold standard of assessing liver fibrosis is liver biopsy which is invasive and not without risk. Therefore searching for noninvasive serologic biomarkers for liver fibrosis is an importantly clinical issue. Methods A total of 16 healthy volunteers and 45 patients with chronic hepatitis C virus HCV were enrolled F0 n 16 F1 n 7 F2 n 17 F3 n 8 and F4 n 13 according to the METAVIR classification . Three serum samples of each fibrotic stage were analyzed by two-dimension difference gel electrophoresis 2D-DIGE . The differential proteins were identified by the cooperation of MALDI-TOF TOF and MASCOT then western blotting and Bio-Plex Suspension Array were used to quantify the protein levels. Results Three prominent candidate biomarkers were identified alpha 2 macroglobulin A2M is up regulated vitamin D binding protein VDBP and apolipoprotein AI ApoAI are down regulated. The serum concentration of A2M was significantly different among normal mild F1 F2 and advanced fibrosis F3 F4 p . The protein levels of VDBP and ApoAI were significantly higher in normal mild fibrosis when compared to those in advanced fibrosis both p . Conclusions This study not only reveals three putative biomarkers of liver fibrosis A2M VDBP and ApoAI but also proves the differential expressions of those markers in different stages of fibrosis. We expect that combination of these novel biomarkers could be applied clinically to predict the stage of liver fibrosis without .