Targeted Proteomics Applied for the Diagnosis of Hepatitis B Virus Infected Patients

• Main Authors: Hsing-Fen Tsai, 蔡幸芬
• Other Authors: He-Hsuan Hsiao
• Format: Others
• Language: zh-TW
• Published: 2015
• Online Access: http://ndltd.ncl.edu.tw/handle/61489810060828752753

碩士 === 國立中興大學 === 化學系所 === 103 === Hepatitis B virus infection is one of the world''s most common and serious infectious diseases, especially in Asia. Chronic hepatitis and liver cirrhosis are two of the leading top ten causes of death and hepatocellular carcinoma is the second of ten most common cancers. There is more than 300 million people have hepatitis B virus infection. Hepatitis B virus infection may cause chronic hepatitis, liver cirrhosis and hepatocellular carcinoma and therefore the study of hepatitis B virus infection is very important. The goal of this research is to develop a MS-based stable isotope-labeled technology for the quantitation of hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) that could help the doctor to take the preliminary diagnosis for hepatitis B virus infection. First, ten peptides were synthesized by utilizing solid phase peptide synthesis (SPPS) and subsequently purified these peptides by using reversed-phase high-performance liquid chromatography (RP-HPLC). The resulting peptides are exchanged 16O to 18O at their c-terminus by utilizing enzyme-catalytic labeling technology. The 18O-labeled peptides were spiked into human serum as internal standards for the quantification of HBsAg and HBeAg by the use of nanoLC coupling with Q-Exactive mass spectrometer. In this study, P5 peptide of HBsAg was observed in hepatitis B virus patient’s serum by using parallel reaction monitoring (PRM) and amount of P5 peptide was 10.88 fmol calculated from the calibration curve of standard P5 peptide. The 18O-labeled P5 peptide was spiked into hepatitis B virus patient’s serum as internal standards and we also found that the intensity of P5 peptide was approximately 10% of 100 fmol of 18O-labeled P5 peptide. In future, we anticipate this method can be extensively applied for all diagnosis of diseases that are caused by the infection of virus or bacteria.