Production and Characterization of Apolipoprotein A-I

• Main Author: 賴繡文
• Other Authors: 毛仁淡
• Format: Others
• Language: en_US
• Published: 2003
• Online Access: http://ndltd.ncl.edu.tw/handle/33378257587234940351

碩士 === 國立交通大學 === 生物科技研究所 === 91 === Coronary artery disease (CAD) is a high mortality heart disorder causing by formation of atherosclerosis and thrombosis. High-density lipoproteins (HDL) in plasma are functioning as to remove the arterial cholesterol from the atheroscleroic lesions via reverse transport. An inverse relationship between the concentrations of HDL and the development of CAD is well established. Since apoA-I is a major protein moiety of HDL (~70%), we have previously demonstrated that apoA-I is a superior marker in patients with CAD. In the present study, monoclonal antibodies 2C7, 2E10 and 3F8 specific to apoA-I were prepared and characterized. Using immunoassay and Western blot analyses, we show that these antibodies recognized different antigenic domains of apoA-I. By chemical modification and limited trypsinization, we further demonstrated that the positively charged groups of apoA-I were involved in maintaining the epitope structure. Since lipid moiety in HDL substantially masks the apoA-I immunoreactivity, we attempted to identify a monoclonal antibody that may react with apoA-I without the interference by HDL lipids. Using the competitive enzyme linked immunosorbent assay (ELISA) we demonstrated that monoclonal antibody 3F8 could determine apoA-I in human plasma without the lipid interference. This 3F8 antibody will be therefore novel for the future clinical determinant of apoA-I.