Title page for etd-0831106-150212


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URN etd-0831106-150212
Author Ming-Lu Hsu
Author's Email Address bioshiner@gmail.com
Statistics This thesis had been viewed 5197 times. Download 1777 times.
Department Biological Sciences
Year 2005
Semester 2
Degree Master
Type of Document
Language English
Title Development of ELISA for measurement of HDGF
Date of Defense 2006-07-14
Page Count 34
Keyword
  • HCC
  • HDGF
  • ELISA
  • MONOCLONAL ANTIBODIES
  • Abstract Hepatocellular carcinoma (HCC) is the most common malignant tumor in Taiwan with more than one million new cases annually in the world. Growth factors play important roles in liver carcinogenesis. Hepatoma-derived growth factor (HDGF), originally isolated from the cultured media of human hepatoma HuH-7 cells, stimulate the growth of fibroblast cells, endothelial cells, and hepatoma cells. Overexpression of HDGF is related to the transformation of human hepatoma, lung cancer and melanoma. Besides, HDGF also exerts strong influence on the prognosis of patients with hepatoma. In this study, recombinant HDGF was expressed and purified with higher purity than 90%. The recombinant protein was used to raise polyclonal HDGF antibodies in rabbits and to generate three lines of HDGF monoclonal antibodies in mice. After antibodies characterization, an in-house, sandwich HDGF ELISA system was established using the purified polyclonal anti-HDGF IgG as the capture antibodies and the monoclonal anti-HDGF IgG as the detection antibodies. By using recombinant HDGF as standard, this ELISA system accurately evaluate the changes of HDGF release in SK-Hep-1 cells after gene delivery. In addition, we also evaluated the effect of anti-HDGF on the growth of hepatoma cells. Application of either polyclonal or monoclonal HDGF antibodies, but not preimmune antibodies, inhibited the proliferation of SK-Hep-1 hepatoma cells in a dose-dependent manner. In summary, the present study generated HDGF monoclonal antibodies for development of HDGF ELISA and application on suppressing HCC progression. Future studies should be carried out to enhance the sensitivity of HDGF ELISA and to evaluate the therapeutic potential of HDGF antibodies for treatment of HCC.
    Advisory Committee
  • Ching-Mei Hsu - chair
  • Chi-Huei Wang - co-chair
  • Ming-Hong Tai - advisor
  • Files
  • etd-0831106-150212.pdf
  • indicate access worldwide
    Date of Submission 2006-08-31

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