The Panning And Preliminary Application Of Novel Homing Peptides For Hepatocarcinoma Targeting Therapy

Human hepatocellular carcinoma (HCC) is one of the most common and deadly cancers in the world. China is one of the most widespread countries of HCC. At least 120,000 people in China were dead from it every year. So it is very important to establish a convenient and reliable system in clinical diagnosis and treatments of hepatocarcinoma.Now the monoclonal antibody and its fragment is still the common used carrier in hepatocarcinoma targeting therapy. Because of the specifically discernment and immunity functional the MAb has become most succeed one in the tumor targeting therapy field. Even some MAb have got the authentication of FDA in USA, the MAb still has its inborn shortcomings: higher molecular weight and immunogenic could induce some bad responses to human body, and these responses will hurt to patients obviously. Then the shortcomings could also decrease the half life of immunotoxin and the therapy effect will also been reduced eventually.In this paper, we will try to screen some specifically binding peptides to hepatocarcinoma tissues by using in vivo phage display technology. Then we can use this peptides which have high specifically and strong binding ability to take the place of monoclonal antibody in traditional tumor targeting therapy. Making the most use of specifically binding peptides' lower molecular weight and immunogenic to make up the shortcomings of MAb and to establish a feasible peptides system for diagnosis and targeting therapy to hepatocarcinoma.This paper can be divided into three parts:A. The panning of specifically binding peptides to hepatocarcinoma by in vivo phage display technology.a) The common used Ph.D.-12 Phage Display Peptide Library Kit which produced by New England Biolabs Inc was chosen to injected into the nude mouse with hepatocarcinoma tumors on their backs, after three rounds panning a lot of candidate specifically binding phages were obtained.b) Accompanying with each round panning, the phages' titering was carried out as the kit manual to inspect the number of phages in each tissues, and the immunohistochemistry assay was also performed to appraise the distribution of phages in each tissue. From the data of titering and immunohistochemistry we can say that after the three rounds panning the specifically binding phages were enriched effectively to the tumor tissue.c) Then the specifically binding clones were picked out for random amplification. There are 46 clones were chosen for sequencing eventually and the sequence of those phage clones were compared and analyzed. From the sequence we can say that also the data are all complex we still...