Peptide Screening For Bladder Cancer Cell Line T24-specific Orientation Of Small Molecules

OBJECTIVE:1. To screen of micromolecule polypeptide specific for bladder carcinoma cell T24 with phagerandom seven peptide library.2. To analysis sequence and investigate the specificity preliminary of small molecule polypeptidewhich was screened.METHODS:In bladder cancer cell lines T24 continuous cultivation period,Screening for bladder cancer cellsT24 with phages random seven peptide library,then amplificating and extracting the twentymonoclonal DNA which were randomly selected from the fourth screening phage repository,lastly measuring DNA sequence, comparing and analyzing the peptide sequence. At the sametime conduct a competitly combined experiment with twenty randomly chosen positive phagemonoclonal and each screening phage library contrast to wild type VcsM13 phage, and theSW620, PC - 3 two different organizational source lines on the specificity of combining analysisto preliminary clear screening for bladder cancer cell lines of monoclonal T24 the specificity ofcombining ability.RESULTS:1.With each combination, elution and amplification screening, the proportion of low combined affinity phageis decline, and the strong affinity phages are gradually amplificate to enrichment, The third and fourth'srecovery increased, but not increase to 10~9 magnitude.2. The competition combined experimental results which contrast to the wild type VcsM13 phage show thatafter four screenings, eighteen monoclonals out of all twenty have higher adhesion, at lest 10 times than thewild type VcsM13 phage. Meanwhile, the four screening phage repository except the first round outside, theadhesion is also 10 times than the wild type VcsM13 phage.3.The specificity of combining analysis results which compared to the two different organizational source linesSW620, PC– 3 show that phages paleo-environments contrast to the other two lines have high specificitycombining ability, the relative combine multiple all reach up to 10 times. 4. After sequencing the 20 monoclonals from the fourth round screening, the results show that Four peptidesequence's frequency is higher,they are -SNARGTE-,-VSEKNRQ-,-DSIYNAR-,-DWSGACS-, and three ofthem have–SNR-.CONCLUTIONCONCLUTION:By the phage random seven peptide library for bladder cancer cell lines T24 to carry out the whole cellscreening can get a phage with bladder cancer cells peptide T24 specifically. And that phage maybe as a carrierof bladder cancer diagnosis and guide targeted drug research.