递呈BAC_5单抗识别位点的M_(13)噬菌体克隆系的建立

The Establishment of M_(13) Phage Clones Displayed Epitope Recognized by BAC_5 McAb

【目的】获得可递呈抗低分化和未分化癌细胞单克隆抗体 (BAC5单抗 )抗原表位的M13 噬菌体克隆系。【方法】以BAC5单抗为靶抗体 ,对由M13 噬菌体构建的随机 12肽文库进行生物淘洗。用抗体竞争方法从阳性克隆中选出与BAC5单抗结合率较高的克隆系。通过ELISA方法检测BAC5单抗对上述克隆的选择性识别。【结果】经过 3轮淘洗 ,获得的阳性克隆率为77% (35 /4 5 )。来自C2、C17和C2 9号克隆的噬菌体对外加BAC5单抗的结合率分别为 71 6 %、49 4%和 6 4 0 % ,高于其它阳性克隆。BAC5单抗与来自上述 3个克隆的噬菌体呈阳性反应 ,与来自辅助型M13 株的噬菌体 (VCSM13 )和淘洗前的文库噬菌体(RPLM13 )呈阴性反应。【结论】来自C2、C17和C2 9号克隆的噬菌体有可能递呈与BAC5单克隆抗相关的抗原表位。

Objectivel To obtain M 13 phages displayed antigenic epitope recognized by BAC 5 McAb, a kind of monoclonal antibody located on poorly or undifferentiated squamous carcinoma cells. Methods Using BAC 5 McAb as selected target, a 12 mers RPL was biopanned. The positive clones were selected by antibody competition assay. The clones combincd with BAC 5 McAb were demonstrated by ELISA. Results 77%(35/45) of positive clones from eluted phage clones were obtained by 3 rounds of biopanning. The combination rates of the phages from clone C2, C17, C29 with added BAC 5 McAb were 71 6%,49 4% and 64 0% respectively much higher than those of phages from other positive clones. BAC 5 McAb had positive reaction to M 13 phages from C2,C17,C29 clones as compared with those from helper M 13 phage strain, VCSM 13 and no biopanning RPL M 13 phages(P/N>2 1). Conclusion The M 13 phages from C2,C17,C29 clones may display antigenic epitope recognized by BAC 5 McAb.