ZHOU Yanª²Chun, HOU Yiª²Feng, CHEN Linª²Fei, ZHENG Xiaoª²Xuan, HE Shaoª²Heng

    Allergy and Inflammation Research Institute, Medical College, Shantou University, Shantou 515041, China

    ¡¾Abstract¡¿ AIM: To predict the B cell epitope of human mast cell carboxypeptidase (hMCª²CP)and identify the region of B cell epitope recognized by monoclonal antibody (mAb)against hMCª²CP. METHODS: The B cell epitopes were predicted by secondary structure (SOMPA, selfª²optimized prediction method with alignment), hydrophilicity(Hopp&Woods), antigenic index (Jamesonª²Wolf) and surface probability plot (Emini). The cDNA encoding the N terminal 1¡«111(P1P2), 97¡«202(P3P4) and 1¡«202(P1P4)Aa of hMCª²CP was amplified by PCR and cloned into prokaryotic expression vector pETª²44 Ek/LIC. After induction with IPTG, the recombinant fusion protein was expressed and analyzed by SDSª²PAGE. For the epitope analysis, mAb 2A9 was used to react with the fusion protein by Western blot. RESULTS: The predicted B cell epitopes were probably located in N terminal 1¡«202 Aa. mAb 2A9 could react with fusion proteins which contained P1P4 and P3P4 in Western blot analysis, but not react with fusion proteins containing P1P2. CONCLUSION: Prediction of the B cell epitope for hMCª²CP can provide a base for the studies of structure and function of hMCª²CP. The epitope recognized by mAb 2A9 is located in 112ª²202 Aa of hMCª²CP. ......