人甲状旁腺激素N端肽中一个含有B细胞新表位的hPTH^(13-27)片段的研究  

Analysis of a new B cell epitope on hPTH^(13-27) in N-terminal human parathyroid hormone

作　　者：赵丹慧[1] 张艳[1] 王娜[1] 袁越[1] 王芊[1] 李成文[1] 

机构地区：[1]北京积水潭医院北京创伤骨科研究所,100035

出　　处：《中华微生物学和免疫学杂志》2006年第9期812-816,共5页Chinese Journal of Microbiology and Immunology

基　　金：北京市自然科学基金(No.7012010);北京市科委骨与关节疾病研究中心资助(951890700);北京市人才基金资助

摘　　要：目的研究并证明人甲状旁腺激素(hPTH)N端肽内的hPTH13-27片段是一个新的B细胞抗原表位。方法计算机辅助分析预测hPTH1-37和hPTH1-84多肽片段内hPTH13-27抗原表位的特征;制备大鼠抗hPTH13-27-KLH多克隆抗体;酶联免疫测定(ELISA)研究hPTH13-27抗原表位和抗体的免疫特征。结果蛋白质结构分析预测在hPTH13-27区域含有B细胞抗原表位。hPTH13-27单表位多克隆抗体制备成功。ELISA分析证明hPTH13-27片段含有B细胞抗原表位,与抗体的结合效果依次为:抗hPTH13-27抗体>抗hPTH1-84抗体>抗hPTH1-37抗体。hPTH13-27片段与固相板结合对免疫原性影响不大。抗hPTH13-27抗体与含hPTH13-27讲片段的多肽结合良好;与不含hPTH13-27片段的多肽无交叉反应。结论hPTH13-27可能是制备hPTH N端肽抗体的核心表位片段。Objective To analyse and study a new B cell epitope of hPTH^13-27 in N-terminal human parathyroid hormone. Methods The secondary structure and solvent accessibility of hPTH^13-27 in hPTH^1-84 and in hPTH^1-37 were predicted by the net soft-ware ＂predict protein＂. The hPTH^13-27-KLH were injected into rats to raise antibodies. The immune feature hPTH^13-27 were analysed by ELISA. Results The structure analysis predict that there was a B cell epitope formed by residues 13-27 in hPTH N-terminal. The single epitope and polyclonal antibodies of hPTH^13-27 were produced successfully. The restths of ELISA indicated： the ability of hPTH^13-27 to combine with antibodies were as follows： anti-hPTH^13-27 〉 anti-hPTH^1-84 〉 anti-hPTH^1-37 antibodies; the combination of hPTH^13-27 with microtiter plate had no obvious influences on its antigenicity; anti-hPTH^13-27 antibodies could combine well with hPTHs including hPTH^13-27 and did not show crees immune reaction with non-hPTH^13-27 epitopic region of hPTHs. Conclusion hPTH^13-27 may be a main B cell epitope to produce antibodies in hPTH N-terminal.

关 键 词：蛋白质结构软件分析 hPTH^13-27 B细胞抗原表位 抗hPTH^13-27抗体 ELISA 

分 类 号：R392[医药卫生—免疫学]