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人绒毛膜促性腺激素β亚基酶解片段免疫性研究

The Immunological Charactristics of The Enzymatic Fragmentsof Human Chorioic Gonadotropin B-subunit

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【作者】 石湛萍杜国光李伟雄刘新军李受增徐一树王悦

【Author】 Shi, Zhan-ping Du, Guo-guang Li, Wei-xiong Liu, Xin-junLi, Shou-zeng Xu, Yi-shu Wang, Yue(Department of Biochemistry, Beijing Medical University, Beijing)

【机构】 北京医科大学生物化学教研室北京医科大学生理教研室北京医科大学第一附属医院中国科学院动物研究所北京医科大学生物化学教研室 北京北京北京

【摘要】 以嗜热菌蛋白酶将人绒毛膜促性腺激素(HCG)β亚单位进行酶解,用HPLC分离酶解产物。酶解片段经SDS-聚丙烯酰胺凝胶电泳鉴定口壬二条带。表观分子量分别为7600及20000。对照氨基酸序列分析,前者为近C端30肽(含糖链),为主要片段;后者为近N端片段。将此酶解片段混合物免疫家兔,用完整HCG作测试抗原,抗体滴度达1:204,800(Avidin-Biotin Micro-ELISA法),与促黄体激素(LH)交叉反应甚弱。表明该片段仍保留有完整HCG的主要抗原决定簇,有较强的免疫原性,可望作为免疫避孕疫苗。

【Abstract】 A β-subunit of human chorionic gonadotropin (HCG) was degraded by protease-thermolysin.The enzymatic fragments were partially purified by HPLC(TSK SIL), analysed on PAGE and the amino acid sequence was performed. Two bands have been identified on SDS-PAGE. The main band, a fragment of Mr 7600, consisted of 28 amino acid residues from Phe115 to IIe142 with its 4 sugar chains still reserved. The other band was a N-terminal fragment of Mr 20000.Three rabbits were immunized with the mixture of these fragments.The anti-HCG titer was obtained up to 1:204800 by Avidin-biotin-MicroELISA.weak cross-reactivity with LH still existed. It indicated that these fragments kept the main antigenic determinant groups of intact HCG and remained most of its immunogenicity.It can be considered as a potential candidate for development of immunological contraceptive vaccine.

【基金】 国家自然科学基金
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