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。
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
关键词
HCG
酶解片断
免疫性
Human Chorionic Gonadotropin, Enzymatic fragment, Immunological characteristic
