人抑制素βA亚基片段的合成及抑制素α亚基、βA亚基单克隆抗体的制备  

SYNTHESIS OF HUMAN INHIBIN βA FRAGMENTS, PREPARATION AND GENERATION OF MONOCLONAL ANTIBODIES AGAINST HUMAN INHIBIN α AND βA SUBUNITS

作　　者：孙颖[1] 鲁桂琛[1] 雷平生[1] 夏辉明[2] 高晓东[2] 黄新[2] 

机构地区：[1]中国医学科学院,中国协和医科大学药物研究所,北京100050 [2]河南医科大学病理生理研究室,河南郑州450052

出　　处：《药学学报》2000年第6期426-430,共5页Acta Pharmaceutica Sinica

摘　　要：目的　合成人抑制素α亚基 1个片段和 βA亚基两个片段 βA(1 2 8) (GLECDGKVNICCKKQFFVSFK DIGW NDW ) ,βA(82 114) (VPTKLRPMSMLYYDDGQNIIKKDIQNMIVEECG)并制备了相应的单克隆抗体。 方法应用固相多肽合成法人工合成了 1个抑制素α亚基片段和 2个 βA亚基片段 ,以化学合成的片段作为半抗原 ,经抗原和单抗的制备获得 7株杂交瘤细胞株。测定了单抗的滴度、特异性和相对亲和力。结果　单抗的鉴定及特异性分析实验表明这些单克隆抗体具有特异性、敏感性和实用性。AIM To synthesize fragments of inhibin subunits βA(1 28) (GLECDGKVNICCKKQFFVSFKDIGWNDW), βA(82 114) (VPTKLRPMSMLYYDDGQNIIKKDIQNMIVEECG) and to obtain their monoclonal antibodies. METHODS The fragments of inhibin subunits α(37 65), βA(1 28) and βA(82 114) have been synthesized manually by stepwise solid phase procedure. These peptide products were conjugated with TG as antigens to prepare McAbs against inhibin α and βA subunit. After immunization, fusion, selection and cloning procedures, seven hybridoma cell lines were obtained. Ascites titer, specificity and relative affinity were identified. RESULTS These McAbs were shown to be highly special, sensitive and useful in detection tests. Initial immunohistochemistry test gave some important information for clinical utility. CONCLUSION The method of detecting serum inhibin level will be established on basis of the present research.

关 键 词：抑制素 固相多肽合成 单克隆抗体 

分 类 号：R392－33[医药卫生—免疫学]