Preparation of monoclonal antibodies to the beta A subunit of ovarian inhibin using a synthetic peptide immunogen.
Preparation of monoclonal antibodies to the beta A subunit of ovarian inhibin using a synthetic peptide immunogen.
复制标题
使用合成肽免疫原制备卵巢抑制素 β A 亚基的单克隆抗体。
DOI：
10.1089/hyb.1991.10.309
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发表时间：
1991
期刊：
影响因子：
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通讯作者：
M. Lawrence
中科院分区：
文献类型：
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作者：
Nigel Groome;M. Lawrence
制备了7种不同的合成肽，对应于32 KDa人卵巢癌细胞结合素β A亚基的区域，预测其含有可能的连续B细胞表位。这些抗体与作为载体的结核菌素偶联，并用于免疫先前给予引发剂量的人结核疫苗的小鼠。这些肽中只有一种对应于序列82-114，通过ELISA始终产生与完整的32 KDa牛胰蛋白酶反应的抗体的良好滴度。从用该肽免疫的小鼠之一制备六个稳定的杂交瘤。其中5个分泌IgM，第6个（克隆E4）分泌IgG 2b抗体。在用十二烷基硫酸钠和巯基乙醇处理后，对卵泡液浓缩物进行的免疫印迹实验显示，来自克隆E4的抗体仅与对应于先前描述的β A亚基形式的约13 kDa和58 kDa的条带具有强反应性。这种单克隆抗体，以及本实验室以前用合成肽制备的α亚基抗体，应证明是进一步研究β-氨基丁酸和激活素的有用试剂。
Seven different synthetic peptides were prepared corresponding to regions of the beta A subunit of 32 KDa human ovarian inhibin predicted to contain possible continuous B cell epitopes. These were coupled to tuberculin as a carrier and used to immunize mice previously given a priming dose of human tuberculosis vaccine. Only one of these peptides, corresponding to sequence 82-114, consistently gave good titres of antibodies reactive with intact 32 KDa bovine inhibin by ELISA. From one of the mice immunized with this peptide, six stable hybridomas were prepared. Five of these secreted an IgM and the sixth (clone E4) secreted an IgG2b antibody. Immunoblotting experiments on follicular fluid concentrates, after treatment with sodium dodecyl sulphate and mercaptoethanol, showed strong reactivity of the antibody from clone E4 only with bands of about 13 kDa and 58 kDa corresponding to forms of the beta A subunit previously described. This monoclonal antibody, and an antibody to the alpha subunit previously made in this laboratory using synthetic peptides, should prove useful reagents for further study of inhibins and activins.
使用针对合成肽片段产生的抗血清检测和纯化抑制素。
影响因子：
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作者：
Vaughan,JM;Rivier,J;Corrigan,AZ;McClintock,R;Campen,CA;Jolley,D;Voglmayr,JK;Bardin,CW;Rivier,C;Vale,W
通讯作者：
Vale,W