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抗牛甲状旁腺激素单克隆抗体:制备与特性鉴定

Monoclonal antibodies to bovine parathyroid hormone: production and characterization.

作者信息

Vieira J G, Federico P, Matsueda G, Neer R M

机构信息

Mineral Metabolism Unit, Massachusetts General Hospital, Boston 02114.

出版信息

Braz J Med Biol Res. 1988;21(5):1005-11.

PMID:3248228
Abstract
  1. This paper describes the production and characterization of monoclonal antibodies against bovine parathyroid hormone (bPTH)-(1-84). 2. Spleen cells from A/J mice successfully immunized with bPTH-(1-84) were fused with SP2/O myeloma cells using PEG 4000 as fusogen. The screening method employed microtiter plates coated with sheep antimouse IgG and the presence of specific monoclonal antibodies was demonstrated by the binding of 125I-bPTH-(1-84). 3. A detailed study of the specificity of the three viable monoclonals with highest affinity showed that two (6FH6 and 6CD4) were amino-terminal specific and the other (5BG9) carboxyl-terminal specific. The two amino-terminal monoclonal antibodies appear to recognize the same antigenic site. 4. The monoclonal antibodies produced are potentially useful reagents for the development of new methods for the measurement of PTH in biological fluids, studies on the interaction of PTH with its receptor, as well as localization of PTH producing cells.
摘要
  1. 本文描述了抗牛甲状旁腺激素(bPTH)-(1-84)单克隆抗体的制备及其特性。2. 用bPTH-(1-84)成功免疫的A/J小鼠的脾细胞与SP2/O骨髓瘤细胞,以聚乙二醇4000作为融合剂进行融合。筛选方法采用包被羊抗小鼠IgG的微量滴定板,通过125I-bPTH-(1-84)的结合来证明特异性单克隆抗体的存在。3. 对三种亲和力最高的存活单克隆抗体的特异性进行的详细研究表明,两种(6FH6和6CD4)是氨基末端特异性的,另一种(5BG9)是羧基末端特异性的。两种氨基末端单克隆抗体似乎识别相同的抗原位点。4. 所产生的单克隆抗体可能是用于开发生物体液中甲状旁腺激素测量新方法、甲状旁腺激素与其受体相互作用研究以及甲状旁腺激素产生细胞定位的有用试剂。

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