Groome N, Hancock J, Betteridge A, Lawrence M, Craven R
School of Biological and Molecular Sciences, Oxford Polytechnic, Headington.
Hybridoma. 1990 Feb;9(1):31-42. doi: 10.1089/hyb.1990.9.31.
A monoclonal antibody was made after immunization of mice with the 1-32 amino terminal peptide of the alpha subunit of 32K human ovarian inhibin. The IgG2a mouse antibody reacted 6 times better with bovine 1-32 peptide than it did with 32K bovine inhibin. By contrast sheep polyclonal antibodies made by a similar method had a 29 fold bias in reactivity towards the immunizing peptide. Relative to homologous 1-32 peptide standards, the monoclonal antibody measured apparently higher amounts of immunoreactive material(s) in human (13.5 fold) and bovine (27 fold) follicular fluids than did the polyclonal anti 1-32 peptide antibodies. Immunochemical studies revealed that the epitope recognized by the monoclonal antibody was different from the major epitope recognized by the polyclonal antibodies. The monoclonal antibody reacted much better with human inhibin 1-32 sequences than with bovine (73 fold) or porcine (23 fold). Although the 32K form of human inhibin has not yet been purified, it can be inferred that the monoclonal antibody would be able to detect as little as 2 ng/ml of 32K human inhibin in competitive radioimmunoassays. The antibody must also react with some of the multiple molecular forms of inhibin found in human follicular fluids, and it was shown to function well in the quantitative immunoaffinity extraction of inhibin-like immunoreactivity from follicular fluid. It seems likely that this monoclonal antibody will prove a useful tool for research on human inhibin.
用 32K 人卵巢抑制素α亚基的 1 - 32 氨基酸末端肽免疫小鼠后制备了一种单克隆抗体。该 IgG2a 小鼠抗体与牛 1 - 32 肽的反应性比与 32K 牛抑制素的反应性高 6 倍。相比之下，用类似方法制备的绵羊多克隆抗体对免疫肽的反应性有 29 倍的偏差。相对于同源的 1 - 32 肽标准品，单克隆抗体在人卵泡液（13.5 倍）和牛卵泡液（27 倍）中检测到的免疫反应性物质含量明显高于多克隆抗 1 - 32 肽抗体。免疫化学研究表明，单克隆抗体识别的表位与多克隆抗体识别的主要表位不同。单克隆抗体与人抑制素 1 - 32 序列的反应性比与牛（73 倍）或猪（23 倍）的反应性好得多。虽然人抑制素的 32K 形式尚未纯化，但可以推断，在竞争性放射免疫测定中，该单克隆抗体能够检测低至 2 ng/ml 的 32K 人抑制素。该抗体还必须与在人卵泡液中发现的抑制素的多种分子形式发生反应，并且已证明它在从卵泡液中定量免疫亲和提取抑制素样免疫反应性方面表现良好。这种单克隆抗体似乎有望成为研究人抑制素的有用工具。
