Groome N
School of Biological and Molecular Sciences, Oxford Polytechnic, Headington, U.K.
J Immunol Methods. 1991 Dec 15;145(1-2):65-9. doi: 10.1016/0022-1759(91)90311-3.
Monoclonal antibodies produced to synthetic peptides of the alpha and beta A subunits of inhibin were used to develop the first highly sensitive two-site assays for recombinant human inhibin-A and activin-A. The assay for inhibin-A could detect as little as 5 pg/ml. Activin-A gave a 5.2% cross-reactivity in the inhibin assay. The assay for activin-A could detect 0.1 ng/ml and inhibin-A showed a 5.3% cross-reaction in the activin assay. Determination of inhibins and activins in biological fluids is complicated by the presence of multiple molecular forms and an excess of free alpha subunits. The present study demonstrates the potential of antibodies raised to synthetic peptides to configure more specific two-site assays for inhibins and activins which could facilitate research on these molecules. The monoclonal antibody used as the labelled antibody in these studies was made by immunizing mice with a peptide corresponding to an internal sequence of the beta A subunit. It appears likely that similar high affinity anti-peptide monoclonals suitable for use in ultrasensitive two-site assays could be made to other molecules if a rigorous screening of the fusion supernatants on the native molecule was carried out. The use of anti-peptide monoclonal antibodies in the development of ultrasensitive immunoassays may be more widely applicable than commonly realised.
针对抑制素α和β A亚基的合成肽产生的单克隆抗体，被用于开发首批针对重组人抑制素A和激活素A的高灵敏度双位点检测方法。抑制素A检测方法的检测下限低至5 pg/ml。激活素A在抑制素检测中显示出5.2%的交叉反应性。激活素A检测方法可检测到0.1 ng/ml，抑制素A在激活素检测中显示出5.3%的交叉反应。生物体液中抑制素和激活素的测定因多种分子形式和过量游离α亚基的存在而变得复杂。本研究证明了针对合成肽产生的抗体在构建更特异的抑制素和激活素双位点检测方法方面的潜力，这可能有助于对这些分子的研究。在这些研究中用作标记抗体的单克隆抗体是通过用与β A亚基内部序列相对应的肽免疫小鼠制备的。如果对融合上清液进行严格的天然分子筛选，似乎有可能针对其他分子制备适用于超灵敏双位点检测的类似高亲和力抗肽单克隆抗体。抗肽单克隆抗体在超灵敏免疫测定开发中的应用可能比通常意识到的更广泛。
