Pockley A G, Williams S, Reid S D, Bowles M J
Professorial Surgical Unit, St Bartholomew's Hospital, West Smithfield, London, UK.
J Immunol Methods. 1995 May 11;182(1):81-4. doi: 10.1016/0022-1759(95)00025-6.
This study describes the development of a sandwich enzyme immunoassay (ELISA) for rat soluble IL-2 receptors (sIL-2R) using a combination of monoclonal antibodies reactive with different epitopes on the rat IL-2R. Coating plates with NDS61 and NDS64 monoclonal antibodies produced similar dose-response curves when incubated with a standard sIL-2R preparation followed by biotinylated OX39, streptavidin-alkaline phosphatase and the substrate. Although normal rat serum inhibited the assay, the effects were more profound when NDS64 was used as the capture antibody and subsequent development of the assay was performed using NDS61. The intra- and interassay variations were typically less than 5%. This assay will be valuable for monitoring immune activation status in a variety of experimental models.
本研究描述了一种用于大鼠可溶性白细胞介素-2受体(sIL-2R)的夹心酶免疫测定法(ELISA)的开发,该方法使用了与大鼠白细胞介素-2受体上不同表位反应的单克隆抗体组合。用NDS61和NDS64单克隆抗体包被酶标板,与标准sIL-2R制剂孵育,随后加入生物素化的OX39、链霉亲和素-碱性磷酸酶和底物,产生了相似的剂量反应曲线。虽然正常大鼠血清会抑制该测定,但当使用NDS64作为捕获抗体并随后使用NDS61进行测定时,这种抑制作用更为显著。批内和批间变异通常小于5%。该测定法对于监测各种实验模型中的免疫激活状态将具有重要价值。