Peuravuori H, Korpela T
Clinical Research Laboratory, Turku, Finland.
Clin Chem. 1993 May;39(5):846-51.
Human IgE was purified to near homogeneity by a two-step procedure consisting of immunoaffinity chromatography and high-performance liquid chromatography. Mouse hybridoma cell lines secreting antibodies against IgE were generated. Monoclonal and polyclonal antibodies were used to develop a "sandwich"-type ELISA for determining total IgE in human serum. Inorganic pyrophosphatase (EC 3.6.1.1), an enzyme having a high turnover number, was used as the label. The mean analytical recovery of our ELISA was 95.2% and the results showed good linear correlation with an established RIA of IgE. We found pyrophosphatase to be a good alternative label for use in immunoassays.
通过由免疫亲和色谱和高效液相色谱组成的两步法将人IgE纯化至接近均一。产生了分泌抗IgE抗体的小鼠杂交瘤细胞系。使用单克隆和多克隆抗体开发了一种“夹心”型ELISA，用于测定人血清中的总IgE。无机焦磷酸酶（EC 3.6.1.1），一种具有高周转数的酶，用作标记物。我们的ELISA的平均分析回收率为95.2%，结果与已建立的IgE放射免疫测定法显示出良好的线性相关性。我们发现焦磷酸酶是免疫测定中一种很好的替代标记物。
