Knopf H P, Duffy F, Papoian R
Department of Biopharmaceutics, Sandoz Pharma Ltd., Basel, Switzerland.
Ann Clin Biochem. 1993 Jan;30 ( Pt 1):69-71. doi: 10.1177/000456329303000111.
We describe a time-resolved fluoroimmunoassay (TRFIA) for recombinant human interleukin-3 (IL-3). The assay design uses two different monoclonal anti-IL-3 antibodies, giving a two-site immunometric assay. The TRFIA for IL-3 is a direct adaptation from our existing ELISA for IL-3. By using the same assay configuration and exchanging a europium-streptavidin conjugate for alkaline phosphatase-streptavidin, we were able to minimize the time required to develop a TRFIA, but still increase the working range of the TRFIA over the ELISA by 20-fold. The limit of detection is 5 ng/L, with the limits of quantification set at 5 and 30,000 ng/L human plasma.
我们描述了一种用于重组人白细胞介素-3（IL-3）的时间分辨荧光免疫分析（TRFIA）方法。该分析方法设计使用两种不同的抗IL-3单克隆抗体，构成双位点免疫分析。IL-3的TRFIA是直接改编自我们现有的IL-3酶联免疫吸附测定（ELISA）。通过使用相同的分析配置，并将铕-链霉亲和素缀合物换成碱性磷酸酶-链霉亲和素，我们能够将开发TRFIA所需的时间降至最短，但仍使TRFIA的工作范围比ELISA扩大了20倍。检测限为5 ng/L，人血浆中的定量限设定为5和30,000 ng/L。
