Berthold W, Merk W, Adolf G R
Arzneimittelforschung. 1985;35(1A):364-9.
A sensitive nonradioactive immunoassay based on monoclonal antibodies was developed. A number of monoclonal hybridomas secreting antibodies against human leukocyte interferon (IFN alpha) were generated using mice immunized with purified lymphoblastoid IFN. Although the binding of antibody to IFN alpha was used as one criterium of selection, all antibodies found can neutralize its antiviral activity. A pair of antibodies binding to different regions of IFN alpha was identified. These were incorporated into sensitive sandwich assays of IFN alpha. A microtiter plate assay - using horse radish peroxidase as marker enzyme - is able to detect IFN alpha at a concentration of 30 IU/ml within 5 h. An overnight tube assay can detect approximately 30 pg IFN alpha 2 or 3 IU per ml solution. The 5-h ELISA (enzyme-linked immunosorbent assay) is well suited for the monitoring of the recovery of rIFN alpha from recombinant organism, during purification and refinement of the protein to a therapeutic drug.
开发了一种基于单克隆抗体的灵敏非放射性免疫测定法。使用用纯化的淋巴母细胞样干扰素免疫的小鼠，产生了许多分泌抗人白细胞干扰素（IFNα）抗体的单克隆杂交瘤。尽管抗体与IFNα的结合被用作选择标准之一，但发现的所有抗体都能中和其抗病毒活性。鉴定出一对与IFNα不同区域结合的抗体。这些抗体被用于构建灵敏的IFNα夹心测定法。一种微量滴定板测定法——使用辣根过氧化物酶作为标记酶——能够在5小时内检测到浓度为30 IU/ml的IFNα。过夜管测定法每毫升溶液可检测到约30 pg IFNα2或3 IU。5小时酶联免疫吸附测定（ELISA）非常适合监测重组生物体中rIFNα的回收情况，以及在将蛋白质纯化和精制为治疗药物的过程中。
