Gallati H, Pracht I, Schmidt J, Häring P, Garotta G
F. Hoffmann-La Roche & Co. Ltd., Central Research Units, Basle, Switzerland.
J Biol Regul Homeost Agents. 1987 Jul-Sep;1(3):109-18.
A rapid and sensitive enzyme-immunoassay for native and recombinant human interferon gamma is described. The test is performed in one step at room temperature and is based on the sandwich principle. The IFN gamma preparation is distributed with horse radish peroxidase-labeled monoclonal antibody to IFN gamma in microtiter plates previously coated with a second mab against IFN gamma. The amount of the IFN gamma mab sandwich fixed in the microtiter plate wells is proportional to the color developed after the addition of peroxidase-specific substrate. The two mab's used in the test neutralize IFN gamma and are directed against the same epitope. For this reason they can only detect the biologically active dimeric form of IFN gamma. The IFN gamma-ELISA works in phosphate buffer as well as in tissue culture medium or human serum. As the assay is routinely performed in 2 hours, the limit of detection is 3 U/ml of IFN gamma (0.3 ng/ml). If the assay is performed in 16 hours, the limit of detection decreases to 0.5 U/ml IFN gamma (0.06 ng/ml). The conditions to preserve the activity of IFN gamma preparation as standard are discussed.
本文描述了一种用于检测天然和重组人γ干扰素的快速灵敏的酶免疫测定法。该检测在室温下一步完成，基于夹心原理。将γ干扰素制剂与辣根过氧化物酶标记的抗γ干扰素单克隆抗体一起加入预先包被有另一种抗γ干扰素单克隆抗体的微量滴定板中。固定在微量滴定板孔中的γ干扰素单克隆抗体夹心的量与加入过氧化物酶特异性底物后产生的颜色成正比。检测中使用的两种单克隆抗体可中和γ干扰素，且针对相同表位。因此，它们只能检测到具有生物活性的二聚体形式的γ干扰素。γ干扰素酶联免疫吸附测定法在磷酸盐缓冲液、组织培养基或人血清中均有效。由于该检测通常在2小时内完成，检测限为3 U/ml的γ干扰素（0.3 ng/ml）。如果检测在16小时内完成，检测限降至0.5 U/ml的γ干扰素（0.06 ng/ml）。文中还讨论了将γ干扰素制剂作为标准品保存活性的条件。
