Use of monoclonal antibodies as sensitive and specific probes for biologically active human gamma-interferon.
Use of monoclonal antibodies as sensitive and specific probes for biologically active human gamma-interferon.
复制标题
使用单克隆抗体作为具有生物活性的人γ-干扰素的敏感和特异性探针。
DOI：
--
复制
发表时间：
1984
影响因子：
11.1
通讯作者：
J. Le
中科院分区：
文献类型：
--
作者：
T. Chang;S. McKinney;V. Liu;P. Kung;J. Vilček;J. Le
小鼠单克隆抗体B1和B3对天然和大肠杆菌衍生的重组人γ-干扰素（IFN-γ）具有特异性。这两种抗体识别IFN-γ分子的不同表位，并且不竞争彼此的结合。我们已经使用这两种抗体来构建人IFN-γ的固相夹心免疫放射测定。将纯化的抗体B1包被在直径为0.64 cm的聚苯乙烯珠上，用作固相免疫吸附剂，抗体B3用125 I标记，用作示踪剂。该测定可在约4小时内完成，并且能够检测人血清或组织培养液中低至0.1 NIH参考单位/ml的IFN-γ水平。来源于大肠杆菌的重组人IFN-γ。在0.02ng/ml浓度下可检测到大肠杆菌。该测定似乎对IFN-γ的生物活性形式具有特异性，因为在暴露于pH 2、37 ℃或56 ℃后，免疫放射测定中的生物活性和反应性平行降低。免疫放射测定可用于分析人IFN-γ分子的结构特征。
Mouse monoclonal antibodies B1 and B3 are specific for natural and Escherichia coli-derived recombinant human gamma-interferon (IFN-gamma). The two antibodies recognize different epitopes of the IFN-gamma molecule and do not compete with each other's binding. We have used these two antibodies to construct a solid-phase, sandwich immunoradiometric assay for human IFN-gamma. Purified antibody B1 was coated on polystyrene beads (0.64 cm in diameter) and used as the solid-phase immunoadsorbent and antibody B3 was labeled with 125I and used as tracer. This assay can be completed in about 4 hr and is capable of detecting IFN-gamma levels in human serum or tissue culture fluids as low as 0.1 NIH reference unit/ml. Recombinant human IFN-gamma derived from E. coli was detectable at a concentration of 0.02 ng/ml. The assay appears to be specific for the biologically active forms of IFN-gamma, since after exposure to pH 2, 37 degrees C, or 56 degrees C, biological activity and reactivity in the immunoradiometric assay decreased in parallel. The immunoradiometric assay can be employed for the analysis of the structural characteristics of the human IFN-gamma molecule.