Frei Y, Lambris J D, Stockinger B
Basel Institute for Immunology, Switzerland.
Mol Cell Probes. 1987 Jun;1(2):141-9. doi: 10.1016/0890-8508(87)90022-3.
We have generated a monoclonal antibody with specificity for the fifth component of mouse complement (C5). This antibody precipitates the two chains of C5 from normal mouse serum and inhibits C5-dependent hemolysis in a functional complement test. In this study we describe its application in an enzyme-linked immunoadsorbent assay (ELISA assay) for the detection of anti-C5 antibodies in serum. Monoclonal anti-C5 coupled to wells of an ELISA plate specifically binds C5 from unfractionated normal mouse serum. This subsequently serves as antigen to bind anti-C5 serum antibodies. By this approach we have circumvented the need for extensive purification of C5 from serum which would be required if C5 was directly coupled to ELISA plates as antigen. Serum antibodies from C5-immunized mice bound with high avidity to wells containing normal serum as antigen source in amounts representing 1 microgram to 250 ng C5. There was no antibody binding to wells containing C5-deficient serum as antigen source. The immune reaction was detected by development with enzyme-coupled goat-anti mouse Ig antibodies specific for various mouse Ig subclasses. This method allows the qualitative characterization of immune responses to mouse C5 which is an ideal model for a natural self antigen in studies of immunological tolerance.
我们制备了一种对小鼠补体第五成分（C5）具有特异性的单克隆抗体。该抗体可从正常小鼠血清中沉淀出C5的两条链，并在功能性补体试验中抑制C5依赖性溶血。在本研究中，我们描述了其在酶联免疫吸附测定（ELISA测定）中用于检测血清中抗C5抗体的应用。与ELISA板孔偶联的单克隆抗C5特异性结合未分级正常小鼠血清中的C5。这随后用作抗原以结合抗C5血清抗体。通过这种方法，我们避免了如果将C5直接作为抗原偶联到ELISA板上时从血清中大量纯化C5的需求。来自C5免疫小鼠的血清抗体以高亲和力结合含有正常血清作为抗原源的孔，其量相当于1微克至250纳克C5。没有抗体与含有C5缺陷血清作为抗原源的孔结合。通过用针对各种小鼠Ig亚类的酶偶联山羊抗小鼠Ig抗体显色来检测免疫反应。该方法允许对小鼠C5的免疫反应进行定性表征，小鼠C5是免疫耐受研究中天然自身抗原的理想模型。
