Werblin T, Mage R
J Immunol Methods. 1977;16(4):337-50. doi: 10.1016/s0022-1759(97)90004-1.
A radioimmunoassay which can measure serum levels of antibody and antigen when both species are immuloglobulins (Igs) is described. The technique is based on competition between insolubilized Ig (bound to Sepharose) and serum Ig for radiolabeled Ig. For measurement of anti-Ig antibodies, labeled Ig antigen is added to a mixture of antibody-Sepharose and unknown or standard antibody solution. For measurement of antigen, labeled, purified antibody is added to a mixture of antigen-Sepharose and unknown or standard antigen solution. Although both assays can detect either Ig antigen or Ig antibody in a given unknown, the antibody assay requires 10-20 times as much antigen as antibody to achieve the same degree of inhibition, and the antigen assay is 100 times more sensitive for antigen than for antibody. By titrating the same unknown in both assays, one can determine whether inhibitory activity is due to antigen or antibody. The routine assay readily detects 1 microgram/ml of antibody or of antigen. A modified assay was also developed which detects levels of antibody as low as 5-10 ng/ml. The technique is simple and allows rapid screening of hundreds of serum samples in a short period of time. The assay can also be modified for measurement of anti-idiotype antibodies.
本文描述了一种放射免疫测定法,当抗体和抗原均为免疫球蛋白(Ig)时,该方法可用于测量血清中抗体和抗原的水平。该技术基于固定化Ig(结合到琼脂糖凝胶上)与血清Ig竞争放射性标记Ig的原理。为了测量抗Ig抗体,将标记的Ig抗原加入抗体-琼脂糖凝胶混合物以及未知或标准抗体溶液中。为了测量抗原,将标记的纯化抗体加入抗原-琼脂糖凝胶混合物以及未知或标准抗原溶液中。尽管两种测定法均可在给定的未知样品中检测Ig抗原或Ig抗体,但抗体测定法需要比抗体多10至20倍的抗原才能达到相同程度的抑制效果,并且抗原测定法对抗原的敏感性比对抗体高100倍。通过在两种测定法中对相同的未知样品进行滴定,可以确定抑制活性是由抗原还是抗体引起的。常规测定法能够轻松检测出1微克/毫升的抗体或抗原。还开发了一种改进的测定法,可检测低至5-10纳克/毫升的抗体水平。该技术简单,可在短时间内快速筛选数百份血清样品。该测定法还可经修改用于测量抗独特型抗体。