Oscarsson S, Chaga G, Porath J
Biochemical Separation Centre, Uppsala University, Sweden.
J Immunol Methods. 1991 Oct 25;143(2):143-9. doi: 10.1016/0022-1759(91)90039-i.
Three types of agarose derivatives have been prepared and investigated as adsorbents for radioimmunoassay and ELISA analysis. The analytical systems were evaluated using beta 2 microglobulin as a model. After a competitive reaction between the immunocomponents in solution, the formed immune complexes were adsorbed onto the adsorbent in the presence of 0.5 M potassium sulfate in 0.1 M Tris, pH 7.5. The binding constant between the interaction site on human IgG and the adsorbent 3-(2-pyridylthio)-2-hydroxypropylagarose (Py-S-gel) was determined to be 1.5 x 10(7) M-1 and the binding capacity was 20 mg/ml gel. The immune complex was desorbed by deleting potassium sulfate from the buffer, and only 0.5% of the total applied protein remained after washing the adsorbent with 0.5 M NaOH. The same adsorbent can be used repetitively with different systems.
已制备并研究了三种琼脂糖衍生物作为放射免疫分析和酶联免疫吸附分析的吸附剂。以β2微球蛋白为模型对分析系统进行了评估。在溶液中的免疫成分之间发生竞争性反应后，在0.1 M Tris（pH 7.5）中含有0.5 M硫酸钾的情况下，形成的免疫复合物被吸附到吸附剂上。测定人IgG上的相互作用位点与吸附剂3-(2-吡啶硫基)-2-羟丙基琼脂糖（Py-S-凝胶）之间的结合常数为1.5×10⁷ M⁻¹，结合容量为20 mg/ml凝胶。通过从缓冲液中去除硫酸钾使免疫复合物解吸，用0.5 M NaOH洗涤吸附剂后，仅0.5%的总加样蛋白残留。相同的吸附剂可在不同系统中重复使用。
