Weltman J K, Frackelton A R, Szaro R P, Rotman B
J Allergy Clin Immunol. 1976 Sep;58(3):426-31. doi: 10.1016/0091-6749(76)90123-8.
We report here the development of a galactosidase-immunosorbent test (GIST) for immunoglobulin E (IgE) antibodies in which the amount of galactosidase adsorbed to a cellulose disc is a single valued function of IgE concentration in human serum. Rabbit anti-IgE immunoglobulin insolubilized on cellulose discs is incubated sequentially with human serum, sheep anti-IgE serum, and a covalent conjugate of rabbit antisheep immunoglobulin with the enzyme beta-D-galactoside galactohydrolase (E.C.) 3.2.1.23). Colorimetric assay of enzyme conjugate adsorbed to discs permits quantitation of 1.0 to 25 ng of IgE per test. Concentrations of IgE in 48 sera as measured by the GIST gave a linear correlation coefficient of 0.97 with IgE concentrations as determined by radioimmunoassay. Preliminary studies indicate that the GIST makes possible nonisotopic measurement of ragweed-specific IgE antibiotics in human serum. The GIST for IgE is simple to perform and requires neither short-lived radioisotopes, expensive scintillation detection equipment, nor scarce, purified IgE.
我们在此报告一种用于检测免疫球蛋白E(IgE)抗体的半乳糖苷酶免疫吸附试验(GIST)的开发,其中吸附到纤维素圆盘上的半乳糖苷酶的量是人类血清中IgE浓度的单值函数。固定在纤维素圆盘上的兔抗IgE免疫球蛋白依次与人血清、羊抗IgE血清以及兔抗羊免疫球蛋白与β-D-半乳糖苷半乳糖水解酶(E.C. 3.2.1.23)的共价缀合物孵育。对吸附在圆盘上的酶缀合物进行比色测定可对每次检测中1.0至25 ng的IgE进行定量。通过GIST测定的48份血清中的IgE浓度与通过放射免疫测定法测定的IgE浓度的线性相关系数为0.97。初步研究表明,GIST使得对人血清中豚草特异性IgE抗体进行非同位素测量成为可能。用于IgE的GIST操作简单,既不需要短寿命放射性同位素、昂贵的闪烁检测设备,也不需要稀缺的纯化IgE。