T_3夹心ELISA测定方法的研究

SANDWICH ENZYME IMMUNOASSAY OF SMALL MOLECULES T3

使用抗Ｔ３抗体、Ｔ３－地高辛（ＤＩＧ）结合物及抗ＤＩＧＦ（ａｂ′）２－ＨＲＰ首次建立了一种新型的Ｔ３小分子的夹心ＥＬＩＳＡ测定方法，探讨了最佳实验条件，并对一步法和二步法作了比较，结果表明二步法优于一步法，一步法有其固有的缺陷。此方法测定Ｔ３的敏感性为０．０８９ｎｇ／ｍｌ。

 new sandwich enzyme immunoassay method for the assay of the T3 has been developed. The method is based on the formation and estimation of the complex Ab1·H1H2·Ab2 where Ab1 and Ab2* are immobilised and labeled antibodies respectivly and H1H2 is a synthetic conjugate of the hapten H1 and the hapten H2. The amount of the complex formed is inversely related to the concentration of free hapten H1 present. Using the T3DIG conjugate and a immobilized polyclonal rabbit antiT3 antibody and a labeled polyclonal rabbit antidigoxin F(ab′)2, a sensitivity of 0.089 ng/ml was obtained for T3. Physiological T4 concentration do not interfere with the T3 assay.