摘要
目的提高小分子酶免疫测定的敏感性。方法以兔抗地高辛(Dig)为固相抗体、Dig-人血清白蛋白(HSA)结合物作为中介物以及抗 Dig F(ab′)_2-HRP 为酶标抗体,建立小分子 Dig 夹心酶免疫方法,并与普通酶联免疫吸附法(ELISA)比较。结果所建立的 Dig 夹心酶免疫测定方法具有良好的敏感性和特异性,测定灵敏度(0.064μg/L)较普通竞争 ELISA 提高3倍。结论该夹心法的模式对各种小分子具有通用性,且避免了小分子的直接酶标,试剂的制备较为简单,为小分子高灵敏度免疫测定探索了一条新途径。
Objective We established a new sandwich enzyme immunoassay for small molecules us- ing digoxin(Dig)as a model hapten,so as to improve the sensitivity of enzyme immunoassay of small molecules.Methods The sandwich enzyme immunoassay for Dig was developed with rabbit anti-Dig as solid phase antibody,Dig-human serum albumin(HSA)conjugate as the intermediate and anti-Dig F_((ab)z)-HRP as the reporter.Its sensitivity was compared with that of the conventional ELISA.Results The sensitivity(0.064μg/L)of this method was about three-folds higher than that of the conventional assay(0.20μg/L).Conclusions The protocol of this method is suitable for detection a variety of small molecules,Moreover,it avoids direct enzyme labeling of the small molecules,and the reagent prepara- tion is simple compared to conventional assays.
关键词
ELISA
地高辛
小分子
Enzyme-linked immunosorbent assay Digoxin Small molecules
