Pazzagli M, Kohen F, Sufi S, Masironi B, Cekan S Z
Reproductive Endocrinology Research Unit, Karolinska Hospital, Stockholm, Sweden.
J Immunol Methods. 1988 Nov 10;114(1-2):61-8. doi: 10.1016/0022-1759(88)90154-8.
A solid-phase immunometric assay of human lutropin (hLH) is described. Two different anti-hLH antibodies were utilized as capture antibodies, and anti-IgG antibodies covalently coupled to magnetic particles and horseradish peroxidase, respectively, served as 'universal' detection reagents. An anti-hLH antibody raised in rabbits was incubated with a goat anti-rabbit IgG covalently bound to magnetic particles. The resulting complex was added to a separately incubated mixture of hLH and monoclonal anti-hLH antibody. Following incubation, the immunocomplex was sedimented in a magnetic field and the supernatant discarded. Finally a sheep anti-mouse antibody (F(ab')2 fragment) conjugated to horseradish peroxidase as label was added. Following a further incubation, the particles were sedimented in the magnetic field and washed. The hLH content of the sample was quantitated by measuring 'enhanced chemiluminescence'. The sensitivity of the assay was 2.5 +/- 0.9 IU/l (mean +/- SD), the within-run variation ranged from 7.9 to 11%, the between-run variation from 12.9 to 19.8%. Cross-reaction with hFSH or hTSH could not be detected, but was approximately 0.1% with hCG. The results correlated well with those obtained by radioimmunoassay (r = 0.84).
本文描述了一种人促黄体生成素（hLH）的固相免疫测定法。使用两种不同的抗hLH抗体作为捕获抗体，分别与磁性颗粒和辣根过氧化物酶共价偶联的抗IgG抗体用作“通用”检测试剂。将兔源抗hLH抗体与共价结合到磁性颗粒上的山羊抗兔IgG一起孵育。将所得复合物加入到单独孵育的hLH和单克隆抗hLH抗体的混合物中。孵育后，免疫复合物在磁场中沉淀，弃去上清液。最后加入与辣根过氧化物酶偶联作为标记的羊抗小鼠抗体（F(ab')2片段）。进一步孵育后，颗粒在磁场中沉淀并洗涤。通过测量“增强化学发光”对样品中的hLH含量进行定量。该测定法的灵敏度为2.5±0.9 IU/l（平均值±标准差），批内变异范围为7.9%至11%，批间变异范围为12.9%至19.8%。未检测到与hFSH或hTSH的交叉反应，但与hCG的交叉反应约为0.1%。结果与放射免疫测定法获得的结果相关性良好（r = 0.84）。
