RESUMEN
Two monoclonal antibodies to thyrotropin prepared in our laboratory were employed for the development of two-site immunometric assays in two modifications for the estimation of human thyrotropin in dried blood spots designed for the screening of neonatal hypothyroidism. The immunoradiometric assay using the second antibody labelled with 125I is simple and fast (one incubation step lasting 2 h). The detection limit of 1 mU/l and the absence of the hook effect up to a concentration of more than 1000 mU/l are optimal for neonatal screening; the presence of other glycoprotein hormones does not interfere with the assay. In the luminescence immunoenzymometric assay the second antibody is labelled with peroxidase. In spite of the two-step configuration the method is fast (4.5 h) and enables specific determination of thyrotropin levels in the range of 2.4-1100 mU/l. The conditions and properties of both immunometric assays described are comparable with the time-resolved immunofluorometric assay widely used in Europe.
Asunto(s)
Hipotiroidismo/diagnóstico , Tamizaje Neonatal/métodos , Tirotropina/sangre , Anticuerpos Monoclonales , Recolección de Muestras de Sangre/métodos , Humanos , Técnicas para Inmunoenzimas , Recién Nacido , Mediciones Luminiscentes , Radioinmunoensayo/métodos , Tirotropina/inmunologíaRESUMEN
The influence of an initial salt concentration, phi(0), on the gradient separation of proteins using hydrophobic interaction chromatography on Separon HEMA 1000 was investigated. The results obtained were compared with the retention times and peak widths calculated according to a mathematical model.