Тип публикации: статья из журнала
Год издания: 2012
Идентификатор DOI: 10.1021/ac300444w
Ключевые слова: Gel chromatography, Good correlations, High sensitivity, Photoproteins, Reproducibilities, Time resolution, Body fluids, Chromatography, Immunology, Phosphorescence, Bioluminescence, calcium, immunoglobulin G, prolactin, animal, article, blood, blood analysis, chemistry, human, immunoassay, luminescence, metabolism, methodology, time, Animals, Blood Chemical Analysis, Humans, Luminescent Measurements, Time Factors
Аннотация: Novel dual-analyte single-well bioluminescence immunoassay (BLIA) for total and IgG-bound prolactins was developed on the base of Ca2+-regulated photoprotein obelin mutants with altered color and kinetics of bioluminescence signal as reporters. The mutants W92F-H22E and Y138F were chemically conjugated with monoclonal mouse anti-hPПоказать полностьюRL and anti-hIgG immunoglobulins and thus displayed signals from total prolactin and IgG-bounded prolactin (macroprolactin) correspondingly. Bioluminescence of the reporters was simultaneously triggered by a single injection of Ca2+ solution and discriminated via bioluminescent signal spectral and time resolution. The developed microplate-based immunoassay allows detection of two prolactin forms in crude serum without additional manipulations (e.g., gel chromatography or PEG-precipitation). Total prolactin bioluminescence immunoassay in standard, control, and clinical sera offers high sensitivity and reproducibility. The BLIA results show good correlation with those obtained by RIA and immunoassay after gel chromatography.
Журнал: ANALYTICAL CHEMISTRY
Выпуск журнала: Vol. 84, Is. 7
Номера страниц: 3119-3124
ISSN журнала: 00032700
Место издания: WASHINGTON
Издатель: AMER CHEMICAL SOC