Simultaneous Bioluminescent Immunoassay of Serum Total and IgG-Bound Prolactins

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, Calcium, Humans, Immunoassay, Immunoglobulin G, Luminescent Measurements, Prolactin, 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-hPRL 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.

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Издание

Журнал: ANALYTICAL CHEMISTRY

Выпуск журнала: Vol. 84, Is. 7

Номера страниц: 3119-3124

ISSN журнала: 00032700

Место издания: WASHINGTON

Издатель: AMER CHEMICAL SOC

Авторы

  • Kudryavtsev A.N. (Siberian Federal University)
  • Krasitskaya V.V. (Siberian Federal University)
  • Frank L.A. (Siberian Federal University)
  • Petunin A.I. (DIAS Ltd.)
  • Burakov A.Y. (Krasnoyarsk Regional Hospital No. 1)

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