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Optimisation of a heterogeneous non-competitive flow immunoassay comparing fluorescein, peroxidase and alkaline phosphatase as labelsстатья
Информация о цитировании статьи получена из
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Дата последнего поиска статьи во внешних источниках: 18 июля 2013 г.
- Авторы: Lindgren A., Emneus J., Marko-Varga G., Irth H., Oosterkamp A., Eremin S.
- Журнал: Journal of Immunological Methods
- Том: 211
- Номер: 1-2
- Год издания: 1998
- Издательство: Elsevier BV
- Местоположение издательства: Netherlands
- Первая страница: 33
- Последняя страница: 42
- DOI: 10.1016/S0022-1759(97)00169-5
- Аннотация: Off- and on-line strategies for a non-competitive heterogeneous flow immunoassay were developed comparing three different labels. The samples, containing the model compounds digoxin or digoxigenin, were either pre-incubated off-line or on-line in a mixing coil with excess of labelled anti-digoxigenin Fab-fragments. The excess of Fab-fragments was then separated from the digoxin bound Fab-fragments by passing the sample through a column with immobilised digoxin. The off-line immunochemical detection system is suitable for sensitive high through-put screening of the analytes, whereas the on-line system is more suitable for coupling as a post-column detection unit to liquid chromatography. The digoxin and digoxigenin content in the sample were quantified using fluorescein (F) and enzyme (peroxidase (POD), alkaline phosphatase (AP)) labelled Fab-fragments. The fluorescein label was directly measured with the fluorescence detector, whereas a fluorescent enzyme product was measured in the two enzyme based systems, using 3-(p-hydroxyphenyl)-propionic acid (HPPA) and hydrogen peroxide for POD and, and 4-methylumbelliferyl phosphate (4-MUP) for AP. The highest sensitivity and lowest limit of detection (LOD) was obtained with the Fab-POD system with LODs for digoxin and digoxigenin in the off-and on-line configurations of 0.025 and 0.01 nM, respectively. The sample through-put for the off and on-line systems were 43 and 32 samples per hour, respectively. (C) 1998 Elsevier Science B.V.
- Добавил в систему: Еремин Сергей Александрович