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Implementation of affinity solid-phases in continuous-flow biochemical detection
Division of Analytical Chemistry, Leiden/Amsterdam Center for Drug Research, Leiden University, Leiden, Netherlands.
Division of Analytical Chemistry, Leiden/Amsterdam Center for Drug Research, Leiden University, Leiden, Netherlands.
Division of Analytical Chemistry, Leiden/Amsterdam Center for Drug Research, Leiden University, Leiden, Netherlands.
Division of Analytical Chemistry, Leiden/Amsterdam Center for Drug Research, Leiden University, Leiden, Netherlands.
1997 (English)In: Journal of Chromatography A, ISSN 0021-9673, E-ISSN 1873-3778, Vol. 776, no 2, 169-178 p.Article in journal (Refereed) Published
Abstract [en]

A continuous-flow biochemical detection system is presented which allows the use of solid-phase immobilised affinity proteins. The biochemical detection is performed by mixing analyte with a labelled ligand followed by the addition of solid-phase immobilised affinity protein. After a reaction time of 85 s, free and bound label are separated by means of a hollow fibre module. Quantitation of the free label is performed with a conventional flow-through fluorescence detector. Total assay time amounts to less than 2 min. Biotin was chosen as the model compound using a range of streptavidin-coated solid-phases and an antibody-coated solid-phase as affinity material, and fluorescein–biotin as low-molecular-mass label. The relative standard deviation for twenty repetitive injections was 10.9%. A calibration curve was constructed in the concentration range between 20 and 400 nmol l−1 leading to a correlation coefficient of 0.994. A limit of detection of 8 nmol l−1 was obtained. © 1997 Elsevier Science B.V.

Place, publisher, year, edition, pages
Amsterdam: Elsevier B.V , 1997. Vol. 776, no 2, 169-178 p.
Keyword [en]
Detection (LC), Affinity-based detection, Fluorescein, Biotin, Streptavidin
National Category
Analytical Chemistry
Identifiers
URN: urn:nbn:se:hh:diva-24127DOI: 10.1016/S0021-9673(97)00354-3ISI: A1997XT07000001Scopus ID: 2-s2.0-0030859255OAI: oai:DiVA.org:hh-24127DiVA: diva2:679532
Available from: 2013-12-16 Created: 2013-12-09 Last updated: 2015-03-03Bibliographically approved

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CiteExportLink to record
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Citation style
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