Journal
JOURNAL OF CHROMATOGRAPHY A
Volume 895, Issue 1-2, Pages 1-7Publisher
ELSEVIER
DOI: 10.1016/S0021-9673(00)00590-2
Keywords
capillary electrophoresis immunoassay; hydrogels; immunoassays; morphine
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In this paper thermally reversible hydrogel used as a replaceable packed material for capillary electrophoresis was examined. A simple and rapid method of detecting morphine was developed, which demonstrated the potential of strong affinity antibodies as a selector for immunologically-based separations in serum by capillary electrophoresis. Polyclonal antibodies were linked to hydrogel and applied to the separation of free fluorescein isothiocyanate (FITC)-labeled antigen and bound FITC antigen. The separation was monitored with laser-induced fluorescence detection. Different separation conditions were studied. The results indicated that poly-N-isopropylacrylamide hydrogel (PNIPA) is a kind of steady, replaceable gel. The specific determination of morphine did not require a long incubation time and PNIPA hydrogel-modified antibodies can be stockpiled at 4 degreesC before assay. It can be used to determine morphine with good precision and a detection limit lower than 8.5 ng/ml. Details of the preparation of hydrogel cross-linked polyclonal antibody and of typical separations of bound and free antigen are presented. (C) 2000 Elsevier Science B.V. All rights reserved.
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