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Clinical Chemistry 43: 2384-2389, 1997;
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(Clinical Chemistry. 1997;43:2384-2389.)
© 1997 American Association for Clinical Chemistry, Inc.

Articles

Kinetics and proposed mechanism of the reaction of an immunoinhibition, particle-enhanced immunoassay

John C. Thompson1, Alan R. Craig1, Carol L. Davey2, David J. Newman2, Michele L. Lonsdale1, William J. Bucher1, Patrick D. Nagle1 and Christopher P. Price2,a

1 Dade International, Glasgow Business Community, Newark, DE 19714-6101.

2 The Department of Clinical Biochemistry, St. Bartholomew's and the Royal London School of Medicine and Dentistry, Turner St., London E1 2AD.
a Author for correspondence. Fax 44 171 377 1544; e-mail c.p.price{at}mds.qmw.ac.uk

We report kinetic studies on the reaction of a latex agglutination immunoassay used to quantify phenytoin in serum. In this assay, polystyrene particles with a covalently attached analog of phenytoin react with an antiphenytoin monoclonal antibody to form light-scattering aggregates, with the rate of this reaction being decreased by addition of phenytoin from sample. In the absence of free (sample) phenytoin, this reaction did not exhibit a maximum rate of agglutination in the presence of excess antibody, i.e., an equivalence point. Furthermore, agglutination was inhibitable by free phenytoin even when the latter was added after agglutination of particles with antibody had begun. Most significantly, the immunoagglutination proceeded in an identical fashion with monovalent F(ab) fragment. These data are consistent with low-affinity immunospecific particle–antibody complexation, which then induces colloidal aggregation, without requiring immunospecific bridging by antibody molecules. The described mechanism is not generalizable to all latex agglutination immunoassays, although disturbance of colloidal stability may be a component in most assays.




The following articles in journals at HighWire Press have cited this article:


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Y. Umebayashi, Y. Miyamoto, M. Wakita, A. Kobayashi, and T. Nishisaka
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P. Englebienne, A. Van Hoonacker, and J. Valsamis
Rapid Homogeneous Immunoassay for Human Ferritin in the Cobas Mira Using Colloidal Gold as the Reporter Reagent
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S. Guaita, J. M. Simo, N. Ferre, J. Joven, and J. Camps
Evaluation of a Particle-enhanced Turbidimetric Immunoassay for the Measurement of Immunoglobulin E in an ILab 900 Analyzer
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T. Q. Wei, V. P. Chu, A. R. Craig, J. E. Duffy, D. M. Obzansky, D. Kilgore, I. S. Masulli, C. M. Sanders, and J. C. Thompson
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