title: Kinetics and proposed mechanism of the reaction of an immunoinhibition, particle-enhanced immunoassay.
abstract: 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.
authors.affiliation: Dade International, Glasgow Business Community, Newark, DE 19714-6101, USA.
authors.forename: J C
authors.initials: JC
authors.lastname: Thompson
authors.forename: A R
authors.initials: AR
authors.lastname: Craig
authors.forename: C L
authors.initials: CL
authors.lastname: Davey
authors.forename: D J
authors.initials: DJ
authors.lastname: Newman
authors.forename: M L
authors.initials: ML
authors.lastname: Lonsdale
authors.forename: W J
authors.initials: WJ
authors.lastname: Bucher
authors.forename: P D
authors.initials: PD
authors.lastname: Nagle
authors.forename: C P
authors.initials: CP
authors.lastname: Price
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D007700:Kinetics; D007841:Latex Fixation Tests; D010672:Phenytoin / Q000097:blood* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D010672:Phenytoin