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Immunoreagents based on polymer dispersions for immunochemical assays
Author(s) -
Lukin Yu. V.,
Pavlova I. S.,
Generalova A. N.,
Zubov V. P.,
Zhorov O. V.,
Martsev S. P.
Publication year - 1998
Publication title -
journal of molecular recognition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.401
H-Index - 79
eISSN - 1099-1352
pISSN - 0952-3499
DOI - 10.1002/(sici)1099-1352(199812)11:1/6<185::aid-jmr419>3.0.co;2-7
Subject(s) - detection limit , chromatography , chemistry , immunoassay , latex fixation test , ferritin , antibody , biochemistry , biology , immunology
Immunoreagents based on polymer dispersions consisting of unimodal polyacrolein (PAL) microspheres with diameters in the range 0.3–2.0 µm have been prepared and evaluated by various immunoassay techniques such as immunoradiometric assay of ferritin and microtitre particle agglutination and immunofiltration dot assay of group‐specific polysaccharide of S. pyogenes (A‐PS) in comparison with conventional carriers and methods. The antibodies were covalently or indirectly bound to the PAL. The coupled antibodies to ferritin retained a high average affinity ( K a = 4.5 × 10 9 M −1 ). In comparison with microcrystalline cellulose‐based immunosorbent, more than an order‐of‐magnitude lower amount of PAL–IgG was necessary for the analysis of ferritin. Use of PAL–IgG gave a higher sensitivity of assay with a detection limit of 0.7 × 10 −13 M l −1 and a wider concentration range of antigen detection (about four orders of magnitude) without manifestation of the high‐dose hook effect. Particle agglutination assay of A‐PS in microtitre plate was shown to be a simple, demonstrative and highly sensitive one‐step analytical method with a detection limit of 0.05 ng A‐PS/ml or 10 4 cells/ml. The sensitivity of immunofiltration assay using both enzyme and latex markers was shown to be approximately the same (50 ng A‐PS/ml) and the duration of the assay was 3–5 min. No cross‐reactions of latex conjugates with non‐A Streptococcus cell lysates were observed. Copyright © 1998 John Wiley & Sons, Ltd.