Immunoreagents based on polymer dispersions for immunochemical assays

✍ Scribed by Yu. V. Lukin; I. S. Pavlova; A. N. Generalova; V. P. Zubov; O. V. Zhorov; S. P. Martsev

• Publisher: John Wiley and Sons
• Year: 1998
• Tongue: English
• Weight: 56 KB
• Volume: 11
• Category: Article
• ISSN: 0952-3499
• DOI: 10.1002/(sici)1099-1352(199812)11:1/6<185::aid-jmr419>3.0.co;2-7

✦ Synopsis

Immunoreagents based on polymer dispersions consisting of unimodal polyacrolein (PAL) microspheres with diameters in the range 0.3-2.0 mm 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 Â 10 9 M À1 ). 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 Â 10 À13 M l À1 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.