Immunoreagents based on polymer dispersions for immunochemical assays

J Mol Recognit. 1998 Winter;11(1-6):185-7. doi: 10.1002/(SICI)1099-1352(199812)11:1/6<185::AID-JMR419>3.0.CO;2-7.

Abstract

Immunoreagents based on polymer dispersions consisting of unimodal polyacrolein (PAL) microspheres with diameters in the range 0.3-2.0 microns 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 (Ka = 4.5 x 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 x 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-reaction of latex conjugates with non-A Streptococcus cell lysates were observed.

Publication types

  • Comparative Study

MeSH terms

  • Acrolein*
  • Agglutination Tests / methods
  • Animals
  • Ferritins / analysis
  • Ferritins / immunology
  • Humans
  • Immunoassay / methods*
  • Immunoblotting / methods
  • Immunochemistry
  • Immunoradiometric Assay / methods
  • Indicators and Reagents
  • Microspheres
  • Particle Size
  • Polymers*
  • Polysaccharides, Bacterial / analysis
  • Polysaccharides, Bacterial / immunology
  • Rabbits
  • Streptococcus pyogenes

Substances

  • Indicators and Reagents
  • Polymers
  • Polysaccharides, Bacterial
  • polyacrolein
  • Acrolein
  • Ferritins