Principle of a new immunoassay based on electrophoretic mobility of poly(styrene/alpha-tert-butoxy-omega-vinylbenzyl-polyglycidol) microspheres: application for the determination of helicobacter pylori IgG in blood serum.
The principle of a novel latex diagnostic test for the determination of antibodies against Helicobacter pylori in blood sera is described. The test is based on the measurement of the electrophoretic mobility of the microspheres with immobilized H. pylori antigens. The electrophoretic mobility of these microspheres depends on the concentration of the antibodies against H. pylori in suspending medium. Particles with hydrophilic polyglycidol in the surface layer were used for the test. The microspheres were obtained by an emulsifier-free emulsion copolymerization of styrene and alpha-tert-butoxy-omega-vinylbenzyl-polyglycidol macromonomer (D(n) = 220 nm, diameter polydispersity factor D(w)//D(n) = 1.02). Activation of polyglycidol hydroxyl groups with cyanuric chloride allowed for covalent immobilization of H. pylori antigens. The fraction of H. pylori not specifically adsorbed onto the microspheres was negligible. Changes of the electrophoretic mobility of the microspheres with the surface concentration of the covalently immobilized H. pylori antigens Gamma = (1.6 +/- 0.3) . 10(-3) g m(-2) were suitable for the detection of the antibodies in the sera of patients with titer in the range (determined by the indirect ELISA test) from 1:500 to 1:32 000.
Basinska
Teresa
T
Center of Molecular and Macromolecular Studies, Polish Academy of Sciences, ul. Sienkiewicza 112, 90-363 Lodz, Poland. basinska@bilbo.cbmm.lodz.pl
Wisniewska
Monika
M
Chmiela
Magdalena
M
D000941:Antigens / Q000737:chemistry; D000942:Antigens, Bacterial / Q000737:chemistry; D001709:Biotechnology / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D016480:Helicobacter pylori / Q000378:metabolism*; D007118:Immunoassay / Q000379:methods*; D007074:Immunoglobulin G / Q000097:blood* / Q000737:chemistry*; D008027:Light; D046911:Macromolecular Substances; D008863:Microspheres; D008956:Models, Chemical; D011100:Polyglycolic Acid / Q000737:chemistry*; D011137:Polystyrenes / Q000737:chemistry*; D012542:Scattering, Radiation
D000941:Antigens; D000942:Antigens, Bacterial; D007074:Immunoglobulin G; D046911:Macromolecular Substances; D011137:Polystyrenes; D011100:Polyglycolic Acid