Novel medical diagnostic test based on electrophoretic mobility of microspheres

Teresa M. Basinska 1Stanislaw Słomkowski 1Magdalena Chmiela 2Monika Wisniewska 2

1. Polish Academy of Sciences, Center of Molecular and Macromolecular Studies (CMMS-PAS), Sienkiewicza 112, Łódź 90-363, Poland
2. University of Łódź, Department of Infectious Biology, Inst. of Microbiol. and Immunol., Łódź 90-237, Poland

Abstract

Suspensions of colloidal polymer microspheres found variety of applications in medical diagnostics. Recently, we developed a model diagnostic test for determination of anti-HSA in blood serum based on simultaneous control of size of formed latex particle aggregates and electrophoretic mobility of latex particles. This method is based on an assumption that charge of particles with bound antigens changes after binding of complementary antibodies. In this work we report on the model test for determination of Helicobacter pylori antigens in patients sera.
For this purpose monodisperse poly(styrene/alpha-tert-butoxy-omega-vinylbenzyl-polyglycidol) particles (P(S/PGL), Dn=220 nm, Dw/Dn=1.02) containing hydrophilic polyglycidol segments with hydroxyl groups in the surface layer were synthesized. The hydroxyl groups were used for covalent binding of H. pylori antigens via hydroxyl groups of polyglycidol activated with cyanuric chloride. It was found that fraction of adsorbed antigen was negligible and essentially the whole amount of attached antigen was covalently bound onto P(S/PGL) microspheres with the maximal surface coverage equal 1.61 mg/m2. H. pylori antigens were isolated from glycine acid extract obtained from H. pylori strain NCTC 11637 (CCUG 17874). Sera with and without H. pylori antibodies were obtained from children patients.
We found that the presence of anti-H. pylori antibodies in the blood serum added to suspension of P(S/PGL)-H. pylori microspheres results in significant change of electrophoretic mobility of particles and leads to some particle aggregation. The aggregation is visualized by a rapid increase in the size of microspheres (from 372.9 nm to 432.4 nm). The degree of these changes depends on concentration of the anti-H. pylori antibodies. The novel diagnostic test allowing quantitative determination of anti-H. pylori antibodies in blood serum can be based on these effects.

 

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Presentation: oral at E-MRS Fall Meeting 2004, Symposium B, by Teresa M. Basinska
See On-line Journal of E-MRS Fall Meeting 2004

Submitted: 2004-04-26 10:12
Revised:   2009-06-08 12:55