Poly(styrene/alpha-tert-butoxy-omega-vinylbenzylpolyglycidol) microspheres for immunodiagnostics. Principle of a novel latex testbased on combined electrophoretic mobility and particle aggregation measurements

Teresa M. Basinska 1Stanislaw Słomkowski Izabela Radomska-Galant 2

1. Polish Academy of Sciences, Center of Molecular and Macromolecular Studies (CMMS-PAS), Sienkiewicza 112, Łódź 90-363, Poland
2. Technical University of Łódź, Department of Biotechnology and Food Science, Stefanowskiego 4/10, Łódź 90-924, Poland

Abstract

Principle of a novel latex agglutination test based on changes in electrophoretic mobility of particles with immobilized antigen in the presence of antibody is described. Poly(styrene/alpha-tert-butoxy-omega-vinylbenzylpolyglycidol) microspheres (P(S/PGL)) were synthesized by a one step soap-free emulsion polymerization of styrene and alpha-tert-butoxy-omega-vinylbenzylpolyglycidol macromonomer with number average molecular weight Mn=2700 and polydispersity Mw/Mn=1.10. The monodisperse latex particles with number average diameter Dn=220 nm and the surface fraction of polyglycidol equal f=0.42 mol% were obtained. Adsorption and covalent immobilization of human serum albumin (HSA) onto these particles were investigated. The hydroxyl groups of polyglycidol chains required activation with 1,3,5-trichlorotriazine, prior to protein attachment. It has been found that human serum albumin (HSA) was efficiently covalently bound with the surface of latex particles. The maximal surface concentration of covalently immobilized HSA was equal 1.2 mg/m2.
A new type of antibody-antigen detection system based on combined measurement of changes in zeta potential and size of aggregates formed caused by immunoreactions of HSA immobilized on latex particles with anti-HSA (as antibody) was designed. In the model immunodiagnostic assay for anti-HSA involving P(S/PGL) particles with covalently bound HSA (P(S/PGL)-HSA), electrophoretic mobility and aggregation of microspheres were simultaneously measured. This approach allowed detection of anti-HSA in the serum in the range of anti-HSA concentrations from 0.1 to 200 μg/ml. The highest changes in electrophoretic mobility values were registered for P(S/PGL)-HSA-anti-HSA aggregates when microspheres with surface concentration of covalently immobilized HSA equal Gammacov=0.92 mg/m2 have been used.

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Presentation: oral at E-MRS Fall Meeting 2003, Symposium E, by Teresa M. Basinska
See On-line Journal of E-MRS Fall Meeting 2003

Submitted: 2003-06-25 17:08
Revised:   2009-06-08 12:55
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