Chloroactivated latex particles for covalent coupling of antibodies. Application to immunoassays.
The aim of the present work is to prepare and characterize a functionalized latex with chloromethyl groups on the surface and to perform the covalent coupling of anti-human serum albumin (a-HSA) IgG protein. The chloromethyl-styrene latex (CMS) was synthesized by means of a core-shell emulsion polymerization in a batch reactor. The monodisperse-obtained latex was characterized by determining the diameter (TEM and PCS), the surface charge density (conductometric and potentiometric titration), the amount of chloromethyl groups on the surface (hydrolysis reaction), and the stability vs electrolyte concentration (turbidity measurements). Electrokinetic characterization was also performed (electrophoretic mobility versus pH and ionic strength). IgG was chemically bound to the latex particles under different sensitization and block-stabilization conditions. Colloidal stability of complexes was studied to select an immunolatex suitable for the development of latex immunoassays. The final part of this work consists of a study of the immunoreactivity of the IgG-latex complexes at different pH and ionic strength, in particular under physiological conditions. The results show that chemically bound IgG to chloromethyl latex provides an IgG-latex complex suitable for application in immunodiagnosis tests.
Miraballes-Martínez
I
I
Departamento de Física Aplicada, Facultad de Ciencias, Universidad de Granada, Spain.
Martín-Rodríguez
A
A
Hidalgo-Alvarez
R
R
D002021:Buffers; D002713:Chlorine / Q000737:chemistry*; D003102:Colloids / Q000737:chemistry; D004337:Drug Carriers / Q000592:standards*; D004586:Electrophoresis; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007118:Immunoassay / Q000379:methods*; D007074:Immunoglobulin G / Q000378:metabolism*; D007840:Latex / Q000378:metabolism*; D007841:Latex Fixation Tests; D009391:Nephelometry and Turbidimetry; D009994:Osmolar Concentration; D011108:Polymers; D012709:Serum Albumin / Q000276:immunology*; D013343:Styrenes / Q000138:chemical synthesis* / Q000378:metabolism
D002021:Buffers; D003102:Colloids; D004337:Drug Carriers; D007074:Immunoglobulin G; D007840:Latex; D011108:Polymers; D012709:Serum Albumin; D013343:Styrenes; C027142:vinyltoluene; D002713:Chlorine