On line immunoaffinity assay- capillary electrophoresis using magnetic nanobeads for the determination of IgG anti-Helicobacter pylori in human serum

PATRICIA W. STEGE; JULIO RABA; GERMÁN A. MESSINA

ELECTROPHORESIS

WILEY-V C H VERLAG GMBH

Año: 2010 vol. 31 p. 3475 - 3481

0173-0835

About two-thirds of the world’s population is infected with Helicobacter pylori (H. pylori).This Gram-negative bacterium is the most important etiological agent of chronic activetype B gastritis and peptic ulcer diseases. Conventional methods such as gastric biopsy,ELISA and culture, require a long time for the determination of H. pylori infections.Moreover, the antibodies in human serum sample are capable to react immunologicallywith the purified H. pylori antigens immobilized on different kinds of support likemagnetic nanobeads. In this study, we have developed an online immunoaffinity assay-CE to determine the concentration of anti-H. pylori IgG using magnetic nanobeads as asupport of the immunological affinity ligands and an LIF as a detector. The separationwas performed in 0.1M glycine–HCl, pH 2, as the background electrolyte. The linearcalibration curve to predict the concentration of H. pylori-specific immunoglobulin Gantibodies in serum was produced within the range of 0.12–100 U/mL. The linearregression equation was i5492.86196.03Canti-H. pylori, with the linear regressioncoefficient r250.999. The LOD calculated by fluorescence detection procedure was of0.06 U/mL. The whole assay was done in no more than 35 min and it was entirelyautomatized. The development of immunoaffinity assay-CE in this study demonstratesthat there is a large possibility to introduce nanotechnology in several fields withsignificant advantages over the classic methodologies. Our proposition comprises thediagnosis and screening field.