Immuno-column for on-line quantification of human serum IgG antibodies to Helicobacter pylori in human serum samples.

LUIS MOLINA; GERMÁN ALEJANDRO MESSINA; PATRICIA WANDA STEGE; ELOY SALINAS; JULIO RABA

TALANTA

Año: 2008

0039-9140

This study report an human serum IgG antibodies to H. pylori quantitationprocedure based on the multiple use of an immobilized H. pylori antigen on an immunocolumnincorporated into an a flow-injection (FI) analytical system. The immuno-adsorbentcolumn was prepared by packing 3-Aminopropyl-modified controlled-pore glass (APCPG)covalently linking H. pylori antigens in a 3 cm of teflon tubing (0.5 i.d.). Antibodies in theserum sample are allowed to react immunologically with the immobilized H. pyloriantigen, and the bound antibodies are quantified by Alkaline phosphatase (AP) enzymelabeledsecond antibodies specific to human IgG. p-aminophenyl phosphate (pAPP) wasconverted to p-aminophenol (pAP) by AP and an electroactive product was quantified onglassy carbon electrode (GCE) modified with multiwall carbon nanotubes (MWCNT)(GCE-CNTs) at 0.30 V. The total assay time was 25 min. The calculated detection limitsfor amperometric detection and the ELISA procedure are 0.62 and 1.8 U mL-1, respectively.Reproducibility assays were made using repetitive standards of H. pylori specific antibodyand the intra and inter-assay coefficients of variation were below 5%. The immuno-affinitymethod showed higher sensitivity and lower time consumed, demonstrate its potentialusefulness for early assessment of human serum immunoglobulin G (IgG) antibodies to H.pylori.