Authors: Sirley V Pereira Julio Raba Germán A Messina
Publish Date: 2010/03/12
Volume: 396, Issue: 8, Pages: 2921-2927
Abstract
In the present article a novel microfluidic immunosensor coupled with electrochemical detection for antigliadin IgG antibody quantification is proposed This device represents an important tool for a fast simple sensitive and automated diagnostic for celiac disease which is carried out through detection of antigliadin IgG antibodies present in human serum samples Celiac disease CD is an autoimmune disease generated by gluten protein fractions called prolamins This pathology affects about one in 250 people around the world produces intestinal inflammation villous atrophy and crypt hyperplasia which causes a range of symptoms including altered bowel habits malnutrition and weight loss Our immunosensor consists of a Plexiglas device coupled to a gold electrode with a central channel containing 3aminopropylmodified controlled pore glass APCPG The quantification of antigliadin IgG antibodies was carried out using a heterogeneous noncompetitive enzymelinked immunosorbent assay ELISA in which IgG antibodies bound to gliadin protein immobilized on APCPG were determined by alkaline phosphatase AP enzymelabeled second antibodies specific to human IgG The paminophenyl phosphate pAPP was converted to paminophenol pAP by AP and the electroactive product was quantified on a gold electrode at 0250 V The calculated detection limits for electrochemical detection and the ELISA procedure were 052 and 272 UR mL−1 respectively and the within and betweenassay coefficients of variation were below 58 The optimized procedure was applied to the determination of antigliadin IgG antibodies in human serum samples
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