Authors: Denise Huber Judith Rudolf Parisa Ansari Brigitte Galler Manuela Führer Christoph Hasenhindl Sabine Baumgartner
Publish Date: 2009/02/27
Volume: 394, Issue: 2, Pages: 539-548
Abstract
Blocking is an important step before an enzymelinked immunosorbent assay ELISA can be performed It reduces nonspecific binding to the microtiter plate to a minimum For detecting food allergens by means of ELISA the problem with protein blocking solutions is obvious The blocker might interfere with the antibodies of the assay and leads to false positive results Therefore other blocking solutions are greatly needed There are some alternatives like synthetic blockers or carbohydrates Comparisons of these different blocking agents namely proteins carbohydrates and synthetic blockers were made at different reaction conditions The incubation periods and temperatures were varied as well as the pH The best combinations were evaluated and compared in respect of their blocking efficiency The two best nonproteinaceous blockers ie polyvinylalcohol and Ficoll were subsequently applied to ELISA tests for the determination of αcasein and peanut The study showed that Ficoll and PVA did as well as BSA in buffer solution Therefore they can be considered as alternative blocking reagents for ELISA especially for the detection of food allergensThe authors thank the Christian Doppler Research Association and RomerLabs for funding the Christian Doppler PilotLaboratory “Rapid Test Systems for Allergenic Food Contaminants” We also thank Dr Marcela Hermann Institute of Medical Biochemistry Department of Molecular Genetics University Vienna for the immunisation of rabbits and chicken
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