Production and evaluation of specific antibodies from egg yolk (IGY Abs) of immunized hens are attracting scientific trends, as indicated from the extensive IGY literature. Considering animal care, high productivity and special suitability for certain diagnostic purposes, hen's eggs represent an alternative that offers some advantages over mammals. Even though, IGY
antibodies still represent an underused resource which can be due to the lack of information concerning the possibility of production, application and IGY isolation from the egg yolk. The main purpose of the present work was to produce IGY specific to Helicobacter pylori (H. pylori), the bacteria of peptic ulcer, chronic gastritis and gastric cancers. The produced
antibodies are to be further employed for diagnostic or therapeutic purposes. Polyclonal anti-H. pylori IGY antibodies were successfully produced, purified and titrated in this study. This was accomplished through immunization of laying hens with the bacterial cell lysate and isolation of the anti-H. pylori IGY by ammonium sulphate precipitation and affinity chromatography. The purified IGY was proved to be reactive with the bacterial cells and with high titre as indicated by ELISA and microplate agglutination.This article comes in accordance with the European Centre for the Validation of Alternative Methods (ECVAM) concerning alternative methods of value to the bioscience and medical fields that call for refining or replacing the use of laboratory animals.