Submitted to: Cytotechnology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/24/2005
Publication Date: 3/25/2005
Citation: Shinmoto, T.T., Yamagishi, K.K., Suzuki, T., Maleki, S.J. 2005. Generation of mouse-human hybridomas secreting antibodies against peanut allergens. Cytotechnology. 46:19-23.
Interpretive Summary: To analyze structures of food allergens, we have produced human antibodies to food allergens. Our method employed the fusion of human B cells with mouse myeloma cells to establish stable human-mouse hybrids (referred to as hybridomas). We have already developed over 5000 multi-clone, human B cell library stocks from thirteen healthy donors. The library contains human B cells that secrete antibodies against soybean, peanut, wheat, milk proteins, egg-white proteins, and some pollen allergens. These antibodies can be used to identify and characterize the allergens in these foods. In this study, we established two hybridomas secreting human monoclonal antibodies that react with a major peanut allergen, Ara h 1, and we also determined the specific antibody binding sites on Ara h 1, using synthetic overlapping Ara h 1 peptides.
Technical Abstract: Five clones of mouse-human hybridomas secreting antibodies to peanut allergens were generated from peripheral blood lymphocytes transformed with Ebstein-Barr virus, followed by cell fusion with mouse myeloma cells. Western blot analysis showed that two antibodies reacted to the Ara h 1 peanut allergen. Epitope analysis with overlapping peptides synthesized on a multi-pin apparatus, revealed two antibody-binding sequences of the Ara h 1 protein.