Abstract: Production of Monoclonal antibodies for assay development
Mentor: Dr. Donald Lightfoot, Department of Biology
Allergenic antigens have several uses, including detecting and quantitating an individual's sensitivity to specific allergens. In addition, dilute solutions of allergens are used for injection to desensitize persons who have mid to severe reaction to allergies. The purpose of making monoclonal antibodies to specific grass allergens is to develop an assay that will identify specific grass pollen allergen components.
The work involves immunization of mice, collection of B lymphocytes, maintenance of mouse myeloma cells, fusion of myeloma cells and spleen lymphocytes, cloning of positive hybridomas and ELISA (Enzyme linked Immuno-Sorbent Assays).
The fusion's of mice immunized with Kentucky Bluegrass and Orchard grass have been completed and the results of ELISA were high in titer. Ten of the best producing cell lines were cloned and frozen. The resulting supernatant (cell culture media) was collected and used for assay development. Eight of the grass extracts were tested for specific binding to antibodies. All eight of the grasses tested positive to the antibodies. Further studies involving SDS polyacrylamide and Western blotting will be studied.
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