Characterisation of antibodies to bovine interleukin 2 and the use of these antibodies in development of a sensitive Elisa

Abstract

Development of effective bovine immunoassays is an important step towards the understanding of the kinetics cytokine production in cattle during infections because at present only one bovine cytokine ELISA (boIFN-) is available commercially. Biological and biochemical characteristics of monoclonal antibodies (mAbs) and polyclonal antibodies raised against bovine IL-2(bolL-2) were described as well as their use in designing a specific and sensitive ELISA for the measurement of the cytokine. Four murine anti boIL-2mAbs and 2 rabbit anti boIL-2 polyclonal antibodies were obtained and their binding specificities judged by direct ELISA, Western blotting and flow cytometry. An ELISA for boIL-2 was been developed using one mAb (28-6) as capture antibody and R1 polyclonal antibody as detector after coupled with horseradish peroxidase (HRP). The minimum level of boIL-2 detectable by this ELISA was 49pg/ml. This assay provides a rapid, reliable alternative to the biological assay generally used to quantitate boIL-2.