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. 1992 Jun;76(2):305–309.

Selection of BoCD25 monoclonal antibodies by screening mouse L cells transfected with the bovine p55-interleukin-2 (IL-2) receptor gene.

J Naessens 1, M Sileghem 1, N MacHugh 1, Y H Park 1, W C Davis 1, P Toye 1
PMCID: PMC1421522  PMID: 1634253

Abstract

The bovine interleukin-2 receptor-alpha (IL-2R alpha) gene has been isolated and a rabbit antiserum against a fusion protein of the gene has been produced. However, the antiserum does not inhibit IL-2-dependent proliferation. Since a panel of monoclonal antibodies (mAb) to bovine activation antigens was available, we transfected the gene into mouse L fibroblasts, selected stable transfectants with the rabbit antiserum, and screened for antibodies that bound the transfected cells but not the untransfected cells. Three mAb were selected and all three precipitated a molecule of M(r) 55,000 (under reducing conditions) from activated cells, as expected from homology with mouse and human IL-2R alpha (CD25, Tac). One of the three mAb was a strong inhibitor of IL-2-dependent proliferation of bovine lymphocytes. Thus, the availability of transfected cells allowed us to establish quickly and unequivocally the antigenic specificity of a number of antibodies.

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Selected References

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