[Preparation and characterization of porcine IL-2 monoclonal antibodies].

Porcine interleukin-2 (pIL-2) is a key cytokine that coordinates cellular and humoral immune responses. To obtain active protein and highly specific tool antibodies for use in porcine immunological research and disease diagnosis, this study cloned the full-length pIL-2 gene into the pcDNA3.1 vector to construct the recombinant plasmid pcDNA3.1-pIL-2. The rpIL-2 was expressed and purified via the CHO eukaryotic expression system, and BALB/c mice were immunized with rpIL-2. Through cell fusion, indirect ELISA, and limiting dilution subcloning, two monoclonal antibodies with strong specificity and good reactivity against pIL-2 were selected, named 1C3 and 4G8, with subtypes IgG2b and IgG1 and antibody titers reaching 1:2 048 000 and 1:1 024 000, respectively. Western blotting and indirect immunofluorescence results confirmed that these two monoclonal antibodies had good reactivity with pIL-2. The successful expression of rpIL-2 and the preparation of highly specific monoclonal antibodies provide essential biological materials for subsequent research on porcine immune function and vaccine efficacy evaluation.